CHEMISTRY RESEARCH AND APPLICATIONS

FLOCCULATION

PROCESSES AND APPLICATIONS

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CHEMISTRY RESEARCH AND APPLICATIONS

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 CHEMISTRY RESEARCH AND APPLICATIONS

FLOCCULATION

PROCESSES AND APPLICATIONS

ELEONORA VOLLAN
EDITOR
Copyright © 2019 by Nova Science Publishers, Inc.

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 CONTENTS

Preface vii
Chapter 1 Flocculant Polysaccharides Mainly from Plants 1
Priscilla B. S. Albuquerque, Weslley F. Oliveira,
Priscila M. S. Silva, Maria T. S. Correia
and Luana C. B. B. Coelho
Chapter 2 Coagulation and Flocculation with Plant Extracts 47
Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta
Chapter 3 The Process of Water Treatment with
Aluminum Sulphate Associated with the
Application of the Cactus Opuntia Cochenillifera 69
Higgor Henrique Dias Goes, Rita de Cássia Pereira de Souza
and Joseane Débora Peruço Theodoro
Chapter 4 Flocculation: Mechanisms and Applications
for Wastewater Treatment 81
Elvis Carissimi, Cristiane Oliveira Rodrigues,
Dounia Elkhatib and Vinka Oyanedel-Craver
Chapter 5 Flocculation of AOM in Water Treatment 107
Martin Pivokonský, Jana Načeradská, Kateřina Novotná,
Lenka Čermáková and Petra Vašatová
Chapter 6 Comparison of Natural Coagulant and Chemistry in Tanning
Wastewater Treatment Using the Flocculation Process 143
Edilaine Regina Pereira, Gustavo da Silva Souza
and Joseane Débora Peruço Theodoro
vi Contents

Chapter 7 Moringa oleifera Seed Use in Salina Solution in

Water Treatment in Lentic Bodies 155
João Carlos Belisário Junior, Edilaine Regina Pereira
and Joseane Débora Peruço Theodoro
Chapter 8 Evaluating New Biopolyelectrolytes for the Meat Processing
Wastewater Treatment via Coagulation-Flocculation 169
E. A. López-Maldonado and M. T. Oropeza-Guzmán
Chapter 9 Treatment of Residual Waters of the Brewery Industry
through the Flocculation Process with the Use
of Inorganic and Organic Coagulants 179
Fellipe Jhordã Ladeia Janz, Edilaine Regina Pereira,
Thaís Ribeiro, Dandley Vizibelli and Julio Cesar Angelo Borges
Chapter 10 Application of Electrocoagulation with Voltage
Variation to Obtain Potable Water 187
Ingrid Cardozo Botelho, Thiago Andre Bezerra Higuchi
and Joseane Débora Peruço Theodoro
Chapter 11 An Evaluation of the Performance of the
Coagulation/Flocculation Process with Aluminum
and Ferric Salts on the Removal of Algal Toxins 197
Ayşe Büşra Şengül
Chapter 12 Treatment of Leachate from the Ouled Berjal Landfill
in Morocco by Coagulation Flocculation:
A Study of the Effect of Order for Reagent Introduction 217
Hajar Bakraouy, Salah Souabi, Khalid Digua and Levent Yilmaz
Chapter 13 OMW Pretreatment by Assisted Sedimentation
Methods: Coagulation/Flocculation 259
Gassan Hodaifa and Cristina Agabo García
Chapter 14 Chitosan Based Flocculants for the
Removal of Heavy Metal Ions 277
Sayaka Fujita and Nobuo Sakairi
Chapter 15 Determination of the Kinetic Coefficient of Aggregation and the
Kinetic Coefficient of Rupture in the Turbidity Removal Process 295
Joseane Debora Peruço Theodoro,
Paulo Sergio Theodoro and Rosangela Bergamasco
Index 303
 PREFACE

Flocculation: Processes and Applications opens by approaching current trends in

preparation and chemical modification of flocculant polysaccharides derived from plants
and their flocculation performance. In addition, aspects including mechanisms of
flocculation, chemical modification, the effect of physicochemical factors on flocculating
activity, and recent applications of flocculant polysaccharides are reviewed.
The authors go on to propose plant extracts which can efficiently perform
coagulation and flocculation operations without the environmental risk of residual sludge
with high concentrations of aluminum or iron.
A separate study aimed to use the organic polymer from Opuntia cochenillifera
cactus associated with the addition of aluminum sulfate to treat the water of a lentic body
applying coagulation, flocculation, sedimentation and filtration processes.
The authors propose that the design and operation of flocculators is crucial for the
process efficiency and largely dependent on the following features: floc characteristics,
flocculation kinetics, and engineering aspects of flocculation.
This compilation also discusses current knowledge on algal organic matter (AOM)
flocculation, the impact of AOM on the removal of other compounds and links AOM
composition and character to the efficiency of flocculation, the reaction conditions and
mechanisms and finally, to the properties of flocs.
Additionally, the performance of natural coagulant tanin compared to chemical
coagulants aluminium sulphate and ferric chloride commonly used in the treatment of
raw wastewater from tannery, by means of the physicochemical processes of coagulation,
flocculation and sedimentation are examined.
Through physical and chemical parameters, the efficiency of the coagulation/
flocculation/sedimentation/filtration processes using organic coagulants in the treatment
of water from a lentic system in Brazil are examined as well.
viii Eleonora Vollan

Later, the physicochemical performance of chitosan and mesquite gum as coagulant

flocculent agent for the treatment of residual water of the cutting and packing of meat
products factory is presented.
The brewing industry generates effluent that can cause serious environmental impacts
when not treated properly due to high loads of organic matter in its composition. Thus, in
view of the growing emergence of breweries in Brazil and consequent increase in effluent
production, alternatives are sought for the auxiliary treatment using coagulants and their
efficiency is analyzed.
Urban development also contributes to increasing water pollution, therefore the
authors perform water treatment (through the electrocoagulation process) to calculate the
cost of the operation.
Eutrophication is one of the most prevalent water quality problems in the United
States as well as other parts of the world. It has led to excessive growth of algal blooms,
which not only cause the death of aquatic plants and animals, but also produce high levels
of toxins and odorous compounds. The authors examine the performance of the
coagulation/flocculation process using aluminum and ferric salt coagulants for the
removal of microcystins.
One study focuses on the coagulation flocculation of young leachate from the Kenitra
city landfill. Tests were carried out by adding ferric chloride mixed with three
flocculants, namely: the chitosan, the Superfloc SD2065 and the Himoloc.
The authors outline researches about combining assisted sedimentation with other
operations such as oxidation processes in order to evaluate the solids removal of the
complete designed wastewater treatment focusing on OMW treatment.
The penultimate chapter focuses on the preparation and characterization of the
chitosan based flocculant for removal of heavy metal ion prepared from chitosan by N-
acylation with ethylenediaminetetraacetic acid monoanhydride.
The concluding study aims to apply the Bratby method in the characterization of the
turbidity removal process, through the determination of the kinetic aggregation
coefficient (KA) of the flocs and the kinetic coefficient of rupture (KB) of the flocs.
Chapter 1 - Natural polymers are biocompatible, low-cost, and easily available
materials of innate origin. These polymers are increasingly preferred over synthetic
materials for industrial applications due to their intrinsic properties; they also are
alternative sources of raw materials with characteristics of biodegradability, biosafety,
and sustainability. Polysaccharides are polymers extracted from plants, algae, animals,
fungi or obtained via fermentation, applied on a wide range of uses, from food to
biomedical industries. Nowadays, they have been attracting considerable attention as
viable alternatives to harmful synthetic flocculating agents for the removal of
contaminants from water and wastewater. Then, a great deal of dedicated effort improved
the production and performance of natural flocculants based on polysaccharides. The aim
of this chapter is to approach current trends in preparation and chemical modification of
 Preface ix

flocculant polysaccharides derived from plants and their flocculation performance. In

addition, aspects including mechanisms of flocculation, chemical modification, the effect
of physicochemical factors on flocculating activity, and recent applications of flocculant
polysaccharides, also derived from non-plant sources are reviewed.
Chapter 2 - The treatment of water in a conventional way includes the operations of
coagulation and flocculation as a fundamental part in the removal of solids and other
substances that are mixed with water, especially organic and inorganic solids with sizes
less than 0.2 mm and densities similar to those of water. This forms a perfect mixture
difficult to separate by natural sedimentation; in these cases the addition of coagulants is
needed, which breaks the stability of the mixture and segregates the particles in the form
of flocs, so that they can be separated by density difference. Reactive substances are used
for this reason, such as aluminum sulfate, ferric chloride and aluminum polychloride;
these substances are derived from industrial chemical reactions, which entail the use of
natural resources and energy to obtain them. In addition to the environmental cost of the
production processes, there is the problem of the final disposal of the thickened sludge,
because due to its aluminum or iron content, the sludge can be harmful to the
environment, especially for plants and animals. To reduce this impact, it is necessary to
carry out these operations with substances that do not represent a danger to the
environment; in this case, it has been proven that there are plant extracts that can generate
the same reaction of segregating the solids and other substances of the water, by means of
the reaction of the natural biopolymers of some plants, within which the Moringa
oleifera, Melocactus sp, Opuntia sp, Cicer arietinum L, Aloe sp and others with
destabilizing activity of particles can generate turbidity and color in water. These plant
extracts can efficiently perform the coagulation and flocculation operations without the
environmental risk of residual sludge with high concentrations of aluminum or iron.
Chapter 3 - The present study aimed to use the organic polymer from Opuntia
cochenillifera cactus associated with the addtion of aluminum sulfate to treat the water of
a lentic body (Igapó II Lake - Located in Londrina in the state of Paraná and Brazil)
applying coagulation, flocculation, sedimentation and filtration processes. The tests were
performed with static a Jar-test reactor, aiming to evaluate the removal of parameters of
turbidity and of apparent color, besides monitoring the pH variation. Regarding the
physico-chemical characteristics, there was little influence of the Opuntia cochenillifera
cactus in the pH parameter tending to neutrality. Therefore, the study showed that the use
of an organic coagulant together with aluminum sulphate is effective for the removal of
parameters of apparent color and of turbidity (95 and 94%, respectively).
Chapter 4 - Flocculation is considered the main physico-chemical treatment prior any
solid-liquid separation unit in water and wastewater treatment plants. Suspended
particles, algae, microorganisms are aggregated in order to increase their density for a
subsequent step of sedimention or flotation (in case of injection of air bubbles) and/or
filtration. Design and operation of flocculators are crucial for the process efficiency, and
x Eleonora Vollan

are largely dependent on the following features: floc characteristics, flocculation kinetics,
and engineering aspects of flocculation. Finally, examples of engineering applications
using flocculation are described.
Chapter 5 - Global proliferation of algal blooms and subsequent deterioration of
water quality by organic compounds that are being produced (algal organic matter –
AOM) pose new challenges to water treatment technologies. Flocculation/coagulation
using primarily Al- and Fe-based coagulants is widely employed as an essential process
in removal of various impurities at drinking water treatment plants and is also
irreplaceable in the case of AOM elimination. This review chapter discusses current
knowledge on AOM flocculation, the impact of AOM on the removal of other
compounds and links AOM composition and character to the efficiency of flocculation,
the reaction conditions and mechanisms and finally, to the properties of flocs. In general,
the removal efficiencies of dissolved AOM are lower compared to intact phytoplankton
cells and usually reach maximum under slightly acidic pH values. The strong pH-
dependence of flocculation is attributed to the fact that the involved mechanisms are to a
great extent determined by the charge ratios in the coagulating system. Furthermore,
substantial differences in flocculation behaviour were observed between diverse AOM
constituents, i.e., between peptides/proteins versus non-proteinaceous matter and high
versus low molecular weight organics. The latter (specifically AOM < 10 kDa) are
reluctant to flocculate and would therefore require other treatment techniques. AOM has
also been reported to influence flocculation of other common impurities, both of organic
and inorganic nature. Mutual interactions have been proven, while their influence on
flocculation efficiency can be either positive or negative, depending on the AOM
character, pH conditions and on the ratio between AOM, the other polluting agents and
coagulants. Finally, AOM also appeared to alter the properties of flocs, with an impact on
the subsequent separation steps. In further research, a particular emphasis should be put
on AOM components that are difficult to coagulate, the interactions of AOM with other
impurities and on elucidation of the relationship between AOM and floc properties.
Chapter 6 - The leather tanning industry uses a large amount of toxic substances and
water. Thus, it generates effluents with high polluting load. Due to the large amount of
effluent generated by tanneries and the difficulty in their treatment, several studies have
been proposed to minimize the environmental impacts caused by inappropriate disposal
of this effluent. This study investigated the performance of natural coagulants Tanin
compared to chemical coagulants aluminium sulphate and ferric chloride commonly used
in the treatment of raw wastewater from tannery, by means of the physicochemical
processes of coagulation, flocculation and sedimentation. Using jar tests methods,
different concentrations for the coagulants (chemical and natural) were applied to the
concerned effluent and it was assessed their efficiency in removing certain parameters
such as apparent color COD, and turbity besides the behavior of electrical conductivity
and pH. The results showed that for pH and electrical conductivity parameters, there was
 Preface xi

no significant variation after application of coagulant in relation to the raw effluent. By

comparing the coagulants for colour parameter it was observed that the natural coagulant
tanin had greater efficiency compared to the chemical ones, reaching 56.9% of removal,
however, the aluminium sulphate was more efficient at removing COD, with 95% of
removal. Therefore, it is noticed that the chemical coagulant aluminium sulphate was
more successful for the majority of the analysed parameters in comparison to other
coagulants.
Chapter 7 - The proposal of this paper was to evaluate through physical and chemical
parameters the efficiency of the coagulation/flocculation/ sedimentation/filtration
processes using organic coagulant (Moringa oleifera) in the treatment of water from a
lentic system (Igapó II Lake), located in Londrina – Paraná - Brasil. It was made the
water collection at two points of the lake, in the entrance (point 1) and at the exit (point
2). The treatment occurred using the jar-test equipment with the same conditions of fast
mixing and slow mixing used in the Water Treatment Station (ETA). The sedimentation
time was 30 minutes. The granulometry of the sand used in the filters was the same, in
the range of 0.600 to 0.850 mm. The concentrations of saline solution of the organic
coagulant to be applied were: C1 = 3 mg.L-1, C2 = 6 mg.L-1 and C3 = 9 mg.L-1. In addition,
it was necessary to correct the pH of the samples in order to evaluate the behavior of the
organic coagulant in different pH levels. After the assays, a study was carried out on the
removal of turbidity and apparent color after the coagulation / flocculation /
sedimentation processes and after the filtration process. It was verified for the parameters
of turbidity and of apparent color, the concentration C2 (6 mg.L-1) and the pH around 7
(neutral) presented the highest removal efficiency, being it above 99%. For the pH, all
concentrations did not present great variations when compared to the crude sample, in
addition the results prove the efficiency of the coagulant in different pH levels. Only
assay 2 in the entrance point, did not meet the maximum values allowed by the potability
ordinance 2,914/11 of the ministry of health for parameters of apparent color and of
turbidity, all other assays are in accordance. The results proved the efficiency of the
organic coagulant extracted from Moringa oleifera seeds for the treatment of water in
lentic systems.
Chapter 8 - The solid-liquid separation processes are of great scientific and
technological relevance in the area of food, beverages, agrochemicals, drugs, mineral
extraction, ceramics and wastewater treatment. Nowadays, various chemical substances
are used that act as destabilizing agents or stabilizers according to their field of
application. One of the strategies to understand the coagulation-flocculation processes is
to use the zeta potential measurements as an electrochemical tool to approximate the
interface phenomena and establish the mechanisms that predominate at the molecular
level. One of the trends is the use of various biopolyelectrolytes (BPE) that can be
obtained, modified and applied strategically in various fields of study. In this chapter, the
physicochemical performance of six biopolyelectrolytes in the coagulation of flocculation
xii Eleonora Vollan

of wastewater with a high content of solids and fats and oils was evaluated. The ones that
presented a better performance for this type of wastewater are mesquite gum and
chitosan. The BPE that showed a better performance in the coagulation-flocculation test
of the wastewater with high content of fats and oils at pH 5.2, was the chitosan, with a
dose of 68 mg/L of chitosan the treated water has the following values of BOD5 = 30 mg
O2/L, fats and oil = 1 mg/L, turbidity = 2 FAU, TSS = 2 mg/L and COD = 50 mg/L.
Chapter 9 - The brewing industry presents a great generation of effluent that can
cause serious environmental impacts when not treated in a regular way since it has a high
load of organic matter in its composition. Thus, in view of the growing emergence of
breweries in Brazil and consequent increase in effluent production, alternatives are
sought for the auxiliary treatment of this effluent using coagulants. In this way, the paper
aimed to analyze the efficiency the usage of two organic and one inorganic coagulant in
the treatment of effluent from the brewing industry. The assay was performed in Jar Test,
thus simulating the coagulation, flocculation and sedimentation processes, and in
sequence the filtration process was carried out. During this process the parameters of
apparent color, electrical conductivity, pH and turbidity were monitored. Three different
treatments were used: Aluminum Sulfate (T1), Tannin (T2) and Moringa oleifera (T3).
The results showed values of apparent color removal and turbidity of more than 90% for
all treatments, but the treatment with Tanino presented the highest values of removal,
being 98.8% for turbidity removal and 99.6% for removal of apparent color. The pH
parameter showed a rise throughout the test for all treatments presenting final values
close to the neutral value, demonstrating compliance with CONAMA Resolution No.
430/2011. The electrical conductivity showed a decrease in all treatments especially in
the treatment where Tanino was used being 0.32μS.m-1 the lowest value obtained. The
use of the coagulants in question showed positive responses in the analyzed parameters in
the treatment of the effluent of the brewing industry, showing more emphasis for the
Tanino that obtained the best values. It’s an option sustainable and is possible to mix this
process with the comum process that there are in brewing industry.
Chapter 10 - Urban development contributes to increasing water pollution, so we
must develop alternative ways to treat water. Therefore, the objective of this paper was to
perform the water treatment - through the electrocoagulation process - and to calculate
the cost of the operation. The paper was done according to predefined experimental
planning that has nine samples with duplicate plus a pair of samples in central point. The
assays were carried out using a bench-level acrylic reactor (2 liters), a magnetic stirrer, a
voltage source and a monopolar electrode system arranged by means of 4 equidistantly
connected aluminum plates in parallel. The studied parameters were final pH and electric
conductivity, tested in two different voltages (5V and 11V). Sampling time were 40, 60
and 80 minutes. At the end of the trials, treatment costs were calculated to ensure that it
was applicable. The analysis of the results concluded that the electrocoagulation process
assists in the neutrality of the solution, brings the acid and basic samples near to the
 Preface xiii

neutral pH (7) and the electric conductivity does not have significant change. Through the
work it was possible to observe that the eletrocoagulation process accomplished with the
continuous voltage of 5V was the one that presented the lowest cost, considering that the
energy consumption and the use of the electrode were smaller 0.03325kWh and
1.23949µg respectively.
Chapter 11 - Eutrophication, or the excessive richness of nutrients in a lake or other
body of water, is one of the most prevalent water quality problems in the United States as
well as other parts of the world. It has led to excessive growth of algal blooms, which not
only cause the death of aquatic plants and animals, but also produce high levels of toxins
and odorous compounds, thereby posing serious risks to safe drinking water and human
health. Microcystis aeruginosa is well known to be one of the most dangerous and most
common bloom-forming cyanobacteria that produces hepatotoxic microcystins. This
study evaluated the performance of the coagulation/flocculation process using aluminum
and ferric salt coagulants for the removal of both microcystins (intracellular and
extracellular) and turbidity. The effects of coagulant dosage, pH, and coagulant-aid
dosage on microcystins and turbidity removal were also evaluated. All results
demonstrated that the optimum dosage of aluminum sulphate, ferric chloride, and ferrous
sulphate for the removal of both microcystins and turbidity was found to be 60, 60, and
20 mg/L, respectively. With optimized coagulant dosages, the optimum pH value for both
microcystins and turbidity removal occurred with a pH of 7, 6, and 7 for aluminum
sulphate, ferric chloride, and ferrous sulphate, respectively. On the other hand, for all
coagulants, the optimum polyelectrolyte dosage for the removal of both microcystins and
turbidity was found to be 0.2 mg/L. Among these coagulants, aluminum sulphate (65.2%)
was found to be more effective for both microcystins and turbidity removal compared to
ferric chloride (56.3%) and ferrous sulphate (30.9%). Last, the concentration of turbidity
decreased to below 5 NTU, which is the maximum turbidity level in drinking water
determined by the World Health Organization (WHO). After coagulation/flocculation,
the total microcystin concentration (4.09 µg/L) in the water remained above the WHO
guideline value of 1 µg/L for drinking water, but the release of toxins in the water was
not observed. Therefore, both intracellular microcystin and turbidity could be removed
through the coagulation/flocculation process, whereas extracellular microcystin is still of
particular concern in drinking water treatment due to its difficult removal.
Chapter 12 - In Morocco, like all the countries of the world, socio-economic
activities coupled with population growth and changes in consumption patterns generate
a significant production of municipal solid waste. This is accompanied by a significant
increase in leachates produced during the fermentation of waste. These leachates have a
considerable impact on the environment. Moroccan regulations require leachate treatment
to reduce environmental impacts. Several techniques are currently used for the
decontamination of leachate such as (coagulation flocculation, biological treatment,
filtration, oxidation, ...). Indeed, the majority of the physicochemical treatments used for
xiv Eleonora Vollan

the purification of the leachates intervenes as pretreatment or finishing stage to complete

the treatment chain, or to eliminate a specific pollutant. The addition of a polymer as a
flocculant improves the effectiveness of coagulation by inorganic coagulants. In fact,
these flocculants make it possible to obtain large flocs, which settle rapidly and which
resist shear forces (Bratby, 2006). According to Bratby (2006), when a liquid is already
destabilized after the addition of an inorganic coagulant, the polymers increase the rate of
orthokinetic flocculation by the formation of larger flocs. This is done through a
sufficient absorption affinity between the polymers and the flocs. The choice of polymers
must be made judiciously because their effectiveness depends on several parameters,
namely: the nature of the polymer, the pH, the temperature, the type of inorganic
coagulant, the nature and size of the pollutant to be eliminated, the concentration of the
coagulant and flocculant (Zemaitaitiene et al., 2003, Bolto and Gregory, 2007). The first
part of this work concerns the coagulation flocculation of young leachate from the
Kenitra city landfill. The tests were carried out by adding ferric chloride mixed with three
flocculants, namely: the chitosan, the Superfloc SD2065 and the Himoloc. The dose of
coagulant was fixed at 6 g/L (determined from preliminary tests), while the doses of
flocculants ranged from 3,3 to 20 mL/L. The evaluation of treatment efficiency as well as
the optimal doses of the flocculants was determined by the monitoring of the removal of
turbidity, COD, phenol, color, Absorbance at 254 nm and the volume of sludge collected
after 24 hours. The purpose of the second part reveals the effect of the order of
introduction of the reagents on the efficiency of leachate treatment by coagulation
flocculation. The reagents used are ferric chloride and the three flocculants previously
mentioned. The authors varied the order of introduction by adding:

 Ferric chloride first followed by flocculant.

 The flocculant first followed by the coagulant;
 The coagulant and the flocculant at the same time.

The effect of the order of introduction of the reagents was evaluated in terms of
elimination of turbidity, COD, color, Abs at 254 nm, phenol, detergents, ammonium ions,
nitrite, nitrate and total phosphorus.
Chapter 13 - Natural and assisted particles sedimentation are crucial operations
during the treatment of wastewater generated in many industrial processes, above all in
the wastewater treatment field. This operation is usually used as an effluent pre-
treatment; normally essential for reducing suspended solids, organic load, turbidity and
colour so as to achieve the correct clarification of wastewater. This chapter introduce the
coagulation and flocculation concepts as well as the influence of operational conditions
on assisted sedimentation. At industrial level, the factual use of both processes has been
proved on different wastewater. In this sense, this chapter outline researches about
combining or comparing assisted sedimentation (coagulation, electrocoagulation,
 Preface xv

neutralization, etc.) with other operations such as oxidation processes in order to evaluate
the solids removal of the complete designed wastewater treatment focusing on OMW
treatment. In addition, in this work new trends about multifunctional and novel eco-
friendly bio-degradable flocculants and coagulants were reviewed and suggested for
OMW pretreatment.
Chapter 14 - This chapter focuses on the preparation and characterization of the
chitosan based flocculant for removal of heavy metal ion prepared from chitosan by N-
acylation with ethylenediaminetetraacetic acid monoanhydride (EDTAM). In this study, a
series of chitosan flocculants with various degree of substitution were prepared by
changing the molar ratio of EDTAM to chitosan in the preparation. The structural
characterization of the chitosan flocculants by FTIR and 13C NMR spectroscopy were
performed. The newly introduced functional group provided properties such as being a
strong chelating reagent and an amphoteric polyelectrolyte. It was found that chitosan
flocculants had good water solubility in both acidic and basic regions and precipitated in
a narrow pH region, which is close to its isoelectric point. The chitosan flocculants has an
ability to remove heavy metal ion from aqueous solution by controlling pH or initial
concentration of the flocculant, and removed Cu(II) almost completely. The flocculation
mechanism was investigated using Cu(II) and found that flocculation occurred by charge
neutralization was sensitive both pH conditions and amount of chelated Cu(II) on
chitosan flocculants. The complexation stoichiometry of Cu(II) and the EDTA residues of
chitosan flocculant determined by UV-vis titration. As a result, the chitosan flocculants
can easily remove Cu(II) from aqueous solution by only flocculation/precipitation
process, which indicated that the chitosan flocculants could be applicable for remediation
and treatment system of wastewater.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 1

FLOCCULANT POLYSACCHARIDES
MAINLY FROM PLANTS

Priscilla B. S. Albuquerque*, Weslley F. Oliveira2,

Priscila M. S. Silva2, Maria T. S. Correia2
and Luana C. B. B. Coelho2,*
1
Centro de Tecnologias Estratégicas do Nordeste-CETENE,
Recife, PE, Brazil
2
Departamento de Bioquímica, Centro de Biociências,
Universidade Federal de Pernambuco, Recife, PE, Brazil

ABSTRACT

Natural polymers are biocompatible, low-cost, and easily available materials of

innate origin. These polymers are increasingly preferred over synthetic materials for
industrial applications due to their intrinsic properties; they also are alternative sources of
raw materials with characteristics of biodegradability, biosafety, and sustainability.
Polysaccharides are polymers extracted from plants, algae, animals, fungi or obtained via
fermentation, applied on a wide range of uses, from food to biomedical industries.
Nowadays, they have been attracting considerable attention as viable alternatives to
harmful synthetic flocculating agents for the removal of contaminants from water and
wastewater. Then, a great deal of dedicated effort improved the production and
performance of natural flocculants based on polysaccharides. The aim of this chapter is to
approach current trends in preparation and chemical modification of flocculant
polysaccharides derived from plants and their flocculation performance. In addition,
aspects including mechanisms of flocculation, chemical modification, the effect of

*
Corresponding Author Email: [email protected].
2 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

physicochemical factors on flocculating activity, and recent applications of flocculant

polysaccharides, also derived from non-plant sources are reviewed.

Keywords: agglomeration, aggregation, coagulation, contamination, flocculation,

grafting, polymerization, polysaccharide, wastewater treatment

1. INTRODUCTION

The flocculation phenomenon is related to the clumping of particles with consequent

destabilization and coming out of the aggregates from suspension. In a chemical
perspective, the flocculation process is essentially physical, occurring due to the contact
and adhesion of aggregates by the formation of large-size clusters called flocs, which are
excluded of the suspension. Commonly, one can observe flocculation being widely used
as synonymous of agglomeration, aggregation, and coagulation (Santos et al., 2014).
Flocculating agents are useful in different industrial fields, such as tapwater and
wastewater treatment, dredging, textile, mining, cosmetology, pharmacology, food and
fermentation industries and downstream processes (Salehizadeh, Yan and Farnood,
2017). Especially in food and beverage industries, flocculant agents are widely used with
the important aim to remove microscopic particles that affect water taste, appearance and
texture (Santos et al., 2014).
Flocculants are generally classified into three groups: inorganic flocculants, organic
synthetic flocculants, and natural flocculants (or bioflocculants). Although the first two
groups are most commonly used due to their effective flocculating performance and low
cost, they could be associated to serious environmental and health problems. In order to
circumvent the above concerns, natural flocculants have been attracting more attention in
utilization (Salehizadeh and Yan, 2014).
Polysaccharides are biopolymers and particularly attractive as natural flocculants due
to their inherent properties; they represent one of the most abundant industrial
biomaterials usually reported by several studies due to their sustainability,
biodegradability and biosafety. Even more, polysaccharides are abundant in nature and
commonly found in many higher plants, being frequently produced as a protection
mechanism following plant injury (Albuquerque et al., 2017; Rana et al., 2011).
The most common polysaccharide bio-based flocculants are alginate, chitosan,
cellulose, and starch; however, microorganisms could also be used as source of raw
material (Lin et al., 2014; Teh et al., 2016; Yang et al., 2016). Moreover, polysaccharides
may be chemically modified to improve their flocculation performance (Liu et al., 2017b;
Lou et al., 2017). Broad industrial application of bioflocculants based on polysaccharides
depends on the use of low-cost substrates and the development of more efficient
fermentation and recovery processes, as well as the application of cost-effective chemical
 Flocculant Polysaccharides Mainly from Plants 3

modification of the extracted polysaccharides for improved performance (Salehizadeh,

Yan and Farnood, 2017).
This chapter describes the progress in polysaccharide flocculants with special
emphasis for those derived from plants. In particular, aspects summarized include
extraction, purification, modification, characterization, and the broad range of
applications in industry.

2. SHORT SUMMARY ON POLYSACCHARIDES

The polysaccharide term gathers collectively quite diverse large carbohydrates

composed by monosaccharides, being named homopolysaccharides if they have only one
kind of repeating units (for example starch and cellulose), or formed by two or more
different monomeric units, named heteropolysaccharides and exemplified as agar,
alginate, and carrageenan. Polysaccharides are considered neutral, anionic or cationic
depending on its electric charge; in addition, the conformation of the polymer chains is
markedly dependent not only on the ionic strength of the medium, but also on the pH,
particularly in the case of the polyelectrolytes, and the temperature and the concentration
of certain molecules. Several anionic and cationic polysaccharides are widely available in
nature and have gained keen interest in food and pharmaceutical fields (Prajapati et al.,
2014).
Gums and mucilages are the constituents of polysaccharides, possessing natural
source and particular differences: gums readily dissolve in water, while mucilages form
viscous masses; in addition, their similarities are related to their broad range of
physicochemical properties, which are widely used for applications in cosmetics, paper,
pharmacy, textile, adhesive, inks, lithography, paint, explosive, and smoking products
(Albuquerque et al., 2017). The preference of polysaccharides when compared to
synthetic materials is closely related to their biological and chemical properties, including
biocompatibility, biodegradability, polyfunctionality, high chemical reactivity, chirality,
chelation and adsorption capacities (Hossain and Mondal, 2014).
Regarding the excellent adsorption behaviour of polysaccharides, it is important to
point out characteristics such as high hydrophilicity (due to hydroxyl groups of the
monomeric units), presence of a vast number of functional groups (acetamide, primary
amino, and/or hydroxyl groups) with high chemical reactivity, and flexible structure of
the polymer chain. In what concerns these advantageous characteristics, the development
of new products based on polysaccharides is an attempt to circumvent the issues
associated to synthetic polymers and a better way of, rationally, using renewable bio-
resources (Hossain and Mondal, 2014; Oladoja et al., 2017).
The global awareness of the society for minimizing the use of synthetic polymers has
been growing and gaining special attention in important environmental campaigns
4 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

because of the increasing environmental damage of this century. In addition, the scientific
community try to develop technological alternatives for this issue. The last two decades
have been marked by a crescent interest in public and scientific communities about the
use and development of biopolymers, which can be obtained from renewable sources,
displaying the important characteristic of biodegradability, with the desired
physicochemical properties of conventional synthetic materials (Albuquerque and
Malafaia, 2017; Rhim, Park and Ha, 2013).
The majority of biopolymers have rather low activity under environmental
conditions. The primary task for scientific researches, in what concerns the
polysaccharides, is to create a methodology for the synthesis of functional materials
derived from them, which have significant properties for the enhancement of their
utilization on a practical scale. This is expected to open the possibility of a real
knowledge of the nature of intermolecular interactions in aqueous solutions of
biopolymers (Oladoja et al., 2017).
The optimal choice of a particular technology for engineering of polysaccharides can
be either by chemical, physical or biological modifications. Physical (ultrasonic
disruption and microwave exposure) and biological (enzymatic degradation) changes
allow modifications of the polysaccharide molecular mass, while the chemical-one could
change the substituent types of groups, number and position (Ghimici and Nichifor,
2018).
Polysaccharide bio-based flocculants contain natural polysaccharides that are
suspected to exhibit excellent selectivity towards aromatic compounds and metals in the
absorption of particles, for example pollutants found in wastewaters (Crini, 2005).
However, it is well known that their feasibility is restricted by their physicochemical
properties, moderate flocculating activity and short shelf life. In recent years, grafted
bioflocculants have been developed and claimed due to their remarkable flocculating
ability and biodegradability. They are covalently modified by inclusion of synthetic
monomers onto their backbone to synthesise the high molecular weight grafted
copolymers that exhibit improved flocculating properties (Crini, 2005; Lee et al., 2014;
Lee et al., 2012).
The main methods for polysaccharide modification in aqueous solution can be
categorized in: (1) conventional method, (2) microwave initiated grafting method, and (3)
microwave assisted grafting method. In the conventional method, a chemical-free radical
initiator is added under an inert atmosphere to produce free radical sites on the polymer
in order to allow the addition of monomer to form the graft chain. This method is not
suitable for an industrial scale of polysaccharide production due to its low
reproducibility. The use of microwave irradiation emerges as a promising alternative; in
the microwave assisted grafting method, ions are produced by the addition of external
redox initiators to the reduction mixture, thus, the free radical initiators facilitates the
grafting reaction. In microwave initiated grafting reactions, in turn, no initiators are
 Flocculant Polysaccharides Mainly from Plants 5

added and a small amount of hydroquinone is required to inhibit the grafting reactions. It
is important to mention that factors including pH, reaction temperature, reaction time,
and reactants/initiator/crosslinker dosages and ratios influence the flocculation
performance of the modified polysaccharide bio-based flocculants (Kumar, Setia and
Mahadevan, 2012; Salehizadeh, Yan and Farnood, 2017; Sen et al., 2009).

3. GENERAL ASPECTS OF THE FLOCCULATION PROCESS

The most commonly used flocculants in industry today are inorganic and synthetic
organic flocculants; their effective flocculation activity and low cost are still preferable
over the bioflocculants, however, the question of their toxicity to human health and
environmental pollution has been a major concern (Okaiyeto et al., 2016).
Inorganic flocculants include alum, polyaluminium chloride (PAC), aluminium
chloride, aluminium sulfate, ferric chloride, ferrous sulfates or composites obtained by
the mixture of these salts. The flocculation process occurs between the salt of these
metals and the negatively charged suspended particles in a solution. This interaction leads
to a reduction in surface charge and the formation of microflocs, which in turn aggregates
to form larger flocs that can easily settle out of solution (Figure 1). Besides, its
application has caused problems of increased metal concentration or residual aluminium
in treated water, which may presents human health implications and produces large
quantity of sludge whose disposal is another problem (Lee et al., 2014).

Figure 1. Flocculation mechanism by charge neutralization.

Synthetic organic flocculants include polyacrylamide (PAA), polyethylene amine,

and poly(diallyl dimethyl ammonium chloride) (DADMAC). They are commonly derived
from oil-based or non-renewable raw materials, usually have a high molecular weight,
6 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

and present polyelectrolytes in their molecular chain, which enhance their flocculating
effectiveness (Lee, Robinson and Chong, 2014; Okaiyeto et al., 2016; Suopajärvi et al.,
2013). However, problems associated to Alzheimer´s disease (Campbell, 2002) and
carcinogenic and neurotoxic risks (Rudén, 2004) to humans were already reported for
aluminium and acrylamide salt flocculants, respectively. In addition, these type of
flocculants have other limitations related to their relatively high dosage requirement, high
pH sensitivity, and poor efficiency for the coagulation of very fine particles (Sharma,
Dhuldhoya and Merchant, 2006).

3.1. Mechanisms of Polysaccharide Bio-Based Flocculant

Bio-based flocculants (bioflocculants) are natural organic flocculants, i.e., products

based on natural polymers considered of great interest due to their environmentally
friendly behaviour and the promise alternative to replace conventional flocculants (Lee,
Robinson and Chong, 2014).
Natural polysaccharides from different sources have long been described and widely
applied in different fields. In recent decades, there has been an increased interest in the
utilization of these biomolecules, particularly the bioactive ones, for various novel
applications owing to their inherent properties. In fact, they have high potential to be
applied not only in food and fermentation processes, pharmaceutical, cosmetic,
downstream processing, but also in water and wastewater treatment (Liu, Willför and Xu,
2015).
The kinetics of flocculation by polymeric flocculants starts with a suitable
concentration of the flocculant, which is fed into the suspension; then, the
macromolecular flocculant makes contact with the suspended colloids by adsorption
through electrostatic interactions, hydrogen bonding, van der Waals forces, etc. This
leads to a rearrangement of the conformation of the adsorbed polymer such that the
adsorbed suspended particles aggregate to form large flocs that finally settle down
effectively (Bolto and Gregory, 2007; Yang et al., 2016).
The flocculation mechanisms that direct the activities of various polymeric
flocculants, including polysaccharides, can be categorized as charge neutralization,
charge patching, bridging, and sweeping. Acting as bioflocculants, polysaccharides could
destabilise the colloidal particles by increasing the ionic strength and giving some
reduction in the zeta potential and thus a decreased thickness of the diffuse part of the
electrical double layer. In the other hand, they could specifically adsorb counterions to
neutralise the particle charge because they have particular macromolecular structures
with a variety of functional groups, such as carboxyl, hydroxyl, amino, and sulphate,
which can interact with contaminants. For many years, chitosan, tannins, cellulose,
 Flocculant Polysaccharides Mainly from Plants 7

alginate, gums and mucilage have been basing flocculants and attracting wide interest of
researchers (Lee, Robinson and Chong, 2014).
It is predicted that some of the active ingredients in the mucilage are responsible for
the flocculating property. Therefore, extraction becomes the essential step to isolate the
active components that exhibit the flocculating activity from the plants. There are two
methods for the production of plant-derived bioflocculants: (1) solvent extraction and
precipitation and (2) drying and grinding. Briefly, the solvent extraction and precipitation
method starts with the cleaning of the plant materials and then extraction with distilled
water overnight, followed by filtration of the mucilaginous extract and precipitation using
alcohol. In turn, the drying and grinding method starts with the cleaned materials being
dried at high temperature and then grounded and sieved to obtain the bioflocculants.
Bioflocculants prepared under solvent extraction and precipitation conditions displayed
excellent flocculating ability in the treatment of wastewater with direct flocculation
process where no coagulant and pH adjustment are required. Thus, these results suggest
that the extraction step is closely related with flocculating efficiency and plays the major
role to extract the active constituents with high flocculating activity from the plant
materials. In addition, the method used to evaluate the flocculating efficiency of the
plant-derived bioflocculants and optimise the flocculation process is called Jar Test (Lee,
Robinson and Chong, 2014; Salehizadeh, Yan and Farnood, 2017).
The most common flocculation mechanisms related for polysaccharide plant-derived
bioflocculants are charge neutralization (including electrostatic patch effects) and
polymer bridging. These mechanisms are intrinsically dependent on the adsorption of
flocculants on particle surfaces; if there is some affinity between polymer segments and a
particle surface, the adsorption process occurs (Bolto and Gregory, 2007; Lee, Robinson
and Chong, 2014; Salehizadeh, Yan and Farnood, 2017). As already mentioned in section
3, the flocculation mechanism of charge neutralisation is only applicable when the colloid
suspended particles and the added flocculants are of opposite charge. In this case, the
particle surface charge density is reduced by adsorption of the flocculant polysaccharide;
the electrostatic repulsions are reduced to a minimum. This mechanism has been found to
be quite effective for low molecular weight polysaccharide flocculants that tend to adsorb
and neutralize the opposite charges on the particles (Yang et al., 2016).
Considering the great number of neutral polysaccharides acting as bioflocculants, it is
suggested that charge neutralisation is not the mechanism responsible for flocculation. In
fact, bridging is the major mechanism in like-charged or neutral polysaccharide
flocculants, especially when they extends from the particle's surface into the solution for
a distance greater than the distance over which the interparticle repulsion occurs. In this
case, segments of the polysaccharide flocculant are adsorbed onto the particle surface
resulting in loops and tails extending into solution with the possibility of attachment of
dangling polysaccharide segments onto other adjacent particles to form flocs. Bridging
could be due to van der Waals force, static, hydrogen bonds or even chemical reaction
8 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

between some radical groups of the polysaccharide molecule and the particle; in addition,
this mechanism is known to be especially effective for high molecular weight
polysaccharide flocculants (Salehizadeh, Yan and Farnood, 2017).
Electrostatic patch and sweeping are mechanisms that contribute to the process of
flocculation. The first one is caused by polymer flocculants of high charge density
interacting with oppositely charged colloidal particles of low charge density. The net
residual charge of the polymer patch on one colloidal particle surface can adsorb onto the
oppositely charged colloidal particle. The second one forms a bulky precipitate that
enmeshed the colloidal particles, which are then either settled out or flocculate together
with the precipitate (Salehizadeh, Yan and Farnood, 2017).
Without doubts, flocculation is one of the most important and widely used treatment
process of industrial wastewaters. Many authors already highlighted the simplicity and
effectiveness of flocculation-coagulation processes (Lee, Robinson and Chong, 2014;
Okaiyeto et al., 2016; Salehizadeh, Yan and Farnood, 2017; Teh et al., 2016). In what
concerns the use of polysaccharides as bioflocculants, some characteristics could limit
their application for certain purposes. For example, chitosan is soluble in acidic media
and most of polysaccharides are water-soluble in their native form, therefore they cannot
be used as insoluble sorbents (Crini, 2005). In the flocculation process, the
polysaccharide biodegradability can lead to the loss of flocs stability and strength (Singh
et al., 2000). Also, some polysaccharides have reduced/or no bioactivity in certain
conditions (Salehizadeh, Yan and Farnood, 2017). During the time, much attention has
been paid to overcome these disadvantages by modifications of polysaccharides.

3.2. The Effect of Physical-Chemical Factors on Flocculating Activity

The chemical composition, physicochemical properties and flocculation activity of

flocculants depend on the source of the flocculant and its production method.
Characteristics such as charge density and grafting ratio are important in determining the
flocculant performance of the substance; even more, earlier studies indicate that
flocculation efficiency could be enhanced by optimizing the grafting ratio (Salehizadeh,
Yan and Farnood, 2017).
Besides the flocculant charge density, the flocculation mechanism can be mainly
affected by other properties, including dosage, biopolymer molecular weight, the nature
of the colloids and aspects such as concentration, pH, ionic strength, composition of
culture media, and temperature, especially for production of bioflocculants from bacteria
(Chaisorn et al., 2016; Salehizadeh and Yan, 2014).
The effect of pH on flocculating activity is one of the most important external factors
affecting bioflocculant production. Several works reported the flocculating activity of
different bioflocculants being pH-stable in a wide range of pH values, usually from 3 to
 Flocculant Polysaccharides Mainly from Plants 9

10 (Aljuboori et al., 2013; Huang et al., 2015; Li et al., 2013; Li et al., 2014; Liu et al.,
2014b).
The metabolism of microorganisms has direct relationship with cultivating
temperature. Maximum enzymatic activation can be obtained at optimal temperatures
(Zhang et al., 2007). In this way, temperature is an important parameter for bioflocculant
production and cell growth, especially considering bacteria and fungi sources.
Thermotolerant microorganisms have an optimum temperature for growth below 45 ºC,
but ability to grow at higher temperatures; they are also defined as microorganisms able
to grow at 60 ºC and below 30 ºC (Chaisorn et al., 2016).
The flocculant dosage of the polysaccharide is a main aspect for the determination of
the flocculating activity of a colloid system. When the polysaccharide dosage is lower
than the optimum, the degree of flocculation is insufficient in a colloid system that results
in stabilization and charge reversal of the colloidal particles. Thus, there is no
bioflocculant enough to adsorb onto the colloidal particle surfaces to bridge between
these particles. An overloaded dosage of the polysaccharide flocculants increases the
electrostatic repulsion forces between the colloidal particles and increases the distance
between the particles to inhibit floc formation and precipitation (Salehizadeh, Yan and
Farnood, 2017).

Figure 2. Percentage of publication in the last 10 years about the scientific literature dealing with
polysaccharides and flocculants.

4. PLANT FLOCCULANT POLYSACCHARIDES

Polysaccharide flocculants can be classified into three main groups based on their
source: marine, microbial, and plant flocculant polysaccharides. Their natural origin
could be related to crustacean shell wastes, seaweeds, agricultural/forestry feedstocks,
10 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

and microorganisms including bacteria, yeast, fungi, and algae. Figure 2 depicts the
percentage of published articles reporting polysaccharides and flocculants in the last 10
years.
Considering the main flocculants derived from plant-polysaccharides, we
summarized below their properties and the current researches about them.

4.1. Cellulose

Cellulose is a linear polysaccharide consisting of β (1→4) D-glucose; it represents

the most abundant biopolymer on earth, with a global economic importance. The annual
production rate of cellulose is estimated at 1011–1012 ton/year, being this polysaccharide
the major structural constituent of the cell wall of plants, but also derived from different
origins, such as animals and microorganisms. Cellulose has many advantages, including
superior thermal and mechanical properties, in addition to biocompatibility,
biodegradability, and cost-effectiveness (Albuquerque et al., 2017; Salehizadeh, Yan and
Farnood, 2017).
In what concerns the different pathways to obtain cellulose, it is possible to highlight
four: the first one is the most popular and industrially important pathway for isolating
cellulose from plants, which includes chemical pulping, separation, and purification
processes to remove lignin and hemicelluloses. The second pathway is related to the
biosynthesis of cellulose by different types of microorganisms, such as unicellular algae,
fungi, and bacteria. The third pathway is the enzymatic in vitro synthesis starting from
materials such as cellobiosyl fluoride, while the last one is a chemical synthesis, whose
production of cellulose occurs through a ring-opening polymerization of the benzylated
and pivaloylated derivatives (Chen, Cho and Jin, 2010).
The polymeric structure of cellulose was first demonstrated by Staudinger in 1920. In
fact, cellulose is a simple polysaccharide with no branching or substituents in its
homogeneous backbone. The morphological hierarchy of this biopolymer is composed by
elementary fibrils, packaged into microfibrils, and finally assembled into fibres. Within
the cellulose fibrils, there are regions of crystallites, where the cellulose chains are
arranged in a highly ordered structure, and amorphous regions of disordering. Five
allomorphic forms of cellulose have been known based on the location of hydrogen
bonds between and within strands. Natural cellulose, called cellulose I, has two different
crystalline structures (Iα and Iβ). Cellulose may be also found with other crystal
structures including cellulose II, III and IV, being cellulose II the most stable structure,
which can be obtained by alkali treatment of cellulose I (Brinchi et al., 2013; Moon et al.,
2011; Roy et al., 2009).
Cellulose obtained from plants are usually composed of hemicellulose, pectin, lignin,
and other substances, while bacterial cellulose is completely pure with much higher water
 Flocculant Polysaccharides Mainly from Plants 11

content and higher tensile strength (Klemm et al., 2005). The hydroxyl groups are the
most targeted reactive groups on the cellulose main chain, which can fully or partially
react with chemical agents to obtain various derivatives with different substitution
degrees. The obtained derivatives of cellulose are been applied in food, cosmetic,
biomedical, and pharmaceutical industries, however, the application of cellulosic material
is limited due to the difficulty in processing. The high crystallinity degree and rigid
intra/intermolecular hydrogen bonds result in its insolubility in water and most organic
solvents. In addition, cellulose becomes amorphous in water at 320 °C and 25 MPa and
can be converted chemically into its monomeric units by reacting with concentrated acids
at high temperatures (Deguchi, Tsujii and Horikoshi, 2006; Liu, Willför and Xu, 2014;
Zhang, Lin and Yao, 2015).
Problems associated to processing cellulose and its limited solubility are observed for
other natural polymers since they present increasingly application in the industrial
technology. Most polymers do not present biological activity until modifications are
made, so it is important to mention that numerous attempts are being performed to
minimize these certain drawbacks with chemical modifications in the polymeric structure
(Albuquerque et al., 2017). Cellulose can be considered an efficient alternative to
produce environmentally friendly functional materials and chemicals such as
bioflocculants due to its inherent physicochemical characteristics, which are improved by
chemical modifications (Salehizadeh, Yan and Farnood, 2017).
The most recent researches on bioflocculant production are reporting isolated strains
(Lee and Chang, 2018; Shahadat et al., 2017). The unique potential of bioflocculants
produced from microorganisms was first investigated in Levure casseeuse yeast by Louis
Pasteur, and a similar trend in bacterial culture was also observed by Bordet in 1899
(Shahadat et al., 2017). More recently, it was reported that some bacteria can produce
bioflocculants by utilizing biomass from raw materials: a lignocellulose-degrading strain
Cellulosimicrobium cellulans L804 isolated from corn farmland soil presented the ability
to produce bioflocculants by the degradation of lignocellulosic biomass (Liu et al., 2015).
Liu et al. (2017a) reported an alkaliphilic strain Bacillus agaradhaerens C9 bioflocculant
producer by using untreated rice bran as carbon source, and Guo et al. (2018) reported a
novel cellulase-free xylanase-producing bacterium G22 with the potential ability to
directly convert various biomasses to bioflocculants.
The scientific literature dealing with polysaccharide flocculants derived from plants
is still scarce when compared to the works reporting bioflocculants produced from
microorganisms, or even those one reporting commercial polysaccharide bioflocculants.
However, the importance of these kind of studies rises with the increasingly serious
environmental problems especially associated to the discharge of effluents. For example,
Liu et al. (2014a) developed an efficient and eco-friendly flocculant from Phyllostachys
heterocycle bamboo pulp cellulose grafted with polyacrylamide for an effluent from
paper mill. After that, Zhu et al. (2016) optimized the method and employed the co-
12 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

polymer with the metal ions Fe3+, Al3+ or Ca2+ to treat the effluent from a surfactant
manufacturer. Liimatainen et al. (2012) evaluated the effectiveness of a flocculation
treatment based on alum and soluble or nanoparticular anionic derivatives of dialdehyde
cellulose derived from the bleached birch chemical wood pulp obtained in dry sheets of
Betula verrucosa and B. pendula, by studying the removal of colloidal material in a
model suspension containing kaolin.

4.2. Starch

Starch is a linear, biodegradable, inexpensive polysaccharide synthesized in a

granular form by green plants for energy storage over long periods. Starch granules
consist of two major components called amylopectin and amylose, which are composed
of α-D-glucopyranose residues, forming α-1,4-glucosidic bonds in linear structure of
amylose and additional α-1,6-glycosydic branches in amylopectin molecules. Minor
constituents such as lipids, proteins, and minerals are present in starch and the levels vary
with the origin, whose botanical sources include wheat, rice, corn, barley, sorghum,
millet, rye, legumes, banana, mango, potato, cassava and so on. Cassava and maize were
known as the major sources of starch on a commercial scale for a long time (Albuquerque
et al., 2017; Ashogbon and Akintayo, 2014; Salehizadeh, Yan and Farnood, 2017).
Starch is soluble in hot water, but insoluble in cold water, alcohol, or other solvents.
The differences between amylose and amylopectin structures have indeed significant
variance in their properties. Amylose is much more prone to crystallization process,
called retrogradation, and can produce tough gels and strong films, while amylopectin
retrogrades much slower due to its dispersion in water, which results in soft gels and
weak films (Liu, Willför and Xu, 2015; Pérez and Bertoft, 2010).
Starch is an excellent material for biotechnological applications due to its
biodegradable, available, low cost, and versatile characteristics; however, its direct
applications are limited by its poor processability and intrinsic properties, such as
thermal, mechanical, and biological properties. Thus, various chemical, physical, and
enzymatic modifications or blending with other materials has supplied solutions to
achieve properties that are more desirable (Albuquerque et al., 2017).
Comparing starch and other high-performance natural-polymeric material, it is much
cheaper than chitosan (Liu et al., 2017b). Similar to the cellulose, conventional chemical
modifications of starch could be performed based on its primary and secondary hydroxyl
groups, and can be obtained by esterification, etherification, oxidation, and graft
copolymerization (Liu, Willför and Xu, 2015).
Considering that cationic moieties aid flocculation and sterilization through effective
charge attractions, various cationic starch-based flocculants have been developed using
different modification methods and even considering the lack of commercial synthesis
 Flocculant Polysaccharides Mainly from Plants 13

methods and high cost of the cationic monomers. For example, three different starch-
based flocculants, all of them containing the strongly cationic quaternary ammonium salt
groups but at different positions, have been designed and prepared through etherification,
graft copolymerization, or their combination. The effects of chain architectures and
charge properties on flocculation of humic acid (HA) and its floc properties have been
investigated in terms of several environmental parameters including pH, dose, and initial
HA concentration (Wu et al., 2016). Posteriorly, authors obtained four variations with
different cationic contents of two versions of the starch-based flocculants reported in the
previous work. The environmental parameters pH and flocculant dose were evaluated
with the effects of structural factors, i.e., cationic group contents and distributions, on the
flocculation of the hairwork effluents (Du et al., 2017). These starch-based flocculants
were also evaluated by the efficient flocculation of real secondary textile dyeing
effluents. Starch-graft-poly[(2-methacryloyloxyethyl) trimethyl ammonium chloride]
(STC-g-PDMC) and starch-3-chloro-2-hydroxypropyl trimethyl ammonium chloride
(STC-CTA) have been systematically investigated and compared with that of the
traditional inorganic coagulant polyaluminum chloride, and the obtained results are of
significance in guiding the design and selection of a suitable polymeric flocculant in
treating target wastewater (Wu et al., 2017a).
Liu et al. (2017b) prepared a series of cationized starch-based flocculants (ST-CTA)
containing various quaternary ammonium salt groups on the starch backbone by using a
simple etherification reaction. They observed an effective performance for the
flocculation of kaolin suspension, two bacterial suspensions (of Escherichia coli and
Staphylococcus aureus), and two contaminant mixtures (kaolin and each bacterium)
under most pH conditions. Wang et al. (2013) prepared a cationic grafted starch (ST-g-
PDMC) with high flocculation performance for kaolin suspensions and efficient
dewatering of anaerobic sludge. More recently, Huang et al. (2017) reported that the St-g-
PDMC was efficient for the flocculation of kaolin suspensions and for the inhibition of E.
coli by introducing new quaternary ammonium groups. A grafted amphoteric starch-
based flocculant (carboxymethyl-starch-graft-aminomethylated-polyacrylamide, CMS-g-
APAM) was efficiently developed by Huang et al. (2016), in which the cationic groups
were randomly distributed on the polyacrylamide branched chains using Mannich
reaction. A series of cationic starches were developed by incorporating a cationic moiety
(STC-CTA) onto the backbone of starch in presence of NaOH. The flocculation
characteristics of these starches were evaluated in silica suspension and compared with
various commercially available flocculants by jar test (Pal, Mal and Singh, 2005).
It is important to mention that the majority of researches about starch flocculants deal
with the commercial presentation of the polysaccharide. Well-prepared natural
flocculants, with or without chemical modifications, owing to their superior performance
and environmental friendliness, have wide-ranging uses in wastewater treatment,
especially when more effective production techniques are developed and optimized in the
14 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

near future. Additionally, it is important to highlight the volume of the sludge produced
from the treatment associated to them and the considerable reduction in the cost of sludge
disposal.

4.3. Pectin

Pectins are ubiquitous plant polysaccharides. They are present in the cell walls
located in the middle lamella, and primary and secondary cell wall. The chemical
structure of this water-soluble gum is heterogeneous, depending on the origin, location in
the plant and extraction method, being composed at least of 65% of galacturonic acid
units plus rhamnose, arabinan, galactan and arabinogalactan (Müller-Maatsch et al.,
2016; Tamnak et al., 2016). Actually, the uronic acid residues linked through a-1-4-
glycosidic bonds make the main structural polysaccharide motif, while there exist three
pectin’s domains: α-(1-4)-linked linear homogalacturonic backbone (HG) alternating
with two types of highly branched rhamnogalacturonans regions called RG-I and RG-II.
The first region (RG-I) is substituted with side chains of arabinose and galactose units,
while the second one (RG-II) has a highly conserved structure, consisting of the HG
backbone branched with eleven different monosaccharides, including some rare sugars
such as apiose, aceric acid, 2-O-methylxylose, 2-O-methylfucose, 2-keto-3-deoxy-d-
manno-octulosonic acid, and 3-deoxy-d-lyxo-2-heptulosaric acid. In all natural pectins,
some of the carboxyl groups exist in the methyl ester form (Albuquerque et al., 2017;
Liu, Willför and Xu, 2015).
Pectin can be categorized as anionic polysaccharides mainly derived from the cell
wall of plants, and also from food-industrial wastes of fruits, whose main sources are
citrus peels (e.g., lemon, lime, orange, and grapefruit), apple pomace, and sugar beet pulp
(Babbar et al., 2015; Marić et al., 2018). Moreover, pectin from non-conventional sources
has been evaluated, for example, from cocoa husks (Chan and Choo, 2013), mulberry
branch bark (Liu, Jiang and Yao, 2011), jackfruit peel (Xu et al., 2018), faba bean hulls
(Korish, 2015), sisal waste (Yang et al., 2018), watermelon rind (Maran et al., 2014),
pomegranate peels (Pereira et al., 2016), potato pulp (Yang, Mu and Ma, 2018), Ubá
mango peel (Oliveira et al., 2018), pistachio green hull (Chaharbaghi, Khodaiyan and
Hosseini, 2017), pequi peel (Leão et al., 2018), and banana peel (OLiveira et al., 2016).
In the industry, the major pectin component (homogalacturonan) can be obtained using
hot water or chemically by acid extraction, but some innovative extraction approaches
have been developed to improve extraction process and pectin quality (Salehizadeh, Yan
and Farnood, 2017).
Pectin structure determines in a great manner its physicochemical properties and
applications; for example, pectin is efficiently used by the food industry due to their
ability to form gels under certain circumstances and to increase the viscosity of drinks. It
 Flocculant Polysaccharides Mainly from Plants 15

is also widely applied as stabilizers in acid milk products, and some may have other
pharmaceutical uses (Müller-Maatsch et al., 2016).
Pectin has many applications because of its low-cost, read availability, harmless and
green nature. However, it may not provide the proper emulsifying activity because of its
hydrophilicity. The following modification techniques have been recommended for the
improvement of pectin physicochemical and functional properties: chemical modification
(such as saponification, distillation, and esterification), enzymatic modification (by pectin
methylesterases), and physical modification (by heat treatment and microwave) (Tamnak
et al., 2016).
Yokoi et al. (2002) were one of the first authors reporting pectin as an efficient
flocculant for various suspensions. They found that pectin had a flocculating activity,
however, in that time; there has so far been no report about flocculating activity of pectin
and its applicability as a flocculating agent. Their results demonstrated that pectin had
flocculating activity in a kaolin suspension, and this activity was enhanced by the
addition of Al3+ and Fe3+ to the suspension; in relation to organic suspensions such as
cellulose and yeast, pectin acted as flocculant when 0.1–0.2 mM Fe3+ was present in the
suspensions. Other inorganic suspensions of activated carbon and acid clay were
flocculated by pectin in the presence of Al3+ or Fe3+.
A commercial citrus pectin with 60% esterification and the common organic
synthetic flocculant polyacrylamide were characterized and used to optimize the
treatment processes of both flocculants in synthetic turbid waste water. The results
reported by Ho et al. (2010) were important for the industrial application of pectins since
the main concern for industry is to use low flocculant concentrations to achieve
maximum results. In this case, the usage of pectin achieved that goal and proved to be
effective at a low concentration of 3 mg/L. The influence of four commercial citrus
pectins, with different degrees of esterification, and pectin extracted from pomelo (Citrus
maxima) on the stability of indomethacin suspension was extensively investigated by
Piriyaprasarth and Sriamornsak (2011). The results demonstrated that the extracted pectin
had comparable activity to the commercial ones. Moreover, the use of low concentration
of pectin and ferric ions allowed obtaining indomethacin suspensions with suitable
stability and redispersibility. More recently, pectin extracted from Nopal (Opuntia ficus-
indica) was tested to treat synthetic waste water contaminated with metallic ions (Ibarra-
Rodríguez et al., 2017). Authors well-characterized the pectin and then demonstrated its
effectiveness in the removal of the heavy metals by coagulation-flocculation treatment.
It is possible to observe that flocculation properties of pectin have been confirmed by
the scientific literature. In fact, pectin can be utilized as a harmless bioflocculant, since it
is biodegradable, and edible and non-toxic toward humans and the environment;
however, the publications are still discreet when compared to the most popular
polysaccharide bio-based flocculants.
16 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

4.4. Other Polysaccharide´ Flocculants Derived from Plants

Other polysaccharide bio-based flocculants derived from different and non-traditional

plant sources have been reported by the scientific literature (Lee et al., 2014) reported
that all of them have mucilaginous texture, due to the main polysaccharide composition,
and a neutral pH.
Plantago psyllium seed husk is known as Isabgol husk in India and widely used as
laxative. This anionic polysaccharide is extensively studied due to its easy availability
and a very economical cost. Mishra et al. (2002) reported for the first time the
flocculation efficiency of this polysaccharide with different effluents. Their results
demonstrated that this mucilage was found to be a very effective flocculant, capable of
removing almost 85 and 95 percentages of suspended solid (SS) from sewage and tannery
wastewater samples, respectively. The chemical modification of P. psyllium was done by
grafting polyacrylamide (PAM), resulting in a graft-copolymer (Psy-g-PAM) and very
effective flocculant, capable of removing more than 93% of SS, 72% of total dissolved
solids (TDS) and 15.24% of colour from a textile waste water (Mishra et al. 2004a). The
same group (Mishra and Bajpai, 2005) assessed P. psyllium for the removal of dyes from
model textile wastewater containing golden yellow (C.I. Vat Yellow 4) and reactive black
(C.I. Reactive Black 5). The mucilage reduced dye concentration by flocculation and
settling, suggesting a preference of this bioflocculant for colour removal because of its
low capital cost, as well as the lower operating costs when compared to other
technologies (Al-Hamadani et al., 2011) evaluating the psyllium husk from seeds of P.
ovata plant. Psyllium was more effective as coagulant aid when used with aluminum
chloride for the treatment of landfill leachate.
The mucilage extracted from tamarind (Tamarindus indica) seeds was used for the
removal of vat and direct dye in aqueous solution (Mishra and Bajpai, 2006). After the
promising results obtained in this study, Mishra et al. (2006) used ceric ion used initiated
polymerization to obtain a grafted mucilage with acrylamide. The grafted copolymer
(Tam-G-PAM) showed better results for dye removal. The optimal flocculant
concentration was independent of dye concentration within the concentration range
examined. Both the grafted and ungrafted flocculants were more efficient for removal of
azo dyes than for reactive and basic dyes. The application of tamarind seed kernel
powder for the sedimentation of clay slurry was examined in association or not with
starch. Initial qualitative results indicated that tamarind can be successfully used to
reduce the turbidity of clay slurry (Chakrabarti et al., 2008).
Naturally occurring Cassia angustifolia (CA) seed gum was evaluated against
polyaluminium chloride for its coagulation-flocculation ability to remove colour from
synthetic dye solutions. The results demonstrated that the gum was found to be a good
working substitute alone or in conjunction with a very low dose of polyaluminium
chloride for decolourization of acid and direct, but not for reactive dye solutions (Sanghi,
Bhatttacharya and Singh, 2002).
 Flocculant Polysaccharides Mainly from Plants 17

Fenugreek, botanically known as Trigonella foenumgraecum, is a leguminous plant

found in long pods and used for various medicinal purposes. This mucilage was reported
for the very first time by (Mishra, Agarwal and Yadav, 2003), proving to be a very
effective flocculant for sewage treatment and capable of removing almost 97% of SS and
20% of TDS. The flocculation efficiency of this mucilage was also reported as an
efficient flocculant for tannery effluent treatment (Mishra et al., 2004b).
Anastasakis, Kalderis and Diamadopoulos (2009) studied optimal dose, contact time
and flocculation efficiency of Malva sylvestris (mallow) and Hibiscus esculentus (Okra)
mucilages to remove turbidity from synthetic wastewater and biologically-treated
effluent. In the pharmaceutical field, tablets prepared using the mucilages of Hibiscus
rosasinensis Linn. and Abelmoschus esculentus Linn. (Okra). Authors observed that the
higher concentration level of Okra mucilage show a slow and sustained release, and can
be considered as an alternative natural excipient in the modified drug delivery systems.
They also reported that Hibiscus mucilage could be used as a platform for prolonged
release if its binder concentrations are increased (Ameena et al., 2010). Fenugreek (from
T. foenumgraecum) and okra (from H. esculentus) were tested as flocculants for treatment
of textile wastewater. Results showed that the mucilages were capable of removing more
than 90% of SS, 30% of TDS, and 30% of dye using a very low concentration of
polysaccharide (Srinivasan and Mishra, 2008).

5. FLOCCULANT POLYSACCHARIDES OF NON-PLANT SOURCED

Despite the considerable number of polysaccharide bio-based flocculants derived

from plants, especially cellulose and starch, other sources of polysaccharides could be
derived from seaweeds, arthropods, and microorganisms, which might be chemically
modified to improve their flocculation performance.
Natural flocculant polysaccharides from microorganisms, microalgae, seaweeds and
animals (Table 1) are also attractive substitutes for synthetic flocculants due to their
ecofriendly nature, biodegradability, stability, cost-effective production process, and
large availability and yield (Salehizadeh, Yan and Farnood 2017). Microorganisms such
as bacteria and fungi are able to produce extracellular polymeric substances (EPS) with
flocculating activity, including polysaccharides, proteins and glycoproteins.
Bioflocculants from these sources are also potent alternatives for usage in wastewater
treatment, textile, pharmacology and cosmetology industries. This section focuses in
recent advances in polysaccharides bioflocculants from marine and microbial
origins, considering their features, production, flocculant mechanisms and emerging
applications.
 Table 1. List of flocculant polysaccharides from bacteria, fungi, marine sources and arthropods

Species Source Structure Reference

Bacteria
Alcaligenes aquatilis AP4 Palm-oil mill efluent Glycoprotein Adebayo-Tayo and
Adebami, 2017.
Bacillus cereus SK Wastewater treatment plant Glycoprotein Busi et al., 2016
Bacillus subtilis F9 Wastewater sludge Polysaccharide and protein Giri et al., 2015
Bacillus subtilis WD161 Recycled activated sludge of a seafood processing plant Glycoprotein Chaisorn et al., 2016
Bacillus thuringiensis Aquaculture and Nutritional Laboratory, Pearl River Polysaccharide Wu et al., 2017b
Fishery Research Institute, Chinese Academy of Fishery
Sciences, Guangzhou, China
Rhodococcus rhodochrous R- Fungal Culture Collection of Lublin Polysaccharide and protein Czemierska et al., 2017
202
Pseudomonas sp. GO2. Paper mill sludge Polysaccharide, protein and Guo et al., 2017
nucleic acid
Anabaena sp. BTA992 National repository for cyanobacteria and microgreen Polysaccharide and protein Khangembam et al., 2016
algae (freshwater) of the Institute of Bioresources and
Sustainable Development, Imphal, Manipur, India
Paenibacillus elgii B69 Soil Polysaccharide Li et al., 2013
Paenibacillus mucilaginosus Soil Polysaccharide Tang et al., 2015
Klebsiella sp. Activated sludge of a secondary sedimentation tank in a Polysaccharide, protein and Liu et al., 2014b
wastewater treatment plant (WWTP) nucleic acids
Streptomyces sp. MBRC-91 Marine sediment Polysaccharide Manivasagan et al., 2015
Achromobacter xylosoxidans Activated sludge of Bongaigaon oil refinery Glycoprotein Subudhi et al., 2016
Fungi
Curvularia sp. DFH1 Water Polysaccharide and protein Abu-Elreesh and Abd-El-
Haleem, 2014
Aspergillus flavus S44-1 Microbial Culture Collection Unit (UNiCC), Laboratory Polysaccharide and protein Aljuboori et al., 2013
of Industrial Biotechnology, Institute of Bioscience,
University, Putra Malaysia
SGMP1 and SGMP2 White rot Polysaccharides Patel et al., 2014
Aspergillus niger Soil surface Polysaccharide Pu et al., 2018
Species Source Structure Reference
Microalgae/Seaweeds
Scenedesmus obliquus AS-6- Water Polysaccharide Guo et al., 2013
1
Seaweed Commercial Agar bacteriological Cationized agarose Prado et al., 2011
Brown seaweed Commercial sodium alginate Polymethyl methacrylate grafted Rani et al., 2013
sodium alginate
Brown seaweed Commercial sodium alginate Alginate graft polyacrylonitrile Salisu et al., 2016
beads
Gyrodinium impudicum Seawater Sulfated polysaccharide Yim et al., 2007
KG03
Animals
Brachyura Commercial chitosan powder from crab shells Chitosan Xu et al., 2013
Animal source Commercial chitosan 3-chloro-2-hydroxypropyl Agbovi and Wilson, 2018
trimethylammonium chloride
grafted onto carboxymethyl
chitosan
Animal source Commercial chitosan Phenylalanine-modified-chitosan Du et al., 2018
20 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

5.1. Flocculant Polysaccharides in Bacteria

Bacteria have been reported to produce polymeric bioflocculants such as

glycoproteins and exopolysaccharides, as metabolites. Bioflocculants with varied sugar
composition have been isolated from bacteria and they have showed strong flocculating
activity to kaolin clay, sewage water and heavy metals, being a promise for industrial and
biotechnological applications
The bacterium Alcaligenes aquatilis AP4 produces a flocculant glycoprotein which
was isolated by first time from Palm-oil mill effluent (Adebayo-Tayo and Adebami,
2017). The isolated bioflocculant showed a high flocculating activity of approximately
90% and potential for large-scale production.
An exopolysaccharide produced by Bacillus cereus SK showed flocculating activity
to kaolin of 83.4% at 12 mg, and no considerable toxicity to zebra fish in vivo, revealing
the potential of this EPS as a bioflocculant in industrial and biotechnological field (Busi
et al., 2016). Other study revealed a flocculant glycoprotein produced by B. subtilis
WD161, composed of mainly protein and sugar, with flocculating activity to precipitate
suspended solid in palm-oil mill effluent increased by 35% (Chaisorn et al., 2016). A
potent bioflocculant was also purified from B. subtilis F9, showing a composition of
protein and mainly sugar (88.3%). The bioflocculant was able to flocculate small
particles, although its low molecular weight, being considerate an excellent bioflocculant
for industrial applications (Giri et al., 2015). A strain of B. thuringiensis highly-producer
of flocculant was isolated from the biofloc in aquaculture waters. The bioflocculant was a
solid substance with white colour and composition mainly of polysaccharides (Wu et al.,
2017b).
The flocculation potential of the exopolymer R-202 produced by Rhodococcus
rhodochrous was investigated (Czemierska et al., 2017). With a composition of
polysaccharides and proteins, and negatively charged, the exopolymer R-202 promoted
the flocculation of a kaolin suspension with and without the addition of cations, being a
promise for applications in flocculation industrial process.
An exopolysaccharide composed of glucose, glucuronic acid, mannose and xylose
isolated from bacterium Paenibacillus elgii B69 showed high flocculant activity when
tested with kaolin clay, dyeing pigment, heavy metal ion and real wastewater (Li et al.,
2013). A study about polysaccharide-based flocculants (PSBs) produced by P.
mucilaginosus GIM1.16 revealed that the production and feasibility of these
bioflocculants can be improved with the presence of metal ions in culture medium. The
presence of Ca2+, Mg2+ and Fe3+ increased the production and flocculating activity of
PSBs (Tang et al., 2015).
M-C11 is a bioflocculant isolated from Klebsiella sp., composed of sugar (91.2%),
protein (4.6%) and nucleic acids (3.9%). This bioflocculant showed high pH and thermal
stability in kaolin suspension in the pH range of 4.0 to 8.0 and temperature of 20 to 60°C,
 Flocculant Polysacharides Mainly from Plants 21

and a sludge dewatering performance more efficient than inorganic flocculants (Liu et al.,
2014b).
An exopolysaccharide produced by cianobacterium Anabaena sp. BTA992 during its
photoautotrophic growth was evaluated and it was observed bioflocculant activity with
potential properties for commercial production and industrial applications (Khangembam
et al., 2016).
A biomass-degrading bacterium Pseudomonas sp. GO2 was able to produce a low-
cost bioflocculant from untreated corn stover through directly hydrolyzing biomasses.
Biochemical analysis showed a composition of polysaccharides (59%) with uronic acid
(34.2%), protein (32.1%) and nucleic acid (6.1%), and a high flocculation potential
flocculating to harvest the green microalgae Chlorella zofingiensis and Neochloris
oleoabundans (Guo et al., 2017).
Achromobacter xylosoxidans TERI L1 isolated from oil refinery waste produces an
exopolysaccharide bioflocculant encompassed of various functional charged groups with
flocculating activity for dispersed kaolin clay particles in suspension and to adsorption of
multi metals from environment (Subudhi et al., 2016). Thus, this flocculant is a good
candidate for heavy metal removal in contaminated waste-water.
Green synthesis of silver nanoparticles was a method developed based in a
polysaccharide flocculant produced by the marine actinobacterium Streptomyces sp.
MBRC-91. The biosynthesized silver nanoparticles showed strong antibacterial potential
in sewage water, being useful for wastewater treatment (Manivasagan et al., 2015). A
flocculant exopolysaccharide from Arthrobacter sp. B4 was also used for green synthesis
of silver nanoparticles, resulting in nanoparticles highly stable, with antimicrobial activity
and low phytotoxicity (Yumei et al., 2017). This represents the use of microbial
flocculants as a potential tool in many fields, including the medical therapies.

5.2. Flocculant Polysaccharides in Fungi

Conventionally, fungi play valuable roles in environment and are involved in global
biotechnological processes, including the production of enzymes, polysaccharides, lipids
and pigments. Some fungal species have been reported as bioflocculant polysaccharide
producers with high flocculating activities.
An oleaginous fungal Curvularia sp. strain DFH1 demonstrated its potential to
coproduce lipids and an exopolymer with a powerful flocculant activity of 95% (Abul-
Elreesh and Abd-El-Haleem, 2014). The chemical analysis of the exopolymer shows a
composition of protein and polysaccharide. An analysis of infrared spectrum indicates the
presence of carboxyl and hydroxyl groups, typical in sugar derivatives.
A polysaccharide bioflocculant named IH-7 was produced and isolated from
Aspergillus flavus fermentation medium (Aljuboori et al., 2013). The bioflocculant
22 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

consisted basically of protein (28,5%) and sugar (69,7%), including neutral sugar, uronic
acid and amino sugar, and showed a good flocculating activity in kaolin suspension
without cation addition.
Two bioflocculant exopolysaccharides were isolated from fungal cultures named
SGMP1 and SGMP2 found in soil samples collected from New Vallabh Vidyanagar,
Gujarat (India). The flocculating activity was 99%, and the fungal isolates could remove
the cations Al3+ and Fe3+ in ideal concentrations for application in bioremediation of
heavy metals (Patel et al., 2014).
A bioflocculant (MBFA18) produced by Aspergillus niger (A18) isolated from soil
sample using potato starch wastewater as nutrients showed strong flocculant efficiency,
less dosage, sludge amount and moderate treating condition comparable to the chemical
flocculants. A flocculating rate of approximately 90% was achieved for kaolin clay under
optimal cultivation condition (Pu et al., 2018).

5.3. Flocculant Polysaccharides in Algae

Exopolysaccharides from marine algae have been explored due to their potential
applications, including as bioflocculant agent, considered more efficient that commercial
flocculants.
Alginates are polysaccharides firstly isolated from brown seaweeds (Phaeophyta),
and recently identified as produced also by two genera of soil bacteria, Pseudomonas and
Azotobacter (Hay et al., 2013). This polysaccharide is constituted to over than 200
different types varying in length and array of β-D-mannuronic acid (M) and α-L-
guluronic acid (G) monomer units (Albuquerque et al., 2017).
Alginates are biocompatible, biodegradable, and non-immunogenic biopolymer
polyelectrolytes (Yang, Xie and He, 2011). The physical properties in aqueous medium
for alginates depend not only on the M/G ratio, but also on the distribution of M and G
units along the chain. At this moment, it is important to highlight that the main property
of alginates is their ability to retain water. Because of their linear structure, and high
molecular weight, alginates form strong films and good fibres in the solid state. Their
gelling and stabilizing properties are also very important characteristics for alginates; the
stiffness of the alginate chains and the complex with counterions could be attributed to
the composition (M/G ratio) and distribution of M and G units in the chains. The higher
content of G units form stable crosslinked junctions with divalent counter ions (for
example, Ca, Ba, and Sr, unless Mg), so the crosslinked network can be considered a gel.
In addition, the low pH also forms acidic gels stabilized by hydrogen bonds
(Albuquerque et al., 2017; Rinaudo, 2008; Yang, Xie and He, 2011).
 Flocculant Polysacharides Mainly from Plants 23

As an anionic polymer, alginate forms electrostatic complex when mixed with a

cationic polymer. The stability of the formed complex depends on the pH and salt
concentration, but even in the best conditions, the complex is insoluble and thus allows
the formation of fibres, films, and capsules. They have been applied widely in
biomedical, pharmaceutical and biotechnology fields due to its desirable characteristics,
versatility and biocompatibility (Albuquerque et al., 2017; Hay et al., 2013; Salehizadeh,
Yan and Farnood, 2017).
Several studies already reported the excellent flocculation and decolourization
capabilities of alginates (Dao, Cameron and Saito, 2016; Rani, Mishra and Sen, 2013;
Salisu et al., 2016; Yuan and Jia, 2013). Considering the use of alginates for
copolymerization, many works described the preparation and characterization of these
complexes, also proposing many applications to them, especially for the most
investigated polyelectrolyte complex based on alginate and chitosan (Ortiz et al., 2018;
Yang et al., 2016).
The sulfated exopolysaccharide p-KG03 produced by the red-tide marine microalga
Gyrodinium impudicum KG03 showed bioflocculant activity more than 90% in kaolin
suspension, and a flocculation rate higher than commercial flocculants as polyacrylamide
and zooglan (Yim et al., 2007).
The microalga Scenedesmus obliquus AS-6-1 is able to carry out flocculation and
self-flocculation due to its cell wall-associated polysaccharides, composed by glucose,
galactose, rhamnose and fructose with molar ration of 8:5:3:2:1. Fast flocculating action
was observed on suspended cells of S. obliquus and Chlorella vulgaris in the presence of
0.6 mg/L flocculating agent (Guo et al., 2013).
Agarose is a polysaccharide obtained of certain red seaweed of easy availability.
Cationized agaroses were synthesized through the reaction of agarose with 3-chloro-2-
hydroxypropyltrimethylammonium chloride in alkaline medium, and showed flocculation
activity in assays with colloid kaolin suspensions comparable with commercial cationic
polyacrylamides, representing an alternative for water treatment (Prado et al., 2011).

5.4. Flocculant Polysaccharides in Animals

5.4.1. Chitosan
According to Wu et al. (2012), chitin is the second most abundant polymer after
cellulose. It is widely synthesized by many living organisms such as fungi, yeasts, algae
and squid pen, being found in exoskeletons of crabs, lobsters, shrimp and crab shells, as
well as insect cuticles. Depending on its source, chitin occurs mainly as two allomorphs,
namely α (the most abundant) and β forms. The chemical structure (poly-β-(1→4)-N-
acetyl-D-glucosamine) of chitin can be partially degraded by acid to obtain a series of
24 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

oligomers namely oligochitins. Under alkaline conditions, a partial deacetylation of chitin

results in one of the most chitin derivative in terms of applications: chitosan, biobased
polymer with many bioactivities, including antitumoral, antimicrobial, fungicidal and
immunotherapeutic properties, eliciting chitinase, and regulating organism growth
(Albuquerque et al., 2017; Salehizadeh, Yan and Farnood, 2017).
Chitosan is a linear, cationic polysaccharide composed of randomly repeating units of
β(1→4)-linked 2-acetamido-2-deoxy-D-glucopyranose (A-unit; the neutral sugar unit
GlcNAc) and 2-amino-2-deoxy-D-glucopyranose (D-unit, the positively charged sugar
unit GlcN). The amount of D-units in chitosan is often more than 60%. It shows different
degrees of deacetylation (DD) between 40–90% and is available commercially in various
molecular weights ranging from 50,000 to 2,000,000 Da. The contents of the A-units in
chitosan can vary from 0.7 (70% acetylated) to 0 (0% acetylated, all units charged).
Therefore, chitosan can be considered as ampholyte (enriched in A-units) or
polyelectrolyte (enriched in D-units). Chitosan is the only commercially available water-
soluble cationic polymer, property that emerges as an advantage when compared to the
marked insolubility of chitin in all usual solvents. The water solubility of chitosan is
related to the positive charges on its primary amino groups, which allows the interaction
of this cationic biopolymer with anionic molecules such as glycosaminoglycans (GAG)
and proteoglycans (Albuquerque et al., 2017). Moreover, chitosan may be considered one
of the most promising natural substitutes for commercial synthetic polymer flocculants
due to its unique flocculation properties, also associated to the presence of these primary
amino groups, as well as the potential to employ chemical modifications in this
carbohydrate polymer.
Chitosan was used as flocculant agent for harvesting of the microalga Chlorella
sorokiniana, with flocculation efficiency reaching over 99% at dosage of 10 mg chitosan
per gram algal biomass and reducing the volume by 20-50 folds (Xu et al., 2013). Recent
studies reported by the scientific literature show the production, characterization, and
application of microwaves (Cui et al., 2017), grafted-copolymers (Wang et al., 2016) and
other hibrids (Dharani and Balasubramanian, 2015; Jia et al. 2016; Sun et al., 2017) of
chitosan and, for example, lignin, ions, antibiotics, acrylamide. Chitosan was
functionalized by graft with 3-chloro-2-hydroxypropyl trimethylammonium chloride,
generating a new flocculant agent with potential for wastewater treatment (Agbovi and
Wilson, 2018). Other chitosan-based flocculant denominated phenylalanine-modified
chitosan showed higher removal potential of the antibiotics norfloxacin and tylosin in
turbid water than commercial flocculants, being a potent contaminant removal (Du et al.,
2018). Great flocculant activity was reported to a terpolymer based on chitosan and lignin
(chitosan-acrylamide-lignin), which showed maximum percentage removal of 99.3% and
67.0% for the dyes reactive orange C-3R and methyl orange, respectively (Lou et al.,
2018; Lu et al., 2017).
 Flocculant Polysacharides Mainly from Plants 25

6. POLYSACCHARIDE FLOCCULANTS FOR

WASTEWATER APPLICATIONS

Wastewater from different sources can be treated with flocculating polysaccharides,

as shown in Figure 3. Contaminant particles dispersed in wastewater, such as dye
molecules, can form flocs with these polysaccharides.

Figure 3. Flocs formation in wastewater by flocculant polysaccharide.

6.1. Dyes Removal

Different types of industries, such as textiles and plastic, use dyes to colouring their
products, generating large volumes of colored wastewater. However, dyes are substances
capable of absorbing ultraviolet light and consequently decrease this absorption by
photosynthetic organisms and reduce dissolved oxygen content of aquatic environment;
characterizing, thus, a serious environmental pollutant (Rangabhashiyam et al., 2013;
Bello et al., 2017). Coagulation-flocculation has been considered a cost effective
technology with excellent color removal ability (Verma et al., 2012).
A polysaccharide with cationic behaviour that is widely used for removal of
negatively charged dyes in wastewater is the chitosan; anionic dyes can be adsorbed
electrostatically with the protonated amine groups of chitosan (Lee et al., 2014; Vakili et
al., 2014). For example, about 99% of the acid blue 92 dye was removed by chitosan,
allowing recovery of the biopolymer from formed flocs with the dye using 0.1 M NaOH
and with subsequent reuse of chitosan in acetic acid solution (Szygula et al., 2009).
Additionally, modifications in chitosan structure as well as its association with other
polymers have been increasingly studied to improve flocculant action in dye removal.
A chitosan-based cationic polymer was elaborated through the modification of this
polysaccharide by (3-chloro-2-hydroxypropyl) trimethylammonium chloride; such
26 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

chitosan modification was useful for dye melanoidin removal from industrial wastewater
(Momeni et al., 2018). In an attempt to elaborate amphoteric flocculants, capable of
eliminating both anionic and cationic dyes, Yang et al. (2013) prepared grafting
flocculants, formed by carboxymethyl chitosan-graft-polyacrylamide (CMC-g-PAM).
These authors have found that the ability to remove methyl orange (anionic dye) and
basic bright yellow (cationic dye) initially occurs by charge neutralization between
produced flocculant and dyes, but flexibility of PAM graft chains allowed formation of
large and insoluble flocs with net-like structure, allowing an easy sedimentation in water
treatment (Yang et al., 2013). The synthetic monomer of acrylamide was used to graft
chitosan and lignin (CAMCL) forming a ternary graft copolymer, since these two
polymers together provide more functional groups to attract dyes, such as orange C-3R,
methyl orange and acid black-172. In addition, flocculating mechanism of CAMCL
occurred by the charge neutralization, bridging and sweeping effects (Cui et al., 2017;
Lou et al., 2018).
Cellulose has a regular structure and matrix formed by hydroxyl groups presenting a
high degree of polymerization and susceptible to undergo chemical modifications making
it a relevant flocculant (Kono and Kusumoto, 2015; Ferreira et al., 2016). Quaternized
celluloses and ampholytes of this polysaccharide in different degree of cationic
substitution, for example, had flocculating capacity against anionic dyes, such as acid red
13 and acid blue 92 (Kono and Kusumoto, 2015; Kono, 2017). However, carboxymethyl
cellulose undergoing the grafting process with hydrolyzed polyacrylamide, generating
CMC-g-HPAM, was able to remove the methylene blue cationic dye. The highest
removal efficiency occurred under alkaline conditions where the carboxyl groups of
CMC-g-HPAM were deprotonized, forming -COO- groups, allowing electrostatic
interactions with methylene blue, favoring such flocculation (Cai et al., 2013).
Gum polysaccharides also went through the grafting process, such as the ghatti-
crosslinked-polyacrylamide (Gg-cl-PAAM) hydrogel synthesized capable of adsorbing
and removing cationic (rhodamine B and brilliant green) and anionic (methyl orange and
congo red) dyes (Mittal et al., 2018). Xylan, a xylose polymer, is another polysaccharide
which, when phosphorylated, acted as a flocculant removing more than 95% of cationic
ethyl violet dye (Liu et al., 2018). A macromolecule originated using corn ethanol
wastewater, named compound biopolymer flocculant (CBF), has in its composition more
than 80% polysaccharide; CBF presented methylene blue removal, whose flocculation
process occurs primarily by adsorption bridging and charge neutralization by its polar
functional groups (Xia et al., 2018). The exopolysaccharide (EPS) produced and purified
from Paenibacillus elgii B69, was able to promote high rate of decolourization for
methylene blue and red X-GRL positive dyes, while less than 50% for anionic and
neutral dyes (Li et al., 2013).
 Flocculant Polysacharides Mainly from Plants 27

6.2. Cleaning of Heavy Metal Ions

Heavy metal ions, such as lead, copper and zinc, can contaminate water due to
natural processes, since such elements can be stored in the soil and transported by surface
waters. In addition, anthropogenic sources contribute to this type of pollution, such as
industrial activities including mining and coal combustion (Kobielska et al., 2018). These
ions are extremely toxic and can cause serious human health problems, such as damage to
the neurological system, blood disorders, toxicity in different organs and even death
(Shtenberg et al., 2015). In front of this serious scenario of contamination with heavy
metal ions, to remove these contaminants in wastewater is very important to leave it free
or in non-toxic concentrations of these ions; such removal can be carried out through
bioflocculants, which include polysaccharides (Salehizadeh and Yan, 2014).
Some polysaccharides, such as xanthan gum and starch, possess many –OH groups
and act as metal ion coordination sites; while other polysaccharides, for example,
chitosan with –NH2 groups and sodium alginate apart from –OH groups –COO- can bind
to the metal ions (Kolya and Tripathy, 2013). Chitosan, for example, promoted
simultaneous adsorption of heavy metal ions (iron, nickel and copper) and salt anion
(sulfate). At pH range 5 to 6, chitosan has protonated and unprotonated amine groups, in
which the latter group form chelate complexes with salt cations and salt anions
are adsorbed through electrostatic interaction on chitosan (Mende et al., 2016).
Polysaccharide inulin underwent a carboxymethylation process to synthesize
carboxymethyl inulin (MIC), which removed metal ions in a kaolin suspension, reducing
the total iron, chromium VI and manganese (II) content. The mechanism proposed for
this chelation is the interaction with the cations by oxygen of the ether linkage and by
oxygen of the carboxylate group through five membered ring formation (Rahul et al.,
2014). A pectin, extracted from O. ficus-indica had affinity to the different metallic ions
(Ca2+, Cu2+, Zn2+, Cr3+, Ni2+, Pb2+ and Cd2+); since pectin is a linear molecule composed
of galacturonic acid, whose hydroxyl and carboxylic groups can be responsible for this
affinity with the metal ions studied (Ibarra-Rodríguez et al., 2017).
Flocculating power of polysaccharides produced by different bacteria also has been
determined. EPS produced by bacteria, such as cyanobacteria, has negative charges and
can act as a chelating agent of heavy metal ions with positive charge (Bhunia et al.,
2018). The EPS bioflocculant purified from P. elgii B69 also had high capacity to adsorb
Al3+, followed by Pb2+, Cu2+ and Co2+ (Li et al.; 2013). A flocculant system was made
using EPS from Rhizobium sp. and polyethyleneimine (PEI), which was able to remove
Cu2+. It was proposed that such process occurred firstly by binding Cu2+ to negatively-
charged groups, mainly carboxylate, of the EPS followed by link of the EPS-Cu2+
complex with PEI resulting in firm and large flocs with easy removal (Escobar et al.,
2015). While more than 90% of heavy metals (Zn2+, Cd2+, Pb2+, Cu2+ and Ni2+) were
adsorbed to the EPS of glycoprotein nature produced by Achromobacter xylosoxidans
28 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

when grown in culture medium supplemented with these ions (Subudhi et al., 2016). A
polysaccharide molecule (MSI021) was purified from Bacillus cereus; and characterized
as a bioflocculant heavy metal remover. A solution containing heavy metal salts (HgCl2,
ZnCl2 and CuSO4) with a bioluminescent bacterium, Vibrio harveyi, was treated with
MSI021. The presence of MSI021 in the solution allowed the V. harveyi growth, which
continued with its luminescent phenotypic expression, differently from the absence of
MSI021, in which there was a decrease in the luminescence of this bacterium; such
phenomenon was possible due to this bioflocculant molecule that mitigated heavy metal
toxicity (Sajayan et al., 2017).

6.3. Treatment of Pulp and Paper Industry Effluents

About 42% of global industrial wastewater is produced by pulp and paper industry.
The processes in the paper manufacturing involve pulping, bleaching and papermaking;
in fact, in all these steps effluents composed of different chemical species are generated
(Toczyłowska-Mamińska, 2017). Wastewater from pulp and paper industry have a
diversity of organic and inorganic contaminants, such as resin acids, fatty acids,
adsorbable organic halide (AOX) and total suspended solids (TSS) besides being able to
have high content of biological oxygen demand (BOD) and chemical oxygen demand
(COD) (Ashrafi et al., 2015; Kumar et al., 2015). These undiluted untreated effluents are
toxic to aquatic organisms and their treatment can be carried through the flocculation
physicochemical method, which includes the flocculating polysaccharides (Kumar et al.,
2015).
Thus, a copolymer flocculant prepared by grafting (2-methacryl-oyloxyethyl)
trimethyl ammonium chloride onto chitosan (chitosan-g-PDMC) has been used to treat
pulp mill wastewater. This novel cationic chitosan-based flocculant promoted efficient
removal of COD, turbidity and lignin as well as water recovery. Grafting of chitosan
showed in favour of the double electric layer compression, charge neutralization and
improved the sweep-floc effects (Wang et al., 2009). In another work, raw and undiluted
pulp and paper mill effluent was treated with Cassia obtusifolia seed gum, which
removed COD and TSS an up to 36.2 and 86.9%, respectively; and the flocs formed after
the treatment had a morphology highly fibrous-like and aggregate (Subramonian et al.,
2014). Polysaccharides from potato starch have been modified by benzylation and
insertion of hydroxypropyl-trimethyl ammonium (HPMA) moieties of different
substitution degrees, generating benzyl 2-hydroxypropyl-trimethylammonium starch
chloride (BnHPMAS). Flocculation performance was tested for BnHPMAS in a model of
paper industry wastewater; these amphiphilic starch derivatives removed contaminants
from recycled paper processing, so named stickies, including total organic carbon (TOC)
(Genest et al., 2015). A cationic bioflocculant has been produced through polymerizing
 Flocculant Polysacharides Mainly from Plants 29

(2-methacryloyloxyethyl) trimethyl ammonium chloride and xylan, generating xylan-

DMC, removed COD, lignin, sugar, BOD and turbidity from pulp mill wastewater; the
flocculation of particles occurred by bridging mechanism (Chen et al., 2018).
Natural dicarboxyl cellulose flocculant (DCC) has been synthesized and presented
great flocculation activity to the effluent from paper mill; DCC combined with CaCl2
reduced turbidity, COD and BOD through the process coagulation-flocculation of the
effluent suspensions (Zhu et al., 2015). Turbidity reduction from paper mill effluent can
allow the recovery of cellulose fibers. Mukherjee et al. (2014) tested flocculant action of
guar gum, xanthan gum and locust bean gum polysaccharides and found that guar gum
reduced the turbidity and recovered more fibers than the other carbohydrates of a
synthetic effluent. In addition, guar gum-based nanocomposite, so called g-GG/SiO2, has
been synthesized by grafting polyacrylamide chains on a guar gum (g-GG) followed by
in situ nano silica incorporation on g-GG surface. This nanocomposite was able to reduce
pollutant contents of paper industry, such as TSS and turbidity, and the flocculation in
acidic environment occurs due to electrostatic and patching mechanism; under alkaline
conditions the flocculation process happens mainly by bridging (Pal et al., 2015).

6.4. Removal of Agricultural and Food Contaminants

More than 300 million ton of organic waste are produced each year by agricultural
and food processing industries in the United States; sustainable treatment of such waste
has become a worldwide challenge (Sheets et al., 2015). These organic substances found
in wastewater from such industries comprise TSS, COD, organic colloids, sludge,
oil/grease, beyond dissolved inorganics (Teh et al., 2016). In addition, nitrogenous
compounds, especially ammonium nitrogen, can be present in large quantities in food
industry wastewater (Zhukova et al., 2011). Phosphate is a common constituent of
agricultural fertilizers and such chemical specie can contaminate effluents; excess of
phosphorus in the aquatic environment can contribute to the process of eutrophication
(Adesoye et al., 2014). Therefore, agricultural and food contaminants in wastewater need
to be removed, and such process can be performed by flocculant polysaccharides.
Wastewater is generated in potato starch process that is considered one of the most
polluted in food industry. Potato starch wastewater (PSW) has been treated with a
bioflocculat complex denominated MBF917, produced by Rhizopus sp. M9 and M17,
such macromolecule is composed of more polysaccharide than protein. MBF917
removed the turbidity and COD from the PSW (Pu et al., 2014). A bioflocculant has been
extracted from Paenibacillus polymyxa, and such compound showed to be thermostable
with indicative of its main backbone being formed by polysaccharide; this bioflocculant
was still used to treat PSW by reducing COD and turbidity efficiently (Guo et al., 2015a;
Guo et al., 2015b). Another bioflocculant of polysaccharide nature has been produced
30 P. B. S. Albuquerque, W. F. Oliveira, P. M. dos Santos Silva et al.

and isolated in the fermentation liquor of Rhodococcus erythropolis. This bioflocculant

was able to reduce COD, ammonium and total phosphorus concentrations of the PSW
medium (Guo et al., 2018).
Rice starch has been used to perform the treatment of agroindustrial wastewater from
palm oil mill; this carbohydrate (at dosage 0.55 g/L) promoted removal of almost 90%
TSS from palm oil mill effluent. Flocs produced with this treatment had a compact
structure with smoother surface, designating formation of denser and larger flocs by
bridging flocculation mechanism (Teh et al., 2014). Removal of orthophosphate (Pi), a
very common contaminant in water bodies by agriculture, was obtained using the
carboxymethyl chitosan grafted with 3-chloro-2-hydroxypropyl trimethylammonium
chloride, CMC-CTA. The CMC-CTA flocculant efficiently removed Pi and the turbidity
under acidic conditions of a simulated wastewater (Agbovi and Wilson, 2018). Therefore,
it can be inferred that different polysaccharides are been used for a great versatility in
wastewater applications. The composition of EPS molecules produced by
microorganisms has been revealed which usually are of glycoprotein nature, and had their
flocculating performance well established. While polysaccharides with structure already
characterized have undergone chemical changes to improve their flocculant capacity.

CONCLUSION

The purpose of this review was to approach scientific literature dealing with
polysaccharide bio-based flocculants with special emphasis for those derived from plants.
Moreover, this review emphasized aspects such as extraction, purification, modification,
characterization, and the broad range of applications of natural flocculants in industry.
The most current scientific publications demonstrate the industrial concern for the
use of new substrates, besides the reduction in the costs of bioflocculants´ production and
the constant awareness about the substitution of inorganic flocculants for the innovative
ones. It was remarkable to note the preference of bioflocculants in comparison with
inorganic flocculants due to their low cost, availability, biodegradability and biosafety; in
addition, they can be obtained from renewable resources and their application is directly
related to the improvement of quality of life.
Several studies investigated the flocculating activity of polysaccharide bio-based
flocculants in wastewater treatment; they are technically promising as flocculants with
high removal efficiency of suspended solids, total dissolved solids, turbidity, colour and
dye. However, its development is limited with variation of flocculating efficiency, short
shelf life, and high production cost.
The scientific literature dealing with flocculants polysaccharides derived from plants
is still scarce when compared to bioflocculants produced from microorganisms, or even
those reporting commercial polysaccharide bioflocculants. Regarding issues such as
 Flocculant Polysacharides Mainly from Plants 31

biosafety and the sake of ecology, more qualitative and quantitative research are
necessary for further exploitation and applications of polysaccharide bio-based
flocculants in different industries.

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BIOGRAPHICAL SKETCH

Name: Priscilla Barbosa Sales de Albuquerque

Affiliation: Centro de Tecnologias Estratégicas do Nordeste (CETENE), Recife, PE,

Brazil.

Education: PhD

Research and Professional Experience: Biochemistry and Biotechnology

 Flocculant Polysacharides Mainly from Plants 45

Professional Appointments: Extraction and characterization of biomolecules derived

from natural sources; experience with polysaccharides, immobilization of
biomolecules, drug delivery systems and potential applications in pharmaceutical,
food and cosmetic fields.

Publications from the Last 3 Years:

1. https://www.sciencedirect.com/science/article/pii/S0141813016324394
2. https://www.ncbi.nlm.nih.gov/pubmed/28433769
3. https://www.ncbi.nlm.nih.gov/pubmed/28916381
4. https://www.ncbi.nlm.nih.gov/pubmed/28987799
5. https://www.ncbi.nlm.nih.gov/pubmed/28545372
6. https://www.ncbi.nlm.nih.gov/pubmed/26840177
7. https://www.ncbi.nlm.nih.gov/pubmed/29632933
8. https://www.ncbi.nlm.nih.gov/pubmed/26428171
9. http://www.journals.ufrpe.br/index.php/JEAP/article/view/1701
10. http://www.sciencedomain.org/abstract/13497
11. http://www.aimspress.com/article/10.3934/molsci.2016.3.386
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 2

COAGULATION AND FLOCCULATION

WITH PLANT EXTRACTS

Jesús Manuel Epalza Contreras1 and Johan Jaramillo Peralta2

1
Environmental Engineering, Universidad de Santander,
Bucaramanga, Colombia
2
Universidad de Valladolid, Valladolid, España

ABSTRACT

The treatment of water in a conventional way includes the operations of coagulation

and flocculation as a fundamental part in the removal of solids and other substances that
are mixed with water, especially organic and inorganic solids with sizes less than 0.2 mm
and densities similar to those of water. This forms a perfect mixture difficult to separate
by natural sedimentation; in these cases the addition of coagulants is needed, which
breaks the stability of the mixture and segregates the particles in the form of flocs, so that
they can be separated by density difference. Reactive substances are used for this reason,
such as aluminum sulfate, ferric chloride and aluminum polychloride; these substances
are derived from industrial chemical reactions, which entail the use of natural resources
and energy to obtain them. In addition to the environmental cost of the production
processes, there is the problem of the final disposal of the thickened sludge, because due
to its aluminum or iron content, the sludge can be harmful to the environment, especially
for plants and animals. To reduce this impact, it is necessary to carry out these operations
with substances that do not represent a danger to the environment; in this case, it has been
proven that there are plant extracts that can generate the same reaction of segregating the
solids and other substances of the water, by means of the reaction of the natural
biopolymers of some plants, within which the Moringa oleifera, Melocactus sp, Opuntia
sp, Cicer arietinum L, Aloe sp and others with destabilizing activity of particles can
generate turbidity and color in water. These plant extracts can efficiently perform the
coagulation and flocculation operations without the environmental risk of residual sludge
with high concentrations of aluminum or iron.
48 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

Keywords: biopolymer, coagulation, flocculation, water treatment

INTRODUCTION

The coagulation and flocculation processes are decisive in the treatment of water,
both in purification and in wastewater, due to their efficiency of removing suspended
particles from the water. To carry out the coagulation, it is necessary to add substances
called coagulants, which come from chemical reactions that have been industrialized.
The most important task is to generate drinking water, which can help the inhabitants
of underdeveloped countries to improve their living conditions, as it is established in the
main policies of the United Nations; but reducing the environmental impact of these
activities and improving the coagulation and flocculation processes with coagulants of
organic origin are easy to produce in agriculture and without metals that can affect the
ecosystem from the ground.
This document shows in a general way the advances in the processes of coagulation
and flocculation with plants, and an emphasis is made on showing those made in recent
years.
The presented study is based on experiences with biopolymers derived from plants,
especially from Melocactus sp, Opuntia sp, Stenocereus griseus, Cereus forbesii, Aloe
arborescens, Aloe vera and Cicer arietinum. Melocactus sp, Stenocereus griseus, and
Cereus forbesii have not been studied very much by other authors.
Conventional water treatment processes include coagulation and flocculation,
especially that which is chemically assisted, for both drinking water and wastewater; the
removal capacity of these operations are effective and possess a speed that could have
been used, especially with waters that have high concentrations of organic matter, and for
this purpose reagents have been used such as aluminum sulfate, which is usually obtained
from the reaction of aluminum hydroxide with sulfuric acid. Another reagent used is
ferric chloride, which is obtained via the reaction of chlorine gas on heated iron, and an
additional reagent that has been used in recent years is the aluminum polychloride that is
obtained through a reaction of aluminum with hydrochloric acid in an aqueous solution.
The reactions associated with the coagulants are determined by some parameters such
as the pH, the temperature of the water, and the concentrations of the solids that are going
to be complexed to form flocs that can be separated by density difference inside the
mixture. When the flocs are separated from the mixture, sludge is generated that must be
thickened and then disposed of within the parameters of a waste management plan. These
residues will have a high concentration of aluminum and iron, respectively, according to
the type of coagulant used, whether it be aluminum sulfate, aluminum polychloride or
ferric chloride. The final disposal of this sludge usually comes with some difficulties,
 Coagulation and Flocculation with Plant Extracts 49

because due to their load of aluminum or iron they are considered toxic for soil in high
concentrations.
The disposal problems of this sludge, which generally has a high concentration of
organic matter, generates environmental impacts when they are discharged into soils or
bodies of water, changing the natural microbiota and affecting the species that have
contact with these high concentrations of aluminum and iron. Because of this, different
products of vegetable origin have been studied, which have properties similar to those of
aluminum or iron compounds and generate coagulation and flocculation with organic
compounds. Some examples of these cases include Melocactus sp, Opuntia dilleni,
Stenocereus griseus, Cereus forbesii, Aloe arborescens, Aloe vera and the Kabuli
Chickpea (Cicer arietinum L); these plants have shown activity for the flocculation of
substances with small particle sizes (below 0.2 mm), which generally cannot be separated
by natural sedimentation.
The sludge derived from the coagulation and flocculation processes with plant
extracts has a completely organic composition, which means that they can be digested by
microorganisms and transformed into carbon, nitrogen and phosphorus substances that
can be incorporated into the corresponding biogeochemical cycles, along with the
absence of toxic metals for the soil, or with safe concentrations for this vital resource; this
technological alternative transforms water treatment into a less aggressive process with
the environment, taking into account that most of the waste generated in drinking water
and domestic waste treatment is sludge.
The extraction systems of plant biopolymers have different methodologies, which are
easy to apply, proven and are part of already standardized unit operations. Taking into
account that different parts are harvested from each plant, we must understand that for
most of the plants their use concerns the majority of the biomass, whereas when we speak
of Cicer arietinum, we are using only their seeds. This diminishes its use, taking into
account the weight ratio of the plant and the mass used for the preparation of the
coagulant.
The operations developed to determine the efficiency of each plant extract in the
coagulation and flocculation operations are defined within the established jar tests, and
some of them have Z potential measurements (a measure of the magnitude of the
repulsion or attraction between the particles).

CONVENTIONAL COAGULANTS

Coagulants are substances with the capacity to break the stability of aqueous
mixtures, causing suspended particles to be grouped in flocs, to be later segregated by
gravity or by air injection separators.
50 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

Coagulation is a process that allows us to separate a quantity of microorganisms in

water due to the agglomeration in the flocculation process, which attracts bacteria and
protozoa, agglomerates them and precipitates them in the process of sedimentation.
The substances commonly used to perform coagulation are aluminum sulfate, ferric
chloride and aluminum polychloride, which are the results of different reactions.

 Aluminum sulfate [Al2 (SO4) 3.x18 H2O]: This substance is an inorganic salt
resulting from the controlled mixing of sulfuric acid and aluminum or bauxite,
according to the purity and concentration desired for the final product.
 Ferric chloride [FeCl3. x6 H2O]: It is an inorganic salt, which is a product of the
reaction between metallic iron and dilute commercial hydrochloric acid; this
reaction is generally carried out with the industrial leftovers of hydrochloric acid,
resulting in a high concentration ferric chloride.
 Aluminum Chloride [Al2 (OH) 3Cl or Al2 (OH) 3Cl3]: This salt is the result of
the reaction of aluminum, bauxite or other minerals with high concentrations of
aluminum and hydrochloric acid; from which different types of aluminum
polychloride are produced, such as those that are named as PAC or PAFC,
depending on their specification. Their alumina content may be between 13% and
17%.

The use of these salts has an environmental impact, especially regarding the
generation of large quantities of sewage sludge with high concentrations of metals,
usually either aluminum or iron.
The final disposal of this sewage sludge is complicated, as their high aluminum
content makes them toxic to the soil and plants that will grow in this site (Casierra Posada
Fanor, 2007).
Less commonly, other coagulating substances may be used, such as:

 Ferric sulfate [Fe2 (SO4) 3.x H2O].

 Ferrous sulfate [FeSO4.x 7 H2O].
 Magnesium carbonate [MgCO3 x 3H2O].
 Aluminate of sodium [NaAlO3].

PLANTS WITH COAGULATION CAPACITIES

Production of coagulants with plants is a technique used in a traditional way (Moa

Megersa, 2014), but the possibility has recently been raised of using several plant species
 Coagulation and Flocculation with Plant Extracts 51

to extract the coagulant biopolymer, preferably in a similar way to the one that performs
with conventional coagulants.
The plants studied for the raw material of this process are diverse; within them, we
have Moringa oleifera, Opuntia spp, Jatropha curcas, Cicer arietinum, Melocactus spp,
Stenocereus griseus, Cereus forbesii, Aloe arborescens, Aloe vera, Cicer arietinum and
Coccinia indica; these plants have presented good performance to coagulate and
flocculate raw water, leading to better sanitary conditions.
In the case of this text, we explored the species Melocactus sp, Opuntia sp,
Stenocereus griseus, Brain forbesii, Aloe arborescens, Aloe vera and Cicer arietinum L,
which were tested in their efficiency for coagulation and flocculation with raw water.

SELECTION OF PLANTS WITH THE POTENTIAL TO PRODUCE

BIOPOLYMERS FOR COAGULATION AND WATER FLOCCULATION

Plants with the capacity to generate biopolymers with coagulants and flocculants
have been studied over the last few decades, especially Moringa olerifera, Opuntia spp,
Cicer arietinum, and others that have demonstrated coagulant capacity as part of the
traditional empirical knowledge of indigenous communities.

Source: http://cactiguide.com/cactus/?genus=Stenocereus&species=griseus.

Figure 1. Stenocereus griseus.

The selected plants took coincided with those referenced and others present in semi-
arid regions in Colombia, such as La Guajira in northern Colombia and the banks of the
Chicamocha River in the northeastern region; the species not studied are Stenocereus
griseus (Figure 1), Cereus forbesii (Figure 2), Aloe arborescens, and Aloe vera; and the
one already studied was Cicer arietinum (Moa Megersa, 2014).
In the case of Opintia sp, they are present in the Colombian regions already named
and sufficiently studied, in the same way Aloe vera and Aloe arborecens were selected
52 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

(Figure 3); the Kabuli Chickpea (Cicer arietinum) is reviewed with studies
already elaborated on by other authors (Hildebrando Ramírez Arcila, 2015). It
should be noted that some species of Opuntia spp are used as part of the animal and
human diet in communities of northeastern Colombia in semi-desert areas
(Fernández, 2002).

Source: http://www.kakteensammlungholzheu.de/en/cereus_forbesii.html.

Figure 2. Cereus forbesii.

Figure 3. Aloe arborescens.

To have a better selection, we reviewed the massive presence of these plants and they
were not part of the list of plants in danger of extinction.

BIOPOLYMER EXTRACTION METHODOLOGIES

Extraction of the Plants Melocactus sp, Opuntia dilleni, Stenocereus griseus,

Cereus forbesii, Aloe arborescens and Aloe vera

The extractions of each plant have particularities, taking into account the usable parts
in the search of their coagulating capacity; the extractions are segregated into two types:
The plants that are used in all their foliage are Melocactus sp (Figure 6), Opuntia dilleni
 Coagulation and Flocculation with Plant Extracts 53

(Figure 4), Stenocereus griseus, and Cereus forbesii, which belong to the family
Cactaceae while the other part corresponds to Aloe arborescens, and Aloe vera, which
belongs to the Xanthorrhoeaceae family, with superior Aloe classification.
For the cactus and aloe species, an extraction methodology was used with different
operations that are:

 The selection of parts of the plant for cutting: The operation of cutting parts of
the plant is done by taking into account the mature parts, with the presence of
thorns in the case of cacti and a hard external surface, similar to the criteria used
for the animal or human consumption of Opuntia spp species. In the case of the
Aloe species, the maturity of the leaves is considered, with the presence of
perimeter spines, as this shows the possibility of isolating the crystals of the
plant. Figure 4 shows a part of the penca or cladodes of a catus Opuntia sp with
skin and thorns, but the part to be used is the vascular tissue of the plant,
eliminating the skin and spines. For the sampling of the species of Aloe (Figure
5) garden plants were considered, which are cultivated in a homemade way,
taking into account the age of the plant, as it must have enough leaves with
enough crystals, and it must not present any evidence of contamination or
parasites, especially the characteristics of the green color of the leaves, absence
of external insects and total absence of organisms associated with diseases of the
plant.
 Cut and transport to the laboratory. The cutting of the parts of the plant takes
measurements (Figure 7) for transportation to the site of the coagulant; the
transport must be carried out in refrigeration to avoid possible contamination
with environmental fungi or other organisms that can significantly change the
composition of the parts obtained.

Source: http://www.fichas.suculentas.es/Almacenfichas/903/903.html

Figure 4. Opuntia sp.

54 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

Source: http://www.fichas.suculentas.es/Almacenfichas/903/903.html.

Figure 5. Aloe vera.

 Weighing of the gross material. The weighing of the material is carried out on a
25 kg scale to determine the weight of the sample taken and then determine its
performance according to its humidity.
 Cutting of thorns and removal of the bark. To perform the extraction of the
coagulant, the cut parts are taken and the thorns are removed (Figure 8) along
with the skin of the cacti, also called the epidermis, which is the external hard
part of the pads or cladodes in the case of cacti and leaves in the case of Aloe
species.

Source: http://www.tephroweb.ch/kuas/melo.htm.

Figure 6. Melocactus sp.

 Cutting of clean material. In order to follow the extraction process, the tissue of
the plants is cut; this is done to improve the loss of moisture in the plant,
increasing the contact surface with the atmosphere.
 Drying the pieces. The drying of the material is carried out outdoors (Figure 10),
and it is important to note that the region in which these operations are carried
out are from a warm tropical climate with low humidity. In these areas, the
 Coagulation and Flocculation with Plant Extracts 55

temperature ranges between 20°C and 35°C with relative humidity between 50%
and 70% on average.

Source: authors.

Figure 7. Opuntia sp and Cereus forbesi samples.

Source: authors.

Figure 8. Removal of thorns.

Source: authors.

Figure 9. Cutting of the plants.

56 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

 Weighing of dehydrated material. The material is kept outdoors, taking care that
it is not hydrated by rain, and the drying time is between 48 and 96 hours
depending on the temperature and relative humidity of the place where they are
dried.
 Grinding of the material. The grinding of the dried material is done with a food
processing device (Photo 1) and then it is passed through a mill that pulverizes
the material. The final characteristics of the material are similar to the raw
materials used in the coagulation and flocculation process, that is, a presentation
similar to the presentation of type A and B aluminum sulfate and aluminum
polychloride in their solid presentations. The liquid presentation is not sought
because it is an organic material with nutritional characteristics for filamentous
fungi and bacteria, which would require a procedure for its sterilization. For any
application of excessive heat, a degradation of the biopolymers will be carried
out, which is a condition that impairs its performance in coagulation.
 Material screening. The material after being macerated is taken to a 1 mm sieve
(Figure 11), which separates the thick parts that cannot be diluted efficiently in
an aqueous solution; this characteristic is based on the solubility of complex
organic substances, such as the vascular tissue of these cacti and Aloe plants.
 Weighing of the material of interest. The material obtained from the plants
Melocactus sp, Opuntia dilleni, Stenocereus griseus, Cereus forbesii, Aloe
arborescens and Aloe vera is weighed to have a reference of its performance in
relation to the weight, with respect to its use.

Source: authors.

Figure 10. Desiccation of the specimens.

 Coagulation and Flocculation with Plant Extracts 57

Figure 11. Material screening.

EXTRACTION OF CICER ARIETINUM COAGULANT

The seeds of the anionic coagulant Kabuli Garbanzo (Cicer arietinum L) were
selected with the following procedures being performed.

 Washed. The seeds selected without evidence of the presence of fungi or yeasts were
washed with large amounts of water to eliminate impurities related to bulk handling.
In terms of their fractionation and packaging, they can take other materials such as
small sand stones or other grain waste.
 Drying. After washing, they were dried for two days in the sun, taking into account
that they cannot be wetted by rain or other water sources.
 Crushed. The material was crushed using a mixer (Oster) and the resulting powder
was sieved with a No. 200 sieve to obtain a very fine powder for storage in plastic
containers to avoid hydration and subsequent use in the preparation of the solutions
of the coagulant.

TESTS OF THE BIOPOLYMERS OBTAINED

After obtaining the biopolymers, we proceed to prepare solutions of these extracts, to

perform jar tests and thus determine the coagulant and flocculant capacity of natural
biopolymers.
The tests are carried out with water of natural characteristics with different types of
solids, which include natural tannins, solids smaller than 0.2 mm, organic matter and
other substances typical of raw or residual waters.
58 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

Preparation of the Solutions for Coagulation and Flocculation of Melocactus

sp, Opuntia dilleni, Stenocereus griseus, Cereus forbesii, Aloe arborescens and
Aloe vera

After obtaining the biopolymers of the plants Melocactus sp, Opuntia dilleni,
Stenocereus griseus, Cereus forbesii, Aloe arborescens and Aloe vera, the preparation of
the solutions is carried out to realize the tests of the jars, which prove the action of the
coagulants and flocculants.
The flocculant preparation process was weighed for 1 gram of a biopolymer; then,
1000 ml of distilled water was added and manual agitation was carried out until it was
completely diluted.

Preparation of Coagulation Solutions of Cicer arietinum L

1% of the solutions was prepared by adding 10 g of the anionic coagulant in 1000 ml

with Milli-Q® water, obtaining a solution of 10,000 mg / l, after which stirring was
carried out for one hour to homogenize the mixture. In this solution, the tests were carried
out to know the adequate doses to treat the synthetic water prepared in the laboratory.
To prepare the synthetic water, laboratory clay was used for the preparation of
samples of turbid water for all the experiments. Twenty grams of clay were added to one
liter of distilled water. The suspension was gently stirred for 1 hour on a magnetic stirrer
in order to achieve a uniform dispersion of the clay particles. The suspension was
allowed to stand for 24 hours to achieve complete hydration of the clay. This clay
suspension served as a stock solution using distilled water to prepare water samples with
a turbidity of 200 NTU.

Control Parameters in Rockrose Tests

To test the coagulating and flocculating effect of the biopolymers of Melocactus sp,
Opuntia dilleni, Stenocereus griseus, Cereus forbesii, Aloe arborescens, Aloe vera and
the Kabuli Chickpea (Cicer arietinum L), jug tests were performed as provided in ASTM
D2035: 08
The control parameters normally used in the efficiency of a coagulant are pH,
turbidity and color, which are governed by standardized methodologies and which
determine the ability to remove solids from water based on their behavior. pH and its
spectrophotometric absorbance in the case of turbidity and color at different wavelengths
were studied.
 Coagulation and Flocculation with Plant Extracts 59

In accordance with the regulations in force in Colombia, the tests were carried out
taking into account the technical standards adopted by the country for each test.
The removal obtained is processed in terms of percentages for the comparison of pH.
It is observed that it complies with the provisions of the national standard for this
parameter; and in the case of turbidity and color, the percentage of removal is taken into
account, which is directly proportional to the decrease in absorbance in each test.

RESULTS OF JAR TESTS WITH NATURAL COAGULANT BIOPOLYMERS

For the results of the tests of the biopolymers, natural coagulants are collected in 4
main tests: The first ones refer to the pH, the turbidity and the color, and the last one
takes the coagulants with good performance and measures the Z potential.
Turbidity and color in water is related to the presence of substances or
microorganisms, which is directly related to its quality to be consumed or used in other
ways; the results offer an overview of the potentialities of all the plants for the
coagulation and flocculation processes, with different degrees of efficiency.
To control the efficiency of coagulation and flocculation as a function of pH, the pH
was taken after coagulation, taking into account that the initial pH of the water is 7.2. The
pH results are shown in Graph 1, showing that the coagulant that most affected the final
pH was the biopolymer of Melocactus sp, which brought the pH up to 6.2. The data show
a standard deviation of 0.3
Graph 1 shows that the only one that did not affect the pH in the jar test was
Stenocereus spp, while the others lowered the pH moderately to values of 7 or 6.8.
The turbidity results (Graph 2) showed that the best biopolymers to remove these
solids associated with turbidity were Melocactus spp and Cicer arietinum, which showed
turbidity removals greater than 95% and 97% respectively. The data show a standard
deviation of 3.1.
The results of the other biopolymers showed a removal capacity greater than 88%
and up to 92%, which shows the effectiveness of these biopolymers with water that has a
neutral pH.
All the biopolymers tested showed effective action in the removal of turbidity, in a
range between 88% and 97%, with some differences and affectations to the pH of the
sample at the final moment of the jar test.
For the case of the color results (Graph 3), we can see a good activity of all the
biopolymers, taking into account that the one that showed the best performance in the
removal of the color was the biopolymer of Melocactus spp, with a performance greater
than 96%, which was then appreciated for the two species of Aloe sp with a removal
greater than 95%. Opuntia sp, Stenocereus sp and the Kabuli Chickpea (Cicer arietinum
L) displayed performances greater than 94%, and the last one was evidenced
60 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

by Cereus forbesi with a performance greater than 92%. The data show a standard
deviation of 1.4.

Source: authors.

Graph 1. Results of the pH test for studied coagulants.

Source: authors.

Graph 2. Turbidity removal with each coagulant studied.

 Coagulation and Flocculation with Plant Extracts 61

Source: authors.

Graph 3. Color removal with each coagulant studied.

Table 1. Potential Z of Melocactus sp and Kabuli Chickpea

(Cicer arietinum L)

pH Zeta Potential (mv) Melocactus sp Zeta potential (mV) Kabuli

Chickpea
3 -1,7 15,1
4 -3,9 -2,5
5 -12,9 -11,8
6 -19,6 -19,8
7 -22,9 -21,9
8 -22,9 -21,9
9 -27,1 -24,5
10 -29,4 -27,1
Source: authors.

The results show a greater efficiency in the removal of color; the best
performance of these tests lies in the Melocactus, which was the best in removing
both turbidity and color, but also had the highest incidence of pH. The
other biopolymer with a high performance is the Kabuli Chickpea (Cicer
arietinum L), whichshowed a good turbidity and color removal capacity with very little
pH affectation.
The zeta potential of the biopolymers shows similar values in the range of pH 4 and
pH 10, which may indicate a similar activity with solid particles of a small size, such as
those that generate turbidity and color in the water.
62 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

The zeta potential measurements were made for the biopolymers with better
performance, which showed the following results.
Table 1 shows the results of the Z potential of Melocactus sp and the Kabuli
Chickpea (Cicer arietinum L), with a pH between 3 and 10.
It is important to note that all plants used have the potential to treat water; the
efficiency differences can be associated with the affinity for different particles and their
extraction form.

CONCLUSION

All the extracts showed turbidity and color removal with efficiencies higher than
88%, which indicates that the extraction methodologies conserve the coagulant and
flocculant capacity of each plant.
The biopolymers of Melocactus sp, Opuntia sp, Stenocereus griseus, Cereus forbesii,
Aloe arborescens, Aloe vera and Cicer arietinum have an activity for coagulation and
water flocculation.
The plants with the best performance in the removal of turbidity and color were
Melocactus sp and Cicer arietinum, with the best percentages of elimination for solids of
small size in water.
The potential zeta measurements for the extracts of Melocactus sp and Cicer
arietinum have similar values in the range of pH 4 to 10, which shows a similar activity
for the suspended particles of the water used in the tests.
The biopolymers of the plants Melocactus sp, Opuntia sp, Stenocereus
griseus, Cereus forbesii, Aloe arborescens, Aloe vera and Cicer arietinum can be
a viable alternative for the treatment of drinking and residual water, in terms
of the replacement of sulfate for aluminum, aluminum polychloride and
ferric chloride; this would allow for decreasing the amounts of dissolved metals in the
drinking water of humans and animals, especially with the aluminum residues associated
with diseases such as autism and Alzheimer’s.

ACKNOWLEDGMENTS

We are very grateful to the University of Santander in Colombia, which granted a

large part of its resources to complete this book chapter. Additionally, we give thanks to
the University of Valladolid, Spain, for allowing one of our authors to help create this
chapter. We also would like to thank all the people of the universities involved who
personally helped by committing to this work and research.
 Coagulation and Flocculation with Plant Extracts 63

CONFLICT OF INTEREST

The authors declare that there is no conflict of interest regarding the publication of
this document.

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exact, physical and natural sciences,] 353-365.
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Hildebrando Ramírez Arcila, J. J. (2015). Agentes naturales como alternativa para el
tratamiento del agua. Revista de Facultad de Ciencias Básicas [Natural agents as an
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BIOGRAPHICAL SKETCH

Jesús Manuel Epalza Contreras

Affiliation: He has developed as a professor at different universities in the region,

such as the Industrial University of Santander in the Chair of Biotechnology and
Industrial Microbiology and the Antonio Nariño University; He is currently Associate
Professor of the University of Santander UDES - Bucaramanga Colombia, and is in
66 Jesús Manuel Epalza Contreras and Johan Jaramillo Peralta

charge of the subjects of Water Purification, Environmental Remediation and

Ecotechnology, in the Environmental Engineering career.

Education: Microbiologist, from the University of Pamplona, Colombia, MSc

Advanced Energy Systems of the University of Santander UDES, and candidate for MSc
in Science Management, Technology and Scientific Policy with the University of Seville,
Spain

Research and Professional Experience: Recognized by COLCIENCIAS as

Associated Researcher, in the 2017 national call.
Inventor of “System and Method of Wastewater Treatment in Nucleos
Decentralizados” patented in Colombia and recognized with the resolution number 52486
of December 2012, of the Superintendence of Industry and Commerce of Colombia.
Inventor of “System that includes drinking water treatment plant, autonomous,
compact, automated and assisted with photovoltaic solar energy and said plant” filed with
the number 15-196159, of December 21, 2015.
In his professional beginnings, he worked as an analyst of potable and residual water
in different accredited laboratories and has worked as a soil analysis consultant for the
determination of biocorrosion in the petroleum industry.
He has developed work with water treatment, since 2002, being a participant in
different designs of drinking water treatment plants and wastewater treatment plants,
especially in physical, chemical and biological treatment operations, designing anaerobic
and aerobic bioreactors, with the respective culture of microorganisms for the start of the
bioreactors; to making improvements of coagulation, flocculation and sedimentation
systems, of potable and residual water treatment plants in populations of Colombia and in
food industries.
It has also developed wastewater treatment works with anaerobic digestion, with
biogas production, and its use; He has also worked as a consultant and researcher of
residual biosolids in wastewater treatment of the food industry, testing different
technologies for their energy use and also in the improvement of biosolids to be used as
raw material in agriculture and livestock.
He has developed different technological development projects at the University of
Santander, especially in the treatment of drinking and residual water, combining different
disciplines; At the moment, it is differencing projects of masters, the ones that stands out
the development of guidelines of public policy for the recovery of the Simití swamp in
the department of Bolivar, and the provision of the service of potable water in the
municipality of Los Santos in the Santander department, with a Master’s degree in Public
Management and Government from the University of Santander.
 Coagulation and Flocculation with Plant Extracts 67

Professional Appointments: He is part of the Environmental Research Group

GAIA-UDES, as a researcher in the drinking water and basic sanitation line since 2010.
He is part of the group of research professors of Environmental Engineering at the
University of Santander UDES since 2008

Honors: Graduated Distinguished from the Master in Advanced Energy Systems in

2016 by the University of Santander.
Recognized by the University of Santander in 2018, due to the results of the call for
researchers in Colombia of Colciencias in 2017.

Publications from the Last 3 Years:

Evaluation of the efficiency of bio-polimers derived from Desertic plants as flocculation

agents. Chemical Engineering Transactions, 2016.
Obtaining a Cementitious Geopolymer from Gold Mining Tailings with Possible Use in
Engineering Applications. American Journal of Engineering Research (AJER), 2018.
Comparison of the Efficiency of Biopolymer Derived from Melocactus Sp and
Aluminum polichloride (PAC) in the Process of Crude Water Flocculation. Chemical
engineering transactions, 2018.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 3

THE PROCESS OF WATER TREATMENT

WITH ALUMINUM SULPHATE ASSOCIATED WITH THE
APPLICATION OF THE CACTUS OPUNTIA
COCHENILLIFERA

Higgor Henrique Dias Goes1,

Rita de Cássia Pereira de Souza1 and
Joseane Débora Peruço Theodoro2
1
Environmental Engineering Student - UTFPR- Londrina,
Londrina, Brazil
2
Department of Environmental Academic, Federal Technological University of
Paraná, Londrina, Paraná, Brazil

ABSTRACT

The present study aimed to use the organic polymer from Opuntia cochenillifera
cactus associated with the addtion of aluminum sulfate to treat the water of a lentic body
(Igapó II Lake - Located in Londrina in the state of Paraná and Brazil) applying
coagulation, flocculation, sedimentation and filtration processes. The tests were
performed with static a Jar-test reactor, aiming to evaluate the removal of parameters of
turbidity and of apparent color, besides monitoring the pH variation. Regarding the
physico-chemical characteristics, there was little influence of the Opuntia cochenillifera
cactus in the pH parameter tending to neutrality. Therefore, the study showed that the use
of an organic coagulant together with aluminum sulphate is effective for the removal of
parameters of apparent color and of turbidity (95 and 94%, respectively).

Keywords: organic coagulant, aluminum sulfate, water treatment

70 H. H. Dias Goes, R. de C. P. de Souza and J. D. Peruço Theodoro

1. INTRODUCTION

Water intended for human consumption must be available in a potable way, in which
it does not present health risks and denote the control and monitoring of its quality,
according to the drinking standard established by the Ministry of Health Ordinance No.
2,914/2011 (Brazil 2011).
In order to reach the standards established through the water regulatory agencies and
to distribute it to a community, it is necessary to carry out its treatment (Howe et al.
2016). In a conventional manner, the water treatment consists of the following steps,
which are shown in Figure 1.
The clarification of the raw water occurs in the removal of solids present and
associated with the parameter of turbidity, being removed by operations and processes as:
coagulation, flocculation, sedimentation and filtration. At the beginning of the
coagulation process, the coagulant is added under intense stirring in the water to be
treated, ensuring uniform distribution of the product. This phase has the objective of
destabilizing the impurities present in the medium, with the addition of a coagulant,
capable of minimizing or eliminating the repulsive forces present between them. The
most commonly used coagulants are metal salts based on aluminum or iron (COMUSA
2017; SANEP 2018). In order to replace or reduce the use of conventional coagulants, the
natural agents with equal or superior efficiency when compared to metallic ones,
generating a sludge with less toxicity, have a lower cost and greater abundance of plants
as a source of extraction available (Yin 2010; Theodoro et al. 2013).

Sorce: SANEP 2018.

Figure 1. Representative scheme of water treatment.

Derived from a renewable resource and its arrangement promotes simple organic
degradation, mucilage of cacti for extraction of polymers is used to eliminate water
turbidity (Pichler; Alcantar, 2012).
 The Process of Water Treatment with Aluminum Sulphate ... 71

The forage cactus Opuntia cochenillifera, originating in Mexico and spread in the
northeast of Brazil, has several uses, ranging from animal and human feeding, to
landscaping projects and dye extraction. Among its characteristics, it has the xeromorphic
form with a cylindrical stem and its branches known as palms.
Thus, the objective of this chapter was to analyze the water quality of Lake Igapó II,
found in the city of Londrina-PR, according to the parameters of apparent color, of
turbidity and of pH, and then to perform the treatment of these waters through the
coagulation/flocculation/ sedimentation/filtration process using the organic polymer from
the Opuntia cochenillifera cactus associated with the addition of Aluminum Sulphate,
verifying the possibility use of these lentic waters for urban supply.

2. METHODOLOGY

The water samples were taken from a lentic system from Lake Igapó II, located in the
city of Londrina, state of Paraná, Brazil.

Fonte: Goes et al., 2017.

Figure 2. Spatial location of the collection point in Lake Igapó II.

72 H. H. Dias Goes, R. de C. P. de Souza and J. D. Peruço Theodoro

Situated at 550 meters above sea level, one has the coordinates of the collection point
are 23°19'42'' S and 51°10'11'' W. Figure 2 spatially represents the location of the
highlighted point for sample collection. To perform the tests, 72 liters of Lake Igapó II
water were collected and stored in previously sanitized plastic gallons.
The methodology for the extraction of the polymer present in the cactus occurred in
two stages: The first consisted in the removal of the spines and the bark, and thus cuts
were made to reduce the size of the cactus to be used in the liquidification of the contents
inside the palms ("pulp").
In the second, two liters of the extraction solution (required to extract the polymer
from the viscous solution formed after liquefying) were prepared by the attachment of
one liter of distilled water with 4g of 1% sodium chloride solutions and one liter of
distilled water with 10g sodium hydroxide 0.10mol L-1 (Zara et al. 2012).
The ratio was one ml of cactus prepared to 2.5 mL of the extraction solution, the
mixture of which was homogenized on magnetic stirrer Nova Ethics, model 114, for 40
minutes. The resulting viscous complex was packed in glass sanitized vials and stored
under refrigeration at 5°C until assays were performed on the JarTest equipment (Goes et
al. 2017). Some of these steps are shown in Figure 3.
The amount of water collected was divided into three gallons with the same volume,
to carry out the tests at different pH values, being basic, neutral and acidic sample in
order to work with the parameters of color and of turbidity after the operations and
processes in the three tracks. To adjust the pH values with the purpose of making it
acidic, the hydrochloric acid solution (HCl) with1 M concentration was used.
For the basic pH sample, the sodium hydroxide solution (NaOH) with 1 M
concentration. The neutral pH value of the crude sample was also measured.
For the experiment, the dosages of coagulant cacti (Opuntia cochenillifera) were 1mg
L , 4mg L-1 and 7mg L-1 with the addition of Aluminum Sulphate in 1mg L-1, 4mg L-1
-1

and 7mg L-1, respectively.

Source: Goes et al. 2017.

Figura 3. a) Opuntia cochenillifera Palm; b) Removing the shell; c) Cut for further liquefaction; d)
Storage of viscous complex.
 The Process of Water Treatment with Aluminum Sulphate ... 73

Table 1. Gradient of rotation of the mixing rods and time of action

Quick Mix Slow Mix Slow Mix Slow Mix Slow Mix
1 2 3 4
Gradient (s-1) Gradient (s-1) Gradient (s-1) Gradient (s-1) Gradient (s-1)
450 90 52 40 30
Time (mim) Time (mim) Time (mim) Time (mim) Time (mim)
00:10 02:00 02:35 02:40 05:40
Source: Higashi (2016).

In the water treatment trials by the coagulation/flocculation/ sedimentation processes,

the six-proof jar-test equipment was used with a rotating regulator of the mixing rods.
Table 1 shows the parameters used in coagulation/flocculation processes.
For coagulation/flocculation/sedimentation assays, a sedimentation time of 10
minutes was adopted. At the end of the set time for sedimentation, all the samples were
collected and the parameters of apparent color, of turbidity and of pH were measured.
For the filtration test an iron structure adapted to fix the sand filters below the jar-test
was used so that the water leaves the apparatus directly to the filters. The filter beds are
made of Polyethylene Terephthalate (PET) of approximately 10 cm of internal diameter,
forming a fixed bed model with downward flow, with six columns in parallel (Goes et al.
2017), as observed in Figure 4.

Source: Goes et al. 2017.

Figure 4. Filtration process.

74 H. H. Dias Goes, R. de C. P. de Souza and J. D. Peruço Theodoro

The tubes are 25cm long with 15cm filled by sand, 3cm filled with crushed
stone and 2cm filled with cotton. The granulometric of the sand used in the 6 filters
were the same, in the range of 0.60 to 0.85mm. The slope used at the exit of the
jar-test was 70º for two minutes, followed by another two minutes with the
angulation of 60º and another two minutes with 50º (Di Bernardo et al. 2003). The water
to be treated entered the top of the column and its supernatant was removed from the
side.
The grains of the filter bed were retained by a coffee filter together with the cotton
and stone, present at the lower end of the column.
For all variations of cactus concentrations and pH, the parameters of apparent color
and of turbidity were determined according to the Standard Methods of Examination of
Water and Wastewater (APHA 2012). In this work we used a statistical planning with
two factors (independent variables), the concentration of coagulant cacti and three
parameters responses (apparent color, turbidity and pH), the tests were performed in
duplicates.

3. RESULTS

The collected raw water had a neutral pH of 7.65. For acid pH and base, we have the
values obtained of 4.87 and 9.08 respectively.
After the treatment, the calculations were performed to remove the parameters
apparent color and turbidity, thus obtaining data after the coagulation/flocculation/
sedimentation operation/filtration process and the overall process/global operation
(coagulation/flocculation/sedimentation/ filtration), besides the monitoring of pH. The
values found are shown in Table 2.
Figures 5, 6 and 7 show the removal of the parameter apparent color and
turbidity for the coagulation/flocculation/sedimentation processes, only filtration
and for the complete process (coagulation/flocculation/ sedimentation/filtration),
respectively.
Analyzing Figure 5, it can be seen that the best results presented are associated with
the neutral pH values (tests 2, 5, 8, 11, 14 and 17), removing around 30% of the color and
turbidity. It is possible to verify that in tests 1, 3, 4, 6, 9, 10, 12, 15 and 18 the dissolution
of the organic coagulant cactus occurred in the solution, causing the increase of the
dissolved solids and, thus, and, thus, justify the negative values of percentage of removal
of the color parameter.
 Table 2. Numerical organization of the tests for different levels of coagulant concentration and pH with real values and
results of parameters color, turbidity and pH

Assay Cactus pH Color Color Color T T T pH pH

(mgL-1) (C/F/S) % (Fi) % (C/F/S/Fi) % (C/F/S) % (Fi) % (C/F/S/Fi) % (C/F/S) (C/F/S/Fi)
1 1 4,87 -7,0 77,6 76,1 17,2 74,8 79,5 6,5 7,51
2 1 7,85 32,6 66,1 77,2 32,5 62,8 75,1 7,5 7,95
3 1 9,08 -19,4 68,4 62,3 2,1 65,5 66,6 6,92 7,88
4 4 4,87 -11,0 93,7 93,0 9,3 93,1 94,0 7,27 7,54
5 4 7,85 29,6 76,5 83,4 31,3 72,4 81,2 7,12 7,97
6 4 9,08 -21,4 84,1 80,7 6,2 81,0 82,4 7,59 7,71
7 7 4,87 14,5 91,2 92,5 24,0 91,1 93,4 7,86 7,43
8 7 7,85 38,3 82,2 89,0 37,4 79,1 87,0 7,68 7,97
9 7 9,08 -14,2 79,5 76,6 5,4 78,1 79,6 7,24 7,78
10 1 4,87 -7,2 91,4 90,8 5,7 89,8 90,6 6,35 7,58
11 1 7,85 32,9 89,6 93,0 30,8 85,1 89,8 7,46 7,95
12 1 9,08 -8,5 78,1 76,3 4,6 78,9 80,1 7,5 7,93
13 4 4,87 -0,6 95,0 95,0 15,4 92,6 94,0 6,97 7,6
14 4 7,85 33,0 87,2 91,4 33,5 83,4 89,1 7,74 7,8
15 4 9,08 -22,1 89,1 86,6 -0,8 86,2 86,3 7,59 7,82
16 7 4,87 22,3 89,6 91,9 31,2 89,3 92,9 6,1 7,53
17 7 7,85 75,0 63,4 90,9 70,6 62,5 89,1 7,42 7,84
18 7 9,08 -20,2 80,7 76,9 3,8 77,7 78,8 7,65 7,85
OBS: C / F / S = coagulation / flocculation / sedimentation; Fi = filtration;
C / F / S / Fi = coagulation / flocculation / sedimentation / filtration.
Source: Own authorship.
76 H. H. Dias Goes, R. de C. P. de Souza and J. D. Peruço Theodoro

Figure 5. Removal of apparent color and turbidity after Coagulation/Flocculation/ Sedimentation

process.

Figure 6. Removal of apparent color and turbidity in the filtration process.

The filtration process was more efficient for the basic and acid pH values, allowing a
removal of more than 80% in the two analyzed parameters. With the completion of the
complete process, it was noticed that in general the tests obtained values of color removal
and turbidity above 80%. Thus, by analyzing the presented results, it can be concluded
that both the coagulation/flocculation/sedimentation processes (Figure 5) and the
filtration process (Figure 6) are important for the removal of color and of turbidity
present in the raw water. Comparing the values obtained with those required by
Ordinance 2,914/2011 of the Ministry of Health (Brazil 2011), the tests did not meet the
VMP (maximum allowed value) of apparent color and of turbidity. The results of pH
 The Process of Water Treatment with Aluminum Sulphate ... 77

variation after coagulation/flocculation/ sedimentation processes and after the

coagulation/flocculation/ sedimentation/ filtration process using cactus (Opuntia
cochenillifera) are presented in Figures 8 and 9.

Figure 7. Removal of apparent color and of turbidity after Coagulation/Flocculation/

Sedimentation/Filtration process.

Figure 8 demonstrates that after coagulation/flocculation/sedimentation the pH of the

acid and basic assays tend to reach neutrality during the process and the assays that were
in this range remain.

Figure 8. pH after the coagulation/flocculation/sedimentation process.

After also passing through the filtration (Figure 9), the water tends to obtain values
closer to the neutrality.
This occurs due to the ability of the sand particles to adsorb the components present
in the water, neutralizing the pH. Comparing the values obtained with those required by
78 H. H. Dias Goes, R. de C. P. de Souza and J. D. Peruço Theodoro

Portaria 2,914 / 2011 (Brazil 2011) of the Ministry of Health, all the tests met the pH
VMP (6 to 9).

Figure 9. pH after the coagulation/flocculation/sedimentation/filtration process.

CONCLUSION

According to the tests carried out in the study, the coagulant solution obtained from
the Opuntia cochenillifera cactus associated with the addition of aluminum sulphate was
efficient in the treatment of water from the Lake Igapó lentic system, tending to maintain
its pH close to neutrality. The results for color removal and turbidity were promising, but
did not reach compliance with the values required by the Ministry of Health Ordinance
2,914/2011 (Brazil 2011), not meeting the VMP (maximum value allowed) for both
parameters.

REFERENCES

APHA – American Public Health Association. Standard Methods for the Examination of
Water and Wastewater. 22 ed. Washington, 2012.
Brazil, Ministry of Health. 2,914, dated December 12, 2011. Available at:
<http://bvsms.saude.gov.br/bvs/saudelegis/gm/2011/prt2914_12_12_2011.html>.
Accessed on: January, 2018.
COMUSA, Water and Sewage Service of Novo Hamburgo. Water treatment, 2017.
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Accessed on: Feb. 2017.
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Di Bernardo, L.; Mendes, C. G. N.; Brandão, C. C. S.; Sens, M. L.; Pádua, V. L. Water
Treatment for Direct Filtration. Di Bernardo, Luiz (coordinator) - Rio de Janeiro:
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of Opuntia cochenillifera cactus in water treatment. Encyclopedia Biosphere,
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Coagulants (Ferric Chloride) And Organic (Moringa Oleifera) In Treatment Of
Waters Derived From The Lentic System. Technical Scientific Congress of
Engineering and Agronomy - CONTECC'2016, August 29 to September 2, 2016 -
Foz do Iguaçu, Brazil. Available at: <https://www.tratamentodeagua.com.br/wp-
content/uploads/2016/11/Uso-de-coagulantes-qu%C3%ADmico-cloreto-ferrico-e-
org%C3%A2nico-moringa-oleifera-in-treatment-of-water-from-system-l%C3%
AAntico.pdf>. Accessed in Feb. 2018.
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Principles of water treatment, São Paulo, SP: Cengrage, 2016. 624 p.
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mucilage of a common cactus. Water Science and Technology: Water Supply, v. 12,
n.2, 179-186, 2012.
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<http://server.pelotas.com.br/sanep/tratamento/>. Accessed on: Feb. 2018.
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Polymers: Perspectives for the Treatment of Water. Plastic and Polymer Technology
(PAPT), v. 2, Issue 3, September 2013.
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In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 4

FLOCCULATION: MECHANISMS AND APPLICATIONS

FOR WASTEWATER TREATMENT

Elvis Carissimi1,*, PhD,

Cristiane Oliveira Rodrigues2, PhD, Dounia Elkhatib3
and Vinka Oyanedel-Craver3, PhD
1
Department of Sanitary and Environment Engineering,
Federal University of Santa Maria, Santa Maria-Brazil
2
Department of Nutrition, Federal University of Health Sciences of Porto Alegre,
Porto Alegre, Brazil
3
Department of Civil and Environment Engineering,
University of Rhode Island, Kingston, Rhode Island, US

ABSTRACT

Flocculation is considered the main physico-chemical treatment prior any solid-

liquid separation unit in water and wastewater treatment plants. Suspended particles,
algae, microorganisms are aggregated in order to increase their density for a subsequent
step of sedimention or flotation (in case of injection of air bubbles) and/or filtration.
Design and operation of flocculators are crucial for the process efficiency, and are largely
dependent on the following features: floc characteristics, flocculation kinetics, and
engineering aspects of flocculation. Finally, examples of engineering applications using
flocculation are described.

*
Corresponding Author Email: [email protected].
82 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

Keywords: water treatment, wastewater treatment, flocculation, aggregation

1. INTRODUCTION

A comparison of source water quality and the desired finished water quality is
essential for any treatment process selection. However, flocculation is one of the most
common unit operations and is frequently employed for water and wastewater treatment.
Water treatment includes essentially three separated and sequential steps: coagulation
formation, particle destabilization, and interparticle interactions. It is an important step
for the removal of organic (viruses, bacteria, algae, protozoan cysts and oocysts,
microplastics, humic acids, particulate and dissolved organic matter that may be present
as natural organic matter - NOM) and inorganic particles (clay, silt, mineral oxides, and
erosion particles). The removal of particles is very important because they cause turbidity
(reduce water transparency and fotosintetic activities), may increase color to water;
promote infectious agents (microorganisms), and have toxic compounds adsorbed to their
external surfaces. NOM removal is quite important due the possibility of desinfection by-
products formation when chlorine is used for disinfection.
Ferric iron and aluminum salts are the most common salts used to promote the
coagulation and have been largely used in water and wastewater treatment plants.
Polymers may be or not associated in order to increase floc density and floc strength and
improve solid-liquid separation rates. Besides the addition of chemicals flocs formation
are extremely dependent on the hydrodynamic conditions. The understanding of all the
phenomena and mechanisms involved during flocculation is crucial to design efficient
flocculation units that may be applied to water or wastewater treatment. Thus, the main
goal of this chapter is to describe these mechanisms and applications of new flocculation
units for water and wastewater treatment.

2. STABILITY OF PARTICLES IN WATER

The stability of the colloidal systems can be explained in part by the balance between
the London and van-der-Waals forces and the electric forces between the double layer of
particles (repulsion energy) known as DLVO theory, in honor of Derjaguin-Landau and
Verwey-Overbeek, a pair of Russian and Dutch scientists, respectively, who in the 1940s
independently developed this theory. However, from the 1980s onwards, with the
development of more advanced techniques (atomic force microscopy, for example), it
was possible to obtain results of the surface forces in aqueous medium, which
demonstrated (verified and proved the existing theoretical models) the existence of
 Flocculation: Mechanisms and Applications for Wastewater Treatment 83

additional forces of hydration (repulsive force) and hydrophobic forces (attractive force
much greater than the van der der Waals forces), which were not predicted by the
classical theory. The inclusion of energy due to these forces, also known as structural
forces, resulted in a more modern concept named extended DLVO theory, or, simply, X-
DLVO (Yoon and Ravishankar, 1994; Israelachvili, 1992; Lins and Adamian, 2000;
Bratby, 2006; Bolto and Gregory, 2007; Vincent, 2012).
The particles generally have surface charge in aqueous media, which may originate
from the ionization of surface groups or sites, imperfection of the crystalline structure of
the solid surface, specific adsorption of ions and/or differentiated solubilization between
cations and anions. The surface potential of the colloids, the distribution of ions in
solution and the thermal effects lead to the formation of the double electric layer, shown
in Figure 1.
The double electric layer is modeled as being composed of two regions separated by
the Stern Layer (SL). The inner layer is known as the Stern Layer and the outer layer as
Gouy-Chapman or diffuse layer. In the presence of ions that are adsorbed specifically by
the chemical mechanism, the presence of two other planes is defined: the
Internal Helmholtz Layer (IHL), with potential I, and the External Helmholtz Layer
(EHL), with potential E. In the IHL the specific adsorption of ions by the chemical
mechanism occurs. The adsorption of co-ions, with charge of equal signal to
that of the surface of the particle, promotes an increase of the potential of the double
electric layer.
The adsorption of positive ions, more common case, promotes a decrease of the
electric potential or even the reversion of the charge of the particle. On the other hand, in
the EHL the ions are adsorbed by the physical or electrostatic mechanism, which, at the
most, promote neutralization of the electrokinetic potential of the colloid (). Because of
the difficulty of determining the surface electric potential of the particle, it is common
practice to measure the potential in the shear plane between the moving particle and the
surrounding liquid. The potential in this plane is known as Zeta Potential () or
electrokinetic potential.

3. PRINCIPLES OF FLOCCULATION

The destabilization of colloidal systems can be accomplished by the addition of

inorganic electrolytes, flocculating polymers, surfactants and oils, and the aggregation or
agglomeration (in the case of oils) of the ultrafine or colloidal particles (< 1μm) occurring
in larger units.
84 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

Negative ion Positive ion

Negatively charged
particle surface

Stern Layer
Diffuse Layer

Ions in equilibrium
with bulk solution

Figure 1. Schematic representation of the electric double layer model.

Destabilization with Inorganic Salts

Inorganic salt destabilization, also known as homocoagulation or coagulation, occurs

by the compression of the electric double layer that surrounds all the colloidal particles
by the addition of inorganic salts (Weber, 1972). The adsorption of these inorganic ions
occurs in the shear planer (Zeta), neutralizing the potential in this plane and reducing the
energy of repulsion, allowing the attraction forces to act and the aggregation of the
particles. This term is also applied for destabilization by the addition of hydrolyzable
electrolytes, such as Fe+3 and Al+3. The mechanism that occurs with the addition of salts
of the sulfate type or chloride of iron or aluminum differs from the monovalent salts,
because in addition to reducing the double electric layer, these salts form hydrolysis
products that polymerize forming large three-dimensional molecules with active ends.
These chains form spongy masses which, when sedimented, entrain new particles,
causing a sweeping effect.
The empirical relationship of Schulze-Hardy expresses the ratio between the molar
concentration of monovalent, divalent and trivalent ions for the coagulation of the
colloids of a system stabilized by the electrostatic mechanism as: 1: (1/2) 6: (1/3) 6 or
100: 1.6: 0.13. Higher-valence ions have a greater tendency for specific adsorption,
improving coagulation efficiency. The order of effectiveness for ions of the same valence
follows the Hoffmeister series, in which coagulation varies directly with the hydrated ion
radius (Adamson and Gast, 1997; Dobiás e Stechemesser, 2005; Bratby, 2006; Bolto and
Gregory, 2007).
Heterocoagulation is an aggregation process that also involves the interaction
between double electric layers and van der Waals forces, and occurs between particles of
 Flocculation: Mechanisms and Applications for Wastewater Treatment 85

different potential, being more complex than homocoagulation, especially when the
particles have negative surface charge. Thus, electrostatic interaction is the main
aggregative force acting on heterocoagulation.

Destabilization with Surfactants

The destabilization results from the adsorption of surfactants at the solid-liquid

interface making the colloidal particles hydrophobic. The aggregation by surfactants may
be due to one of the following mechanisms:

(a) Hydrophobic effect and hemimicle formation: in which the adsorption of

surfactant reagents at the solid-liquid interface occurs mainly through
electrostatic, chemical interactions (between the polar part and the superficial
sites) and hydrophobic forces, with a decrease in the entropy of the system
(surfactant molecules). When adsorbed the molecules orient themselves in the
form of doublet/triplet, etc. with the tails interacting by hydrophobic forces, these
conformations being known as hemimicles, and the phenomena that operate are
given by the molecular recognition of the hydrophobic fractions;
(b) Neutralization of charges and hydrophobic effect: neutralization of the surface
charge of the dispersed particles can occur producing destabilization by the
action of attraction forces (London, van der Waals and hydrophobic). The
process is spontaneous, reducing the Gibbs free energy of the system by reducing
the solid-solution surface area and the hydrophobic effect;
(c) Shear aggregation or shear aggregation: shear aggregation occurs when
hydrophobicized particles interact with surfactants in a turbulent hydrodynamic
regime through the hydrophobic effect. This phenomenon of shear aggregation is
due to the low energy involved in this interaction, and to aggregate the particles
need to collide with a minimum energy for these hydrophobic forces to operate.

Destabilization by Oily Agglomeration

When aggregation occurs with oils the process is known as agglomeration and,
because the agglomerates acquire spherical shape, it is also termed spherical
agglomeration. Through this mechanism hydrophobic particles present in water or
hydrophobized with residual surfactants may be agglomerated by the addition of a
nonpolar oil. This process consists of two stages:
86 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

(a) Oil-particle interaction: being the oil little soluble in water, the interaction occurs
between the droplets dispersed in water and the surface of the particles. If the
particles are hydrophobic, the oil spreads initially forming a lens and then a
liquid film that recovers its surface, making them more hydrophobic. Therefore,
the interaction is of hydrophobic character (hydrophobic forces) (Israelachvili,
1992);
(b) Capillary effect: when the oil concentration is high, the droplets occupy the entire
internal area available between the particles. At this stage, the capillary effect is
maximal and defines the spherical shape of the agglomerate with the particles
being held together by oily bridges. The formation of oil capillaries between the
particles promotes bonding and an increase in the hydrophobicity of the flocs.

Destabilization with Aquasoluble Polymers

The use of synthetic polymers in the solid-liquid separation rather than the coagulant
electrolytes allows a more effective process, providing more resistant aggregates (flocs),
higher sedimentation rates and more permeable filtration pans (Metcalf and Eddy, 2003;
Sincero et al., 2003; Fleer, 2010). The flocculating polymers employed for colloidal
destabilization include natural and synthetic products. Among the natural ones are
polyacrylamides, starches, proteins, tannins, biopolymers, guar gums and derivatives of
natural products, such as dextrin and sodium alginate (Metcalf and Eddy, 2003;
Schwoyer, 1981; Bratby, 1980). Most commercial polymers fits as synthetic polymers,
such as, for example, polyacrylamides and polyamides, or nonionic polymers such as
ethylene polyoxide (POE) and polyvinyl alcohol (PVA). As for the charge, the
flocculating polymers may be cationic (radical - NH3+), anionic (-COOH- radical),
nonionic (such as ethylene polyoxide), or amphoteric (semi-hydrolyzed polyacrylamides
which have negative and positive charges in the same jail). Most polymers are
hydrophilic, however, the presence of hydrophobic polymers (such as polyethylene oxide
and polyvinyl alcohol) may occur. The polymers may be low (10,000-100,000) and high
(> 100,000) molecular weight, reaching a molecular length of up to 1000 Å (Schwoyer,
1981; Bratby, 2006; Bolto and Gregory, 2007; Fleer, 2010).
The aggregation of the particles by polymeric bridges is called flocculation. The
polymer adsorb at the solid-liquid interface (hydrogen bonds, hydrophobic forces and
electrostatic attraction) by the electrostatic attraction mechanisms, polymer bridges or the
entrapment of the particles in polymer networks.
The kinetics of the formation of the flocs depends on the following steps:

(a) Diffusion of the polymer under turbulent conditions, followed by adsorption at

the solid-liquid interface. The molecule adsorbs on the surface of the particle at
 Flocculation: Mechanisms and Applications for Wastewater Treatment 87

one or more active sites, leaving the rest of the chain free, extended in the
solution;
(b) Conformation of the polymer forming loops, tails and trains. The conformation
of the adsorbed polymers depends on chain size (molecular weight), chain
flexibility, charge density (% hydrolysis), interaction energy between the
polymer and the colloid, the chemical and physical nature of the superficial sites
of the particles and competition between the polymer and other molecules
present in the solution;
(c) Adsorption of loops and tails and formation of polymer bridges. The
strength of the flocs depends on the number of bridges formed, and hence on the
number of loops and tails available. A factor of crucial importance
is the availability of sites in the particles to accommodate the bonds of
neighboring particles;
(d) Growth of the flocs under slow stirring. According to some authors (Arboleda,
1973; Bratby, 1980; Metcalf and Eddy, 2003; Bratby, 2006), after addition of the
destabilizing agent, rapid mixing is required for diffusion to take place in the
solid-liquid suspension and formation of the primary flocs. After the
appearance of the primary flocs, a slow mixing stage is usually required for
growth and formation of larger flocs and subsequent sedimentation.
However, flotation separation does not require the formation of large flocs. The
energy for the aggregation process is provided by the induction of velocity
gradients within the system (orthokinetic aggregation). The main
parameters involved in orthokinetic energy are the velocity gradient applied and
the stirring time.

The main mechanisms involved in flocculation aggregation are shown in Figure 2.

4. PRINCIPLES OF FLOCCULATION
HYDRODYNAMIC MIXING

The effect of hydrodynamic conditions is another relevant factor in the

destabilization of colloidal systems through hydrolyzable electrolytes and/ or flocculating
polymers. In the aggregation of the particles, two processes must occur sequentially.
First, the particles must collide with each other, and then, under the influence of the
colloidal forces, they must be grouped into aggregates. Thus, the global aggregation
equation is given by Equation (1):
88 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

(a)

(d) (c)

Figure 2. Schematic description of the mechanisms of adsorption and flocculation by bridges:

conformation of the polymer in trains, tails and loops after adsorption at the particle/solution interface.
(a) diffusion of macromolecules; (b) adsorption and conformation of the chains; (c) formation of
primary flakes; (d) growth of the flakes (Kitchener, 1972; Rodrigues, 2010).

𝑑𝑁
− 𝑑𝑡
= {𝐶𝑜𝑙𝑙𝑖𝑠𝑖𝑜𝑛 𝑒𝑓𝑓𝑖𝑐𝑖𝑒𝑛𝑐𝑦}𝑥{𝐶𝑜𝑙𝑙𝑖𝑠𝑖𝑜𝑛 𝑓𝑟𝑒𝑞𝑢𝑒𝑛𝑐𝑦} = 𝛼 𝑥 𝐽 (1)

In equation (1) N represents the concentration of particles at a given time t. Also, J is

the collision frequency and represents the number of collisions that occur per unit time.
Finally, the collision efficiency, α, reflects the fraction of the total number of collisions
that result in the formation of aggregates.
Conventionally (especially in flocculation), after the addition of the destabilizing
agent of a system, there follows a rapid mixing step and a slow stirring step. In rapid
mixing, in addition to the diffusion of the reactants, the breakdown of the repulsive
energy barrier between reagents and particles and between particles and particles occurs.
The primary flocs originate, which will have significant importance on the kinetics of the
subsequent processes. After the appearance of the primary flocs in the fast mixing stage,
a slow stirring stage allows the formation of larger flocs. According to Metcalf and Eddy
(2003) the energy for the process of colloid aggregation is provided by two types of
aggregation:

 microflocculation or pericinetic flocculation, and

 macroflocculation or orthokinetic flocculation.

Microflocculation is the term used to denote particle aggregation due to the random
motion of molecules in the fluid. This random spatial motion of the molecules in the fluid
is also known as Brownian motion. The pericinetic aggregation starts immediately after
 Flocculation: Mechanisms and Applications for Wastewater Treatment 89

destabilization and stabilizes within seconds, since it is significant for particles in the
range of 0.001 to 1 μm.
Macroflocculation is the term that refers to the aggregation of particles larger than 1
or 2 μm. Macroflocculation may occur due to the induced velocity gradient or differential
sedimentation. The energy imposed by a mixer is dissipated through velocity gradients
and the energy rate introduced is proportional to the set velocity gradient. The velocity
gradient is symbolized by G and is used to measure the mixing intensity. A high value of
G means an intense mixture, and a low value of G denotes a slow mixture. According to
Thomas et al. (1999), for a given value of G, orthokinetic aggregation is the predominant
mechanism when the particles exhibit uniformity of size, whereas the differential
sedimentation predominates when the particles present significant disparity of sizes. The
balance between the G employed and the mixing time can be expressed by the Camp
number (G.t).
In any unstable state, the composition of the liquid mass varies with time. This is the
case of most reactors used in treatment plants (mixers, flocculators, decanters, etc.) where
at any point it is found that both the velocity and the composition change constantly
because the water does not flow homogeneously, from the inlet to the outlet, i.e., not all
the flow entering the initial time reaches the outlet, exactly at the nominal holding time td
(Arboleda, 1973).
According to Bratby (1980); Bratby (2006) and Bolto and Gregory, 2007, the ideal
mixing type is the plug flow type, where all particles have the same residence time. In
mixers of the full mix type, some particles are short-circuited and others have very long
residence times. Such distribution of residence times is not desirable in the use of
hydrolyzable salts or polymers. In the case of the hydrolyzable salts, a short residence
time does not allow the complete adsorption of the hydrolyzed species on the surface of
the particles. The same occurs in the adsorption of polymers. On the other hand, intense
mixing for a very long time can break the polymer bridges between the particles and even
the polymer.

5. FLOCCULATION PROCESSES

Usually, three types of flocculators are employed in water and wastewater treatment:
hydraulic, mechanical, and pneumatic (Sincero et al., 2003). Hydraulic flocculators take
advantage of the energy that the flow acquires when flowing through a conduit to agitate
the liquid mass. Also, according to the direction of the liquid stream, inside the chambers,
hydraulic flocculators are divided into: horizontal flow, vertical flow and helical
flow.The mechanical flocculators are classified according to the type of movement of the
agitators, and can be alternating or rotating (Arboleda, 1973). Pneumatic flocculators
employ air to promote agitation. Table 1 shows the main classification of flocculators.
90 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

Hydraulic Flocculators

Hydraulic flocculators utilize the kinetic energy that the flow acquires when flowing
through a conduit to agitate the liquid mass. The most commonly hydraulic flocculator
models are the baffles (Arboleda, 1973; Sincero, et al. 2003).

Baffled Flocculators
The baffle flocculators consist of tanks, provided with internal channels, in which
water flows at a fixed velocity, producing some turbulence, at each change of flow
direction, making a 180° turn at the end of each channel, shown in Figure 3.
The most common are horizontal flow and vertical flow. A head loss occurs mainly
because:

(a) Change of direction and turbulence;

(b) By enlargement and contraction of the section;
(c) By friction in the straight stretches.

However, baffle flocculators present some disadvantages, such as:

(i) greater head loss (greater velocity gradient) in the 180° flow turns compared to
the straight stretches;
(ii) in the case of fixed baffles, the velocity is constant for each flow. In case of
variations in flow rate, the speed also changes, being very high or very low.

Outlet

Inlet

Figure 3. Baffled Flocculator unit (http://www.nzdl.org).

 Flocculation: Mechanisms and Applications for Wastewater Treatment 91

Table 1. Usual classification of the flocculators

Type Classification Example

Hydraulic Horizontal Baffles interspersed transversely of the tank
Vertical Baffles interspersed above and below the tank
Helical Tanks with input and output devices in opposite directions
Mechanical Rotating Vertical or horizontal axis rotary vane
Alternates Alternating vanes
Pneumatic Air diffusers

Helical Flow Flocculators

In this type of flocculator (also called tangential flow or spiral flow) the hydraulic
energy is used to generate a helical movement to the water through inlet and outlet
devices located in opposite directions, creating a mechanical torque and imparting a
movement of rotation of the net mass (Arboleda, 1973; Richter and Netto, 1991). Figure
4 shows the design of a helical flow flocculator.
Hydraulic flocculating flocculators are not as widespread as mechanical flocculators
and wall flocculators. Arboleda (1973) cites that the Engineering Institute of the National
Autonomous University of Mexico (UNAM) has developed studies in units of this type,
with the objective of establishing parameters for such flocculators. Oliveira (1979, 1981)
evaluated the performance and orthokinetic characteristics of the helical stream using this
type of flocculator with a cylindrical chamber (avoiding dead zones) in the water
treatment plant of the city of Atlântida (Southern Brazil) and verified the flocculation
efficiency, in terms of turbidity removal, being technically feasible for small treatment
plants (low velocity gradient). Both studies verified the limitation that occurs with the use
of this type of flocculator for relatively large flow rates, and it is necessary to increase the
number of chambers too much, resulting in loss of simplicity and structure savings.

Figure 4. Helical flocculator unit (Richer and Netto (1991)).

92 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

Alabama Flocculator
The Alabama Flocculator is made up of compartments interconnected by the bottom
through 90° curves facing upwards. The flow can be up and down inside the same
compartment. Removable nozzles installed at the exit of the reactor allow adjusting the
speed to the conditions of calculation or operation (Richter and Netto, 1991). According
to Vianna (1997), the flocs brought by the upstream tributary current collide with those
carried by the effluent stream, descending, resulting in the growth of the flocs. Thus,
according to the same author, it allows the existence of fewer chambers than vertical
baffle flocculators. Figure 5 illustrates an Alabama flocculator.

Figure 5. Alabama flocculator unit (Source: Richer and Netto (1991)).

Cox Flocculator
This flocculator was developed by Cox for the Public Health Services Foundation of
Brazil (Richter and Netto, 1991). According to Vianna (1997), this type of flocculator has
a small number of chambers, and the interconnections between the chambers alternate
upper and lower positions, as shown in Figure 6. The main advantage of this type of
flocculator is the small number of compartments and as the main disadvantage, there is
cited the unevenness of the degree of agitation conferred on the liquid mass.
Table 2 summarizes the major advantages and disadvantages of hydraulic
flocculators.
 Flocculation: Mechanisms and Applications for Wastewater Treatment 93

Inlet

Treatment Flux
Outlet

Figure 6. Cox Flocculator (Source: Richer and Netto (1991)).

Table 2. Main advantages and disadvantages of hydraulic flocculators

Advantages Disadvantages
 Flow close to plug flow  There is no flexibility in changing G
 Does not require complementary  Usually occupies a large space
devices
 It does not require electrical energy
 Ideal for small installations

Mechanical Flocculators

Mechanical flocculators require an external power source to move an agitator in a

tank or series of tanks, in which the suspension remains for a theoretical holding time td.
According to the movement of the agitator are classified in rotary or alternating. The first
can be of low speed of rotation (vane) or of high speed of rotation (turbines and
propeller), being able to be of vertical or horizontal axis. The second consists of wooden
beams that move vertically and alternately (walking beams) or oscillating systems like
ribbons (flocculator) that move horizontally inside the tank. In both cases the energy
transferred to the liquid mass is directly proportional to the energy with which the
mechanical element moves within it (Arboleda, 1973).

Vanes
These rotating flocculators have a system of vanes adhered by a horizontal or vertical
axis, which rotates by an electric motor, displacing the water and producing work. These
stirrers may have two, three or four arms, and, on each arm may have two or more vanes
or impellers joined by a central axis, as shown in Figure 7.
94 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

Figure 7. Rotating vanes (Source: Richer and Netto (1991)).

Impellers (Turbine and Propeller)

Turbine and propeller type flocculators are typically more compact and can generate
more energy when operated at high speed. Essentially, turbine-type flocculators consist
of a central axis in which one or more discs with fins are assigned, as shown in Figure 8
(Metcalf and Eddy, 2003). The propeller-type flocculators have fins attached directly to
the central axis of rotation. The speed of rotation of these agitators is greater than in the
vane flocculators. In some plants, turbine or propeller flocculators are employed for the
first tanks (which require a higher velocity gradient) and for vats for the tanks that will
promote the slow stirring stage.

Figure 8. Impellers (http://www.solidscontrolsystem.com/four-mud-agitator-impeller-type/).

 Flocculation: Mechanisms and Applications for Wastewater Treatment 95

Table 3. Main advantages and disadvantages of

mechanical flocculators
Advantages Disadvantages
Formation of dense flocs Production of short circuits or dead zones
Possibility of changing the speed of agitation Equipment dependency
according to the need of treatment Electric power consumption
Constant and homogeneous agitation Need for electro-mechanical maintenance.

Table 3 presents the main advantages and disadvantages of mechanical flocculators.

According to Hudson and Wolfman (1957) cited by Arboleda (1973), the problem of
short circuits generated in flocculators by mechanical agitation is the main impediment in
the comparison of flocculation results obtained in jar tests with the data obtained in the
treatment plants. This is because in the jars test the water and the reactants are retained in
their entirety in the cells during the test time, with no short circuits occurring. And the
opposite occurs in the flocculation chambers, where part of the water passes quickly
while another is held for longer periods.

6. PNEUMATIC FLOCCULATORS OR
AIR-ASSISTED FLOCULATION

The necessary stirring for flocs formation can be accomplished by the introduction of
air into the system. The difference in density between the air bubbles and the water
causes the bubbles to rise to the surface. As the bubbles rise, they cause the water to
move, and consequently the mixture required for flocculation. In this case, the velocity
gradient reached can be controlled by adjusting the airflow. In published pneumatic
flocculation studies, chambers or columns are usually used where air is injected through a
diffuser (McConnachie, 1984; Sincero et al., 2003).
The in-line pneumatic flocculation of a biphasic flow (air-water), promoted by
mixers, can be explained by mechanisms similar to those of pneumatic flocculation in
chambers. In this process, the agitation necessary to promote flocculation is increased by
the injection of air through diffusers into the stream containing the effluent to be treated
and the destabilizing agent. In this way, the air released in the form of bubbles, in
addition to promoting a piston-type mixing in in-line mixers, has a high flocculation
efficiency for the removal of impurities that are suspended (Sholji and Kazi, 1997).
Through the principles of pneumatic flocculation, many authors have suggested a
new proposal for the generation of aerated flocs, in which dispersion of the air injected in
the form of small bubbles that adhere and/or trapped themselves to the flocs during its
formation (Owen et al., 1999; Fan et al., 2000; Da Rosa, 2002; Jameson, 1999; Zhao,
2002). In this way, aerated flocs “float” and are more easily separated later in a
96 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

separation tank. These aerated flocs are structures composed of particles or

colloids flocculated by high molecular weight cationic polymers containing trapped air in
the form of microbubbles. The aerated flocs normally formed are of the order of
centimeters, whose rise rate is significantly greater than the velocity of independent air
bubbles.
Bratby (1980) cites the generation of a local velocity gradient much larger than the
mean gradient as one of the disadvantages of diffused air flocculation. This depends on
the bubble size that is generated by the available diffusers. According to the same author,
the maximum speed gradient generated is given by Equation (2).

𝑔.𝐷𝑏(𝜌𝑙 −𝜌)
𝐺𝑚𝑎𝑥 = 6.𝜇
(2)

where: ρ = density of the liquid (kg.m-3); ρ1 = density of the liquid; ρ = air in the bubble
(kg.m-3); Db = bubble diameter (m); g = acceleration of gravity (9.81 m /s2); μ = absolute
viscosity (10-3 N.s.m-2, for water at 20 ° C).
Table 4 shows the Gmax values for a bubble size range.

Table 4. Gmax values generated by a range of bubble diameters

(water at 20°C)

Bubble Diameter (mm) Gmax (s-1)

0.01 16
0.05 82
0.1 163
0.15 245
0.5 816
1.0 1633
2.0 3265
Source: Bratby, J. Coagulation and Flocculation, p. 226, 1980

Table 5. Main advantages and disadvantages of pneumatic flocculators

Advantages Disadvantages
Possibility of formation of aerated flocs, assisting Need for energy and specific equipment for air
later separation dispersion
Adjusting the stirring by controlling the flow of air.

However, according to Masschelein (1992) cited by Metcalf and Eddy (2003) the
mean bubble diameter formed in this process is 5 mm, in a mean air flow of 10 percent of
the liquid flow. The velocity gradient due to the formation of bubbles in this range ranges
from 200 to 8200 s-1. In the case of extremely small bubbles of the order of micrometers,
the value of Gmax will also be small, becoming practically negligible. Table 5 presents
the main advantages and disadvantages of pneumatic flocculation.
 Flocculation: Mechanisms and Applications for Wastewater Treatment 97

7. FLOCCULATION APPLICATIONS

Considerations about the main applications of the flocculation considering water and
wastewater treatment are discussed considering a compact helical unit named here as
Flocs Generator Reactor that may be applied for water and wastewater treatment
(emphasis is given for physicochemical processes) and a biological process application
for sewage treatment named here as bioflocculation for nitrogen uptake.

Flocs Generator Reactor (FGR)

Curved configurations of circular tubes are widely used in heat exchangers, chemical
reactors, reverse osmosis units, blood oxygenation membranes, enzyme polymerization,
etc. These helical mixing units present enormous advantages over straight-line systems or
mechanical mixers, mainly due to hydrodynamic characteristics (higher Reynolds
numbers and higher velocity gradients) and secondary flow. This flow presents an action
of centripetal forces, with a movement along the walls and near the center of the tube,
increasing the resistance to the flow (Streeter, 1961; Berger et al., 1983, Agrawal and
Nigam, 2001; Elmaleh and Jabbouri, 1991; Ødegaard et al., 1992; Buchanan et al., 1998;
Carissimi and Rubio, 2005; Carissimi et al., 2007, Carissimi and Rubio, 2015; Carissimi
et al., 2018).
The evaluation of curved systems employed industrially and the analysis of the
different aggregation equipment used in the water and effluent treatment plants cited by
Carissimi and Rubio (2005) allowed the design and the design of a hydraulic aggregation
system in line with a piston regime, called Reactor Generator - RGF. Figure 9 shows the
response curve generated by the instant introduction of a tracer (methylene blue) for
determining the axial flow rate of the RGF. The pulse shown shows a slow tracer
spreading at the feed rate of 3 Lmin-1, and the time of 25 seconds denotes the tracer
residence time inside the RGF.
The FGR consists of an in-line helical mixing reactor for the solid-liquid aggregation
and separation of suspended particles, with a Brazilian Patent (INPI PI 0406106-3). The
necessary stirring for dispersion of the destabilizing agent and generation of the
aggregates is carried out by exploiting the kinetic energy of the hydraulic flow along the
helical tubular reactor.
98 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

Figure 9. Curve of the tracer response along the FGR. Conditions: feed flow =
3 Lmin-1, [AM] = 10000 mgL-1; FGR with a length of 12 m and a diameter of 2.5 cm.

Length
comprimento

microbolhas
Air Outer
diâmetro
de ar
microbubbles diameter
externo

Inlet
entrada
saída
Outlet
efluente + polímero
Water/wastewater + polymer

direção do fluxo
Flow direction

Figure 10. Flocs Generator Reactor (FGR).

One (semi-pilot) unit of the FGR is shown in Figure 10; consisting of a transparent
polyurethane tube with internal diameter of 0.0125 m wrapped in the outside of a fixed
polyvinyl chloride (PVC) column, with an internal radius of 0,05 m, consisting of 32
rings, 12 m volume of 1.2 L, occupying a surface area of 0.60 mx 0.13 m. This model has
the alternative of injecting air microbubbles, generated through an air depressurising
system (such as in the dissolved air flotation process), making the reactor as an aerated
float floater (detailed later).
The use of online aggregation (pipelines, ducts, pipes) is not a common practice,
however, already existing in some industrial plants, either by the practical necessity of
operators, space optimization or by commercialization by companies specialized in water
and effluent treatment.
 Flocculation: Mechanisms and Applications for Wastewater Treatment 99

Bioflocculation in Wastewater Treatment Plants

Bioflocculation combines the primary influent of wastewater and the return activated
sludge from the secondary treatment in order to improve the nitrogen removal efficiency
as well as the nitrogen recovery.
Excess Nitrogen in rivers and streams causes water pollution and hence affects the
public health (Manuel, 2014). Nitrogen effluent discharge from wastewater treatment
plants are therefore becoming increasingly strict (Agency, 2018). The need for effective
nitrogen removal strategies to meet the nitrogen effluent limits while maintaining
minimal operating Wastewater Treatment Plants (WWTP) costs is therefore increasing
(Stare, Vrečko, Hvala, & Strmčnik, 2007). Conventional biological nitrogen removal
processes are usually conducted in a series of anoxic/aerobic reactor (Metcalf Eddy,
2014). The biological processes that remove nitrogen are nitrification and denitrification.
During the nitrification stage, high energy use for aeration is required to oxidize the
ammonia to nitrite then nitrate, making it a cost burden to WWTP. Nitrogen removal
requires 12 kWhel per person per year and significantly contributes to the overall
wastewater energy budget (Verstraete & Vlaeminck, 2011).
Nitrous oxide (N2O), which is a potent greenhouse gas, contributing for 5.3% of the
global anthropogenic greenhouse gas emissions in 2013 (EPA, 2014), can be emitted
during the nitrogen removal process in wastewater treatment. During the biological
nitrogen removal process, N2O is produced by nitrite reduction in the aerated
compartments (Toyoda et al., 2011) and during the anoxic stages as an intermediate
product (Colliver & Stephenson, 2000). The Environmental Protection Agency of the
United States reported that N2O from the wastewater sector accounts for about 3 per cent
of N2O emissions from all sources and ranks as the sixth largest contributor in the United
States (EPA, 2014). The increased presence of this gas in the atmosphere causes a rise in
the equilibrium temperature of the earth, thus arousing climate change. Therefore, the
need of an efficient nitrogen removal system that achieves low N2O emissions while
maintaining the nitrogen removal standards, is required.
The bioflocculation process is a proposed application, where the waste activated
sludge from the secondary treatment is used as flocculent. This process has proven to
enhance the nitrogen removal and recovery during the primary treatment stage. The
removal of nitrogen and organics from the primary effluent could reduce the energy
requirements in the activated sludge process and also reduce the N2O emissions produced
during the nitrogen removal processes. The biogas and the residual solids from the
facility’s anaerobic digester will increase upon digestion of primary sludge, which has
high energy and nitrogen content.
The bioflocculation application was tested using the influent wastewater and the
activated sludge obtained from the Narragansett Bay Commission Wastewater Treatment
Facility, located at Field’s Point in Providence, Rhode Island. The facility has an average
100 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

daily flow of 40 MGD providing services to about 226,000 people. This plant uses the
Integrated Fixed Film Activated Sludge (IFAS) process for the secondary treatment to
promote nitrification.
The influent wastewater used in this experiment was obtained from the outlet of the
primary treatment, while the activated sludge was obtained from the Return Activated
Sludge (RAS), which should have the same characteristics of the waste activated sludge.
The collected samples were tested and analyzed at the environmental engineering
laboratory located at the University of Rhode Island.

Figure 11. Diagram flow for the proposed treatment system.

Table 6. Mixture Conditions used during the Experiment

Volume of Sludge [mL] Volume of Sewage [mL]

0 1000
250 750
500 500
1000 0

In order to apply the concept of bio flocculation process at this existing wastewater
facility with minimal investment costs, a pipe and a pump have to be installed to deliver
the secondary activated sludge back into the primary clarifier.
Figure 11 shows a schematic drawing of the proposed treatment system. The
recirculation of the activated sludge to the inlet primary treatment is shown in red.
The tests were conducted in sealed 1000 mL graduated cylinder. Influent wastewater
(or sewage) and activated sludge were mixed in the cylinders according to the test
conditions summarized in Table 6. The graduated cylinders were shaken to assure a good
mixing of the components, followed by 30 minutes of contact time between the two
materials. Bioflocculation efficiency was determined by the removal percentages of
nitrogen from the influent.
The removal percentage of organic matter takes into account the removal
mechanisms from both the influent and the activated sludge. This method was based on a
research study where settleable solids removal from bioflocculation was assessed
 Flocculation: Mechanisms and Applications for Wastewater Treatment 101

(Araneda, Pavez, Luza, & Jeison, 2017). However, this method was modified for the
purpose of this application to calculate the total removal organic matter from the influent.
The results show that there is a considerable removal efficiency of respectively 59%,
47% and 25% for COD, TKN and NH3 concentrations in the second mixture (500:500),
where the sludge and sewage are mixed in the same ratio. The N2O emissions at the NBC
Field’s Point plant were found to be reduced by 1.4 µmol N2O/m2.s.
Anaerobic digesters are one of the most common waste sludge treatment. In this
process, the biosolids are broken down in the absence of oxygen, and partially converted
into gases, while the remainder is dried and becomes a residual soil-like material (Metcalf
Eddy, 2014). The treated sludge contains useful concentrations of nitrogen, phosphorus
and organic material.
When applying the suggested enhanced primary treatment using the bioflocculation,
the total nitrogen removed from the primary effluent will be transferred to the primary
sludge. The increase of the nitrogen availability in the primary sludge by 47% will
increase the benefits of the treated sludge, as it will have higher nitrogen contents, and
hence more beneficial to the grassland. The organic matter in sludge can also improve the
water retaining capacity and structure of some soils, especially when applied in the form
of dewatered sludge cake (Natural Resources Management and Environment
Department). The nitrogen being transferred to the primary sludge, is hence recovered to
a more useful way than being consumed by microorganisms in the secondary treatment,
which requires high amounts of energy too.
Anaerobic digestion reduces the volume of sludge and produces methane containing
biogas, a renewable energy, as a by-product. Collecting the energy from the biogas
through an efficient combustion process is both economically and environmentally
beneficial. Based on the results of the bioflocculation application, the percent of COD
concentrations in the influent that is transferred to the primary sludge was found to be
59%. The methane production achieved by primary sludge was found by (Mahdy,
Mendez, Ballesteros, & González-Fernández, 2015) to be 266 mL per gram of COD.
And, the methane has a calorific potential of 6.22E-3 kW.h/L CH4 (Metcalf Eddy, 2014).
By using these information, the amount of CH4 produced per day will increase by 76,021
cubic feet, which is around 51% increase of the usual methane production at the NBC
facility.
The bioflocculation application of the influent sewage can be advanced primary
treatment process. Its mechanism for the removal of organics, without any addition of
chemicals, enhances the nitrogen removal and recovery from wastewater treatment
plants. The experiments showed that TKN in the influent was reduced by 46% and the
NH3-N was reduced by 24% as a result of mixing the activated sludge from secondary
treatment with the primary influent, for a flocculation time of 30 min.
The removal performance of the bio flocculation in the primary treatment, has also
shown some energy consumption savings in the treatment aeration process. For the same
102 Elvis Carissimi, Cristiane Oliveira Rodrigues, Dounia Elkhatib et al.

removal percentages mentioned above, the expected reduction in the aeration process was
about 18% for the cost and 36% for the air requirements.
Another main advantage of the bio flocculation process is the recovery of nitrogen
and organics in the primary sludge. The primary sludge, which is rich in nitrogen content,
can be used as fertilizer to supply agriculture soils instead of chemical fertilizers that
require large amounts of energy to produce. Also, the primary sludge which has a higher
energy content than the sludge removed at the secondary treatment due to the high
organics content, produce more biogas during digestion.
Finally, the calculations from the bio flocculation process have proven a reduction of
the nitrous oxide emissions from wastewater treatment due to the reduction of Nitrogen
content. In the enhanced primary treatment where the bio flocculation was applied, the
N2O emissions were reduced by 27%.
In conclusion, the Bio flocculation process has proven to be a new approach in
primary treatment of wastewater that leads to an energy efficient and improved process
design in wastewater treatment sector.

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Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 5

FLOCCULATION OF AOM
IN WATER TREATMENT

Martin Pivokonský*, Jana Načeradská,

Kateřina Novotná, Lenka Čermáková and Petra Vašatová
Institute of Hydrodynamics of the Czech Academy of Sciences,
Prague, Czech Republic

ABSTRACT

Global proliferation of algal blooms and subsequent deterioration of water quality by

organic compounds that are being produced (algal organic matter – AOM) pose new
challenges to water treatment technologies. Flocculation/coagulation using primarily Al-
and Fe-based coagulants is widely employed as an essential process in removal of various
impurities at drinking water treatment plants and is also irreplaceable in the case of AOM
elimination. This review chapter discusses current knowledge on AOM flocculation, the
impact of AOM on the removal of other compounds and links AOM composition and
character to the efficiency of flocculation, the reaction conditions and mechanisms and
finally, to the properties of flocs. In general, the removal efficiencies of dissolved AOM
are lower compared to intact phytoplankton cells and usually reach maximum under
slightly acidic pH values. The strong pH-dependence of flocculation is attributed to the
fact that the involved mechanisms are to a great extent determined by the charge ratios in
the coagulating system. Furthermore, substantial differences in flocculation behaviour
were observed between diverse AOM constituents, i.e., between peptides/proteins versus
non-proteinaceous matter and high versus low molecular weight organics. The latter
(specifically AOM < 10 kDa) are reluctant to flocculate and would therefore require other
treatment techniques. AOM has also been reported to influence flocculation of other

*
Corresponding Author Email: [email protected].
108 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

common impurities, both of organic and inorganic nature. Mutual interactions have been
proven, while their influence on flocculation efficiency can be either positive or negative,
depending on the AOM character, pH conditions and on the ratio between AOM, the
other polluting agents and coagulants. Finally, AOM also appeared to alter the properties
of flocs, with an impact on the subsequent separation steps. In further research, a
particular emphasis should be put on AOM components that are difficult to coagulate, the
interactions of AOM with other impurities and on elucidation of the relationship between
AOM and floc properties.

Keywords: algae, algal organic matter, AOM characterization, coagulation,

coagulation mechanism, flocculation, floc properties, water treatment

1. INTRODUCTION

Increasing worldwide occurrence of algal blooms in surface waters is an ongoing

challenge in water treatment. In addition to the presence of algal and cyanobacterial cells,
these are sources of a wide range of dissolved organic compounds, recognized as algal
organic matter (AOM). AOM often comprises a substantial part of natural organic matter
(NOM) occurring in water bodies that supply water treatment plants, where it adversely
affects water quality and subsequent treatment processes (Zhang et al., 2010; Pivokonsky
et al., 2016).
AOM has gained a great attention due to the possible content of harmful
cyanobacterial toxins (Carmichael, 1992; Dixon et al., 2011; Pearson et al., 2016) and
compounds that give undesirable tastes and odours (Paerl et al., 2001; Huang et al., 2007;
Zhang et al., 2010; Li et al., 2012). Further, AOM also serves as precursors of hazardous
disinfection by-products (DBPs) that are being formed as a result of chlorination or
chloramination of organic compounds (Nguyen et al., 2005; Fang et al., 2010; Li et al.,
2012; Goslan et al., 2017). For example, trihalomethanes (THMs; e.g., trichloromethane
(chloroform), dichlorobromomethan, dibromochloromethane or tribromomethane) and
halogenated acetic acids (HAAs; e.g., monochloroacetic acid, monobromoacetic acid,
dichloracetic acid or bromochloroacetic acid) are among the most pronounced DBPs
(Fang et al., 2010; Li et al., 2012; Goslan et al., 2017).
Apparently, AOM needs to be effectively removed in order to produce safe drinking
water of satisfactory quality. Nevertheless, diverse AOM components act dissimilarly
during water treatment processes and might pose specific complications in the treatment
technology, such as interference with flocculation/coagulation1, membrane fouling or

1
The terms ‘flocculation’ and ‘coagulation’ commonly appear in the field of water treatment. These processes
result in clumping of particles/molecules and subsequent formation of flocs, while ‘coagulation’ rather stands
for the destabilization of particles (involving charge neutralization) and ‘flocculation’ for the aggregate (floc)
formation. However, the terminology is not consistent among different studies and the terms are being used in
 Flocculation of AOM in Water Treatment 109

competing with target pollutants regarding adsorption onto activated carbon (Pivokonsky
et al., 2016). Flocculation followed by separation of the formed aggregates
(sedimentation, flotation, filtration) is a fundamental step in water treatment. It is
extensively employed at many drinking water treatment plants worldwide and is
irreplaceable due to its operational benefits and cost-effectiveness. It also improves
performance of potential downstream treatment processes, such as membrane filtration or
adsorption onto activated carbon (Zhang et al., 2014). Flocculation has a potential to
remove different polluting agents. Good removal efficiencies (94-99%) of algal and
cyanobacterial cells were obtained under optimized flocculation conditions (Henderson et
al., 2010; Baresova et al., 2017). However, AOM is generally much more difficult to
coagulate, and additionally, AOM also influences flocculation of other compounds
(Bernhardt et al., 1985; Henderson et al., 2010; Safarikova et al., 2013; Pivokonsky et al.,
2015, 2016; Baresova et al., 2017).

2. WHAT IS AOM

2.1. Origin of AOM

Algal blooms are formed by various phytoplankton species, belonging to

cyanobacteria, green algae, diatoms, dinoflagellates or cryptophytes (Paerl et al., 2001).
The concentrations of algal/cyanobacterial cells within the algal bloom have been
reported to reach up to the millions of cells per mL in natural surface waters (Dixon et al.,
2011). Consequently, considerable amounts of AOM are being produced.
A part of the AOM is generated extracellularly, resulting from the metabolic
processes of living algal/cyanobacterial cells, and is referred to as extracellular organic
matter (EOM). For example, in laboratory cultures, EOM concentrations in a stationary
growth phase reached approximately 20, 40 and 60 mg L-1 DOC (dissolved organic
carbon2) for cell concentrations of 12·106, 12·104, and 18·106 mL-1 (different species),
respectively (Pivokonsky et al., 2014).
Furthermore, when the phytoplankton cells die and decompose, a mixture of
compounds named cellular organic matter (COM3) is being formed. Especially when
phytoplankton blooms decay, a sudden release of large amounts of COM may rapidly

similar meaning. Thus, the terms ‘flocculation’ and ‘coagulation’ are considered equal in this chapter and the
term ‘flocculation’ is preferred.
2
As AOM is a mixture of various organic compounds, its concentrations are commonly being expressed as DOC
(dissolved organic carbon) in mg L-1 DOC.
3
Also the term ‘IOM – intracellular organic matter’ is sometimes being used in this sense (Pivokonsky et al., 2006;
Fang et al., 2010; Li et al., 2012). Additionally, ‘SOM – surface-retained organic matter’ is distinguished in
some studies (Takaara et al., 2010).
110 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

alter overall water quality (Zhang et al., 2010). Additionally, COM can also occur as a
result of water treatment processes that induce cell rupture, e.g., pre-oxidation (Ma et al.,
2012b; Coral et al., 2013).

2.2. Composition of AOM

Algal organic matter contains a wide range of organic compounds, including

proteins, peptides, amino acids, mono-, di-, oligo-, and polysaccharides, lipids, fatty
acids, nucleic acids, amino sugars, lipopolysaccharides, aldehydes, etc. (Bertocchi et al.,
1990; Myklestad, 1995; Paerl et al., 2001; Her et al., 2004; Nguyen et al., 2005;
Pivokonsky et al., 2006, 2014; Huang et al., 2007; Henderson et al., 2008a; Markou et al.,
2012; Laurens et al., 2014; Villacorte et al., 2015). AOM might also comprise highly
undesirable compounds such as harmful toxins, e.g., hepatotoxic microcystins, nodularins
and cylindrospermopsins, or neurotoxic anatoxins and saxitoxins (Carmichael, 1992;
Dixon et al., 2010; Pearson et al., 2016). Additionally, substances that deteriorate
organoleptic properties of water (taste and odour compounds) can be also present, e.g.,
geosmin or 2-methylisoborneol (Paerl et al., 2001; Huang et al., 2007; Zhang et al., 2010;
Li et al., 2012).
An exact composition of both COM and EOM is species-dependent and is also given
by a growth stage of the phytoplankton (Pivokonsky et al., 2006, 2014; Henderson et al.,
2008a). In laboratory cultures, four growth phases are being distinguished, i.e., lag,
logarithmic/exponential (exp), stationary (stat) and decline phase, where the amounts and
character of AOM evolve as the growth proceeds (Pivokonsky et al., 2006, 2014;
Baresova et al., 2017). Additionally, environmental conditions, such as nutrient status
(Myklestad, 1995), temperature or light intensity (van der Westhuizen and Eloff, 1985)
affect the production and properties of AOM.
The composition and character of AOM significantly influence its appearance in the
drinking water treatment processes. For example, peptide/protein and non-proteinaceous
fraction of AOM have been proven to behave differently in flocculation experiments
(Pivokonsky et al., 2009b, 2016), as described in detail in section 3.

2.2.1. AOM Peptides/Proteins

Peptides and proteins often form a substantial part of AOM, comprising up to 60-
70% (Pivokonsky et al., 2006, 2009b). In laboratory cultures, the relative portion of
peptides/proteins in EOM was observed to increase during the cultivation of different
phytoplankton species (cyanobacteria Anabaena flos-aquae4 and Microcystis aeruginosa,

4
recognized as Dolichospermum flos-aquae since 2009
 Flocculation of AOM in Water Treatment 111

diatom Fragilaria crotonensis, green algae Chlamydomonas geitleri and Scenedesmus

quadricauda). DOCP (DOC constituted by peptides/proteins) in the stationary growth
phase was the highest for cyanobacteria (up to 47%) and the lowest for green algae (max
28%) (Pivokonsky et al., 2006, 2014). However, the greatest content of peptides/proteins
was determined as a part of COM, of which DOCP accounted for 60-66% for M.
aeruginosa (Pivokonsky et al., 2006, 2009b, 2012, 2014; Safarikova et al., 2013), 53%
for F. crotonensis (Pivokonsky et al., 2014), 51% for A. flos-aquae (Pivokonsky et al.,
2006), 43% for Merismopedia tenuissima (Baresova et al., 2017), 33% for C. geitleri
(Pivokonsky et al., 2014) and 29% for S. quadricauda (Pivokonsky et al., 2006).

2.2.2. AOM Carbohydrates

Besides peptides/proteins, carbohydrates are also often contained in AOM in
significant amounts (Myklestad, 1974, 1995; Laurens et al., 2014; Villacorte et al., 2015).
For example, the carbohydrate proportion of AOM dry weight was reported to be up to
90% in case of marine diatoms (Myklestad, 1974) and up to approximately 50% in green
algae (Chlorella sp., Scenedesmus sp.). Similar to the content of peptides/proteins, the
carbohydrate content of AOM has been observed to change during the growth of the
culture (Laurens et al., 2014). AOM carbohydrates are usually composed of mainly
arabinose, fucose, galactose, glucose, mannose, rhamnose, ribose, or xylose (Bertocchi et
al., 1990; Myklestad, 1995; Nicolaus et al., 1999; Huang et al., 2007; Markou et al.,
2012). Additionally, uronic acids (glucuronic acid, galacturonic acid), aminosugars
(glucosamine, galactosamine, mannosamine) or methyl-sugars might be also present
(Bertocchi et al., 1990; Nicolaus et al., 1999). A substantial part of algal carbohydrates is
often comprised of polysaccharides formed by some of these subunits (Myklestad, 1995).
The polysaccharides are species-specific, including e.g., glycogen in cyanobacteria or
starch in green and red algae (Markou et al., 2012).

2.3. Properties of AOM Related to Water Treatment

Due to its varying compositions, algal organic matter differs in several characteristics
that alter its flocculation behaviour. Apart from the division into peptides/proteins and
non-proteinaceous matter, properties of AOM that are substantially important from the
perspective of water treatment include (1) the degree of hydrophilicity/hydrophobicity,
(2) molecular weight (MW) distribution and (3) surface charge (Pivokonsky et al., 2016).

2.3.1. Hydrophilicity/Hydrophobicity
Algal organic matter is usually divided into three groups according to the level of its
hydrophobicity, i.e., into hydrophilic (HPI), hydrophobic (HPO) and transphilic (TPI)
fraction (Her et al., 2004; Henderson et al., 2008a; Pivokonsky et al., 2014; Baresova et
112 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

al., 2017). Additionally, HPI compounds can be further distinguished as neutral

hydrophilic (NHPI) and charged hydrophilic (CHPI) (Li et al., 2012).
It was revealed that AOM is predominantly hydrophilic, with the HPI fraction
ranging from 54-90% among various species (Anabaena flos-aquae, Aphanizomenon
flos-aquae, Asterionella formosa, Chlamydomonas geitleri, Chlorella vulgaris,
Fragilaria crotonensis, Merismopedia tenuissima, Microcystis aeruginosa, Scenedesmus
subspicatus). Besides the species-dependence, the portion of HPI material was found to
differ in EOM versus COM, while the cellular material was determined to be more
hydrophilic. Small variations in the HPI/HPO/TPI proportions were also observed
between EOM acquired in exponential and stationary growth phases (Her et al., 2004;
Henderson et al., 2008a; Li et al., 2012; Pivokonsky et al., 2014; Baresova et al., 2017;
Goslan et al., 2017). Percentage representation of hydrophilic, hydrophobic and
transphilic fractions in EOM and COM of selected species is shown in Table 1. The
highly hydrophilic nature of AOM is supported by low SUVA5 values (ranging from
0.34-1.7 L m-1 mg-1) ascertained for organic matter produced by a number of species (Her
et al., 2004; Henderson et al., 2008a; Fang et al., 2010; Li et al., 2012; Goslan et al.,
2017).

Table 1. Percentage representation of hydrophilic (HPI), hydrophobic (HPO)

and transphilic (TPI) material in exponential-phase EOM (EOMexp),
stationary-phase EOM (EOMstat) and COM of different phytoplankton species.

Species EOMexp proportion (%) EOMstat proportion (%) COM proportion (%)
HPI HPO TPI HPI HPO TPI HPI HPO TPI
A. flos-aquaea - - - 81 8 - - - -
Aph. flos-aquaea - - - 63 18 - - - -
A. formosab 73 14 13 68 19 13 - - -
C. geitleric 71 21 8 73 22 5 89 10 1
C. vulgarisb 63 24 13 72 11 17 - - -
F. crotonensisc 74 17 9 74 19 7 90 8 2
Melosira sp.b - - - 60 32 8 - - -
M. tenuissimad - - - - - - 77 7 16
M. aeruginosac 69 27 4 69 28 3 87 12 1
S. subspicatusa - - - 54 26 - - - -
a
Goslan et al., 2017; bHenderson et al., 2008a; cPivokonsky et al., 2014; dBaresova et al., 2017.

The HPI fraction of organic matter include carbohydrates, amino acids and other
organic acids, aldehydes, ketones, etc. (Edzwald, 1993). HPO compounds (determined to
form approximately 8-32% of EOM and 7-12% of COM) include humic and fulvic acids,
aromatic acids and amines, and high-MW alkyl carboxylic acids (Edzwald, 1993;

5
SUVA (specific UV absorbance) corresponds to the aromaticity of a sample. It is defined as the UV absorbance at
a given wavelength (254 nm) normalized for DOC concentration.
 Flocculation of AOM in Water Treatment 113

Pivokonsky et al., 2014; Baresova et al., 2017; Goslan et al., 2017). AOM
peptides/proteins contain both HPI and HPO parts because they are composed of
hydrophilic as well as hydrophobic amino acids. However, in aqueous solutions, the HPO
segments tend to cluster together, leading to protein folding, while the HPI regions are
exposed to the surroundings (Creighton, 1993).

2.3.2. Molecular Weight Distribution

AOM contains components of molecular weights (MW) ranging from several
hundred daltons to hundreds of kilodaltons (Pivokonsky et al., 2006, 2014; Henderson et
al., 2008a; Fang et al., 2010; Li et al., 2012). Low-MW fractions (< 10 kDa) include
substances such as aldehydes, amines, amino acids, peptides, and mono-, di- and
oligosaccharides (Nguyen et al., 2005; Huang et al., 2007). Compounds with medium
MW (10-100 kDa) are, for example, polypeptide molecules, such as enzymes and their
components (Pivokonsky et al., 2014) and high-MW fraction (> 100 kDa) comprises of
biopolymers like proteins and polysaccharides. These polymers can form a significant
portion of AOM (Myklestad, 1995; Henderson et al., 2008a; Pivokonsky et al., 2014).
For example, proteins > 100 kDa accounted for 25 and 22% of EOM and COM, resp., for
Microcystis aeruginosa stationary-phase peptide/protein fraction. Non-proteinaceous
matter of several species (Chlamydomonas geitleri, Fragilaria crotonensis, M.
aeruginosa) contained 20-27% (EOM) and 22-35% (COM) with MWs of > 100 kDa
(Pivokonsky et al., 2014). Henderson et al. (2008a) quantified even AOM fraction > 500
kDa in M. aeruginosa and determined it to reach 25%.

Figure 1. Molecular weight fractionation of (a) COM peptides/proteins and (b) non-proteinaceous
cellular organic matter of different phytoplankton species (Chlamydomonas geitleri, Chlorella vulgaris,
Fragilaria crotonensis, Merismopedia tenuissima and Microcystis aeruginosa). Adapted from
Pivokonsky et al. (2014), Baresova et al. (2017) and Naceradska et al. (2018).
114 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

By contrast, substances with very low MW can also form a substantial part of AOM
(Hoyer et al., 1985; Henderson et al., 2008a). E.g., 30% of Chlorella vulgaris AOM, 38%
of M. aeruginosa AOM, 53% of Melosira sp. AOM and even 81% of Asterionella
formosa AOM was determined to be < 1 kDa (Henderson et al., 2008a). More
information on the proportion of high- and low-MW organics in AOM of different
species is shown in Figure 1, where the difference between AOM peptides/proteins and
non-proteinaceous matter is emphasized.

Figure 2. Molecular weight distributions of EOM and COM peptides/proteins of (a) Chlamydomonas
geitleri, (b) Fragilaria crotonensis, (c) Merismopedia tenuissima and (d) Microcystis aeruginosa. EOM
MWs were determined at exponential and stationary growth phases (EOMexp, EOMstat, resp.), COM was
obtained at the stationary growth phase. Adapted from Pivokonsky et al. (2014) and Baresova et al.
(2017).

The MW distribution differs not only between the diverse phytoplankton species, but
also depends on the age of the culture and AOM fraction (i.e., EOM vs. COM). When the
MWs of AOM peptides/proteins were analysed, COM was found to contain a much
 Flocculation of AOM in Water Treatment 115

broader range of MW values compared to EOM in case of several phytoplankton species

(Anabaena flos-aquae, C. geitleri, F. crotonensis, M. aeruginosa, Scenedesmus
quadricauda), while some of the peptides/proteins were COM-specific (Pivokonsky et
al., 2006, 2014). Additionally, changes in MW distribution were observed
between exp and stat EOM, where the proportion of higher MW compounds usually
increased with time, due to the exclusion of low-MW substances (Pivokonsky et al.,
2014). Examples of AOM MW distribution of four different species are depicted
in Figure 2.

2.3.3. Charge
The charge of AOM is strongly pH dependent and is attributable to the presence of
diverse functional groups that can release or accept proton, depending on the pH
conditions (Creighton, 1993). It was found that AOM is negatively charged throughout a
wide range of pH values (Bernhardt et al., 1985; Henderson et al., 2008a), e.g.,
Henderson et al. (2008a) determined negative zeta potential values of EOM derived from
various species in the pH range 2-10. Nevertheless, the charge density varies among the
species and their growth stage (Bernhardt et al., 1985; Paralkar and Edzwald, 1996;
Henderson et al., 2008a). For example, the charge density of M. aeruginosa EOM at pH 7
decreased from 0.2 meq per 1 g DOC at the exponential growth phase to 0.1 meq per 1 g
DOC at the stationary growth phase. On the contrary, the charge density of C. vulgaris at
pH 7 increased from 0.9 to 3.2 meq per 1 g DOC from exp to stat phases (Henderson et
al., 2008a).
Additionally, distinct AOM constituents differ in their charge properties. AOM
peptides/proteins are amphoteric due to the content of diverse functional groups (both
acidic and basic, i.e., -OH, -COOH, -SH, -NH3+, =NH2+) (Creighton, 1993).
Peptides/proteins with isoelectric points (pI) ranging from 4.8-8.1 were identified in
COM of M. aeruginosa (Pivokonsky et al., 2012; Safarikova et al., 2013). Nevertheless,
negative charge of peptides/proteins prevails within pH ranges relevant for water
treatment (Pivokonsky et al., 2012). The charge of polysaccharides stems from the
presence of uronic acids, containing weakly acidic -COOH groups (Hoyer et al., 1985).
However, the non-proteinaceous fraction is generally considered to bear less ionisable
groups and thus is less charged. For example, COM peptides/proteins produced by M.
aeruginosa were reported to contain 100 mmol titratable functional groups per 1 g DOC
(Safarikova et al., 2013), while non-proteinaceous COM of Chlorella vulgaris possessed
about 14 mmol titratable functional groups per 1 g DOC (Naceradska et al., 2018).
Similarly, alginate, an anionic polysaccharide, appeared to involve approx. 10 mmol -
COOH groups per 1 g DOC (Gregor et al., 1996).
116 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

3. FLOCCULATION OF AOM

A variety of chemicals have been trialled for coagulating algae and AOM including
traditional metal coagulants aluminium sulphate (alum), ferric sulphate (FS),
ferric chloride (FC) and inorganic polymers, for example polyaluminium chloride (PACl)
and polyferric sulphate (PFS). Natural polymers, such as chitosan or cationic
starch, have also been tested for coagulation of cell + AOM mixtures (Cheng et al., 2011;
Vandamme et al., 2012, 2014; Garzon-Sanabria et al., 2013). In addition,
alkaline flocculation, i.e., by chemical co-precipitation with calcium and magnesium salts
of the culture medium under alkaline pH values, is used for algal biomass harvesting for
biofuel production (Vandamme et al., 2012; González-Fernándes and Ballesteros, 2013;
Vandamme et al., 2016; Branyikova et al., 2018).
Most of the investigations into the flocculation of algae-laden waters have focused on
algal and cyanobacterial cells (Henderson et al., 2008b, 2010; Cheng et al., 2011;
Vandamme et al., 2012; Wyatt et al., 2012; Garzon-Sanabria et al., 2013; Gonzalez-
Torres et al., 2014; Baresova et al., 2017). They found that algal cells easily combine
with commonly used Al-based or Fe-based coagulants through electrostatic bridging
between negatively charged algal cell surface and positively charged coagulant hydroxide
precipitates at about neutral pH values (Henderson et al., 2008b; Wyatt et al., 2012;
Gonzalez-Torres et al., 2014; Baresova et al., 2017; see Figure 3). At higher coagulant
doses, sweep flocculation has also been reported to be influential (Wyatt et al., 2012;
Gonzalez-Torres et al., 2014).

Figure 3. Flocculation mechanism of cells and coagulant.

The negative surface charge of algal cells arises from deprotonation of acidic groups
(mostly -COOH) present at the cell surface or in EOM attached at the cell surface
(Henderson et al., 2008b; Wyatt et al., 2012; González-Fernándes and Ballesteros, 2013).
Wyatt et al. (2012) observed efficient flocculation of Chlorella zofingiensis cells even at
alkaline pH values, which may be attributed to interactions between cationic amine
 Flocculation of AOM in Water Treatment 117

groups on the cell surface with anionic coagulant precipitates, or when using sweep
coagulation at higher coagulant doses (Duan and Gregory, 2003) or co-precipitation with
calcium and magnesium salts. When natural polymers are used as coagulants, different
coagulation mechanisms may be expected, as shown by Cheng et al. (2011) for
flocculation of cells of Chlorella variabilis by chitosan.
In this study, flocculation of Chlorella cells improved at pH 8.5 compared to pH 5.5
and 7, suggesting that hydrogen bonds between chitosan (a poly-glucosamine polymer
with an isoelectric point around 6.5) and cell wall polysaccharides may be more
important than electrostatic interactions. Several studies (Bernhardt and Clasen, 1991;
Henderson et al., 2008b, 2010; Wyatt et al., 2012; Zhang et al., 2012) revealed a strong
stoichiometric relationship between cell surface area and coagulant demand
for spherical cells. The presence of EOM usually leads to the higher coagulant demand
(Henderson et al., 2010; Ma et al., 2012b; Zhang et al., 2012), which is further elaborated
in section 4.3.
Compared to algal cells, flocculation of AOM is markedly less investigated and
provides substantially lower removal efficiencies (Bernhardt et al., 1985, 1986, 1991;
Widrig et al., 1996; Pivokonsky et al., 2009a, b; Henderson et al., 2010; Safarikova et al.,
2013; Pivokonsky et al., 2012, 2015, 2016; Baresova et al., 2017; Naceradska et al.,
2017; Tang et al., 2017; Naceradska et al., 2018).
Henderson et al. (2010), for example, observed good cell removal (94-99%) for
Microcystis aeruginosa, Chlorella vulgaris, Asterionella formosa and Melosira
sp. by alum, while their EOM removal efficiency was 46-71%. Tang et al.
(2017) recorded high removal efficiencies (> 90%) for cells of cyanobacterium
Microcystis aeruginosa coagulating with PACl, but low removals for dissolved EOM (<
10%) and for cell-bound EOM (< 40%). Similarly, Baresova et al. (2017) obtained 99%
cell removal efficiencies for cyanobacterium Merismopedia tenuissima and
ferric sulphate, while its COM was reduced by 43-53% depending on the initial COM
concentration.
Similar to cell removal, AOM surface charge that is closely related to pH value is
crucial for flocculation. AOM is usually removed at acidic pH values, at which AOM
negatively charged functional groups interact with positively charged coagulant
hydroxopolymers (Widrig et al., 1996; Hu et al., 2006; Pivokonsky et al., 2009a, b). To
illustrate, Widrig et al. (1996), who examined the removal of EOM derived from green
algae Scenedesmus quadricauda and Dictyosphaerium pulchellum and cyanobacterium
Microcystis aeruginosa by alum and ferric chloride at two pH values, 5 and 8, showed
that overall removals were improved at pH 5 (M. aeruginosa – 20%, S. quadricauda –
25%, and D. pulchellum – 50%) compared to pH 8 (5-10%). Likewise, Baresova et al.
(2017) obtained the highest coagulation efficiencies for M. tenuissima COM at pH
ranging between 5-6.5, while cells were preferably removed at pH 6-7.7, suggesting that
different coagulation mechanisms were employed. Cells probably interacted through
118 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

adsorption onto Fe-precipitates, whereas COM preferred charge neutralisation by Fe-

hydroxopolymers. Pivokonsky et al. (2009a), who coagulated COM of M. aeruginosa by
ferric sulphate and alum, recorded the highest removals, 50% for ferric sulphate and 40%
for alum, at pH 4.5-6.5. However, the lowest coagulant residual concentrations, which
must also be taken into consideration, were observed in different pH range for alum (pH
6.5-7) and narrower pH range for ferric sulphate (pH 6-6.5). Unsurprisingly, these pH
ranges correspond to the lowest solubility of aluminium and ferric hydroxide precipitates
(Stumm and Morgan, 1996). Furthermore, the study demonstrated higher removal
efficiencies for proteinaceous (74% and 50%, respectively) than non-proteinaceous
compounds (12% and 22%, respectively), for ferric sulphate and alum. Higher removal
efficiencies for proteins (75% and 69%, respectively) than for saccharides (30% and
40%, respectively) were achieved also by Cui et al. (2016), who coagulated effluent
organic matter from wastewater treatment plant by aluminium chloride and
polyaluminium chloride (PACl). Several studies have, therefore, focused on coagulation
behaviour of proteinaceous and non-proteinaceous compounds separately (Pivokonsky et
al., 2012, 2015; Safarikova et al., 2013; Naceradska et al., 2018). The study on
coagulation of peptides/proteins contained in COM of Microcystis aeruginosa showed
that peptides/proteins were removed by ferric sulphate in the pH range of 4-6 owing to
charge neutralization of peptide/protein negative surface (given by the presence of -
COOH groups) by positively charged hydrolysis products of ferric coagulant (Figure 4a).
Moreover, adsorption of peptides/proteins onto ferric precipitates at pH 6-8 was observed
at low COM/Fe ratio (COM/Fe < 0.33, Figure 4b). By contrast, at higher COM/Fe ratio
(COM/Fe > 0.33), charge stabilization of Fe precipitates by peptides/proteins bound to its
surface hindered flocculation at pH 6-8 (Figure 4c). In addition, peptide/protein
flocculation was disturbed at pH of about 6.2 as a consequence of formation of dissolved
Fe-peptide/protein complexes. However, this can be avoided by consistent optimization
of pH conditions. The study by Pivokonsky et al. (2015) indicated that alum coagulates
COM peptides/proteins of M. aeruginosa at pH values slightly higher (5-6.5) than in the
case of ferric coagulant (Pivokonsky et al., 2012). Peptide/protein flocculation was also
disturbed by formation of complexes between coagulant and peptides-proteins at pH of
about 7 during the flocculation by alum. The difference in the pH values effective for
coagulation using alum and ferric sulphate is attributed to a difference in the hydrolysis
product distributions of Al and Fe (Stumm and Morgan, 1996).
The only study investigating the flocculation of non-proteinaceous AOM was
undertaken by Naceradska et al. (2018). Non-proteinaceous COM was gained from
Chlorella vulgaris through protein precipitation. Only up to 25% of non-proteinaceous
COM (mostly carbohydrates) was removed by flocculation by either alum or PACl in the
optimized pH ranges 6.6-7.5 for alum and 7.5-9 for PACl. These pH values indicated that
the prevailing coagulation mechanism was adsorption of non-proteinaceous COM onto
aluminium hydroxide precipitates (Figure 5) rather than charge neutralization as stated
 Flocculation of AOM in Water Treatment 119

for COM peptides/proteins (Pivokonsky et al., 2012). The low removals were attributed
to high content of low-MW organics (< 10 kDa) in non-proteinaceous COM, which were
disinclined to flocculate.

Figure 4. Flocculation mechanisms of COM proteins and coagulant: (a) pH = 4-6, (b) pH = 6-8, low
COM/Fe ratio and (c) pH = 6-8, high COM/Fe ratio. Adapted from Pivokonsky et al. (2012).

Figure 5. Flocculation mechanisms of non-proteinaceous COM and coagulant.

120 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

The reluctance of low-MW organics to flocculate was also ascertained for COM
peptides/proteins (< 10 kDa) (Pivokonsky et al., 2012, 2015) and low-MW humic
substances as a part of natural organic matter commonly occurring in surface water
sources (Sillanpää et al., 2017). It should be noted that most cyanobacterial toxins are of
low-MW, for instance microcystins have MW around 1 kDa, and therefore coagulation is
largely ineffective for their removal (de Figueiredo et al., 2004).
When flocculation performance of the most used coagulants is compared, it can be
concluded that aluminium-based and ferric-based coagulants perform very similarly in
AOM coagulation in terms of removal efficiencies under slightly shifted optimum pH
values, given by different distribution diagrams of their hydrolysis products (Widrig et
al., 1996; Pivokonsky et al., 2009a; Safarikova et al., 2013).

4. IMPACT OF AOM ON COAGULATION

OF OTHER IMPURITIES

4.1. Inorganic Particles

Studies that dealt with the impact of AOM on coagulation of inorganic particles
showed that the AOM interacted with the inorganic particles and that the presence of
AOM substantially changed the optimum pH for effective flocculation by ferric or
aluminium coagulants (Bernhardt et al., 1985, 1986, 1991; Safarikova et al., 2013).
Inorganic particles are often present in raw water, where they cause high turbidity, and
they usually bear a negative charge in a wide range of pH values. The interactions
between clay surfaces and organics were demonstrated in soils for lots of organic
compounds, such as mono- and polysaccharides, amino acids, proteins, nucleic acids and
humic substances (Goring and Bartholomew, 1952; Greenland, 1956; Pinck, 1962; Parfit
and Greenland, 1970; Labille et al., 2005). Organic compounds that bear a charge (e.g.,
acidic polysaccharides, proteins) can interact with the negative charge of clays by
electrostatic interactions. These are affected by pH of the solution, which governs
dissociation constants of polar functional groups and thus the amount of charged groups
in both clays and organic compounds (Pinck, 1962). For neutral compounds, several
molecular mechanisms for adsorption via weak bonds were proposed: (1) substitution of
interlayer water molecules solvating exchangeable cations (Parfit and Greenland, 1970),
(2) hydrogen bonds between macromolecule hydroxyl and clay surface oxygens, or (3)
hydrogen bonds between hydroxyl groups on sheet edges and macromolecule (Labille et
al., 2005). Clay-organic interactions were ascertained also in cases of water treatment.
For instance, Bernhardt and co-workers (Bernhardt et al., 1985, 1986, 1991), who
investigated the effect of EOM of MW > 2 kDa excreted by several algal species on the
 Flocculation of AOM in Water Treatment 121

coagulation of inorganic quartz particles, concluded that EOM polymers (neutral and
acidic polysaccharides) were able to attach to the negatively charged surface of quartz
particles by means of hydrogen and covalent bonds. Likewise, a study by Safarikova et
al. (2013) suggested that cellular peptides/proteins of M. aeruginosa interact
electrostatically (though -NH3+ and =NH2+ groups) with the negatively charged surface of
kaolin particles. In both cases, AOM and inorganic particles formed negatively charged
clusters, which were then removable by both ferric and aluminium coagulants in the pH
range, where Fe/Al formed hydrolytic products bearing a positive charge, i.e., at pH
ranges of 4-6.5 in the case of Fe and 4.5-7 in the case of Al (Figure 6).

Figure 6. Flocculation mechanism of kaolin, COM proteins and coagulant. Adapted from Safarikova et
al. (2013).

By comparison, in the absence of AOM, inorganic particles are flocculated at higher

pH values through adsorption onto Fe/Al hydroxide precipitates provided by electrostatic
interactions, exchanging reactions and hydrogen bonds (Shin et al., 2008). For instance,
optimum pH values for kaolin removal are between 6 and 8 for ferric coagulants and
between 7 and 8.5 for aluminium ones. It should be noted that kaolin interacted with
Fe/Al hydroxopolymers also at lower pH values, but flocculation efficiency was low
(Ching et al, 1994; Kim and Kang, 1998; Safarikova et al., 2013). In the presence of
AOM, the negative charge in the system increased with increasing pH to alkaline values
and electrostatic repulsions between negatively charged particles led to system
stabilization. The key factor for flocculation of inorganic particles and AOM is the ratio
of coagulant to organic matter or more precisely, the ratio of charges in the system
(Bernhardt et al., 1985, 1991; Safarikova et al., 2013). Bernhardt et al. (1991) found that
a critical mass Fe/DOC ratio for EOM removal was within the range of 2-3 mg (0.036-
0.054 mmol) Fe per 1 mg DOC depending on species. Above this ratio, a partial coating
of the quartz particles surface by EOM polymers can lead to particle aggregation through
polymer bridging and aid flocculation. Below this ratio, large coverage of quartz particles
122 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

by EOM polymers enhances the stability of the dispersed system by steric and charge
stabilization (Bache and Gregory, 2007) and impairs flocculation. Similar results were
obtained also in the case of alginic acid, an polyuronic acid composed of two uronic acids
(poly-D-mannuronic acid and L-guluronic acid), which was used as a model compound
for EOM acidic polysaccharides (Bernhardt et al., 1985, 1986, 1991). Similar to the
flocculation of AOM solely (section 3.), Safarikova et al. (2013) ascertained that high-
MW peptides/proteins were removed by flocculation, while low-MW ones were reluctant
to flocculate.

4.2. Humic Substances

In many regions, humic substances (HS), comprising humic and fulvic acids (HA and
FA), constitute the majority of natural organic matter in surface waters and may also
occur together with AOM (Knauer and Buffle, 2001). While a lot of emphasis has been
put on removal of HS, very little attention has been paid to possible HS-AOM
interactions and their simultaneous removal by flocculation. One of the rare
investigations was done by Jiang et al. (1993) who flocculated cells of diatom
Asterionella formosa and AOM excreted during its growth by four different inorganic
coagulants (polyferric sulphate, ferric sulphate, aluminium sulphate, polyaluminium
chloride) in the presence of HS. They ascertained that the addition of HS to the algae +
AOM solutions lead to a reduction of flocculation performance with increasing
concentration of HS. With a greater HS concentration, higher doses of coagulants were
required to achieve overall charge neutralization and removals comparable to those in the
absence of HS. On the other hand, several studies have demonstrated that HS is able to
adsorb onto the surfaces of freshwater phytoplankton cells and that the HS-cell
interactions are strongly dependent on pH value (Campbell et al., 1997; Vigneault et al.,
2000; Knauer and Buffle, 2001). Campbell et al. (1997) have proposed two mechanisms
for the adsorption of HS onto the phytoplankton cell surfaces: (1) hydrogen bonds and (2)
the formation of hydrophobic bonds between the cell surface and the hydrophobic
domain of the HS. Interactions between HS and microbial products, such as
polysaccharides and proteins, were also observed during membrane filtration (Myat et al.,
2014). Myat et al. (2014) investigated the impact of interactions between model
compounds, sodium alginate and bovine serum albumin (BSA) as representative
biopolymers, with humic acid on membrane fouling. They detected formation of
aggregates between sodium alginate and BSA with humic acid by liquid chromatography
and they also employed molecular dynamics simulations to provide insights into the
interaction mechanisms.
For the BSA-humic acid system, they showed that electrostatic, hydrophobic and
hydrogen bonding were the dominant types of interactions predicted. For the alginate-
 Flocculation of AOM in Water Treatment 123

humic acid system, the interactions predicted were divalent ion-mediated interactions
only. The existence of interactions between HS and BSA was confirmed by Pivokonsky
et al. (2015). Moreover, they found that HS interact also with cellular proteins of M.
aeruginosa (MA) and that interactions between HS-BSA and HS-MA proteins have very
similar consequences for flocculation. In both cases, the clusters of HS-BSA and HS-MA
proteins were flocculated by alum with DOC removals of 83% and 80%, respectively, in
the pH range 5.5-6.2 through a charge neutralization mechanism, i.e., positively charged
Al-hydroxopolymers interacted with negatively charged functional groups of BSA/MA
proteins and HS (Figure 7).

Figure 7. Flocculation mechanism of humic substances, COM proteins and coagulant. Adapted from
Pivokonsky et al. (2015).

Interestingly, protein-HS interactions lead to significant reductions in coagulant dose.

While optimized coagulant doses were 0.25, 0.40 and 1.10 mg Al mg-1 DOC for BSA,
MA proteins and HS, respectively, Al doses of 0.16 and 0.28 mg Al mg-1 DOC were
sufficient for flocculation of HS-BSA and HS-MA proteins mixtures. Similar to
flocculation of single AOM (section 3.) and AOM together with inorganic particles
(section 4.1.), high-MW proteins underwent flocculation, while peptides of MW < 10
kDa formed residual organics after flocculation.

4.3. Algal Cells

Most studies that dealt with the simultaneous flocculation of AOM and algal cells,
have focused on the flocculation of algal cells in the presence of extracellular organic
matter, either produced by cells during their growth into cultivation media in water
treatment or during the harvest of algal biomass (Bernhardt and Clasen, 1991; Henderson
et al., 2010; Vandamme et al., 2012; Garzon-Sanabria et al., 2013; Wu et al., 2012;
124 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

González-Fernándes and Ballesteros, 2013; Branyikova et al., 2018), or EOM/COM

released by oxidation processes employed prior to flocculation in water treatment (Ma
and Liu, 2002; Plummer and Edzwald, 2002; Henderson et al., 2008b; Ma et al., 2012a,
b; Wang et al., 2013).
It was found that the presence of EOM increased the amount of required coagulant
(Henderson et al., 2010; Vandamme et al., 2012; Garzon-Sanabria et al., 2013). For
example, Garzon-Sanabria et al. (2013) reported that during flocculation of
Nannochloropsis salina, EOM increased demand of various coagulants. For 90%
flocculation efficacies, a 3-fold more of aluminium chloride, a 7-fold dose of synthetic
cationic polymers and almost a 10-fold more of chitosan was needed. Similarly,
Vandamme et al. (2012) showed that a 6-fold dose of aluminium sulphate, a 9-fold dose
of chitosan and about a 4.5-fold dose of cationic starch was required for 85% removals of
Chlorella vulgaris cells in the presence of EOM carbohydrates compared to flocculation
of cells in the absence of EOM carbohydrates. Moreover, Wu et al. (2012) ascertained
that the release of algal polysaccharides reduced alkaline flocculation efficiencies for
algal cells of five species, but this could be mitigated by a further increase in pH values.
On the other hand, Branyikova et al. (2018) reported that COM isolated from C. vulgaris,
which contained a large proportion of non-peptide fractions (95%), showed little
interference in alkaline flocculation of C. vulgaris cells. The study by Henderson et al.
(2010) revealed the stoichiometric relationship between coagulant demand and charge
density of algae + EOM system and further reported that coagulant demand decreased as
molecular mass of EOM increased. Moreover, they pointed out that, in the case of some
algal species (e.g., C. vulgaris), charge density is predominantly associated with the
EOM component.
On the other hand, some studies observed a positive influence due to the presence of
high-MW EOM and/or COM released after pre-oxidation on the flocculation of algal
cells, since algal biopolymers acted as a polymer coagulant aid (Ma and Liu, 2002;
Henderson et al., 2010; Ma et al., 2012a; Wang et al., 2013). High-MW compounds
easily combine with coagulants, promote flocculation and cell removal, most likely
through adsorptive bridging. However, the release of COM and degradation of
EOM/COM may have not only beneficial but also detrimental effects on flocculation,
depending on oxidizer dose, pH value and the properties of EOM/COM (Hoyer et al.,
1987; Paralkar and Edzwald, 1996; Henderson et al., 2008b; Ma et al., 2012a, b; Coral et
al., 2013; Wang et al., 2013). Overdosing of pre-oxidant leads to cell lysis, releasing
undesirable toxins or taste and odour compounds, and to EOM/COM degradation to low-
MW compounds which may then be difficult to flocculate (Hoyer et al., 1987; Henderson
et al., 2008b; Ma et al., 2012a, b). The positive impact of COM compounds on
flocculation of algal cells was also observed by Baresova et al. (2017). In their study,
COM did not originate from pre-oxidation, but was extracted from cells and then dosed
into cell suspensions. They found that during flocculation of Merismopedia tenuissima
 Flocculation of AOM in Water Treatment 125

cells, COM served as a cationic polymer coagulation aid and that the presence of COM
shifted the optimum pH values for flocculation. While the cells were removed by
adsorption onto coagulant precipitates (Fe-oxide-hydroxides) at around neutral pH, the
COM + cell mixtures underwent charge neutralisation by Fe-hydroxopolymers within
moderately acidic pH (5-6.5). Furthermore, coagulant doses for COM + cell mixtures
were even slightly lower than those for single COM flocculation and the achieved DOC
removals for COM + cell mixtures (37-57%) were comparable to the single COM
flocculation (43-53%, rising with the increasing initial COM concentration). The high
cell removals (up to 99%) were obtained in both the presence and absence of COM. The
beneficial effect of COM on flocculation of M. tenuissima cells stemmed from bridging
cells by high-MW COM, mainly on basis of hydrogen bonds and electrostatic
interactions (Figure 8).

Figure 8. Flocculation mechanism of cells, COM and coagulant. Adapted from Baresova et al. (2017).

5. AOM AS A COAGULANT

Natural polymeric materials are increasingly used in water and wastewater treatment
as a coagulant or coagulant aid. The primary flocculation mechanism is ‘bridging’ which
involves adsorption of the polyelectrolyte molecule from solution onto the surface of
suspended particles. The most commonly used natural polymers are chitosan, alginate,
starch and extract from seeds of Moringa oleifera. Since alginates bear a strong
resemblance to algal products, it can be assumed that some compounds contained in
AOM may also serve as coagulants. For instance, Devrimci et al. (2012)
showed that calcium alginate proved to be a successful coagulant for turbid waters
containing clay (predominantly smectite) and that calcium ions eased alginate-clay
interaction.
126 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

As stated in section 4.1., many studies, investigating the processes in soils,

demonstrated that natural organic compounds, such as mono- and polysaccharides, amino
acids, proteins, nucleic acids and humic substances adsorb onto clay surfaces by a range
of mechanisms. To illustrate, Labille et al. (2005) ascertained that bacterial
polysaccharides (dextran, succinoglycan, YAS34, xanthan, MWAP71, rhamsan,
RMDP17) are able to attach to and subsequently flocculate Na-montmorillonite particles
in the presence of CaCl2, which screened electrostatic repulsions between
polysaccharides and Na-montmorillonite. The study further showed that flocculation of
charged polysaccharide succinoglycan, which carries negative charge due
to acidic moieties, was favoured at pH values below 6 due to electrostatic interactions
between polysaccharide carboxyl groups and positively charged sheet edges
of clay. At higher pH values, deprotonated sheet edges of Na-montmorillonite prevented
such interactions and decreased flocculation efficiency was observed.
Only a handful of studies on the effect of AOM as a coagulant have been conducted
(Paralkar and Edzwald, 1996; Safarikova et al., 2013; Pivokonsky et al., 2015; Baresova
et al., 2017). Several studies suggest that autoflocculation of algal cells in algal biomass
harvesting for biofuel may be induced by extracellular polymeric substances (EPS) as
reviewed by González-Fernándes and Ballesteros (2013). Carbohydrates are the main
constituents of EPS, together with proteins in a lesser extent.
Regarding drinking water treatment, a study by Paralkar and Edzwald (1996) pointed
out that EOM produced by diatom Cyclotella sp., green algae Scenedesmus quadricauda
and Chlorella vulgaris can interact with positively charged latex particles during
experiments conducted at pH 7. Nevertheless, only EOM of Cyclotella sp., which
contained the highest percentage of high-MW material, flocculated latex particles into
aggregates that underwent sedimentation.
As stated in section 4.1., Safarikova et al. (2013) observed electrostatic interactions
between M. aeruginosa cellular peptides/proteins and kaolin particles. At pH values
above 4.5, these interactions did not lead to aggregate formation due to electrostatic
repulsions between peptide/protein-kaolin clusters and the addition of positively charged
Al/Fe coagulant was necessary to induce flocculation.
Nevertheless, at pH < 4.5, the negative charge of both kaolin and peptides/proteins
was suppressed by protonation of their functional groups (especially -COOH groups of
peptides/proteins), which enabled more peptide/protein-kaolin interactions and
subsequent flocculation (Figure 9a). The turbidity removal efficiencies remained equal to
those in the presence of coagulant, while DOC removal efficiencies decreased from 70-
80% to 45%. Similarly, Pivokonsky et al. (2015) ascertained that protonation of
functional groups relating to a reduction in negative charge resulted in the flocculation of
M. aeruginosa cellular peptides/proteins and humic substances at pH < 4 in the absence
of coagulant (Figure 9b). DOC reductions of 55-69% were recorded, increasing with
higher protein/humic ratio.
 Flocculation of AOM in Water Treatment 127

Figure 9. Flocculation mechanisms of (a) kaolin and COM proteins (adapted from Safarikova et al.,
2013), (b) humic substances and COM (adapted from Pivokonsky et al., 2015) and (c) cells and COM
proteins (adapted from Baresova et al., 2017).

Furthermore, Baresova et al. (2017) found that the interactions of COM of

Merismopedia tenuissima with its cells (mentioned in section 4.3.) resulted in
flocculation at a pH range 3.5-4.5 with DOC and cell removal efficiencies comparable to
those achieved in the presence of Fe coagulant. COM-cell flocculation was also attributed
to protonation of -COOH groups of both COM and cell surface at low pH values and the
attachment of COM to cells via hydrogen bonds and attractive electrostatic interactions
(Figure 9c).
128 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

6. INFLUENCE OF AOM ON FLOC PROPERTIES

Floc properties, such as size, structure, shape, strength or settling velocity, are
important parameters for the downstream treatment processes, i.e., separation by
sedimentation, flotation and filtration. Each of these technological steps requires different
flocs for their successful separation. Small (generally < 50-60 µm) and dense flocs are
suitable for a single-stage separation by sand filtration (Ngo et al., 1995; Bubakova and
Pivokonsky, 2012). For DAF (dissolved air flotation), Edzwald (1995) recommended
producing strong flocs with particle size distributions in the range 10-30 µm (or 25-50
µm in Edzwald, 2010); however, Vlaški et al. (1997) reported that larger flocs (> 50 µm)
formed by very low shear rate (G = 10 s-1) and hence having low density, resulted in
efficient DAF as well. Finally, sedimentation is generally considered to be suitable for
removal of large (> 100 µm) and dense flocs which have high settling velocities (e.g.,
Edzwald, 1995).
Generally, floc properties are influenced by several basic parameters: the
composition (type) and concentration of floc components, i.e., impurities to be removed
and the coagulant; the shear rate (velocity gradient, G) and the time of its action
(flocculation time). The properties of algal/cellular organic matter flocs have only been
examined marginally thus far; however, some works concerning AOM, sometimes
together with algal/cyanobacterial cells, have been published in recent years (Henderson
et al., 2006; Pivokonsky et al., 2009a, b; Gonzalez-Torres et al., 2014, 2017; Vandamme
et al., 2014; Jiao et al., 2015; Chekli et al., 2017; Filipenska et al., 2018).
Some of them examined either the influence of coagulant dose and type, or pH value
on AOM floc properties. To illustrate, Chekli et al. (2017) evaluated the performance of
TiCl4 and polytitanium tetrachloride (PTC) for the removal of AOM from a marine
diatom Chaetoceros muelleri in comparison with conventional FeCl3 treatment. They
found that PTC and TiCl4 reached higher removal efficiencies (in terms of turbidity,
DOC and UV254) and made larger, stronger and more compact flocs than FeCl3.
Gonzalez-Torres et al. (2014) examined the properties of flocs made of M. aeruginosa
cells and its AOM using Al2(SO4)3 or FeCl3 as coagulants at pH 6 (lower coagulant dose,
charge neutralisation mechanism) and 7 (higher coagulant dose, sweep flocculation
mechanism). Ferric flocs were confirmed to be larger than alum flocs regardless of the
pH value or coagulant dose. Within one coagulant type, the coagulant dose was another
factor influencing the floc size, since a higher dose produced larger flocs. In the case of
the constant both coagulant type and dose, the pH values also affected the size of flocs.
Specifically, pH 7 resulted in larger flocs than pH 6. Floc strength was determined
primarily by the coagulant dose, irrespective of coagulant type. Higher coagulant doses
produced stronger flocs, the strongest of which being produced at the higher dose of Fe at
pH 6.
 Flocculation of AOM in Water Treatment 129

The others evaluated the influence of AOM on the properties of flocs made of other
impurities, such as clay minerals or cells. Unfortunately, a direct comparison of floc
properties reported by different researchers is made difficult because of the varying
experimental conditions (different/unknown shear rates, different coagulant doses or
various AOM compositions and concentrations). Yet some general conclusions can be
made. The principal finding is that the presence of AOM increases the floc size
(Vandamme et al., 2014; Filipenska et al., 2018). For instance, Vandamme et al. (2014)
coagulated Chlorella vulgaris cells with and without AOM using alum, chitosan and
cationic starch as coagulants, electro-coagulation and alkaline flocculation. Generally,
AOM increased the floc size (up to ten times for chitosan). However, flocs formed using
cationic starch remained of the same size irrespective of the AOM presence. A clear trend
was observed by Filipenska et al. (2018) who coagulated kaolinite and COM proteins of
M. aeruginosa (separately and together) using Al and Fe salts as coagulants. In all cases
COM proteins definitely caused the floc size to increase. However, the structure of flocs
was more influenced by the coagulant type than by the presence of COM as the most
compact and regular (in shape) flocs were Al-kaolinite and Al-COM flocs. Henderson et
al. (2006) reported that flocs created from C. vulgaris cells and EOM were larger, but
weaker, than flocs formed by inorganic particles (kaolin) or NOM (humic matter).
Unfortunately, the tests with single cells without EOM have not been made, therefore it is
not possible to evaluate the specific EOM contribution to the resulting floc size.
Moreover, Pivokonsky et al. (2009b), who coagulated COM of M. aeruginosa with ferric
sulphate, reported that COM concentration considerably affected the floc size, with lower
COM concentration producing smaller flocs. The influence of the origin and composition
of AOM on the floc characteristics can be demonstrated by research conducted by
Gonzalez-Torres et al. (2017). They coagulated cells and associated EOM of C. vulgaris
and M. aeruginosa by alum and used reflectance Fourier transform infrared (FTIR)
imaging for biochemical characterisation of the associated flocs. M. aeruginosa flocs
were measurably more uniform and smaller than C. vulgaris flocs and visibly more
dense, which was attributed to the difference in AOM concentration and composition.
The large flocs of C. vulgaris were characterized by a homogenous distribution of
proteins and polysaccharides across the floc, indicative of high glycoprotein content. In
contrast, the smaller but stronger flocs of M. aeruginosa had localized areas of increased
protein concentration at the edge of regions which were absent of proteins and
polysaccharides (and thus potentially comprising coagulant).
Flocculation mechanisms and inter-particle interactions (mentioned in
previous sections) acting within flocs also seem to influence the floc properties.
Filipenska et al. (2018) pointed out an interesting phenomenon. When floc size
130 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

and shear rate were plotted in a log-log plot6, a change in γ constant at approximately G ~
120 s-1 for all types of flocs and an additional change in γ for COM flocs,
between G of 40 to 50 s-1 and 60 to 70 s-1 for Al and Fe, respectively, could be observed
(Figure 10).

Figure 10. Floc size vs. shear rate log-log plots of six different types of flocs. A change in γ constant at
approximately G ~ 120 s-1 for all types of flocs and an additional change in γ for COM flocs, between G
of 40 to 50 s-1 and 60 to 70 s-1 for Al and Fe, respectively, can be observed. Adapted from Filipenska et
al. (2018).

The authors attributed these changes to different intermolecular interactions

(hydrogen bonding or hydrophobic interaction) that are not able to withstand the
hydrodynamic force when shear rate is increased above certain levels.

CONCLUSION

This review shows that flocculation is a useful technology for the treatment of algae-
laden waters. Specific outcomes and further research needs can be summarized as
follows:

6
The dependence of the floc size on the shear rate is described by a decreasing power function (e.g., Parker et al.,
1972) dav/max = CG-2γ, where dav/max is the average or maximal diameter of flocs in the system, G is the shear
rate (global, average, mean) and C and γ are constants that include parameters such as the composition and
concentration of contaminants, type and dose of coagulant and reaction pH (determining the attractive forces
between individual floc components) and the type of flow or energy dissipation (the effect of hydrodynamics).
 Flocculation of AOM in Water Treatment 131

 AOM is a mixture of various organic compounds (e.g., peptides/proteins,

carbohydrates, etc.) that behave differently during flocculation due to their
diverse properties, such as the degree of hydrophobicity, MW distribution and
charge. Determination of these characteristics can contribute to optimizing
removal of AOM and to elucidating the involved mechanisms.
 Flocculation effectively removes algal cells and algae-derived organic substances
of higher molecular weights, but is ineffective in removing those of low-MWs
(including cyanobacterial toxins) and thus other treatment needs to be employed
for their removal. Flocculation seems to be an effective pre-treatment to improve
the performance and reduce the cost of downstream processes, such as adsorption
onto activated carbon or membrane filtration, which are susceptible to the
presence of biopolymers in source water.
 Optimization of flocculation pH is crucial for efficient AOM removal. For
removal of algal cells and whole COM, suitable pH values lie in the range 6-8
and 5-6.5, respectively, for Al-based or Fe-based coagulants. The latter is similar
to COM proteins (pH 4-6.5). For non-proteinaceous matter, the optimum pH lies
in the alkaline range due to a different flocculation mechanism (see Figure 11).
Higher removal efficiencies (70-80%) are achieved for peptides/proteins than for
non-proteinaceous organics (< 25%). Flocculation of non-proteinaceous COM
and also species-specific differences in flocculation behaviour require further
investigation.
 Polymers contained in algal organic matter can act as polymer aids and enhance
the removal of other impurities (inorganic particles, humic substances, cells).
However, at this time studies are limited to a few algal/cyanobacterial species
and extension of this research on more species would be helpful.
 Floc size decreases and density increases with increasing shear rate, ferric flocs
being larger than alum flocs. The presence of AOM results in an increase of the
floc size, where COM concentration has a proportional relationship to floc size.
The floc properties also depend on the AOM origin (e.g., microorganism
species), and are influenced by the intermolecular interactions acting within
flocs. However, very few studies concerning properties of AOM flocs have been
published thus far and it is almost impossible to compare their results because of
diverse input parameters, such as coagulant/AOM concentration ratio. Therefore,
comparative studies would substantially advance the knowledge on AOM floc
properties.
132 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

Removal efficiencies correspond to reductions in AOM concentration (expressed as dissolved organic

carbon concentration).
*Algal cells removal efficiencies correspond to reductions in optical densities.

Figure 11. Optimal pH ranges for efficient removal of AOM, AOM together with other impurities and
AOM removal efficiencies. Based on Pivokonsky et al. (2012, 2015), Safarikova et al. (2013), Baresova
et al. (2017) and Naceradska et al. (2018).

ACKNOWLEDGMENTS

The research project was funded by the Czech Science Foundation under the Project
No. 18-14445S with institutional support RVO: 67985874. The authors acknowledge the
financial assistance on this project.

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134 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

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 Flocculation of AOM in Water Treatment 135

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138 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

plant operation and water quality: A review. Critical reviews in environmental

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140 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

BIOGRAPHICAL SKETCH

Martin Pivokonský

Affiliation: Institute of Hydrodynamics of the Czech Academy of Sciences, Prague,

Czech Republic

Education: Institute for Environmental Studies, Faculty of Science, Charles

University, Prague, Czech Republic

2016 Associate Professor in Environmental sciences

Inaugural thesis: “Characterisation of algal organic
matter and its influence on water treatment processes”
2002 Ph.D. degree in Environmental engineering
Ph.D. thesis: “Influence of the velocity gradient and time
of its application on the size characteristics of the
aggregates formed during water treatment”
1999 M.S. degree in Environmental engineering

Research and Professional Experience: Scientific management of the departments

at the Institute of Hydrodynamics; coordination and management of the research in the
field of water treatment, coagulation, flocculation, hydrodynamics, filtration,
sedimentation, adsorption, natural organic matter (NOM), algal organic matter (AOM)
Supervision and guidance of the M.S. and Ph.D. students of the Institute for
Environmental Studies, Faculty of Science, Charles University, Prague and the
Department of Water Technology and Environment Engineering, Faculty of
Environmental Technology, University of Chemistry and Technology, Prague
Expertise and consultancy activity in the field of the drinking water treatment and
technological audits of the waterworks, design of new technologies for suspension
formation and separation, e.g., PP&A Water Consulting (Pretoria, South Africa); Sv.
Trojice, Želivka, Kouty and Světlá nad Sázavou waterworks; AquaServis, Ltd.; Česká
voda – Czech Water, Ltd. (Czech Republic), EMBA spol. inc.

Professional Appointments:

06/2017 – Present Director

Institute of Hydrodynamics of the Czech Academy
of Sciences, Prague, Czech Republic

01/2014 – 05/2017 Deputy Director

 Flocculation of AOM in Water Treatment 141

Institute of Hydrodynamics of the Czech Academy

of Sciences, Prague, Czech Republic
2010 – Present Scientist – Associate Professor (10/2016 – present),
Research Assistant (2010 – 9/2016)
Institute for Environmental Studies, Faculty of Science,
Charles University, Prague, Czech Republic
2007 – 12/2013 Head of Department of Mechanics of Fluids and
Disperse Systems
Institute of Hydrodynamics of the Czech Academy
of Sciences, Prague, Czech Republic
2005 – Present Scientist
Institute of Hydrodynamics of the Czech Academy
of Sciences, Department of Mechanics of Fluids and
Disperse Systems, Prague, Czech Republic

Publications from the Last 3 Years:

Naceradska, J., Novotna, K., Cermakova, L., Cajthaml, T. & Pivokonsky, M. (2018).
Investigating the coagulation of non-proteinaceous algal organic matter: optimization
of coagulation performance and identification of removal mechanisms. Journal of
Environmental Sciences (submitted).
Filipenska, M., Vasatova, P., Pivokonska, L., Cermakova, L., Naceradska, J. &
Pivokonsky, M. (2018). Influence of COM-peptides/proteins on floc properties
coagulated by metal salts at different shear rate. Langmuir (submitted).
Branyikova, I., Filipenska, M., Urbanova, K., Ruzicka, M. C., Pivokonsky, M. &
Branyik, T. (2018). Physicochemical approach to alkaline flocculation of Chlorella
vulgaris induced by calcium phosphate precipitates. Colloids and Surfaces B:
Biointerfaces, 166, 54-60.
Angst, G., Mueller, C.W., Angst, Š., Pivokonský, M., Franklin, J., Stahl, P.D. & Frouz, J.
(2018). Fast accrual of C and N in soil organic matter fractions following post-
mining reclamation across the USA. Journal of Environmental Management, 209,
216-226.
Cermakova, L., Kopecka, I., Pivokonsky, M., Pivokonska, L. & Janda, V. (2017).
Removal of cyanobacterial amino acids in water treatment by activated carbon
adsorption. Separation and Purification Technology, 173, 330-338.
Baresova, M., Pivokonsky, M., Novotna, K., Naceradska, J. & Branyik, T. (2017). An
application of cellular organic matter to coagulation of cyanobacterial cells
(Merismopedia tenuissima). Water Research, 122, 70-77.
Naceradska, J., Pivokonsky, M., Pivokonska, L., Baresova, M., Henderson, R. K.,
Zamyadi, A. & Janda, V. (2017). The impact of pre-oxidation with potassium
142 Martin Pivokonský, Jana Načeradská, Kateřina Novotná et al.

permanganate on cyanobacterial organic matter removal by coagulation. Water

Research, 114, 42-49.
Pivokonsky, M., Naceradska, J., Kopecka, I., Baresova, M., Jefferson, B., Li, X. &
Henderson, R. K. (2016). The impact of algogenic organic matter on water treatment
plant operation and water quality: A review. Critical Reviews in Environmental
Science and Technology, 46(4), 291-335.
Novotná, K., Barešová, M., Čermáková, L., Načeradská, J. & Pivokonský, M. (2016).
Effect of cyanobacterial peptides and proteins on coagulation of kaolinite. European
Journal of Environmental Sciences, 6(2), 83-89.
Pivokonsky, M., Naceradska, J., Brabenec, T., Novotna, K., Baresova, M. & Janda, V.
(2015). The impact of interactions between algal organic matter and humic
substances on coagulation. Water Research, 84(1), 278-285.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 6

COMPARISON OF NATURAL COAGULANT AND

CHEMISTRY IN TANNING WASTEWATER TREATMENT
USING THE FLOCCULATION PROCESS

Edilaine Regina Pereira1, Gustavo da Silva Souza2

and Joseane Débora Peruço Theodoro1
1
Department of Environmental Academic,
Federal Technological University of Paraná,
Londrina, Paraná, Brazil
2
Environmental Engineer - UTFPR- Londrina,
Londrina, Brazil

ABSTRACT

The leather tanning industry uses a large amount of toxic substances and water.
Thus, it generates effluents with high polluting load. Due to the large amount of effluent
generated by tanneries and the difficulty in their treatment, several studies have been
proposed to minimize the environmental impacts caused by inappropriate disposal of this
effluent. This study investigated the performance of natural coagulants Tanin compared
to chemical coagulants aluminium sulphate and ferric chloride commonly used in the
treatment of raw wastewater from tannery, by means of the physicochemical processes of
coagulation, flocculation and sedimentation. Using jar tests methods, different
concentrations for the coagulants (chemical and natural) were applied to the concerned
effluent and it was assessed their efficiency in removing certain parameters such as
apparent color COD, and turbity besides the behavior of electrical conductivity and pH.
The results showed that for pH and electrical conductivity parameters, there was no
significant variation after application of coagulant in relation to the raw effluent. By
comparing the coagulants for colour parameter it was observed that the natural coagulant
144 E. R. Pereira, G. da Silva Souza and J. D. P. Theodoro

tanin had greater efficiency compared to the chemical ones, reaching 56.9% of removal,
however, the aluminium sulphate was more efficient at removing COD, with 95% of
removal. Therefore, it is noticed that the chemical coagulant aluminium sulphate was
more successful for the majority of the analysed parameters in comparison to other
coagulants.

Keywords: water treatment, organic coagulants, inorganic coagulants

1. INTRODUCTION

In developed and developing countries, most of the environmental impacts generated

are in the industrial sector. The industrial sector consumes a large part of natural
resources and also generates many types of wastes that are normally dumped into the
environment, more specifically into aquatic ecosystems, with or without adequate
treatment.
One of the productive processes that generate a considerable volume of liquid
effluents is the leather tanning industry. These industries have a great potential for
pollution, since the entire productive process of leather tanning consumes a large amount
of water, and the current average total consumption of the Brazilian sector is estimated at
25-30 m3 of water/ton of salty skin, about of 630 liters of water/salt skin on average
(PACHECO, 2005).
As a consequence, a quantity of liquid effluents similar to the total water withdrawn,
as well as solid waste, and the use of certain substances, such as sodium sulphide,
hydrated lime, inorganic acids, aluminum, titanium and chromium, the latter being the
main pollutant in tannery, used as a tanning agent in about 90% of the industry (BAYER,
2005).
In view of the problem of the treatment and inadequate disposal of effluents, which
may leave the quality of river water compromised since this is the destination of the
treated effluents of the industries, an efficient treatment in the stations of the industry is
necessary. An alternative currently under study is the substitution of chemical coagulants
used in the treatment of effluents by natural coagulants, as a way of minimizing the
impacts generated and modifies the composition of the sludge presented, making
proposals more sustainable and efficient.
The tannin has as one of its main advantages to present a reduction of contaminants
present in the treated effluent through the performance in the clarification of effluents.
Therefore, the objective of this research is to make a comparative application of the
efficiency of the natural coagulant tannin and the chemical coagulants aluminum sulphate
and ferric chloride already used in the industry for the treatment of tannery effluent.
 Comparison of Natural Coagulant and Chemistry in Tanning … 145

2. METHODOLOGY

The effluent used in this research came from a leather tanning industry, industry
located in the city of Ibiporã – PR, Brazil. The effluent was collected in the treatment
system of the still raw industry, before the application of the coagulant used by the
industry. After being collected, the effluent was sent for testing and experimental analysis
at the Sanitation Laboratory of the Federal Technological University of Paraná - Campus
Londrina. A pre-test was performed first, aiming at defining the most appropriate
concentrations of the chemical and natural coagulants used. Thus, the following
concentrations were obtained: for Tannin C1 3mgL-1, C2 6mgL-1, C3 9mgL-1; for
aluminum sulfate C1 15mgL-1, C2 20mgL-1, C3 25mgL-1; and for ferric chloride C1
15mgL-1, C2 21mgL-1, C3 28mgL-1.

Table 1. Parameters, equipment and methodology used

in this research

Parameters Equipments Methodology*

Apparent color Spectrophotometer HACH 4000 2120 C
pH pH monitor mPA-210 4500-H+ B
Electric conductivity Conductivity Meter Mca 150 2510 A
Solid series Greenhouse SL 100, Muffle MA 385, Balance 2540 A
AW 220
Chemical Oxygen Demand Biodigestor Thermo Digestor 462, 5220 A
Spectrophotometer HACH 4000

To prepare the coagulants 100 g of each coagulant were weighed and diluted in 1 L
of distilled water using a volumetric flask. After the concentrations of each coagulant
were defined in the pre-test, these concentrations were added to the effluent in a
coagulation/flocculation/sedimentation test using the Jar-test equipment.
The methodology for the test in the Jar Test was adapted from Theodoro (2012). The
parameters analyzed are described in Table 1 and are in accordance with APHA (2012).
The statistical model used to perform the analysis of the values resulting from the
four trials was the completely randomized design, in a factorial scheme. The statistical
model considered the effect of the interaction between Coagulant and Concentration
factors in addition to the effect of the Coagulant factor (in three levels) and the factor
Concentration (in three levels). Thus, the multiplicative statistical model is given by
equation 1:

Yijk    i   j  ij  eijk (1)

146 E. R. Pereira, G. da Silva Souza and J. D. P. Theodoro

On what:

 Yijk = observation corresponding to the kth experimental unit submitted to the i-th
level of the Coagulant factor and the jth level of the factor Concentration;
 : overall average common to all observations;
 i : effect of the ith level of the coagulant factor;
 j : effect of the jth level of the factor Concentration;
 ij: is the interaction effect between the i-th level of the Coagulant factor and
the j-th level of the factor Concentration;
 eijk: component of the random error associated with observation Yijk.

For the analysis of variance and comparison of the means of the three variables, a
level of significance of 10% was considered. The results of the analysis of variance were
grouped in Table 2 as proposed by Martins (2006).

Table 2. Model adopted to organize the results obtained with the

analysis of variance

Source of variation GL SQ QM FC P-value

Coagulant
Concentration
Coagulant X Concentration
Residue

3. RESULTS

The Table 3 shows the values with pH variation during sedimentation times.
For the coagulant Tannin, it was observed that there was not a very significant
variation in relation to the initial pH (12.97), and the greatest variation occurred in the
time of 30 minutes, when the lowest concentration of 300 mgL-1 reached the value of
12.90. The three concentrations obtained similar values and a little below the initial one,
varying from 12.90 to 12.92. The pH can be adjusted for an application of the Aluminum
Sulphate, in this way, the pH must be improved to be removed.
When analyzing the coagulant Ferric Chloride, it was observed that there was the
greatest reduction in the pH value with the highest coagulant concentration, reaching
12.53. After this, it can be observed that there was an increase in pH values for all
concentrations.
When comparing the data obtained with CONAMA Resolution 430 (CONAMA,
2011), it is observed that even after the treatment done in the effluent, it does not fit into
 Comparison of Natural Coagulant and Chemistry in Tanning … 147

the standards of release standard in relation to pH, since the Resolution requires that the
pH value is maintained between 5 and 9 to be released into a receiving body. In this case,
the pH of the tannery effluent should be corrected so that its value becomes less alkaline
and falls within the parameters required by the law for the pH parameter. There was no
significant variation in the pH values for tannery effluent with both natural and chemical
coagulants.

Table 3. pH variation as a function of coagulant versus time

Time [Tanin] = 3 mgL-1 [Tanin] = 6 mgL-1 [Tanin] = 9 mgL-1

(min) R1 R2 Average R1 R2 Average R1 R2 Average
Tanin

10 12.93 12.91 12.92 12.91 12.91 12.91 12.91 12.94 12.92

20 12.93 12.9 12.91 12.91 12.91 12.91 12.91 12.9 12.90
30 12.91 12.89 12.9 12.91 12.92 12.91 12.92 12.91 12.91
40 12.91 12.93 12.92 12.92 12.92 12.92 12.92 12.93 12.92
Time Al2(SO4)3 = 15 mgL-1 Al2(SO4)3 = 20 mgL-1 Al2(SO4)3 = 25 mgL-1
(min) R1 R2 Average R1 R2 Average R1 R2 Average
Al2(SO4)3

10 12.67 12.67 12.67 12.66 12.68 12.67 12.66 12.68 12.67

20 12.63 12.66 12.65 12.68 12.66 12.67 12.68 12.67 12.67
30 12.69 12.69 12.69 12.7 12.69 12.69 12.68 12.68 12.68
40 12.69 12.67 12.68 12.69 12.69 12.69 12.7 12.69 12.69
Time [FeCl3] = 15 mgL-1 [FeCl3] = 21 mgL-1 [FeCl3] = 28 mgL-1
(min) R1 R2 Average R1 R2 Average R1 R2 Average
FeCl3

10 12.66 12.62 12.64 12.65 12.67 12.66 12.63 12.63 12.63

20 12.67 12.67 12.67 12.65 12.68 12.66 12.63 12.65 12.64
30 12.65 12.69 12.67 12.66 12.67 12.66 12.65 12.65 12.65
40 12.69 12.69 12.69 12.67 12.68 12.67 12.63 12.67 12.65

Figure 1 shows the behavior of the coagulants adopted in the work for the percentage
of removal vesus time of the apparent color. The effluent collected in crude form had a
color value of 96.800 mgPtCo L-1.
Figure 1a shows that for the tannin all the concentrations used reached an
apparent color removal greater than 56%. When analyzing the efficiency of
the Coagulant Aluminum Sulphate (Figure 1b) it is observed that the largest removal
was 36%. It is noted that the removal of the chemical coagulant Aluminum Sulphate
was lower than the percentage of removal of the natural coagulant
Tanino which demonstrates the viability of the organic coagulant for the effluent in
question.
In Figure 1c when looking at the Ferric Chloride as a coagulant agent in the removal
of apparent color, it was observed that it is not a promising agent in the removal of color,
because there was an increase in the parameter in all the concentrations used.
148 E. R. Pereira, G. da Silva Souza and J. D. P. Theodoro

(a)

(b)

(c)

Figure 1. Percentages of removal of the apparent color parameter as a function of time for the
coagulants Tanino (a), Aluminum Sulphate (b) and Ferric Chloride (c).
 Comparison of Natural Coagulant and Chemistry in Tanning … 149

In a study carried out by Vaz (2010), a similar result was obtained concluding the
author that when the ferric chloride is added with some excess to the effluent, part of it
does not participate in the coagulation/flocculation reaction, being this in solution having
an increase of the values of the apparent color parameter.

Table 4. Comparisons of means by the Tukey’s test for

the apparent color

Time Coagulant Média Tukey (α=10%)

1 FeCl3 430,500 a
Al2 (SO4)3 402,333 a
Tanin 227,500 b
FeCl3 382,667 a
2 Al2 (SO4)3 357,333 a
Tanin 185,333 b
FeCl3 356,500 a
3 Al2 (SO4)3 340,500 a
Tanin 161,667 b
FeCl3 364,333 a
4 Al2 (SO4)3 354,500 a
Tanin 139,667 b

Statistical analysis showed a P-value of 0.841 > 0.1, indicating that the interaction
between the Coagulant and Concentration factors is not significant, at 10% significance
and it is also noted that the Concentration is not significant (value- P = 0.928 > 0.1), but
the effect of Coagulant was significant (P-value = 0.000 < 0.1). The same occurred for
the second, third and fourth sedimentation times, thus, mean comparisons were
performed only for Coagulant and are presented in Table 4, using the Tukey test.
The organic coagulant Tanino presented in all times of sedimentation statistically
different averages and values statistically smaller than the other coagulants, thus
confirming its greater efficiency in the removal of apparent color in comparison with the
other coagulants.
The Table 5 shows the behavior of the electrical conductivity during the
sedimentation process in the assays for each coagulant. The effluent of tannery had an
electrical conductivity of 21.84 mScm-1 before the treatment done in this work.
For the coagulant Tannin, it is possible to observe that in all the concentrations used
in the test, an increase occurred in the electrical conductivity, where in the time of
sedimentation of 40 minutes the highest value was observed, reaching this 22.57 mScm-1
using the lowest concentration of the coagulant.
150 E. R. Pereira, G. da Silva Souza and J. D. P. Theodoro

Table 5. Behavior of the electrical conductivity as a function

of time for the coagulants

Time [Tanin] = 3 mgL-1 [Tanin] = 6 mgL-1 [Tanin] = 9 mgL-1

(min) R1 R2 Average R1 R2 Average R1 R2 Average
Tanin

10 22.27 21.98 22.12 22.33 21.79 22.06 21.73 22.12 21.92

20 22.46 22.55 22.50 22.51 22.52 22.51 21.63 22.53 22.08
30 22.51 22.59 22.55 22.47 22.52 22.49 22.06 22.56 22.31
40 22.55 22.59 22.57 22.54 22.58 22.56 22.53 22.53 22.53
Time [Al2(SO4)3] = 15 mgL-1 [Al2(SO4)3] = 20 mgL-1 [Al2(SO4)3] = 25 mgL-1
(min) R1 R2 Average R1 R2 Average R1 R2 Average
Al2(SO4)3

10 22.50 21.65 22.07 21.51 21.56 21.53 21.38 21.30 21.34

20 21.32 22.18 21.75 21.31 21.10 21.20 21.25 21.06 21.15
30 21.26 20.96 21.11 21.20 21.21 21.20 20.97 22.23 21.60
40 21.28 22.18 21.73 21.16 22.20 21.68 20.98 20.97 20.97
Time [FeCl3] = 15 mgL-1 [FeCl3] = 21 mgL-1 [FeCl3] = 28 mgL-1
(min) R1 R2 Average R1 R2 Average R1 R2 Average
FeCl3

10 21.50 21.40 21.45 21.85 21.74 21.8 22.00 22.00 22.00

20 21.64 21.54 21.59 21.87 21.69 21.78 22.14 22.12 22.13
30 21.60 21.52 21.56 21.91 21.74 21.82 22.22 21.97 22.10
40 21.68 21.55 21.61 22.08 21.80 21.94 22.33 22.27 22.3

When analyzing the results of the electrical conductivity for the Coagulant
Aluminum Sulphate, it can be said that there was an insignificant variation. For the
coagulant Ferric Chloride, it was observed that there was an increase in the electric
conductivity for the highest concentration of coagulant used. At the lower concentration,
there was a decrease in conductivity values. In general, it is noticed that there was not a
significant change in the electric conductivity after the coagulants application and, in this
way, in relation to the parameter the coagulants do not have any variation of the same.
Figure 2 shows the behavior of fixed and volatile solids in the treatment of tannery
effluent. The crude effluent values for fixed solids and for volatile solids are 60,430 and
38,200 mgL-1, respectively. For the natural coagulant Tanino, it is observed in Figure 2
that the intermediate concentration of 6 mgL-1 was the one that best behaved in the
removal of the solids, reaching a 48% removal for the fixed solids and 59% for the
volatile solids. By observing the behavior of the chemical coagulant Aluminum Sulphate
in the removal of solids, it was observed that it was efficient reaching values above 70%
in all concentrations. The highest removal of fixed solids was 75% and occurred at the
intermediate concentration of 20 mgL-1, the highest removal of volatile solids was 83%
and occurred at the concentration of 28 mgL-1.
In relation to the chemical coagulant Ferric Chloride, as well as the natural coagulant
Tanino, the greatest removal of the solids occurred for the intermediate concentration. At
the concentration of 21 mgL-1 there was a 51% removal of fixed solids and 59% of
volatile solids, indicating values similar to Tannin and a similar behavior for solids
 Comparison of Natural Coagulant and Chemistry in Tanning … 151

removal, however, the chemical coagulant uses a higher concentration to obtain this
removal.

Figure 2. Solids series for the coagulants Tannin (a), Aluminum Sulphate (b)
and Ferric Chloride (c), in percentage values. TC1 = Tannin 3 mgL-1; TC2 = Tannin
6 mgL-1; TC3 = Tannin 9 mgL-1; SC1 = Aluminum Sulfate 15 mgL-1; SC2 = Aluminum Sulphate 20
mgL-1; SC3 = Aluminum Sulphate 25 mgL-1; CCl = Ferric Chloride 15 mgL-1; CC2 = Ferric Chloride
21 mgL-1; CC3 = Ferric Chloride 28 mgL-1.

(a) (b)

(c)

Figure 3. Removal percentage of the Chemical Oxygen Demand parameter for the coagulants Tanino
(a), Aluminum Sulphate (b), and Ferric Chloride (c).
152 E. R. Pereira, G. da Silva Souza and J. D. P. Theodoro

Figure 3 shows the percentage of COD removal obtained for the coagulants
used in the assay. The raw effluent collected at the company had a COD value of
79.850 mgL-1.
For the parameter COD, it is possible to infer that the best results were obtained with
the chemical coagulants, reaching values higher than 90% of COD removal, since no
natural coagulant did not obtain values higher than 80%. Although the coagulants,
especially the chemical ones, present great removal, the effluent still could not be
released in the water body, since it does not meet the standards established by CEMA no.
70/2009 (CEMA, 2009) in annex 7, exceeding the limit value for COD, which is 350
mgL-1 for tanneries.

CONCLUSION

Based on this research, it was verified that the chemical coagulant aluminum sulphate
presented better efficiency in relation to the coagulant tannin. Therefore, the application
of aluminum sulphate would be more feasible with regard to the physical-chemical
treatment of the tanning effluent.

REFERENCES

APHA – American Public Health Association. Standard Methods for the Examination of
Water and Wastewater. 22 ed. Washington, 2012.
Bayer, V. Study of the extraction of hexavalent chromium, by the technique of surfactant
liquid membranes, aiming the treatment of liquid effluents of Tanneries. 2005. 126f.
Dissertation (Master’s Degree in Chemical Engineering - Post-Graduation Program
in Chemical Engineering, Federal University of Minas Gerais, Belo Horizonte, 2012.
Brazil. National Environment Council - Conama. Resolution No. 357, dated March 17,
2005. Provides for the classification of water bodies and environmental guidelines for
their classification, as well as establishing the conditions and standards for effluent
discharge, and other measures. Official Gazette of the Union. Executive Power,
Brasilia, DF, March 18. 2005. Available at: <http://www.mma.gov.br/port/
conama/res/res05/res35705.pdf>. Accessed on: May 25th. 2014.
Brazil. National Environment Council - Conama. Resolution no. 430, dated May 13,
2011. Provides for conditions and standards for the discharge of effluents,
complements and amends Resolution 357 of March 17, 2005, of the National Council
for the Environment - CONAMA. Official Journal of the Union. Executive Branch,
 Comparison of Natural Coagulant and Chemistry in Tanning … 153

Brasília, DF, May 16, 2011. Available at: <http://www.mma.gov.br/

port/conama/legiabre.cfm?codlegi=646>. Accessed on: May 25, 2014.
Martins, G. A. General and Applied Statistics. 3 ed. São Paulo: Editora Atlas S. A, p.
237- 238, 2006.
Theodoro, J. D. P. Study of Coagulation/Flocculation Mechanisms for Obtaining Supply
Water for Human Consumption. 2012. 184f. Thesis (Doctorate in Chemical
Engineering, area of process development) - State University of Maringá, Maringá,
2012.
Vaz, L. G. De L.; Klen, M. R. F.; Veit, M. T.; Silva, E. A. Da; Barbiero, T. A.;
Bergamasco, R. Evaluation of the efficiency of different coagulating agents in the
removal of color and turbidity in electroplating effluent. Eclética Química, São
Paulo, v. 35, n. 4, p. 14-22, 2012. Available at: <http://dx.doi.org/10.1590/S0100-
46702010000400006>. Accessed on: 3 nov. 2014.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 7

MORINGA OLEIFERA SEED USE IN SALINA SOLUTION

IN WATER TREATMENT IN LENTIC BODIES

João Carlos Belisário Junior1, Edilaine Regina Pereira2

and Joseane Débora Peruço Theodoro2
1
Environmental Engineer - UTFPR- Londrina,
Londrina, Brazil
2
Department of Environmental Academic,
Federal Technological University of Paraná,
Londrina, Paraná, Brazil

ABSTRACT

The proposal of this paper was to evaluate through physical and chemical parameters
the efficiency of the coagulation/flocculation/ sedimentation/filtration processes using
organic coagulant (Moringa oleifera) in the treatment of water from a lentic system
(Igapó II Lake), located in Londrina – Paraná - Brasil. It was made the water collection at
two points of the lake, in the entrance (point 1) and at the exit (point 2). The treatment
occurred using the jar-test equipment with the same conditions of fast mixing and slow
mixing used in the Water Treatment Station (ETA). The sedimentation time was 30
minutes. The granulometry of the sand used in the filters was the same, in the range of
0.600 to 0.850 mm. The concentrations of saline solution of the organic coagulant to be
applied were: C1 = 3 mg.L-1, C2 = 6 mg.L-1 and C3 = 9 mg.L-1. In addition, it was
necessary to correct the pH of the samples in order to evaluate the behavior of the organic
coagulant in different pH levels. After the assays, a study was carried out on the removal
of turbidity and apparent color after the coagulation / flocculation / sedimentation
processes and after the filtration process. It was verified for the parameters of turbidity
and of apparent color, the concentration C2 (6 mg.L-1) and the pH around 7 (neutral)
presented the highest removal efficiency, being it above 99%. For the pH, all
156 J. C. Belisário Junior, E. R. Pereira and J. D. Peruço Theodoro

concentrations did not present great variations when compared to the crude sample, in
addition the results prove the efficiency of the coagulant in different pH levels. Only
assay 2 in the entrance point, did not meet the maximum values allowed by the potability
ordinance 2,914/11 of the ministry of health for parameters of apparent color and of
turbidity, all other assays are in accordance. The results proved the efficiency of the
organic coagulant extracted from Moringa oleifera seeds for the treatment of water in
lentic systems.

Keywords: Moringa oleifera, water treatment, lentic systems

INTRODUCTION

The fresh water is divided into lentic ecosystems, lotic ecosystems and flooded
ecosystem. The lentic systems are characterized by low flow or absence of hydrological
flow, being represented by lakes, ponds and tanks. The lotic systems are characterized by
the presence of water flow, such as rivers, streams and springs, on the other hand flooded
systems, have water levels floating up and down during the year, being represented by
swamps and marshes (Soares, 2005).
According to the World Health Organization, about 80% of all diseases that spread in
developing countries come from poor water. Diseases such as typhoid fever, cholera,
diarrhea among others (Richter; Netto, 1991).
The treatment of water aims to reduce impurities and/or correct some aspects of raw
water with the possibility of reuse, or make it drinkable (Freitas, 2011).
Vanacor (2005) states that in addition to making it drinkable, water treatment must
meet the community’s need to have good quality water from a chemical, physical and
bacteriological point of view, and may also serve hygienic purposes (removal of bacteria,
viruses, microorganisms, algae, undesirable substances), aesthetics (color correction,
turbidity, odor and taste) and economic (reduction of corrosivity, hardness, iron,
manganese, etc.).
Coagulation is one of the fundamental processes in surface water treatment systems
for public supply purposes, as it is responsible for water clarification, removal of most
heavy metals, and chemical and microbiological agents (Macedo, 2007).
Coagulation and flocculation are practically simultaneous and interdependent, to the
point that they can be considered a single step: coagulation / flocculation. Coagulation /
flocculation is the process in which the particles agglutinate into small masses (flakes)
with a specific weight higher than that of water. The transport of these particles into the
liquid causes contact, establishing bridges between the particles and forming a three-
dimensional mesh of porous clots (Vanacor, 2005).
 Moringa oleifera Seed Use in Salina Solution … 157

Sedimentation is the physical phenomenon in which, as a result of the gravity action,

the suspended particles present a downward movement in a liquid medium with a lower
specific mass.
The occurrence of sedimentation of the suspended particles is no more
than the separation of liquid and solid phase, this phenomenon provides clarification of
the water while the particles in the bottom form a layer of mud (Di Bernardo;
Dantas, 2005).
Filtration is a solid-liquid separation process, involving physical, chemical, and
sometimes biological phenomena. It aims to remove impurities from the water by passing
through a porous medium. When the velocity at which the water crosses the filter bed is
low, the filter is called the slow filter, when it is high; it is called a fast filter (Ritcher;
Netto, 1991).
The Moringa oleifera`s seed contains the presence of a high molecular
weight dimeric cationic protein, which destabilizes the particles contained in the
water through a process of neutralization and adsorption. Its water-soluble powder
acquires positive charges that attract negatively charged particles such as clays
and silts, forming dense sedimentary flakes (Ndabigengesere, A., Narasiah, K. S.; Talbot,
1995).
According to Arantes et al., (2015), the use of this organic coagulant combined with
filtration in the treatment of water shows good results, reaching 99.0% turbidity in water.
He further states that the filtration step is necessary for the removal of the organic matter
introduced by the coagulant.
With this, the present work aims to evaluate, from Ordinance 2,914 / 11 updated to
consolidation ordinance nº5, from September 28, 2018 both from the ministry of health
(Brazil, 2011, Brazil 2017), according to the parameters color and turbidity, previously
determined, in addition to performing the treatment of these water through the
coagulation / flocculation / sedimentation / filtration process using the saline solution
extracted from the Moringa oleifera seed.

METHODOLOGY

The water samples were taken from a lentic system from Lake Igapó II, located in the
city of Londrina, state of Paraná, Brazil. Water samples of entry (Point 1) and exit (Point
2) were used. Figure 1 spatially represents the location of the collection points of each
sample.
Sampling was carried out at the entrance (Point 1) and exit (Point 2) of Lake Igapó II,
highlighted according to Figure 1 and conditioned in plastic gallons of 50 liters each,
previously sanitized.
158 J. C. Belisário Junior, E. R. Pereira and J. D. Peruço Theodoro

Figure 1. Scheme of the satellite view of the location of Lake Igapó II.

For the preparation of the Moringa oleifera organic coagulant solution, first the
Moringa oleifera seed was peeled and 10g of it was weighed. Also weighed 58.44 g of
NaCl and added to 1 L of distilled water, the solution was stirred well for the salt to
dissolve completely, the salt solution being 1M NaCl. Finally, the 10 g of the seeds were
ground together with the saline solution in the blender. The solution was then filtered
using a cloth strainer and the Moringa oleifera coagulant saline at a concentration of 10
g.L-1 was obtained.
To adjust the pH values to make it acidic, the solution of hydrochloric acid (HCl),
concentration 1 M was used. It was necessary to add 20 mL of HCl to the volume used in
order to achieve the acidic pH set. Meanwhile, the sodium hydroxide solution (NaOH), at
1 M concentration, was used for the basic pH sample. It was necessary to add 20 mL of
the basic solution to achieve the set basic pH.

Table 1. pH values used in the assays

Samples pH Point 1 pH Point 2

Acid 4.5 4.3
Natural 6.8 6.8
Basic 10.13 10.8

Table 2. Mixing rod rotation gradient and time of action

Fast Mixing Slow Mixing Slow Mixing Slow Mixing Slow Mixing
Gradient (s-1) 540 90 52 40 30
Time (min) 00:10 02:00 02:35 02:40 05:40
Source. (Trevisan et al. (2014); Higashi (2016); Goes et al., (2017))
 Moringa oleifera Seed Use in Salina Solution … 159

The final pH values applied in the samples are shown in Table 1. For the experiment,
the dosages found for coagulant at both points were varied following the concentrations 3
mg.L-1, 6 mg.L-1 and 9 mg.L-1.
The assays were carried out in the Jar-test equipment of six tests with regulator of
rotation of the mixer rods. Placing in each compartment of the 2L water equipment with
different pH and its respective concentration of extraction solution and natural coagulant.
After it was held equipment rotation process at different rates (Table 2) for that to happen
the coagulation and flocculation processes.
For the filtration assay an iron structure adapted to fix the sand filters below the jar-
test was used so that the water exited the jar-test directly into the filters. The filter beds
are made of Polyethylene Tereftalo (PET) of approximately 10 cm of internal diameter
configuring a model of fixed bed with downward flow, with six columns in parallel, as
observed in Figure 2.
It was recommended by Di Bernardo et al., (2003) the tubes are 25 cm long with 15
cm filled by sand, 3 cm filled with crushed stone and 2 cm filled with cotton. The
granulometric values of the sand used in the 6 filters were the same, in the range of 0.600
to 0.850 mm. The slope used at the exit of the test jar was 70 ° for 2 minutes, followed by
an additional 2 minutes at an angle of 60 ° and a further 2 minutes at 50 °. A model of the
filter layers is shown in Figure 3.

Figure 2. Filtration columns used in the assay.

160 J. C. Belisário Junior, E. R. Pereira and J. D. Peruço Theodoro

Figure 3. Representative model of filter layers.

The water to be treated entered through the top of the column and was withdrawn
from the bottom. The grains of the filter bed were retained by a coffee filter together with
the cotton and crust layer present at the lower end of the column. After this process, all
parameters were measured.
It is important to say that the assays were performed in duplicate for each pH
value and concentration. For all variations of Moringa oleifera and pH concentrations
the parameters of apparent color, turbidity and pH were determined (Table 3)
according to the Standard Methods of Examination of Water and Wastewater (APHA,
2012).

Table 3. Equipment and methodology for the test battery

Parameter Equipment Methodology

Apparent Color Spectrophotometer HACH DR-5000 2120 C
Turbidity Policontrol Turbidimeter AP-2000 2030 B
pH pH-meter mPA-210 4500 H+ B
Source: APHA, 2012.

Table 4. Permissible values of some water parameters according to the ministry of

health ordinance 2,914/11 and consilidation ordinance no5, of September 28, 2018

Parameter Pattern
Apparent color 15 mgPtCo.L-1
Turbidity 5 NTU
pH 6.0 a 9.0
Source: Ministry of Health, Ordinance 2914/11 PRC September 20, 2017 and consilidation ordinance no5, of
September 28, 2018.
 Moringa oleifera Seed Use in Salina Solution … 161

The results were evaluated and compared with the standard allowed by ordinance
2,914/11 updated to consolidation ordinance no5, of September 28, 2018 both from the
ministry of health (Table 4) and can indicate if the experimental procedure was
satisfactory.
In this work, statistical analysis was used with two factors (independent
variables), the concentration of the Moringa oleifera coagulant, and pH. The assays
had a repeat, thus, performed in duplicate in Table 5 it is possible to verify these
assay values.

Table 5. Numerical organization of the assays for different levels of coagulant

concentration and pH with coded and real values of point 1 and point 2

Assay Duplicate Concentration of pH pH

Coagulant (mg.L-1) Point 1 Point 2
1 1 3 4.5 4.3
2 1 3 6.8 6.8
3 1 3 10.13 10.8
4 1 6 4.5 4.3
5 2 6 6.8 6.8
6 2 6 10.13 10.8
7 1 9 4.5 4.3
8 1 9 6.8 6.8
9 2 9 10.13 10.8
10 2 3 4.5 4.3
11 2 3 6.8 6.8
12 2 3 10.13 10.8
13 2 6 4.5 4.3
14 1 6 6.8 6.8
15 1 6 10.13 10.8
16 2 9 4.5 4.3
17 2 9 6.8 6.8
18 1 9 10.13 10.8

RESULTS

Figures 4, 5 and 6 show the removals of the apparent color parameter for the
coagulation / flocculation / sedimentation processes (Figure 4), for the filtration process
(Figure 5) and for the complete process, coagulation / flocculation / sedimentation /
filtration (Figure 6) with the use of Moringa oleifera in different values of concentration
and pH.
Analyzing the presented results, it can be concluded that both the coagulation /
flocculation / sedimentation processes (Figure 4) and the filtration process (Figure 5) are
162 J. C. Belisário Junior, E. R. Pereira and J. D. Peruço Theodoro

of great importance for the removal of color in the water treatment, both reached removal
values above 80%. The processes together (complete treatment) demonstrated in almost
all tests percent removal of color close to 100% as can be seen in Figure 6.

Figure 4. Removal of the apparent color parameter after the coagulation / flocculation / sedimentation
processes.

Figure 5. Removal of the apparent color parameter after the filtration process.

Figure 6. Removal of the apparent color parameter after the coagulation / flocculation / sedimentation /
filtration process.

In Figure 4 it is possible to verify the assay 2 as the assays with the lowest percentage
of color removal, that is, a concentration of 3 mg.L-1 and a neutral pH of 6.8. However,
 Moringa oleifera Seed Use in Salina Solution … 163

its replica at the same point was the one with the highest removal value, besides that in
point 2 the values of color removal with these same variables were also satisfactory.
Also in Figure 4, assays 9 and 18 in point 2 were the assays with the lowest
percentage of color removal at this point, these assays were replicates of each other, that
is, concentration of 18 mg.L-1 and basic pH equal to 10.8. Comparing with the assays of
point 1 it is verified that for point 1 these tests were also the ones with lower percentages
of removal. With this we conclude that for color removal this combination was the worst.
The other assays are ranging from 81% to 91% color removal after coagulation /
flocculation / sedimentation processes.
In the filtration process (Figure 5) it is also possible to verify a small variation of the
percentages of color removal between 85% and 97%, except test 11 of item 1, which had
removal value of 70%. Its replica at the same point also had below-average value, so it is
understood that for such pH value and concentration the filtration process is slightly
affected.
In general, very satisfactory results were obtained, as can be seen in Figure 6, the
complete treatment provided a reduction of the parameter apparent color of almost 100%
in all the tests, it can be concluded that the processes are complementary and very
efficient for the removal of color in the treatment of water. According to studies by
Okuda et al., (1999), which demonstrate that the percentage of color removal using both
the seed and the aqueous extract of Moringa oleifera was 80 to 99%.
It is also possible to verify that the coagulant Moringa oleifera can be effective in a
wide range of pH and concentration in this case, without significantly altering its results
of color removal with the variation of these variables. According to studies conducted by
Vaz (2009), Moringa oleifera can be used in a pH range between 4.0 and 12.0.
Comparing the values obtained with those required by ordinance 2,914/2011 of the
ministry of health 2017 and consilidation ordinance no5, of September 28, 2018, only one
assay did not meet the VMP of apparent color, assay 2 of point 1. All others are in
accordance with the amount required for the use of water for urban supply.

Figure 7. Removal of the turbidity parameter after the coagulation / flocculation / sedimentation
process.
164 J. C. Belisário Junior, E. R. Pereira and J. D. Peruço Theodoro

Figure 8. Removal of the turbidity parameter after the filtration process.

Figure 9. Removal of the turbidity parameter after the coagulation / flocculation / sedimentation /
filtration process.

Figures 7, 8 and 9 show the removals of the turbidity parameter for the coagulation /
flocculation / sedimentation processes (Figure 7), for the filtration process (Figure 8) and
for the complete process, coagulation / flocculation / sedimentation / filtration (Figure 9)
with the use of Moringa oleifera in different values of concentration and pH.
Analyzing the results presented in Figures 7 and 8, it is possible to verify that both
the coagulation / flocculation / sedimentation processes and the filtration process had a
high percentage of turbidity parameter removal, around 80% in Figure 7 and around 90%
in Figure 8. With this it is possible to understand that the processes together are of
extreme importance for the removal of turbidity in the treatment of water. Confirming the
data presented in Figure 9 where the processes together (complete treatment)
demonstrated in almost all the tests percentage of turbidity removal close to 100%.
This confirms studies carried out by Higashi (2015), where after the treatment of
Lake Igapó II water through the coagulation / flocculation / sedimentation processes with
the use of organic coagulant Moringa oleifera obtained removal results above 80%.
All other assays are ranging from 77% to 88% turbidity removal after coagulation /
flocculation / sedimentation processes.
 Moringa oleifera Seed Use in Salina Solution … 165

In the filtration process (Figure 8) it is also possible to verify a small variation of

turbidity removal percentages between 89% and 95%, except for assays 2 and 11 of point
1 and point 2, which are replicates of each other. These had removal value between 78%
and 84%. Thus it is understood that for such pH value and concentration the filtration
process is slightly affected.
With the data of Figures 7, 8 and 9 it is possible to understand that the Moringa
oleifera coagulant is effective over a wide pH range and also concentration, in this case,
not significantly altering its turbidity removal results with the variation of these variables.
Confirming studies by Cardoso et. al (2008) where the results indicate that the organic
coagulant Moringa oleifera can be used in a wide range of pH.
Comparing the values obtained with those required by ordinance 2,914/2011 of the
ministry of health and consilidation ordinance no5, of September 28, 2018, only one assay
did not meet the turbidity VMP, assay 2 of point 1. All others are in accordance with the
value required for the use of water for supply.

CONCLUSION

The results of the present paper showed that the use of organic coagulant from the
Moringa oleifera seed used in a saline solution (NaCl, 1M) was very efficient in the
removal of the parameters apparent color and turbidity. For the parameters of apparent
color and turbidity, all the assays, at the end of the complete treatment, showed almost
100% removals.

REFERENCES

APHA – American Public Health Association. Standard Methods for the Examination of
Water and Wastewater. 22 ed. Washington, 2012.
Arantes, C. C. et al. Different forms of Moringa oleifera seed in water treatment.
Brazilian Journal of Agricultural and Environmental Engineering. v.19, n.3, p.266-
272, 2015.
Brazil, Ministry of Health. 2,914, dated December 12, 2011. Available at:
<http://bvsms.saude.gov.br/bvs/saudelegis/gm/2011/prt2914_12_12_2011.html>.
Accessed on: January, 2018.
Brazil, Ministry of Health. Consolidation Ordinance No. 5 of September 28, 2017.
Available at: <http://bvsms.saude.gov.br/bvs/saudelegis/gm/2017/prc0005_03_10_
2017.html> Accessed: April 11 2018.
166 J. C. Belisário Junior, E. R. Pereira and J. D. Peruço Theodoro

Di Bernardo, L.; Mendes, C. G. N.; Brandão, C. C. S.; Sens, M. L.; Pádua, V. L. Water
Treatment for Direct Filtration. Di Bernardo, Luiz (coordinator) - Rio de Janeiro:
ABES, RiMa, 2003. Project PROSAB 498 p.
Di Bernardo, L.; Dantas, A.D. B. Methods and Techniques of Water Treatment. 2nd
Edition. São Carlos: Rima, 2005.
Freitas, D. B. Study of improvements in the chlorination systems of the water supply in
the corsan by the installation of chlorine evaporators and gas scrubbers. Porto
Alegre. 2011. Diploma in Chemical Engineering. Federal University of Rio Grande
do Sul.
Goes, H. H. D.; Souza, R. C. P.; Melo, J. M.; Theodoro, J. D. P.; Study of the application
of Opuntia cochenillifera cactus in water treatment. Encyclopedia Biosphere,
Knowing Scientific Center - Goiânia, v.14 n.25; P. 554-563. 2017.
Higashi, V. Y.; Theodoro, J. D. P.; Pereira, E. R.; Theodoro, P. S.; Use Of Chemical
Coagulants (Ferric Chloride) and Organic (Moringa Oleifera) In Treatment Of
Waters Derived From The Lentic System. Technical Scientific Congress of
Engineering and Agronomy – CONTECC’2016, August 29 to September 2, 2016 -
Foz do Iguaçu, Brazil. Available at: <https://www.tratamentodeagua.com.br/wp-
content/uploads/2016/11/Uso-de-coagulantes-qu%C3%ADmico-cloreto-ferrico-e-
org%C3%A2nico-moringa-oleifera-in-treatment-of-water-from-system-
l%C3%AAntico.pdf>. Accessed in Feb. 2018.
Macêdo, J. A. B. Waters & Waters: 3 ed. Belo Horizonte. MG: CRQ-MG, 2007. 1048p.
Ndabigengesere, A.; Narasiah, K. S.; Talbot, B. G. Active agents and mechanism of
coagulation of turbid water using Moringa oleifera. Water Research. V.29, p.703-
710, 1995.
Okuda, T. et al. Improvement of extraction method of coagulation active components
from Moringa oleifera seed. Water Res. V.33, n.15, p.3373-3378. 1999.
Pichler, T., Young, K.; Alcantar, N. Eliminating turbidity in drinking water using the
mucilage of a common cactus. Water Science and Technology: Water Supply, v. 12,
n.2, 179-186, 2012.
Richter, C. A.; Netto, J. M. A. Water treatment: Upgraded technology. São Paulo:
BLUCHER, 1991. 332p.
Soares, D. H. G. Ecosystems in the ecological context. Pernambuco. Monography
(Undergraduate). Center for Higher Education Arcoverde, 2005.
Theodoro, J. D. P.; Lenz, G. F.; Zara, R. F.; Bergamasco, R.. Coagulants and Natural
Polymers: Perspectives for the Treatment of Water. Plastic and Polymer Technology
(PAPT), v. 2, Issue 3, September 2013.
Trevisan, Thales S. Coagulant Tanfloc SG as an alternative to the use of chemical
coagulants in the treatment of water in ETA Cafezal. 2014. 106 f. Course Completion
Work (Undergraduate) - Superior Course in Environmental Engineering. Federal
Technological University of Paraná, Londrina, 2014.
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Vanacôr, R. N. Evaluation of the Organic Organic Coagulant used in a water treatment

plant for public supply. Porto Alegre. 2005. Master’s Degree in Water Resources
Engineering and Environmental Sanitation at the Federal University of Rio Grande
do Sul.
Vaz, L. G. Performance of the coagulation / flocculation process in the treatment of the
liquid effluent generated in electroplating. 2009. 100f. Dissertation (Master in
Chemical Engineering) - Center for Engineering and Exact Sciences, State University
of Western Paraná, Toledo, 2009.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 8

EVALUATING NEW BIOPOLYELECTROLYTES FOR

THE MEAT PROCESSING WASTEWATER TREATMENT
VIA COAGULATION-FLOCCULATION

E. A. López-Maldonado1,* and M. T. Oropeza-Guzmán2

1
Faculty of Chemical Sciences and Engineering,
Autonomous University of Baja California, Tijuana, Mexico
2
Center for Graduates and Research in Chemistry,
Technological Institute of Tijuana, Tijuana, Mexico

ABSTRACT

The solid-liquid separation processes are of great scientific and technological

relevance in the area of food, beverages, agrochemicals, drugs, mineral extraction,
ceramics and wastewater treatment. Nowadays, various chemical substances are used that
act as destabilizing agents or stabilizers according to their field of application. One of the
strategies to understand the coagulation-flocculation processes is to use the zeta potential
measurements as an electrochemical tool to approximate the interface phenomena and
establish the mechanisms that predominate at the molecular level. One of the trends is the
use of various biopolyelectrolytes (BPE) that can be obtained, modified and applied
strategically in various fields of study. In this chapter, the physicochemical performance
of six biopolyelectrolytes in the coagulation of flocculation of wastewater with a high
content of solids and fats and oils was evaluated. The ones that presented a better
performance for this type of wastewater are mesquite gum and chitosan. The BPE that
showed a better performance in the coagulation-flocculation test of the wastewater with
high content of fats and oils at pH 5.2, was the chitosan, with a dose of 68 mg/L of

*
Corresponding Author Email: [email protected].
170 E. A. López-Maldonado and M. T. Oropeza-Guzmán

chitosan the treated water has the following values of BOD5 = 30 mg O2/L, fats and oil =
1 mg/L, turbidity = 2 FAU, TSS = 2 mg/L and COD = 50 mg/L.

Keywords: wastewater, zeta potential, coagulation-flocculation, biopolyelectrolytes, fats

and oil

1. INTRODUCTION

One of the issues of greatest scientific and technological interest is the quality and
use of water, derived from the problems of scarcity and water contamination. The
interaction of water with the environment is influenced by the water quality that both
society and the industrial sector confer on water.
Each type of industry has a particular interest in the care of water quality and its
reuse, which is why day by day, they require new strategies to treat and recycle the
wastewater they generate in the different production processes [1]. Water quality is
affected by various chemical substances that dissolve in water used in each stage of the
manufacturing process.
In general, the main pollutants that are identified in the industrial wastewater are
suspended particles, organic matter, drugs, dyes, heavy metals, agrochemicals, the
hardness of the water and fats and oils [2].
The levels of concentration in which these contaminants or undesirable substances
are present in the wastewater are directly related to the operating conditions of the
productive processes. The presence of these pollutants in the water causes an impact on
the efficiency of the production processes, limits the reuse of water, increases the
consumption of clean water, the discharge of the wastewater generated contaminates the
water bodies, and this implies sanctions to the industry for exceeding the maximum
permissible limits at the effluent discharge point [3].
Currently, there are different technologies to perform the treatment of wastewater,
including adsorption using various adsorbent materials (polymers, carbon, nanomaterials,
clays, zeolites), electrodeposition, membrane filtration (ultrafiltration, nanofiltration and
reverse osmosis), coagulation-flocculation, chemical precipitation with hydroxides,
sulfides and chelating precipitations, advanced oxidation processes, electrocoagulation,
electrodialysis, ion exchange, biological treatment, photocatalysis, and electroflotation.
Each technology has certain advantages and disadvantages. Considering the most
demanding environmental legislation, industries need more efficient wastewater
treatments to eliminate suspended or dissolved metals [5-20].
In the case of coagulation-flocculation process, a very simplest equipment is required
 Evaluating New Biopolyelectrolytes … 171

thus remains among the most popular process for the separation of various
types of contaminants, in which chemical substances are used as synthetic coagulant-
flocculating agents, for example, alum, ferric chloride, polyaluminum
chloride and synthetic organic polymer. The synthetic polyelectrolytes (PE) have also
been reported to pose a number of environmental problems since some of the
derivatives and intermediates are non-biodegradable and reported as hazardous to human
health (their monomers are neurotoxins and carcinogens). One of the promising
alternatives is the use of biopolyelectrolytes (BPE) as flocculating coagulants,
which can be extracted from various sources such as residues from the food industry and
agriculture.
One of the most appreciated characteristics of PE is its efficiency and effectiveness in
the destabilization of colloidal systems or dispersion of particles. In a colloidal system the
suspended particles have a surface charge density, which causes that by
electrostatic interactions the particles repel, preventing their sedimentation by gravity
forces.
Such is the case of the wastewater generated by the factory for cutting and packing of
meat products, which consists of a dispersion of particles of fats and oils from animal
origin. In this chapter, the physico-chemical characterization of meat processing
wastewater is presented. The coagulation-flocculation performance in the wastewater
treatment was evaluated using the two natural biopolyelectrolyte extracted from the
shrimp husks (chitosan) and mesquite gum. The zeta potential was used as a key
electrochemical tool to strategically dose the BPE allowing the destabilization of the
contaminants present in the wastewater.

2. EXPERIMENTAL

2.1. Wastewater Sampling

In this chapter, presents the physicochemical characterization of meat processing

wastewater and how to develop a treatment strategy for water recovery and add value to
the byproducts formed. The wastewater sampling protocol was followed as recommended
by Mexican sampling standard (NMX-AA-003-1980). Residual water samples were
taken from a factory for cutting and packing meat products. Tested parameters were: total
solids (TS), total dissolved solids (TDS), total suspended solids (TSS), turbidity, color,
particle size, electrical conductivity (EC), zeta potential (), total phosphorous (TP),
biological oxygen demand (BOD5), chemical oxygen demand (COD), total organic
carbon (TOC), fats and oils and total nitrogen (TN). Tests were carried out following the
current Mexican standard procedures that are equivalent to those published by EPA
(AWWA standard methods respectively).
172 E. A. López-Maldonado and M. T. Oropeza-Guzmán

2.2. Zeta Potential f (pH) Profiles of the Industrial Wastewater and BPE

Zeta Potential from wastewater and biopolyelectrolyte data was recorded on a

Stabino Particle Charge Mapping (Microtrac). The measurements were done at ambient
temperature in Teflon cuvettes. Influence of pH on the zeta potential behavior of each
biopolyelectrolytes was studied within a pH range of 2-11 with 0.1 M NaOH and 0.1 M
HCl [20].

2.3. Wastewater Coagulation-Flocculation Tests Using BPE

The performance of cationic chitosan biopolyelectrolyte in the coagulation-

flocculation of meat processing wastewater was carried out using 20mL of residual water
at pH 5.2, and in different doses of chitosan extracted from the shrimp shells. After each
addition of chitosan, the mixture of residual water with chitosan was stirred at 200rpm for
2min, and subsequently at 50rpm for 20min.
For the evaluation of the quality of the treated water, a sample of the supernatant was
extracted [21].

3. RESULTS AND DISCUSSION

3.1. Meat Processing Wastewater

Tables 1 show the physicochemical characterization of meat processing wastewater.

The values of the main residual water quality parameters such as BOD5, COD,
alkalinity, hardness, pH, electrical conductivity, fats and oils, content of dissolved and
suspended solids, settleable solids, temperature, turbidity, nitrogen and total phosphorus
are shown. Of these normative parameters the environmental legislation dictates which
ones must comply, considering the type of industry and body of discharge of residual
water. Additionally, other non-regulated parameters, which are fundamental to
understand and operate the coagulation-flocculation process, such as particle size,
turbidity, total organic carbon, biodegradability and zeta potential [21].
The measurement of these parameters is key to implement the design and sequence of
an industrial wastewater treatment train to achieve the best quality of treated water.
In all the regulated parameters exceed the maximum permissible limits, both for their
discharge to the water receiving bodies and for their reuse. Considering the main
interactions that occur between the suspended particles and the coagulant-flocculant
agents, the zeta potential is a key parameter to determine the surface charge density of the
 Evaluating New Biopolyelectrolytes … 173

suspended particles and the biopolyelectrolytes, as well as the optimum dose to perform
the solid-liquid separation of the suspended particles.

Table 1. Physicochemical characterization of Meat Processing Wastewater

Parameter Meat Processing Wastewater

Temperature (oC) 28
Alkalinity (mg/L CaCO3) 98
Fats and oils (mg/L) 15,732
TSS (mg/L) 20,000
TDS (mg/L) 3,500
SS (mL/L) 900
Turbidity (FAU) 750
EC (mS/cm) 2.00
 (mV) -20
Particle size (nm) 750
Color (Pt-Co) 35,083
pH 6.8
COD (mg O2/L) 60,587
TOC (mg C/L) 53,45
BOD5 (mg O2/L) 40,250
Biodegradability (BOD5/COD) 0.67

3.2. f (pH) Profiles of the BPE

Figure 1 the = f (pH) plot shows the charge density variation the BPE of maiz gum
(GMZ), chitosan (Ch), mesquite gum (GMT), guar gum (GG), Tule lignin (LT) and
Coffe lignin (LC) with respect to pH. The change in pH had a distinct effect with each
BPE, because of the difference between the functional groups present in the chains of the
iomacromolecules. Considering the zeta potential value (= -20 mV) of the wastewater
at pH 5.2, the biopolyelectrolyte of chitosan and mesquite gum having positive surface
charge density, were selected to carry out the strategic dosing in the wastewater with a
high content of fats and oils and to evaluate their performance in coagulation-
flocculation.

3.3. Coagulation-Flocculation Tests with Biopolyelectrolytes

Figure 2 shows the variation of zeta potential with respect to the pH of the meat
processing wastewater and chitosan.
174 E. A. López-Maldonado and M. T. Oropeza-Guzmán

30
20
10 BPE-GMZ
0
BPE-Ch
-10
-20 BPE-GMT
 (mV)
BPE-GG
-30
-40
-50 BPE-LT
-60
-70
BPE-LC
-80
-90
2 3 4 5 6 7 8 9 10 11 12
pH

Figure 1. = f (pH) profiles of GMZ, Ch, GMT, GG, LT and LC.

The zeta potential value of the wastewater shows that the suspended particles
have a negative surface charge density at pH 4-12, while at pH <4 zeta potential
values close to neutrality or positive. The = f (pH) profile show that at
pH = 3.0 the wastewater has their isoelectric point ( = 0). The stability of the
particles to remain suspended is due to the value of the negative zeta potential ( = -
25.5mV).
In order to destabilize the dispersion of fats and oil, the addition of a cationic
coagulant-flocculant agent that allows the neutralization of the negative surface charge is
necessary.
The wastewater at pH 5.4 has a  = -25.5 mV and the chitosan = 15.0mV, the
dosage of chitosan at this pH by pure electrostatic interaction ensures its reaction.
As expected, in order to carry out a coagulation-flocculation process in an
efficient way, it is necessary to add a polyelectrolyte with a positive charge to neutralize
the negative charge of meat processing wastewater. The monitoring of the zeta
potential value with respect to the polyelectrolyte dose in the wastewater
allows to construct the coagulation-flocculation operation curves, this ensures
that the operators of the wastewater treatment plants avoid the problems of
overdosing of agents coagulants-flocculants and save on the consumption of chemical
substances [22].
In this chapter, the capacity of two biopolyelectrolytes for the clarification of
wastewater was evaluated. This with the aim of increasing the reuse potential of the
treated water and improving the efficiency of solid-liquid separation processes.
 Evaluating New Biopolyelectrolytes … 175

In Figure 3, it is shown that the zeta potential value of the meat processing
wastewater as the dose of chitosan increases ( = -20.1mV to = -0.1mV), reaching the
isoelectric point at a dose of 68 mg/L chitosan. The water treated at pH 5.4 with chitosan
has a value of BOD5 = 30mg O2/L, fats and oil = 1mg/L, turbidity = 2 FAU, TSS =
2mg/L and COD = 50mg/L, the treated water with these physicochemical characteristics
can be discharged to the municipal sewer system or reused as process water.

OH OH
O
50 O O O
HO n
NH HO NH2
40 O

30 chitosan
20

10
 (mV)

-10

-20
Meat processing
-30
wastewater
-40
2 4 6 8 10 12

pH

Figure 2.= f (pH) profiles of Ch and meat processing wastewater.

20
BPE-Ch (68 mg/L)
15 BPE-Ch
10
BPE-GMT (90 mg/L)
5

0
 (mV)

-5
BPE-GMT
-10

-15

-20

-25
0 20 40 60 80 100 120
BPE dose (mg/L)

Figure 3. = f (BPE dose) profiles of GMT and Ch.

176 E. A. López-Maldonado and M. T. Oropeza-Guzmán

Additionally, Figure 3 shows the behavior of zeta potential with respect to the dose of
mesquite gum, indicating that a higher dose of this biopolyelectrolyte is required to reach
the isoelectric point of the residual water.
It should be noted that in a higher dose than the IEP of wastewater, chitosan can
reestablish particles flocculated, unlike mesquite gum. This is attributed to the fact that
mesquite gum has a lower surface charge density [23-24].
The quality of the treated water obtained with the best dose (90 mg/L) of mesquite
gum does not comply with the maximum permissible limits of COD = 650 mg/L and
BOD5 = 1200 mg/L for discharge to the sewage system, however, it does not stop to be an
attractive alternative as a physicochemical pretreatment stage for the residual water
coupled to an advanced oxidation process that allows the complete mineralization of the
dissolved substances in the treated water.

CONCLUSION

The wastewater generated by industries is becoming more complex and difficult to

treat to restore its quality and reuse it. The coagulation-flocculation process has become
one of the most used technologies to remove suspended particles, dyes and heavy metals,
however, one of the trends consists of the substitution of synthetic coagulant-flocculant
agents with biopolyelectrolytes. This leads to the development environmentally friendly
technologies and take advantage of the waste that contains biodegradable polymeric
materials and with high potential for its application in the elimination of toxic pollutants
from industrial wastewater. In this study, it is presented that mesquite gum, given its
chemical nature, has a different flocculation coagulation mechanism (flocculation
sweep). Unlike chitosan, which has a high cationic charge density, it shows a charge
neutralization mechanism in the colloidal destabilization process of the wastewater,
according to the profiles of = (BPE dose). The quality of the water obtained with
mesquite gum does not comply with the maximum limits allowed for its discharge to the
sewer system, however, the coupling of this coagulation flocculation process to an
advanced oxidation system can help to improve its performance, reducing electrolysis
times and avoiding electrode fouling.

ACKNOWLEDGMENTS

The authors thank the Autonomous University of Baja California for the financing
granted for the development of this project (UABC-PTC-668).
 Evaluating New Biopolyelectrolytes … 177

CONFLICT OF INTEREST
The authors state that there is no conflict of interest.

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[9] dos Santos, A. B., Cervantes, F. J., van Lier, J. B., (2007). Review paper on current
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[11] Ali, I., Khan, T. A., Asim, M., (2011). Removal of arsenic from water by
electrocoagulation and electrodialysis techniques. Sepn. Purfn. Rev., 40, 25 - 42.
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photocatalytic oxidation process for treating waste water containing surfactants.
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from water by adsorption onto banana peel as bioadsorbent. Int. J. GEOMATE., 2,
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Maleic Polyelectrolytes, Environmental Science and Technology., 36, 1856 - 1860.
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applications.1st ed. Fondo editorial de la Pontificia Universidad Catolica del Perú;
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multiparameter colloidal titrations for the determination of cationic poly-
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[21] López-Maldonado, E. A., Oropeza-Guzman, M. T., Jurado-Baizaval, J. L., Ochoa-
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[23] López-Maldonado, E. A., Zavala Garcia, O. G., Cruz Escobedo, K., Oropeza
Guzmán, M. T., (2017). Evaluation of the chelating performance of
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In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 9

TREATMENT OF RESIDUAL WATERS OF

THE BREWERY INDUSTRY THROUGH
THE FLOCCULATION PROCESS WITH
THE USE OF INORGANIC
AND ORGANIC COAGULANTS

Fellipe Jhordã Ladeia Janz1,

Edilaine Regina Pereira2, Thaís Ribeiro1,
Dandley Vizibelli1 and Julio Cesar Angelo Borges1
1
Environmental Engineer - UTFPR- Londrina, Londrina, Brazil
2
Department of Environmental Academic,
Federal Technological University of Paraná, Londrina, Paraná, Brazil

ABSTRACT

The brewing industry presents a great generation of effluent that can cause serious
environmental impacts when not treated in a regular way since it has a high load of
organic matter in its composition. Thus, in view of the growing emergence of breweries
in Brazil and consequent increase in effluent production, alternatives are sought for the
auxiliary treatment of this effluent using coagulants. In this way, the paper aimed to
analyze the efficiency the usage of two organic and one inorganic coagulant in the
treatment of effluent from the brewing industry. The assay was performed in Jar Test,
thus simulating the coagulation, flocculation and sedimentation processes, and in
sequence the filtration process was carried out. During this process the parameters of
apparent color, electrical conductivity, pH and turbidity were monitored. Three different
treatments were used: Aluminum Sulfate (T1), Tannin (T2) and Moringa oleifera (T3).
180 F. J. L. Janz, E. R. Pereira, T. Ribeiro et al.

The results showed values of apparent color removal and turbidity of more than 90% for
all treatments, but the treatment with Tanino presented the highest values of removal,
being 98.8% for turbidity removal and 99.6% for removal of apparent color. The pH
parameter showed a rise throughout the test for all treatments presenting final values
close to the neutral value, demonstrating compliance with CONAMA Resolution No.
430/2011. The electrical conductivity showed a decrease in all treatments especially in
the treatment where Tanino was used being 0.32μS.m-1 the lowest value obtained. The
use of the coagulants in question showed positive responses in the analyzed parameters in
the treatment of the effluent of the brewing industry, showing more emphasis for the
Tanino that obtained the best values. It’s an option sustentable and is possible to mix this
process with the comum process that there are in brewing industry.

Keywords: wastewater treatment, tannin, flocculation.

1. INTRODUCTION

The brewing market has shown a growth in Brazil, and between 2014 and 2017 there
was a 91% increase in the number of establishments registered in the country, from 356
to 679 breweries (Marcusso; Müller, 2017).
The brewing sector in Brazil is considered a relevant sector for the country’s
economy, having produced between 2011 and 2014 a turnover of 70 billion reais a year,
generating 2.2 million jobs and producing 14 billion liters of beer per year, the sector is
responsible for 1.6% of the Brazilian domestic productof the country (CERVBRASIL,
2018).
The effluent generation in the brewery varies from 3 to 6 L of effluent per liter of
beer produced, occurring in several sectors of the industry, and the lavage sector is
responsible for the largest generation of effluent, but this effluent presents low organic
load, and the fermentation and filtration sectors generate little volume of effluent, but
with high organic load (CETESB, 2005).
The effluent generated has high organic load, suspended solids content and presence
of nutrients (phosphorus and nitrogen), which causes this effluent to be considered a high
polluting potential (CETESB, 2005).
Most of the breweries have a treatment system composed of biological and
physicochemical processes, and the physical-chemical process has the function of
reducing the organic load of the effluent that will later undergo a biological process,
being aerobic or anaerobic. The organic load of the brewery effluent is present in solid
form, and its removal can be through the physical-chemical process of coagulation
(Kochenborger, 2012).
Knowing the ability of the coagulation process to remove solid materials and
improve effluent quality, the study aimed to compare the use of organic and inorganic
coagulants in the treatment generated by the brewing industry.
 Treatment of Residual Waters of the Brewery Industry ... 181

2. METHODOLOGY

For the accomplishment of the study three forms of treatments were determined,
being used three different coagulants, so defined: T1): application of the Aluminum
Sulphate; (T2): application of Tannin; and (T3): application of Moringa oleifera, using
the same concentration of solution corresponding to 9mg.L-1 in all treatments.
The preparation of the Moringa oleifera solution was carried out by processing 50g
of peeled seed and 1M NaCl in 1L of distilled water. The Tannin solution was prepared
with homogenization of 1mL of coagulant in 1L of distilled water. For the preparation of
the Aluminum Sulphate solution, 1g of the chemical was homogenized in 1L of distilled
water.
The laboratory test was performed on Jar-Test equipment in order to reproduce the
coagulation, flocculation and sedimentation processes, followed by the filtration process
of the effluent. The equipment was programmed according to Theodoro (2012), initiating
the rotation of the blades at 150rpm for a time of 3 minutes, then taking the reduction to
15rpm for a period of 10 minutes and then ceasing the stirring process. The filtration
process was performed using a downflow sand filter and the filter construction was
performed according to Di Bernardo et al. (2003). The filter bed was built in the lower
part with sand with granulometry less than 0.425 mm and in the upper part with sand
with granulometry in the range of 0.425 to 0.800 mm.
The sample collections during the test were programmed to be performed every 10
minutes during the sedimentation process, and a final sample collection was performed
after the filtration process. The samples were submitted to analysis of the parameters of
apparent color, turbidity, pH and electrical conductivity according APHA (2012).
The laboratory test was performed in duplicate to avoid possible errors and the data
were analyzed statistically through the application of analysis of variance (ANOVA) and
Tukey test with a significance level of 5% with the use of BioEstat 5.0 software.

3. RESULTS

The behavior of the electric conductivity is presented in Figure 1 the behavior of T1

and T2 were similar to each other, and values presented during the sedimentation stage
presented values close to that found in the crude effluent (0.83mSm-1), but the T3 showed
an increase of the electrical conductivity, this occurring due to the presence of Sodium
Chloride in the solution of Moringa oleifera.
After the filtration process, all the processes presented reduction of the parameter,
obtaining values of 0.55 mSm-1 (T1), 0.32 mSm-1 (T2) and 0.39 mSm-1 (T3), lower
182 F. J. L. Janz, E. R. Pereira, T. Ribeiro et al.

electrical conductivity value. The reduction presented by all treatments is due to the
adsorption process of the salts in the filter media used.
The electrical conductivity data were submitted to the ANOVA variance test and the
results obtained are presented in Table 1.
Statistical analysis showed that the value of P is 0.1629, the fact that P is greater than
0.05, shows that the treatments did not present significant variances with each other when
the values obtained after the filtration process were analyzed.
The pH values of all treatments remained between 4.5 and 5 before the filtration
process (Figure 2). Comparing with the value of 4.6 of the crude effluent it is possible to
observe that the coagulants did not influence the variation of this parameter. The values
collected in the samples after the filtration presented an increase of the parameter, where
T1 presented the highest increase in pH, obtaining 6.6.
All treatments presented pH values within the limit established by Resolution No.
430 of 2011 of the National Environmental Council (Brazil, 2011), which determines that
effluent must have pH between 5 and 9 for launch in any water body. The results of the
statistical analysis showed that in the ANOVA test (Table 2) that the treatments presented
differences between them, and after the Tukey test (Table 3) it was evidenced that the
difference occurs in the comparison of T1 and T2.

Figure 1. Mean values of the electrical conductivity obtained in the test.

Table 1. Statistical data obtained through the ANOVA variance test

Variation Source GL SQ MQ F P
Between groups 2 0.058 0.029 3.5358 0.1629
Within the groups 3 0.024 0.008
Total 5 0.082
 Treatment of Residual Waters of the Brewery Industry ... 183

Table 2. Statistical data obtained through the ANOVA variance test

Variation Source GL SQ MQ F P
Between groups 2 0.395 0.198 11.2273 0.04
Within the groups 3 0.053 0.018
Total 5

Table 3. Statistical data obtained through the Tukey test

Tukey Difference Q (p)

T1 and T2 0.6200 6.6092 <0.05
T1 and T3 0.2200 2.3452 Not significant
T2 and T3 0.4000 4.2640 Not significant

Figure 2. Mean values of pH obtained in the assay.

Figure 3. Turbidity removal efficiency values obtained in the test.

184 F. J. L. Janz, E. R. Pereira, T. Ribeiro et al.

When analyzing the turbidity removal efficiency of all treatments before the filtration
process (Figure 3), it is possible to observe that the efficiency of the coagulants used was
related to time, that is, the efficiency increases over time, T3 was the one that presented
the best behavior, but T3 in its first instants after the application showed
increased turbidity, this occurs due to the solution being opaque and the presence of
organic matter.
After the filtration process, better values of turbidity removal efficiency were
obtained for all treatments, which were higher than 90%, with T2 (Tannin) showing the
best turbidity removal efficiency with a result of 98.8%, while T1 and T3 obtained results
of 93.5% and 94.1%, respectively. Statistical results (Table 4 and Table 5) demonstrated
that T2 presented a significant difference with the other treatments, thus highlighting its
positive performance of removal.

Table 4. Statistical data obtained through the ANOVA variance test

Variation Source GL SQ MQ F P
Between groups 2 33.699 16.849 19.9381 0.0178
Within the groups 3 2.535 0.845
Total 5

Table 5. Statistical data obtained through the Tukey test

Tukey Difference Q (p)

T1 and T2 5.2650 8.0996 <0.05
T1 and T3 0.5150 0.7923 Not significant
T2 and T3 4.7500 7.3073 <0.05

Figure 4. Apparent color removal efficiency values obtained in the assay.

 Treatment of Residual Waters of the Brewery Industry ... 185

For the apparent color parameter, it can be seen from Figure 4 that
T1 and T3 treatments presented higher efficiencies than T2 treatment. With
the use of filtration, an improvement in the apparent color removal values of all
treatments was observed, thus presenting removal values above 90%. The T2 treatment
presented an increase of 18.2% (sedimentation) to 99.6% (filtration) denotand
efficiency with the filtration process, thus finalizing the test with the best result in the
parameter.
Statistical analysis showed that the procedures presented a significant difference
when ANOVA (Table 6) and Tukey’s test (Table 7) were performed. This difference was
shown in the comparison of T2 with the other treatments. The treatments T1 and T3 do
not present significant differences if.

Table 6. Statistical data obtained through the ANOVA variance test

Variation Source GL SQ MQ F P
Between groups 2 29.093 14.531 29.9624 0.0098
Within the groups 3 1.455 0.485
Total 5 30.548

Table 7. Statistical data obtained through the Tukey test

Tukey Difference Q (p)

T1 and T2 4.0000 8.1229 <0.05
T1 and T3 1.1300 2.2947 Not significant
T2 and T3 5.1300 10.4176 <0.05

CONCLUSION

The use of the three coagulants presented demonstrated efficiency for the
treatment of the effluent from the brewing industry, besides showing that the
application of organic coagulants presents final results similar or superior to the inorganic
coagulant. All treatments obtained results superior to 90% for removal of apparent color
and turbidity, but the tannin presented greater prominence presenting removal efficiency
of 98.8% of turbidity and 99.6% of apparent color. All treatments were in compliance
with CONAMA 430/2011.

REFERENCES

APHA – American Public Health Association. Standard Methods for the Examination of
Water and Wastewater. 22 ed. Washington, 2012.
186 F. J. L. Janz, E. R. Pereira, T. Ribeiro et al.

Brazil. National Council for the Environment. Resolution No. 430, of May 13, 2011.
Official Gazette of the Federative Republic of Brazil, Brasília, DF, May 16. 2011.
Available at: <http://www.mma.gov.br/port/conama/legiabre.cfm?codlegi=646>.
Accessed on: 31 Aug. 2015.
CERVBRASIL. Brazilian Association of the Beer Industry. Yearbook 2016. Available at:
http://www.cervbrasil.org.br/paginas/index.php?page=anuario-2015. Accessed on:
May 15, 2018.
CETESB. Secretariat of the Environment. Environmental Sanitation Technology
Company - CETESB. Beers and Soft Drinks. São Paulo: CETESB, 2005. Available
at: http://www.cetesb.sp.gov.br. Accessed on: May 17, 2018.
Di Bernardo, L., Mendes, C. G. N., Brandão, C. C. S., Sens, M. L., Pádua, V. L. Water
Treatment for Direct Filtration. Di Bernardo, Luiz (coordinator) - Rio de Janeiro:
ABES, RiMa, 2003. Project PROSAB 498 p.
Kochenborger, G. Physical-chemical treatment for brewery effluent. 2012. 40 f. TCC
(Undergraduate) - Course of Environmental Engineering, Passo Fundo University,
Passo Fundo, 2012.
Marcusso, E. F., Müller, C. V. Beer in Brazil: The Ministry of Agriculture Informing and
Clarifying. Ministry of Agriculture, 2017. Available at: http://www.agricultura.
gov.br/assuntos/inspecao/produtos-vegetal/pastapublicacoes-DIPOV/a-cerveja-no-
brasil-28-08.pdf. Accessed on: May 15, 2018.
Theodoro, J. D. P. Study of coagulation/flocculation mechanisms to obtain potable water
for human consumption. 2012. 185f. Doctoral Thesis (Department of Chemical
Engineering) - Technology Center, State University of Maringá, Maringá, set. 2012.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 10

APPLICATION OF ELECTROCOAGULATION WITH

VOLTAGE VARIATION TO OBTAIN POTABLE WATER

Ingrid Cardozo Botelho1, Thiago Andre Bezerra Higuchi1

and Joseane Débora Peruço Theodoro2
1
Environmental Engineering Student - UTFPR- Londrina, Londrina, Brazil
2
Department of Environmental Academic,
Federal Technological University of Paraná,
Londrina, Paraná, Brazil

ABSTRACT

Urban development contributes to increasing water pollution, so we must develop

alternative ways to treat water. Therefore, the objective of this paper was to perform the
water treatment - through the electrocoagulation process - and to calculate the cost of the
operation. The paper was done according to predefined experimental planning that has
nine samples with duplicate plus a pair of samples in central point. The assays were
carried out using a bench-level acrylic reactor (2 liters), a magnetic stirrer, a voltage
source and a monopolar electrode system arranged by means of 4 equidistantly connected
aluminum plates in parallel. The studied parameters were final pH and electric
conductivity, tested in two different voltages (5V and 11V). Sampling time were 40, 60
and 80 minutes. At the end of the trials, treatment costs were calculated to ensure that it
was applicable. The analysis of the results concluded that the electrocoagulation process
assists in the neutrality of the solution, brings the acid and basic samples near to the
neutral pH (7) and the electric conductivity does not have significant change. Through the
work it was possible to observe that the eletrocoagulation process accomplished with the
continuous voltage of 5V was the one that presented the lowest cost, considering that the
energy consumption and the use of the electrode were smaller 0.03325kWh and
1.23949µg respectively.
188 I. C. Botelho, Th. A. Bezerra Higuchi and J. D. Peruço Theodoro

Keywords: Electrocoagulation, water treatment

1. INTRODUCTION

This paper studied an option of water treatment, the eletrocoagulation, which consists
in the application of potential difference in an aluminum electrode, which undergoes
oxidation at the anode and reduction at the cathode, the oxygen microbubbles formed by
the cathode help that the pollutant particles are suspended (Martins 2017).
According to Adapureddy (2012), the initial pH influences the removal parameters
for water purification or effluent treated by electrocoagulation. His job shows that the
initial pH around 8 was the best results of removal parameters, and he also observed that
at pH ≥ 9 the generation of microbubbles increases which shows more gas production.
The electrical conductivity is an indicator of the amount of dissolved salts, however it
is possible to measure the presence of salts in the collected water since in this work no
salts will be added for an acceleration of the process. Therefore, the objective of this
paper is to analyze the parameters of pH and of conductivity in the water treatment
process by electrocoagulation, and also to calculate the cost of this operation.

2. METHODOLOGY

The sampling was made at the water inlet tank of the treatment station (Figure 1),
located in Paraná, Brazil.

Figure 1. Treatment station.

 Application of Electrocoagulation with Voltage Variation ... 189

Source: Theodoro (2017) and Ferrareze (2016).

Figure 2. Bench level reactor: A) Monopolar electrode system; B) Recipient of the reactor; C) Magnetic
stirrer; and D) Continuous voltage source.

Table 1. Experimental table

Initial pH
Tests Time (min)
5V 11V
1 40 4.06 4.03
2 60 4.06 4.03
3 80 4.06 4.03
4 40 6.31 6.31
5 60 6.31 6.31
6 80 6.31 6.31
7 40 8.80 9.07
8 60 8.80 9.07
9 80 8.80 9.07
10 60 6.31 6.31
11 40 4.06 4.03
12 60 4.06 4.03
13 80 4.06 4.03
14 40 6.31 6.31
15 60 6.31 6.31
16 80 6.31 6.31
17 40 8.80 9.07
18 60 8.80 9.07
19 80 8.80 9.07
20 60 6.31 6.31
190 I. C. Botelho, Th. A. Bezerra Higuchi and J. D. Peruço Theodoro

The bench-level reactor (Figure 2) used to accomplish the electrocoagulation was a

rectangular acrylic pitcher with 12cm of width, 12cm of length and 20 of height; a
magnetic stirrer, a monopolar electrode system and a continuous voltage source witch is
adjustable in 5V and 11V.
In relation to the monopolar electrode system, these were connected in parallel and
fixed by steel screws, the four aluminum plates have 2cm equidistance and dimensions
16cm high x 8cm long x 0.03cm thick, resulting in two pairs of electrodes. The adjustable
voltage source allowed the current to pass through the system and the magnetic stirrer
was used to homogenize the fluid.

Table 2. pH corretion

Initial pH 5V Drops 11V Drops

Neutral 6.31 0 6.31 0
Acidic 4.06 92 de HCl 4.03 50 de HCl
Basic 8.80 35 de NaOH 9.07 36 de NaOH

For the analysis of pH and conductivity, were collected 25mL samples in 40,
60 and 80 minutes; the reading was made straight after collecting them. In the
statistical planning (Table 1) it was performed 9 trials in duplicate and a pair in the
central point.
The methodology and equipment used in the parameter readings pH and
conductivity were, respectively, Potentiometric Method range 1.0 to 13.0 and
pHmetro mPA-210 and LQ Electrometric Method: 0.01μS.cm-1 and Mca 150
Conductor (Standard Methods for the Examination of Water and Wastewater (American
Public Health Association, 2012). The acid and basic pH were corrected using solutions
of hydrochloric acid (1M HCl) and sodium hydroxide (1M NaOH), respectively (Table
2).
In relation to the monopolar electrode system, these were connected in parallel and
fixed by steel screws, the four aluminum plates have 2cm equidistance and dimensions
16cm high x 8cm long x 0.03cm thick, resulting in two pairs of electrodes. The adjustable
voltage source allowed the current to pass through the system and the magnetic stirrer
was used to homogenize the fluid.
On the other hand the use of the electrode (1) and the energy consumption (2) were
calculated by de equations below.

i.t.M
𝑚𝑒 = n.F
(1)

where:
𝑚𝑒 Is an amount of dissolved electrode (g);
 Application of Electrocoagulation with Voltage Variation ... 191

i Is current (A);
t The process time (s);
M The molar ratio of the predominant element of the electrode (g.mol-1);
n Is the number of electrons in the oxidation reaction of the anodic electrode; and F It
is a Faraday constant (F = 9.65 x 104 C.mol-1).

𝑈.𝑖.𝑡
𝐶= (2)
𝑣

where:
𝐶 Energy consumption (Wh.m-³);
𝑈 Applied electric voltage (V);
𝑖 Electric current (A);
𝑡 Time of application of the current or process time (h); and
𝑣 Volume of treated effluent (m³)
Through the results found with the two equations above, the cost of the process was
calculated using equation (3).

𝐶𝑜𝑝 = 𝑎. 𝐶𝑒𝑛𝑒𝑟𝑔 + 𝑏. 𝐶𝑒𝑙𝑒𝑡 (3)

where:
𝐶𝑜𝑝 Total system operating cost (R$.m-3);
𝑎 Cost of electricity (R$.kWh-1);
𝐶𝑒𝑛𝑒𝑟𝑔 Consumption of electric energy (kWh.m-3);
𝑏 Cost of the electrodes (R $.kg aluminum-1); and
𝐶𝑒𝑙𝑒𝑡 Electrode consumption (kg.m-3 water treated).

3. RESULTS

The characterization values of the crude sample are shown in Table 3. In addition, the
results of the experimental design for the electrocoagulation process are
presented in Table 4.

Table 3. The parameter values of the crude sample

Parameters Values
pH 5.99
Conductivity (μS.cm-1) 0.17
192 I. C. Botelho, Th. A. Bezerra Higuchi and J. D. Peruço Theodoro

Table 4. Values founded

Conductivity
Initial pH Final pH
Tests Time (min) (μS.cm-1)
5V 11V 5V 11V 5V 11V
1 40 4.06 4.03 5.47 4.95 0.18 0.17
2 60 4.06 4.03 5.74 5.1 0.18 0.17
3 80 4.06 4.03 5.52 5.47 0.18 0.17
4 40 6.31 6.31 5.59 6.52 0.16 0.16
5 60 6.31 6.31 5.66 6.41 0.18 0.16
6 80 6.31 6.31 5.88 6.87 0.16 0.15
7 40 8.80 9.07 7.25 7.74 0.18 0.20
8 60 8.80 9.07 7.38 7.32 0.18 0.20
9 80 8.80 9.07 7.38 7.13 0.18 0.21
10 60 6.31 6.31 5.79 6.22 0.16 0.17
11 40 4.06 4.03 4.94 5.54 0.20 0.18
12 60 4.06 4.03 4.77 5.16 0.20 0.18
13 80 4.06 4.03 4.87 5.11 0.20 0.17
14 40 6.31 6.31 5.61 6.25 0.17 0.16
15 60 6.31 6.31 6.05 6.63 0.17 0.15
16 80 6.31 6.31 6.02 6.74 0.16 0.15
17 40 8.80 9.07 6.99 7.58 0.18 0.19
18 60 8.80 9.07 6.73 7.45 0.18 0.18
19 80 8.80 9.07 6.64 7.61 0.18 0.18
20 60 6.31 6.31 5.88 6.44 0.18 0.16

Figure 3. Graphic of final pH variation.

 Application of Electrocoagulation with Voltage Variation ... 193

Figure 4. Graphic of electrical conductivity variation.

Table 5. Electrode use by the electrocoagulation process

Tests Electrode wear (µg)

5V 11V
3 1.23949568 9.915965458
6 1.23949568 3.718487047
9 1.23949568 6.197478411
13 1.23949568 8.676469775
16 1.23949568 6.197478411
19 1.23949568 6.197478411

Table 6. Energy consumption by electrocoagulation process

Energy consumption (kWh)

Tests
5V 11V
3 0.03325 0.26600
6 0.03325 0.09975
9 0.03325 0.16625
13 0.03325 0.23275
16 0.03325 0.16625
19 0.03325 0.16625

Figure 3 shows the graphic of final pH variation, the analysis of the results
confirmed what Adapureddy says, that the electrocoagulation process brings
the water near to neutral pH. At the conventional water treatment, after the
addition of the coagulant the pH decrease, thus the neutralization of water is required.
194 I. C. Botelho, Th. A. Bezerra Higuchi and J. D. Peruço Theodoro

However, the study shows that the electrocoagulation process tends to keep the water pH
neutral.
The variation of electrical conductivity represented in Figure 4 had few variations, it
can be interpreted by no salts addition moreover there aren’t many changes at the
conductivity by variation of 5V and 11V voltage.

Table 7. Total cost of the eletrocoagulation process

Total cost (R$)

Tests
5V 11V
3 0.022989 0.127076
6 0.022989 0.047654
9 0.022989 0.079423
13 0.022989 0.111192
16 0.022989 0.079423
19 0.022989 0.079423

The last analysis was to determine the cost of the operation, the beginning of the
electrode use was calculated (Table 5), then the energy consumption (Table 6) and then
with these values and the prices of the material and kWh, the total cost was found
(Table 7).

CONCLUSION

In the analysis of pH and the electrical conductivity behavior, it was observed that the
electrocoagulation assists in the neutrality of the solution, and for electrical conductivity
the process does not show significant changes, maintaining the range of 0.15 to
0.20μS.cm-1.
The operation cost in the applied voltage of 5V is lower, therefore more interesting to
use this voltage to the eletrocoagulation process, which consumes less energy, making the
experiment more satisfactory.

REFERENCES

Adapureddy, Sri Malini; Goel, Sudha. Optimizing Electrocoagulation of Drinking Water

for Turbidity Removal in a Batch Reactor. In: International Conference on
Environmental Science and Technology, 2012, Singapore. Available at:
 Application of Electrocoagulation with Voltage Variation ... 195

<http://www.ipcbee.com/vol30/020--ICEST2012_N30001.pdf> Accessed on: 23

March 2018.
APHA – American Public Health Association. Standard Methods for the Examination of
Water and Wastewater. 22 ed. Washington. 2012.
Ferrareze, C. N. M. Treatment of water through the electrocoagulation process. 107 f.
2016. TCC (Graduation in Environmental Engineering), Federal Technological
University of Paraná. Londrina, 2016.
Martins, Ana L. N. Study of the variation of sodium cjloride concentration in the
electrocoagulation process applied to water treatment. 2017. 69 p. Paper of
completion of course (Environmental Engineering). Federal Tchnological Univerity
of Paraná.
Theodoro, J. D. P. Water treatment through the electrocoagulation process associated
with the use of an organic coagulant (Tanfloc SG). [personal message] 15 August
2017.
In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 11

AN EVALUATION OF THE PERFORMANCE OF THE

COAGULATION/FLOCCULATION PROCESS WITH
ALUMINUM AND FERRIC SALTS ON THE REMOVAL
OF ALGAL TOXINS

Ayşe Büşra Şengül*

Department of Mechanical Engineering, Wichita State University, Wichita, KS, US

ABSTRACT

Eutrophication, or the excessive richness of nutrients in a lake or other body of

water, is one of the most prevalent water quality problems in the United States as well as
other parts of the world. It has led to excessive growth of algal blooms, which not only
cause the death of aquatic plants and animals, but also produce high levels of toxins and
odorous compounds, thereby posing serious risks to safe drinking water and human
health. Microcystis aeruginosa is well known to be one of the most dangerous and most
common bloom-forming cyanobacteria that produces hepatotoxic microcystins. This
study evaluated the performance of the coagulation/flocculation process using aluminum
and ferric salt coagulants for the removal of both microcystins (intracellular and
extracellular) and turbidity. The effects of coagulant dosage, pH, and coagulant-aid
dosage on microcystins and turbidity removal were also evaluated. All results
demonstrated that the optimum dosage of aluminum sulphate, ferric chloride, and ferrous
sulphate for the removal of both microcystins and turbidity was found to be 60, 60, and
20 mg/L, respectively. With optimized coagulant dosages, the optimum pH value for both
microcystins and turbidity removal occurred with a pH of 7, 6, and 7 for aluminum

*
Corresponding Author Email: [email protected].
198 Ayşe Büşra Şengül

sulphate, ferric chloride, and ferrous sulphate, respectively. On the other hand, for all
coagulants, the optimum polyelectrolyte dosage for the removal of both microcystins and
turbidity was found to be 0.2 mg/L. Among these coagulants, aluminum sulphate (65.2%)
was found to be more effective for both microcystins and turbidity removal compared to
ferric chloride (56.3%) and ferrous sulphate (30.9%). Last, the concentration of turbidity
decreased to below 5 NTU, which is the maximum turbidity level in drinking water
determined by the World Health Organization (WHO). After coagulation/flocculation,
the total microcystin concentration (4.09 µg/L) in the water remained above the WHO
guideline value of 1 µg/L for drinking water, but the release of toxins in the water was
not observed. Therefore, both intracellular microcystin and turbidity could be removed
through the coagulation/flocculation process, whereas extracellular microcystin is still of
particular concern in drinking water treatment due to its difficult removal.

Keywords: blue-green algae, cyanobacteria, microcystis aeruginosa, microcystin,

coagulation/flocculation

INTRODUCTION

Eutrophication, or the excessive richness of nutrients in a lake or other body of water,

is one of the most prevalent water quality problems in the United States as well as other
parts of the world. It has led to excessive growth of algal blooms in water, which not only
cause the death of aquatic plants and animals, but also produce higher levels of toxins and
odor compounds, thereby posing serious risks to safe drinking water and human health
(Huang et al., 2015; Zamyadi et al., 2015). These blooms can also be a financial problem
to water treatment plants where cells can clog the filters, increase the chemical demand
and also lead to the formation of disinfection byproducts (Campinas & Rosa, 2010a; Shen
et al., 2011; Zhang et al., 2011).
Microcystis aeruginosa is one of the dominant blue-green algaes found frequently in
eutrophic water bodies (Sun et al., 2013; Dong et al., 2014). Microcystins produced by
Microcystis aeruginosa are hepatotoxins, which have caused several poisonings of
animals and also pose a health hazard for humans (Wang et al., 2015; Ma et al., 2016).
One of the primary effects of microcystins is their cause of liver damage and failure. It
also has toxic effects on gastrointestinal system and kidneys (Hoko & Makado, 2011;
Zamyadi et al., 2015). In addition, chronic exposure to these toxins causes tumor
promotion (Lévesque et al., 2014; Shamsollahi et al., 2015). Although more than 80
variants have been detected in water so far, microcystin-LR is considered one of the most
toxic and abundant microcystins that contains lucien (L) and arginine (R) (Westrick et al.,
2010; D’Anglada & Strong, 2015; Shamsollahi et al., 2015). Therefore, the United States
Environmental Protection Agency (USEPA) has placed microcystins as liver toxins on
the Drinking Water Contaminants Candidate List (USEPA, 2009), and the World Health
 An Evaluation of the Performance of the Coagulation/Flocculation … 199

Organization (WHO) has specified a guideline value for microcystins in drinking water
as 1 µg/L (WHO, 2003).
Generally, microcystins are contained within cells that are classified as cell-bound or
intracellular (Campinas & Rosa, 2010b, c; Dixon et al., 2011). When the cells are
subjected to natural toxin release (natural cell lysis and death, active release) or induced
toxin release (mechanical/chemical stresses), microcystins can be released into
surrounding waters, and these are classified as dissolved or extracellular toxins (Schmidt
et al., 2002; Campinas & Rosa, 2010b; Dixon et al., 2011; Li et al., 2015). Due to the fact
that extracellular microcystins are not removed as easily as intracellular toxins (Sun et al.,
2012; Sun et al., 2013), it is necessary to develop an efficient method to remove
intracellular microcystins without cell damage.
Coagulation/flocculation is one of the most common processes to improve treatment
efficiency and cost effectiveness for the removal of cyanobacterial cells and intracellular
toxins (Pei et al., 2014; Jiang, 2015). A number of researchers have been focusing on the
impacts of the coagulation/flocculation process on cyanobacterial cells and toxin release
(Han et al., 2012; Sun et al., 2012; Han et al., 2013; Sun et al., 2013; Pei et al., 2014; Li
et al., 2015; Ma et al., 2016). However, in the literature, the coagulation/flocculation
process for the uptake of cyanobacterial cells and toxin is shown to be controversial.
Therefore, some research has been reported that the optimized coagulant dosage as well
as mechanical conditions did not damage the membrane of cyanobacterial cells and the
release of cell metabolites (Chow et al., 1998; Chow et al., 1999; Sun et al., 2012; Sun et
al., 2013; Pei et al., 2014; Li et al., 2015). Other research has emphasized that chemical
treatment with maximum dosage and long contact time causes cell damage, thus releasing
large amounts of toxin into the water (Han et al., 2012; Han et al., 2013). These findings
clearly show that the efficiency of the coagulation/flocculation process can be strongly
affected by the type and dosage of coagulant, pH values, applied shear, and
characteristics of the raw water.
Up to now, a limited number of studies have been assessing the effects of the
coagulation/flocculation process for these studies have only focused on the impact of the
coagulant dosage and contact time for the toxin removal. The literature survey has
demonstrated that there has not been a study on intracellular and extracellular microcystin
removal relative to the influence of pH and coagulant-aid dosage in addition to a
coagulant. Therefore, the main objective of this study was to examine the performance of
the coagulation/flocculation process using aluminum sulphate, ferric chloride, and ferrous
sulphate coagulants for the removal of both microcystins (intracellular and extracellular)
and turbidity. The effects of coagulant dosage, pH, and coagulant-aid dosage on
intracellular and extracellular microcystins and turbidity removal were also thoroughly
investigated.
200 Ayşe Büşra Şengül

MATERIALS AND METHODS

Algae Culture

The freshwater blue-green algae Microcystis aeruginosa was selected as a strain in

this study. It was grown in sterilized BG-11 medium with a 12h light:12h dark
photoperiod and constant temperature (25°C). This medium was composed of 1.5 g/L
NaNO3, 0.04 g/L K2HPO4, 0.075 g/L MgSO4·7H2O, 0.036 g/L CaCl2·2H2O, 0.006 g/L
citric acid, 0.006 g/L ferric ammonium citrate, 0.001 g/L EDTA, 0.02 g/L NaCO3, and 1
ml trace elements. Formulation of the trace elements was as follows: 2.86 g/L H3BO3,
1.81 g/L MnCl2·4H2O, 0.22 g/L ZnSO4·7H2O, 0.39 g/L Na2MoO4·2H2O, 0.08 g/L
CuSO4·5H2O, and 0.05 g/L Co (NO3)2·6H2O. Mixing was done daily by hand. The
cultures were harvested on the 21st day of incubation during the stationary phase of
growth.

Natural Water

Natural water was collected from Büyükçekmece Lake (a drinking water source in
Istanbul, Turkey). After filtering through a 0.45 µm filter, cultures in the BG-11 medium
were transferred to the natural water. Characteristics of this natural water are provided in
Table 1.

Table 1. Characteristics of natural water from Büyükçekmece Lake

Parameter Unit Average

pH 8.07
Temperature °C 22.46
Turbidity NTU 5.70
DOC mg/L 5.67
UV254 cm-1 0.058
SUVA254 L/mg.m 1.023
DOC: dissolved organic carbon; UV: ultra-violet absorbance; SUVA: specific ultraviolet absorbance

Chemicals

Aluminum sulphate (Al2(SO4)3.18H2O), ferric chloride (FeCl3.6H2O), and ferrous

sulfate (FeSO4.7H2O) purchased from Sigma-Aldrich were used as coagulants using a 1%
stock solution. Coagulants were added in a range of dosages (20, 40, 60, 80, and 100
 An Evaluation of the Performance of the Coagulation/Flocculation … 201

mg/L) to determine the optimum coagulation conditions. The pH was varied between 5
and 8 by adding 0.1 M sodium hydroxide (NaOH) and 0.1 M hydrochloric acid (HCl). A
coagulant aid, cationic polyelectrolyte, was used as a 1g/L stock solution and added to the
beaker in a range of dosages (0.2, 0.4, 0.6, 0.8, and 1 mg/L).

Coagulation/Flocculation Experiment

Coagulation/flocculation experiments were carried out at room temperature (20 

2°C) in a standard jar test apparatus (FC 6S, Velp Scientifica) and equipped with six
glass beakers of 1 L. The experimental procedures for determining optimum
coagulation/flocculation conditions have been described and discussed in detail in our
previous studies (Şengül et al., 2016; Şengül et al., 2018).

Analytical Method

Turbidity
After settling, turbidity of the supernatant was measured using a turbidimeter (WTW
TURB 550).

Determination of Intracellular and Extracellular Microcystins

Analysis of intracellular and extracellular microcystins was conducted according to
the method described by Chorus and Bartram (1999) with some adaptations. Details of
this analysis were provided in our previous study (Şengül et al., 2018).

RESULTS AND DISCUSSION

Removal of Microcystins and Turbidity by Coagulation/Flocculation

The performance of the coagulation/flocculation process for the removal of both

microcystins and turbidity were evaluated by aluminum sulphate, ferric chloride, and
ferrous sulphate coagulants. First, the effects of coagulant type and dosage on
microcystins and turbidity removal were determined. Then, the effect of pH with an
optimized coagulant dosage under different pH values on microcystins and turbidity
removal was evaluated. Last, coagulation/flocculation experiments were performed using
an optimized coagulant dosage and pH under various polyelectrolyte dosages. At the
202 Ayşe Büşra Şengül

beginning of the experiments, the initial concentration of microcystins was found to be

10.61 µg/L, which was comprised of 10 µg/L intracellular and 0.61 µg/L extracellular
microcystins, respectively. The initial concentration of turbidity was measured as 9 NTU.

Removal of Microcystins and Turbidity by Aluminum Sulphate

The optimum coagulation/flocculation conditions with aluminum sulphate in terms of

microcystins (65.2%) and turbidity (85.1%) removal were determined as a coagulant
dosage of 60 mg/L, pH value 7, and polyelectrolyte dosage of 0.2 mg/L. This was also
determined from our previous studies (Şengül et al., 2016; Şengül et al., 2018). After a
dosage of 60 mg/L aluminum sulphate, no significant turbidity occurred and no further
significant removal of microcystins was achieved. Moreover, no further increase in the
amount of extracellular microcystin was observed.

Removal of Microcystins and Turbidity by Ferric Chloride

Figure 1 shows the results of microcystins and turbidity removal after the
coagulation/flocculation process under various ferric chloride dosages, pH values, and
polyelectrolyte dosages. The removal of intracellular microcystin increased by increasing
the ferric chloride dosage from 0 to 60 mg/L. At a dosage of more than 60 mg/L,
intracellular microcystin concentration slightly decreased, and no further significant
removal was achieved. The concentration of intracellular microcystin decreased from 10
to 5.11 µg/L, and its removal efficiency was 48.9% in the presence of 60 mg/L ferric
chloride. The trend of turbidity removal by coagulation/flocculation was similar to that of
intracellular microcystin removal. Turbidity concentration decreased from 9 to 0.93
NTU, with 89.70% removal efficiency when the ferric chloride dosage was 60 mg/L. On
the other hand, no significant change in the amount of extracellular microcystin was
observed. Therefore, the optimum ferric chloride dosage was determined to be 60 mg/L
for both microcystins and turbidity removal.
As shown in Figure 1(b), removal of the intracellular microcystin increased by
increasing the pH from 5 to 6, but there was no significant change in its removal when
the pH level increased from 7 to 8. Turbidity removal showed a similar trend with
intracellular microcystin. Thus, the high performance of ferric chloride for removing
intracellular microcystin and turbidity was determined to be at pH 6. The concentration of
intracellular microcystin was decreased from 10 to 4.65 µg/L, with 53.5% removal
efficiency in the presence of a 60 mg/L ferric chloride dosage at pH 6. Turbidity
concentration decreased from 9 to 1.21 NTU, with a removal efficiency of 86.6%. In
 An Evaluation of the Performance of the Coagulation/Flocculation … 203

addition, the removal of extracellular microcystin was not impacted when the pH level
was changed. The removal efficiency of intracellular microcystin by ferric chloride
coagulation was increased 8.6% with a change in the pH, whereas the removal efficiency
of turbidity was decreased.
The effect of polyelectrolyte dosage on microcystins and turbidity removal was
evaluated with a 60 mg/L ferric chloride dosage and pH level 6 under various dosages of
polyelectrolytes, as shown in Figure 1(c). The removal of intracellular microcystin and
turbidity increased by increasing the polyelectrolyte dosage from 0 to 0.2 mg/L; however,
no remarkable increase was obtained with a further increment in the polyelectrolyte
dosage beyond 0.2 mg/L. The concentration of intracellular microcystin decreased from
10 to 4.37 µg/L, with 56.30% removal efficiency. Turbidity concentration decreased from
9 to 2.07 NTU, with removal efficiency of 77%. Moreover, no significant changes were
observed in the amount of extracellular microcystin concentration. The removal
efficiency of intracellular microcystin by ferric chloride coagulation was increased 13.1%
with a change in the pH and the addition of a polyelectrolyte, whereas the removal
efficiency of turbidity was decreased.
Until now, only two studies in the literature have evaluated the effect of ferric
chloride coagulation/flocculation on Microcystis aeruginosa cells and
toxin release (Chow et al., 1998; Li et al., 2015), which reported that
coagulation/flocculation with ferric chloride achieved up to 100% removal of
intracellular microcystin. Also, ferric chloride and mechanical action did not damage
cyanobacterial cells and the release of extracellular toxin. In this study, only 56.3%
intracellular microcystin removal was achieved, and no further increase in the amount of
extracellular microcystin was observed, with its concentration measured at 0.61 µg/L
over the experimental period. In general, the predominant removal
mechanism at relatively low doses and slightly acidic pH value of coagulants is charge
neutralization.
However, at higher dosages and approximately neutral-to-alkaline pH values sweep
flocculation occurs (Duan & Gregory, 2003). Therefore, all results clearly showed that
the iron ions can be transformed to Fe(OH)3 by hydrolysis at pH 6 with a dosage of 60
mg/L, and the Fe(OH)3 tends to remove intracellular microcystin via sweep flocculation.
Nevertheless, natural organic matter in aquatic systems may result in lower
intracellular microcystin removal efficiency (Ma & Liu, 2002) and extracellular
microcystin are efficiently removed by charge neutralization more than sweep
coagulation (Ghernaout, 2014). Another reason is that when the initial concentration of
microcystin is around 10 µg/L or higher, there may not be a significant reduction in the
microcystin concentration.
204 Ayşe Büşra Şengül

Figure 1. Uptake of microcystins (intracellular and extracellular) and turbidity after

coagulation/flocculation process under different conditions: (a) ferric chloride dosage, (b) pH value,
and (c) polyelectrolyte dosage.

Removal of Microcystins and Turbidity by Ferrous Sulphate

Figure 2 demonstrates the jar test results of microcystins and turbidity removal after
the coagulation/flocculation process under various ferrous sulphate dosages, pH values,
and polyelectrolyte dosages. The removal of intracellular microcystin increased with a 20
mg/L dosage of ferrous sulphate. Then, the removal of intracellular microcystin gradually
decreased with the continuing increase in ferrous sulphate dosages. The concentration of
intracellular microcystin decreased from 10 to 6.98 µg/L, and its removal efficiency was
 An Evaluation of the Performance of the Coagulation/Flocculation … 205

30.2% in the presence of a 20 mg/L ferrous sulphate dosage. A similar trend was also
observed using ferrous sulphate at a dosage of 20 mg/L. Turbidity concentration
decreased from 9 to 2.53 NTU, with a removal efficiency of 71.9%. No significant
turbidity occurred, after a dosage of 20 mg/l. On the other hand, the amount of
extracellular microcystin was not changing. Hence, 20 mg/L ferrous sulphate dosage was
considered as optimum dosage due to the highest obtained microcystin and turbidity
removal.
As shown in Figure 2(b), removal of the intracellular microcystin was increased by
increasing the pH from 5 to 7, but no significant change was observed for both
microcystin and turbidity removal when the pH level increased from 7.5 to 8. Turbidity
removal showed a similar trend with intracellular microcystin. Thus, the high
performance of ferrous sulphate in removing intracellular microcystin and turbidity was
determined to be at pH 7.
The concentration of intracellular microcystin was decreased from 10 to 5.43
µg/L, with 45.7% removal efficiency in the presence of a 20 mg/L ferrous sulphate
dosage at pH 7. Turbidity concentration decreased from 9 to 3.26 NTU, with a
removal efficiency of 63.78%. In addition, the removal of extracellular microcystin was
not clearly impacted by changing the pH level. The efficiency of removing
intracellular microcystin using ferrous sulphate coagulation was increased
33.9% under different pH levels, unlike the efficiency of removing turbidity, which
decreased.
The effect of the polyelectrolyte dosage on the removal of microcystins and turbidity
was evaluated with an optimized ferrous sulphate dosage and pH under
various polyelectrolyte dosages, as shown in Figure 2(c). The removal of intracellular
microcystin and turbidity increased by increasing the polyelectrolyte dosage
from 0 to 0.2 mg/L. However, results showed that when the polyelectrolyte
dosage was further increased from 0.2 to 1 mg/L, no additional obvious increase was
observed.
The concentration of intracellular microcystin decreased from 10 to 6.91 µg/L, with
30.9% removal efficiency. The turbidity concentration decreased from 9 to 3.62 NTU,
with 59.8% removal efficiency. Moreover, no significant increase in the amount of
extracellular microcystin concentration was observed with the addition of
polyelectrolytes. The removal efficiency of intracellular microcystin and turbidity by
ferrous sulphate coagulation was decreased with a change in the pH and the addition of
polyelectrolytes.
In this study, ferrous sulphate showed poor intracellular microcystin and turbidity
removal efficiency compared to aluminum sulphate and ferric chloride. The maximum
intracellular microcystin removal for ferrous sulphate was only 45.7% in the presence of
a 20 mg/L ferrous sulphate dosage at pH 7. When a higher dosage of ferrous sulphate was
206 Ayşe Büşra Şengül

added, the effectiveness of the processes decreased further. There was no change in the
removal of extracellular microcystin.

(a)

(b)

(c)

Figure 2. Uptake of microcystins (intracellular and extracellular) and turbidity after

coagulation/flocculation process under different conditions: (a) ferrous sulphate dosage, (b) pH value,
and (c) polyelectrolyte dosage.

Jar test experiments were performed with aluminum sulphate, ferric chloride, and
ferrous sulphate coagulants to evaluate the performance of the coagulation/flocculation
process based on microcystins and turbidity removal (Figure 3). The concentration of
intracellular microcystin decreased from 10 µg/L to 3.48, 4.37, and 6.91 µg/L, with 65.2,
56.3, and 30.9% removal efficiency for aluminum sulphate, ferric chloride, and ferrous
sulphate coagulants, respectively. The optimum dosage of aluminum sulphate, ferric
chloride, and ferrous sulphate for maximum microcystins and turbidity removal was
found to be 60, 60, and 20 mg/L respectively. The corresponding turbidity removal
 An Evaluation of the Performance of the Coagulation/Flocculation … 207

efficiencies of aluminum sulphate, ferric chloride, and ferrous sulphate were found to be
85.1, 77.7, and 59.8%, respectively. The concentration of turbidity decreased from 9
NTU to 1.34, 2.07, and 3.62 NTU for aluminum sulphate, ferric chloride, and ferrous
sulphate, respectively.

AS FC FS
AS_T FC_T FS_T

Turbidity (NTU)
Microcystins %
100 10
80 8
Removal
60 6
40 4
20 2
0 0
20 40 60 80 100
Coagulant Dosage (mg/L)
(a)

AS FC FS
AS_T FC_T FS_T
Microcystins %

Turbidity (NTU)
100 10
Removal

80 8
60 6
40 4
20 2
0 0
5 6 7 7.5 8
pH
(b)

AS FC FS
AS_T FC_T FS_T
Turbidity (NTU)
Microcystins %

100 10
80 8
Removal

60 6
40 4
20 2
0 0
0.2 0.4 0.6 0.8 1

Polyelectrolyte Dosage (mg/L) (c)

Figure 3. Removal of microcystins (intracellular and extracellular) and turbidity removal after
coagulation/flocculation process under different conditions: (a) coagulant dosage, (b) pH value, and (c)
polyelectrolyte dosage.

An excessive dosage of coagulant caused the reversal of surface charge and the
restabilization of particles, which led to a decrease in the microcystins and turbidity
208 Ayşe Büşra Şengül

removal efficiency. With the optimized coagulant dosages, the pH for maximum
microcystins removal occurred at a pH of 7, 6, and 7 for aluminum sulphate, ferric
chloride, and ferrous sulphate, respectively, while the pH change with optimized
coagulant dosage led to a decrease in turbidity removal efficiency. Furthermore, the
polyelectrolyte dose increased the removal efficiency of microcystins, whereas it
decreased the removal efficiency of turbidity.
Overall, results indicated that aluminum sulphate was found to be a more effective
coagulant for both microcystin and turbidity removal compared to ferric chloride and
ferrous sulphate coagulants. Also, ferrous sulphate was found to be the least effective of
the other coagulants. Turbidity concentration decreased below 5 NTU, which is the
maximum level of turbidity in drinking water determined by the WHO after the
coagulation/flocculation process with all coagulants. However, microcystin concentration
in the water did not achieve the WHO guideline value of below 1 µg/L for drinking
water. Coagulation/flocculation processes achieved limited ability to remove the
microcystins, possibly because of the natural organic matter and the initial concentration
of microcystins in the water.

LITERATURE REVIEW
Coagulation/flocculation is still one of the essential treatment processes for algae and
cyanobacteria, and the associated removal of their metabolites (Ma & Liu, 2002;
Gonzalez-Torres et al., 2014). This process involves the addition and mixing of
coagulants to destabilize negative charges of particles and to prevent electrostatic
repulsion between particles. Therefore, destabilized particles can tend to agglomerate and
form flocs that are subsequently removed by sedimentation (Merel et al., 2013; Sun et al.,
2013). Coagulation/flocculation in water treatment occurs by several mechanisms: charge
neutralization, adsorption and bridging between particles, electrostatic patch, and sweep
flocculation (Duan & Gregory, 2003; Ghernaout & Ghernaout, 2012; Gonzalez-Torres et
al., 2014). Intracellular toxins can be removed by coagulation, via either charge
neutralization (requires acidic pH values and lower dosages) or sweep flocculation
(requires neutral to alkaline pH values and high dosages), and subsequent
sedimentation and filtration processes (Jiang & Kim, 2008; Wu et al., 2011).
Extracellular toxins are removed more efficiently by charge neutralization than
by sweep flocculation (Ghernaout, 2014). However, the efficiency of intracellular or
extracellular toxin removal through coagulation can be strongly affected by the type and
dosages of coagulant, applied shear, solution pH, and characteristics of water,
particularly natural organic matter in water (Hitzfeld et al., 2000; Ma & Liu, 2002;
Zhang et al., 2014).
 Table 2. Summary of studies investigating performance of coagulation/flocculation process
on cyanobacterial cells and toxin release

Percent Removal
Cyanobacteria Coagulant Mechanical Action Comments References
Intracellular Extracellular
Microcystis CTS, AC, Rapid mix: 97.3 53.08  Best MC removal obtained when CTSAC set to 2.6 (Ma et al.,
aeruginosa and CTSAC 200 rpm 2 min mg/L CTS plus 7.5 mg/L AC. 2016)
Slow mix:  Negligible cell lysis observed during CTS, AC, and
20 rpm 15 min CTSAC coagulation processes.
Settling time:
30 min
Microcystis Ferric Rapid mix: > 80 38.5  Optimum coagulant dosage determined as 50 mg/L. (Li et al.,
aeruginosa Chloride 250 rpm 0.5 min  Coagulant dosage and mechanical actions did not 2015)
Slow mix: cause any damage to cell integrity.
20 rpm 30 min  FeCl coagulant had no effect on extracellular MCs
Settling time: with coagulant dosage below 50 mg/L, but obvious
30 min removal observed at coagulant dosage of 100 mg/L.
 During flocs storage process, number of intracellular
MCs released into supernatant, but cells remained
viable up to 10 d.
Microcystis CTS Rapid mix: 99.1 46.45  Optimum extracellular MCs removal efficiency (Pei et al.,
aeruginosa 226.64 rpm 1.098 obtained with chitosan concentration of 7.31 mg/L. 2014)
min  Chitosan flocculation showed MCs adsorption ability.
Slow mix:  During flocs storage period, MCs release observed
18.86 rpm 11.73 after 4 d.
min
Settling time:
30 min
Microcystis PACI Rapid mix: 94.3 ~ 12.0  Optimum coagulant dosage found to be 4 mg/L. (Sun et al.,
aeruginosa 150 rpm 2 min During floc storage process (after 2 days), PACl caused 2013)
Slow mix: obvious damage to cells and led to large amount of MC
40 rpm 30 min release.
Settling time:
20 min
Microcystis AS – 95 –  Alum treatment with maximum dosage (48 mg/L) (Han et al.,
ichthyoblabe and long contact time (7 days) caused Microcystis 2013)
cell damage, resulting in large amounts of toxic
microcystin released into water.
 Table 2. (Continued)

Percent Removal
Cyanobacteria Coagulant Mechanical Action Comments References
Intracellular Extracellular
Microcystis PAC, Rapid mix: > 99  Coagulation treatment removed Microcystis (Chen
aeruginosa Sepiolite and 200 rpm 1 min aeruginosa cells efficiently, and combined use of et al., 2013)
PAC/ Slow mix: PAC and sepiolite had higher removal efficiency
Sepiolite 50 rpm 10 min compared to each coagulant alone.
Settling time:  Coagulation treatment with PAC (250 mg/L) +
– sepiolite (2.8 mg/L) or sepiolite (7 mg/L) had no
negative effect on Microcystis aeruginosa cells;
however, with PAC treatment, low level of MC
detected in water.
Microcystis AC Rapid mix: > 99 –  All cells removed without damage to membrane (Sun et al.,
aeruginosa 250 rpm 1 min integrity under optimum coagulant dosage (15 mg/L 2012)
Slow mix: AlCl3).
20 rpm 20 min  When flocs were stacked over 6 days, cells lysed and
Settling time: microcystin-LR concentration increased (nearly
30 min 41.7%).
Microcystis AS Rapid mix: – –  Small dosage (7 mg/L as Al) of alum treatment not (Han et al.,
ichthyoblabe 300 rpm 1 min effective in precipitating Microcystis cells, and large 2012)
Slow mix: dosage (14 mg/L as Al) of alum treatment caused cell
50 rpm 5 min damage and released toxins over 80% of initial
Settling time: intracellular.
–
Anabaena AS Rapid mix: 88 –  No significant turbidity occurred after dosage of 65 (Hoko &
circinalis and 200 rpm 1 min mg/l, and no further significant algae removal Makado,
Microcystis Slow mix: achieved after dosage of 90 mg/l. 2011)
aeruginosa 20 rpm 15 min  Algae removal by coagulation found to be higher at
Settling time: lower pH (pH 7).
30 min
Chlorella CTS Rapid mix: 99.03 ± 0.7% –  Optimal chitosan dosage for destabilizing algae cells (Ahmad
spirulina 150 rpm 2 min was 10 ppm with of 99.3 ± 0.7% removal efficiency. et al., 2011)
Settling time:  Concentrations higher than 10 ppm restabilized
20 min microalgae cultures, thus reducing process efficiency.
Microcystis AS, PACl, Rapid mix: – –  Dosage of 0.04 mM, PACl (90%) achieved better (Jiang &
aeruginosa Bent, Na- 400 rpm 1 min performance than AS (75%) with respect to algal Kim, 2008)
Bent, Mont- Slow mix: removal.
KSF, and 35 rpm 20 min  Dosage of 200 mg/L, 100% chlorophyll-a removal
Mont-K10 Settling time: obtained by Bent and Mont-KSF clays.
60 min
 Percent Removal
Cyanobacteria Coagulant Mechanical Action Comments References
Intracellular Extracellular
Microcystis AS Rapid mix: – –  All cells removed without damage to membrane (Chow
aeruginosa 20 rpm 1 min integrity; thus, aluminum sulphate (5.3 mg/l as Al at et al., 1999)
Slow mix: pH 6.7) and mechanical action did not cause any
25 rpm 14 min damage to cultured M. aeruginosa cells.
Settling time:
15 min
Microcystis Ferric – – –  Cultured M. aeruginosa and A. circinalis cells not (Chow
aeruginosa Chloride damaged by ferric chloride treatment; however, ferric et al., 1998)
chloride seemed to stimulate growth of both M.
aeruginosa and A. circinalis.
AC: aluminum chloride; AS: aluminum sulphate; Bent: bentonite; CTS: chitosan; CTSAC: chitosan-aluminum chloride; Mont-K10: montmorillonite; K10Mont-KSF:
montmorillonite KSF; Na-Bent: sodium modified bentonite PAC: polyaluminum chloride; PACI; polyaluminum chloride
212 Ayşe Büşra Şengül

Many studies have investigated the effect of the coagulation/ flocculation process on
cyanobacterial cells and cyanotoxin release during water treatment. Results of several
investigations into the efficiency of this process to remove cyanobacterial
cells have suggested that coagulation is more effective for the removal of
intracellular toxins than extracellular toxins, due to their hydrophilicity. Cyanobacterial
cells generally possess a negative surface charge at most pH levels, and many
common species are less than 10 mm in diameter; therefore, surface charge
can be neutralized by introducing coagulants. These coagulants can easily
flocculate algal cells and form flocs (Henderson et al., 2010; Dong et al., 2014; Fast et al.,
2014).
Nevertheless, effects of the coagulation/flocculation process on cyanobacterial
cell lysis in the literature are debatable. For example, Chow et al. (1998; 1999) evaluated
the effect of coagulation on cells of Microcystis aeruginosa and Anabaena circinalis, and
reported that chemicals and mechanical action did not damage cyanobacterial cells and
the release of cell metabolites. Sun et al. (2012) also reported that coagulant
dose and shear under optimum conditions did not cause the lysis of cells and ensuing
release of microcystin-LR.
Furthermore, Pei et al. (2014), Sun et al. (2013), and Li et al. (2015) documented that
the optimized coagulants as well as mechanical conditions did not damage the
membrane of the cultured M. aeruginosa cells. However, Han et al. (2012; 2013) found
that alum treatment with the maximum dosage and a long contact time caused
Microcystis cell damage, and as a result, large amounts of toxic microcystin were
released into the water.
Table 2 summarizes the effectiveness of different coagulation/ flocculation
conditions in removing intracellular and extracellular toxins of several of the most
important cyanobacteria.

CONCLUSION

This study thoroughly evaluated the performance of the coagulation/flocculation

process using aluminum sulphate, ferric chloride, and ferrous sulphate coagulants for the
removal of both microcystins (intracellular and extracellular) and turbidity. The effects of
coagulant dosage, pH, and polyelectrolyte dosage on intracellular and extracellular
microcystins and turbidity removal were also investigated. The following conclusions can
be drawn.

 The highest intracellular microcystin removal was achieved using the aluminum
sulphate (65.2%) coagulant in comparison with ferric chloride (56.3%) and
ferrous sulphate (30.9%) under the determined optimum conditions.
 An Evaluation of the Performance of the Coagulation/Flocculation … 213

 The highest turbidity removal was found to be 85.1%, 77%, and 59.8% for
aluminum sulphate, ferric chloride, and ferrous sulphate, respectively.
 The pH and addition of polyelectrolyte had less effect on microcystins and
turbidity removal.
 The concentration of turbidity decreased to below 5 NTU, which is the maximum
turbidity level in drinking water determined by the WHO when the
coagulation/flocculation process is applied.
 The total microcystin concentration (4.09 µg/L) in the water remained above the
WHO guideline value of 1 µg/L for drinking water, but the release of toxins in
the water was not observed after the coagulation/flocculation process.
 Overall, results indicate that both intracellular microcystin and turbidity could be
removed through the coagulation/flocculation process, whereas extracellular
microcystin is still of particular concern in drinking water treatment due to its
difficult removal.

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In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 12

TREATMENT OF LEACHATE FROM THE OULED

BERJAL LANDFILL IN MOROCCO BY
COAGULATION FLOCCULATION:
A STUDY OF THE EFFECT OF ORDER FOR
REAGENT INTRODUCTION

Hajar Bakraouy1,*, Salah Souabi1, Khalid Digua1

and Levent Yilmaz2
1
Process and Environmental Engineering Laboratory,
Faculty of Science and Technics,
Hassan II University, Mohammedia, Morocco
2
Department of Civil Engineering, University of Nişantaşı, Turkey

ABSTRACT

In Morocco, like all the countries of the world, socio-economic activities coupled
with population growth and changes in consumption patterns generate a significant
production of municipal solid waste. This is accompanied by a significant increase in
leachates produced during the fermentation of waste. These leachates have a considerable
impact on the environment. Moroccan regulations require leachate treatment to reduce
environmental impacts. Several techniques are currently used for the decontamination of
leachate such as (coagulation flocculation, biological treatment, filtration, oxidation, ...).

*
Corresponding Author Email: [email protected].
218 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Indeed, the majority of the physicochemical treatments used for the purification of
the leachates intervenes as pretreatment or finishing stage to complete the treatment
chain, or to eliminate a specific pollutant.
The addition of a polymer as a flocculant improves the effectiveness of coagulation
by inorganic coagulants. In fact, these flocculants make it possible to obtain large flocs,
which settle rapidly and which resist shear forces (Bratby, 2006).
According to Bratby (2006), when a liquid is already destabilized after the addition
of an inorganic coagulant, the polymers increase the rate of orthokinetic flocculation by
the formation of larger flocs. This is done through a sufficient absorption affinity between
the polymers and the flocs.
The choice of polymers must be made judiciously because their effectiveness
depends on several parameters, namely: the nature of the polymer, the pH, the
temperature, the type of inorganic coagulant, the nature and size of the pollutant to be
eliminated, the concentration of the coagulant and flocculant (Zemaitaitiene et al., 2003,
Bolto and Gregory, 2007).
The first part of this work concerns the coagulation flocculation of young leachate
from the Kenitra city landfill. The tests were carried out by adding ferric chloride mixed
with three flocculants, namely: the chitosan, the Superfloc SD2065 and the Himoloc.
The dose of coagulant was fixed at 6 g/L (determined from preliminary tests), while
the doses of flocculants ranged from 3,3 to 20 mL/L. The evaluation of treatment
efficiency as well as the optimal doses of the flocculants was determined by the
monitoring of the removal of turbidity, COD, phenol, color, Absorbance at 254 nm and
the volume of sludge collected after 24 hours.
The purpose of the second part reveals the effect of the order of introduction of the
reagents on the efficiency of leachate treatment by coagulation flocculation.
The reagents used are ferric chloride and the three flocculants previously mentioned.
We varied the order of introduction by adding:

 Ferric chloride first followed by flocculant.

 The flocculant first followed by the coagulant;
 The coagulant and the flocculant at the same time.

The effect of the order of introduction of the reagents was evaluated in terms of
elimination of turbidity, COD, color, Abs at 254 nm, phenol, detergents, ammonium ions,
nitrite, nitrate and total phosphorus.

Keywords: coagulant, flocculant, ferric chloride, order of introduction, COD, turbidity,

nitrates, ammonium ions, nitrites

INTRODUCTION

Solid waste management remains an issue for countries around the world. In
Morocco, the improvement of the standard of living, as well as the change in production
and consumption patterns have led to an increase in the volume of solid waste, estimated
at 6.9 million tonnes/year according to the third report on the state of of Morocco’s
environment, published in 2015. The quantity produced is of the order of 0.76 kilos per
day per inhabitant in urban areas and 0.3 kilograms per day per inhabitant in rural areas.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 219

This increase affects the quality of the collection and generates a saturation of
landfills. They become a source of contamination for soil, air and human health, instead
of being the ultimate solution for getting rid of waste.
Landfills are sources of methane, which is a greenhouse gas, and toxic liquid
effluents called “leachate” (Renou et al., 2008). The design of a landfill must ensure the
capture of biogas for energy recovery, and the drainage and treatment of leachates before
their release into the natural environment.
In this context, Morocco is part of a proactive approach to waste management, whose
main objective is to improve waste collection, sorting and treatment conditions while
limiting the nuisance caused by methane emissions and leachate production.
The genesis of leachates is the result of a very complex process. Indeed, as soon as
they are deposited, the waste is subject to biodegradation processes that are the result of a
multitude of biological and physicochemical reactions. The water that infiltrates the
layers of waste is the main vector of this biodegradation. Thus, leachate results from the
solubilization of compounds during the percolation of rainwater through the pile of
waste, as well as moisture contained in the waste itself (Souabi et al., 2010). The
composition of leachate depends on several parameters including: The age of the landfill,
the type of waste landfilled and the technique used for landfilling (Chofqi et al., 2007).
The challenge is now significant because it will be necessary to ensure effective
drainage of leachates and their treatment for reuse or rejection in the receiving
environment. This approach will provide a significant water resource for use, particularly
in areas experiencing water shortages and drought.
Several treatment techniques are implemented to reduce and eliminate the pollution
contained in leachates. These techniques are physicochemical, biological or a
combination of both.
Among these methods, we distinguish the technique of coagulation flocculation.
Several researchers have used this process for the pre or post treatment of landfill
leachate, to improve its biodegradability or to eliminate refractory organic matter (Aziz et
al., 2007).
The goal is to eliminate many types of pollutants, namely: COD, BOD5, NH4+,
Phosphorus, color, humic and fulvic compounds, phenol ... (Maranon et al., 2008,
Poznyak et al., 2008; al., 2014).
In a study conducted by Zheng et al. (2009), coagulation flocculation using
Aluminum chloride shown to be an effective treatment for the removal of phthalic acid
esters. The removal of these pollutants reached 32% and 50% respectively for young and
intermediate leachates.
Li et al. (2010) compared the effect of four coagulants, namely: Alumina sulphate
(Al2(SO4)3), ferric chloride (FeCl3), Aluminum polychloride and polyferric Sulphate, for
the treatment of stabilized leachate. At the end of this study, Polyferric Sulfate gave the
220 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

best results by eliminating 93% of suspended solids, 97% of turbidity, 70% of COD and
74% of toxicity. The minimum volume of sludge produced was 32 mL/L.
Abood et al. (2014) studied the effectiveness of coagulation flocculation as a
pretreatment of stabilized leachate from the Chang Shankou landfill in China. They were
able to remove 70,6% of the COD, 49,4% of the BOD5 and 26,6% of NH3-N by adding
1,2 g/L of polyferric Sulphate and 1 mL of 0,1% polyacrilamide flocculant at an optimal
pH of 5.
The objective of this chapter is to determine the type and the dose of the flocculant
suitable for the leachate treatment of Ouled Berjal landfill (Situated at Kenitra City in
Morocco), and to study the effect of the order of introduction of the coagulant and the
flocculant on the effectiveness of coagulation flocculation.

COAGULATION FLOCCULATION PROCESS

Principle of the Process

The goal of coagulation flocculation is to remove suspended solids (SS) and colloids
contained in a liquid (Adamczyk, Z. 2003), which are collected as flocs and removed by
settling or flotation.
This process takes place in two stages:

Coagulation or Destabilization of Particles

Colloids are negatively charged, so they repel each other. They are subject to two
types of forces: Van Der Waals gravitational force and the zeta potential electrostatic
repulsion force. The addition of the coagulant provides multivalent cations that approach
the surface of the colloids and adsorb. The zeta potential becomes zero, thus neutralizing
all nearby charges. This requires a homogeneous distribution of the coagulant by strong
agitation in a short time before the precipitation of the hydroxides formed.

Flocculation or Formation of Flocs

This step consists in the agglomeration of the destabilized particles in large
aggregates, which tend to decant by partition by the hydroxides resulting from the
hydrolysis of the coagulant or by the effect of the cationic polyelectrolytes added to the
medium.
It is then necessary to ensure a slow stirring in a sufficiently high time to favor the
collision of the destabilized particles and thus their aggregation.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 221

Coagulants Used

There are three types of coagulants: Iron or aluminum-based metals Salts, organic
coagulants and bio-coagulants (Renou el al., 2008).

Mineral Coagulants
In general, metal salts based on iron or aluminum are the most commonly used.
Among these salts are: Ferric chloride (FeCl3), Ferric sulphate (Fe2(SO4)3) and ferrous
sulphate (FeSO4), Aluminum sulphate (Al2(SO4)3), Aluminum chloride (AlCl3) and
Sodium Aluinate (NaAlO2) (Maranon et al., 2008, Liu et al., 2012). The effectiveness of
these salts is due to the hydroxides (resulting from their hydrolysis in the medium) whose
solubility is strongly related to pH.

Figure 1. Coagulation diagram of iron (Amirtharajah, 1988).

Figure 1 shows the coagulation diagrams of iron as a function of pH. We notice that
the iron coagulation diagram is divided into three zones:

 The first zone corresponds to the flocculation zone by neutralization,

destabilization, adsorption and precipitation of colloids. In this pH range, the
 cationic species of iron predominate and the coagulation flocculation is optimal.
 The second zone corresponds to the coagulation flocculation of
the colloid carried out by trapping in poorly soluble iron hydroxides
(Fe(OH)3(s)). The colloids are then removed, but less effectively than in the
previous zone.
222 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

 The third zone is that in which the surface of the colloids is modified until the
zeta potential of the particles is reversed, which can lead to the
stabilization of these colloids or the destabilization of the formed flocs.
Flocculation coagulation is no longer effective in this range of pH and iron
concentration.

Organic Coagulants
The use of organic coagulants is more effective when the waters to be treated are
composed of fine suspended matter. They are synthetic cationic polyelectrolytes such as:
melamine-formaldehyde, epichlorohydrin DimethylAmine (EPI-DMA) of chemical
formula C5H12ONCl, and PolyDiAllylDimethylAmmoniumChloride (POLYDADMAC)
of chemical formula C8H16NCl.
According to Mouchet (2000), these coagulants have the advantage of not modifying
the pH of the medium, reducing the volume of sludge produced and also reducing the
dose of reagent necessary for optimum coagulation (less reagent is added).

Bio-Coagulants
Although metal salts are the most used, their negative effect on the environment and
human health is undeniable. As a result, the orientation towards the use of coagulants of
natural origin has become an extreme necessity (Chaibakhsh et al., 2014).
These types of coagulants are known by their biodegradability, low cost and
especially their non-toxicity to humans and the environment. They can be used alone or
mixed with other conventional coagulants (Nwaiwu and Bello, 2011, Awad et al., 2013).
Patel and Vashi (2012) conducted a comparative study of three types of
biocoagulants: Moringa oleifera seeds, corn seeds and chitosan for the treatment of
wastewater from the textile industry. The best results in terms of removal of COD and
color were obtained by the first bio-coagulant, whit respective removal efficiencies of
74,1% 58,5%.
Moreover, Rasool et al. (2016) showed the effectiveness of basil seeds (dried,
crushed and then sieved) mixed with Alumina Sulphate and Aluminum Polychloride for
the pretreatment of landfill leachate. The optimal conditions obtained by the
experimental design methodology are: 15min contact time, neutral pH, and 1:1 alumina
sulphate: Basil ratio. Under these conditions, 64.4% of the COD and 77.8% of the color
were eliminated.

Floculants Used

In the field of water treatment, the addition of a polymer such as a flocculant

improves the effectiveness of coagulation by inorganic coagulants. In fact, these
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 223

flocculants make it possible to obtain large flocs, which settle rapidly and which resist
shear forces (Bratby, 2006).
According to this author, when a medium is already destabilized following the
addition of an inorganic coagulant, the polymers increase the rate of orthokinetic
flocculation by the formation of larger flocs. This is done through a sufficient absorption
affinity between the polymers and the flocs.
The choice of polymers must be made judiciously because their effectiveness
depends on several parameters, namely: the nature of the polymer, the pH, the
temperature, the type of inorganic coagulant, the nature and the size of the pollutant to be
eliminated (Zemaitaitiene et al., 2003, Bolto and Gregory, 2007).
These flocculants may be inorganic or organic. For mineral flocculants, activated
silica is the most used.
Organic flocculants can be of natural origin such as alginates which are extracted
from marine algae (Degrémont, 2005) or synthetic such as polyelectrolytes. These are
either anionic, cationic or nonionic.
They are used to improve the elimination of organic matter (Tatsi et al., 2003) and
also promote the agglomeration of flocs and improve their settling (Renou et al., 2008).
Solberg and Wagberg (2003) have shown that the removal efficiency of a pollutant
depends on the type of polymer, its surface charge density, the concentration and
solubility of the polymer, the chemical affinity of the polymer in thr surface, ionic
strength....
Nowadays, the use of biopolymers is very important because of their ecological
nature. According to Renault et al. (2009), there is no risk of secondary contamination
because the biopolymers do not leave residual metals either in the treated water or in the
sludge produced.

MATERIALS AND METHODS

Sanitary Landfill Leachate Sample

In this study, leachate was sampled from Ouled Berjal landfill located in Kenitra city
in Morocco.
Kenitra is a town of 76Km² located 45Km north of Rabat. It is one of the most
important industrial centers in Morocco (the fourth largest industrial city in the country).
The climate in this city is sub humid to semi-arid with an average rainfall of about
600 mm. According to the urban commune of Kenitra, the population reached in 2014
nearly 1,061,435 habitants.
The rapid evolution of the city of Kenitra is accompanied by increased production of
solid waste, which quantity was estimated at 120,000 tons in 2011.
224 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Table 1. Physical composition of household waste at Kenitra city

(Aouane et al., 2010)

Composants Mean (%)

Organic waste 70
Plastic 12
Paper cardboard 10
Class 2
Metals 4
Various (wood, etc.) 2

The physical composition of household waste at the Kenitra landfill is reported in

Table 1.
The collection and cleaning service for this waste was delegated to two professional
operators: SEGEDEMA and TECMED. While landfilling and treatment at the common
Kenitra landfill has been delegated to SOS.
Figure 2 shows illustrates ouled berjal landfill.

Figure 2. Ouled Berjal landfill.

Leachate is stored in four basins noted B1, B2, B3 and B4 (Figure 3). The samples
were collected in 30L plastic bottles, and then they were transported to the laboratory,
stored at 4°C and subsequently characterized.
The Ouled Berjal landfill covers an area of 20 ha and receives an average of 329 tons
per day of waste. The landfill has:

 Landfill area consisting of a 4 hectare bin which is subdivided into 4 cells;

 Drainage of leachate and rainwater;
 Four large basins to collect leachate.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 225

Figure 3. Leachate storage basins at Ouled Berjal landfill.

2.2. Analytical Methods

The leachates collected were analyzed for many parameters, namely: pH,
conductivity, turbidity, Absorbance at 254nm, color, Chemical Oxygen Demand (COD),
Biochemical Oxygen Demand (BOD5), NH4+, NO3-, phenol, total phosphorus, surfactants,
suspended matter.
Turbidity was measured using a turbidimeter Model 2100N HACH, color and
absorbance at 254 nm with a UV/Visible spectrophotometer Model 9200 and pH by a pH
meter model 6209. Conductivity was measured using a Conductivity pH meter model
YK-2001PH.
The suspended matter was determined by the centrifugation method.
The chemical oxygen demand (COD) was determined using Open Reflux Method
(5220-B). The BOD5 is analyzed according to the manometric method using the
WARBURG respirometer.
For the analysis of surfactants: The anionic surfactants form, with methyl violet, a
complex soluble in toluene and which can be determined by the colorimetric method. The
absorbance of the residue recovered after extraction is spectrophotometrically read at a
wavelength of 615 nm.
The ammonium ions were assayed according to the method stated by Rodier in 1996,
which consists in the treatment of these ions with a solution of sodium nitroprusside. The
spectrophotometric reading is then carried out at a wavelength of 630 nm.
Determination of nitrates was carried out in accordance with the NF EN ISO 78-90
January 1997 (T 90-045) method.
226 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

The total phosphorus was determined according to the NF T 90-023 January 1997
method.
Phenol was measured using the Folin-Ciocalteu reagent method (Macheix et al.,
1990). This reagent develops, in the presence of phenolic compounds, a blue color whose
intensity is read by a spectrophotometer at a wavelength of 725 nm.

Reagents Used

Ferric Chloride
Table 2 shows the physicochemical characteristics of the FeCl3 coagulant.

Polyacrylamide Himoloc
Himoloc DR3000 flocculant is a cationic polyacrylamide of medium molecular
weight, specially adapted for the physico-chemical treatment of wastewater because it
contains a special molecule that facilitates the separation of the colloidal material by
flotation.
This flocculant is soluble in any type of wastewater, regardless of its quality.
The table below lists the main characteristics of the Himoloc DR3000 flocculant.

Chitosan
Chitosan is a substance not widespread in nature. It exists only in the cell walls of
certain fungal microorganisms such as zygomycetes fungi and in mycelium.
Figure 4 shows the chemical structure of chitosan.
The particular properties of this biopolymer are related to the presence of the amine
function carried by the carbon 2. These properties depend strongly on the pH:
At acidic pHs, chitosan carries several positive charges and can bind to negatively
charged molecules. It is considered a good coagulant or flocculant (Fang et al., 2001);
At pH’s exceeding neutral, chitosan loses its positive charges while releasing the
electronic doublets of nitrogen. Thus, chitosan is considered an excellent complexing
agent, particularly heavy metals (Yen et al., 2009).

Table 2. Characteristics of coagulant

Parameter Value
FeCl3 (% by mass) 39,0 – 41,0
Fe3+ (% by mass) 13,4 – 14,2
Density at 20°C (kg/dm3) 1,400 – 1,440
pH at 20°C <1
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 227

Table 3. Caractéristiques du floculant Himoloc DR3000

Parameter Value
Density (g/cm3) 1,2
Viscosity (cP) < 600
pH 3,0 – 4,1

Figure 4. Chemical structure of chitosan.

In this study, we prepared a chitosan solution as follows:

 Weigh 3g of chitosan powder;

 Add 1 mL of Acetic Acid;
 Make up to 1L with distilled water.

The solution must be thoroughly shaken before each use.

Flocculant Superfloc SD2065

The Superfloc SD2065 flocculant produced by Kemire Company is a cationic
polyacrylamide in the form of an emulsion. It is ideal for wastewater treatment and water
clarification in all industries.
The characteristics of this flocculant are summarized in Table 4.

Table 4. Characteristics of the flocculant Superfloc SD2065

Parameter Value
Density (g/cm3) 1,01 - 1,05
Viscosity (cP) 270

RESULTS AND DISCUSSION

Monitoring of Leachate Flow over Time

In order to estimate the volume of leachate produced by the Ouled Berjal

landfill, a monthly monitoring of the collected leachate flow was performed as shown in
Figure 5.
228 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

The leachate flow rate was measured using a submersible pump with an hourly flow
rate of 20 m3/h and operating automatically with floats. We were able to determine the
daily flow rate of the leachate collected by a daily recording of the operating time of the
pump.
The monthly flow of the leachate admits a maximum value of 2900 m3/month and a
minimum value of 640 m3/month. Indeed, maximum flows are presented during the
months of January, February and March, which coincides with the rainy season of the
year. During this period, stormwater increases the volume of the leachate produced,
causing overflows in the storage basins.

Figure 5. Variation of the monthly and daily flow of leachate produced in Ouled
Berjal landfill.

While the months of July and August present the minimum flows, as it is the summer
period known by relatively high temperatures, leading to the evaporation of a large part
of the stored leachates.

Characteristics of Ouled Berjal Landfill Leachate

The characterization of leachate aims to evaluate the risk that the landfill may
represent to the environment.
We carried out 3 sampling campaigns during the months of February, March (Rainy
season) and October (End of summer) 2015.
Figures 6, 7 and 8 report the results of the characterization.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 229

Variation of pH
The leachates stored in the different basins B1, B2 et B3 have a basic pH varying
between 8 and 9. The pH values are strongly related to the existence of the volatile
organic compounds in the leachate. Indeed, during the first phase of waste fermentation
(Acidogenesis phase), leachates contain high levels of volatile compounds, thus inducing
low values of pH, which are generally less than 4. Over time, the landfill ages and the
leachates are depleted of volatile organic compounds, resulting in increased pH values up
to 7 or higher (Kjeldsen et al., 2002).
During the decomposition, waste is subject to chemical and biological reactions
leading to the production of ammonium ions and carbonic acid. The dissociation of the
latter results in the formation of hydrogen cations and bicarbonate anions which have a
direct effect on the pH of the percolation water or leachate. Thus, the pH of the leachate
is strongly influenced by the nature of the dissolved materials existing in the medium. An
acidic pH typically characterizes a young leachate containing a large organic charge
(Naveen et al., 2017). Moreover, an alkaline pH is an indicator of leachate maturity,
which is found to be depleted in organic matter and rich in refractory to biodegradation,
such as humic substances (Renou et al., 2008).

Variation of COD and BOD5

The maximum values recorded for COD and BOD5 were 5877,6 and 4800 mg/L
respectively, while the minimum values were 1152 and 800 mg/L for the same
parameters. These results are close to those reported by Silva et al. (2013).
It should be noted that the COD and the BOD5 of the leachate contained in B1 are
greater than those of the basins B2 and B3. This can be explained by the fact that the
basin B1 collects the young leachates from the landfilling zone, which are mixed with the
leachate already present in the same basin.
We note also that both parameters reach low values during the summer season, which
can be explained by the degradation of organic matter by microorganisms, which
intensify at high temperatures (Khattabi et al., 2002). Moreover, during the winter season,
the temperature of the leachate decreases and partially inhibits bacterial activity.
The BOD5/COD ratio determines the age of the leachate (Young, intermediate or
stabilized) and its biodegradability. According to Renou et al. (2008), BOD5/COD ratios
greater than 0,3 characterize young biodegradable leachate, while aged or stabilized
leachate has a ratio of less than 0,1. These leachates contains low organic load composed
mainly of humic and fulvic compounds.
Millot (1986) showed that an old leachate is rich in humic and fulvic substances and
has a pH above 7,5, a COD < 4000 mg/L and a BOD5/COD ratio < 0,1. The author
concludes that the leachate analyzed is not very biodegradable.
 Table 5. Results of physicochemical analyzes of leachates from basins B1, B2 and B3
(average of three campaigns)

BOD5 (mg/L)
Conductivity

COD (mg/L)

NH4+ (mg/L)
phosphorus

NO3-(mg/L)
Surfactant
Turbidity

(FD=100)
(mS/cm)

SS (g/L)
(FD=20)

(mg/L)

(mg/L)

(mg/L)
Phenol
254nm
Abs at
(NTU)

Color

Total
pH
Basin

B1 8,4 22,1 114,5 0,272 0,300 0,7 4519,2 992,5 1126,6 24,5 24,1 626,4 374
B2 8,7 20,5 73,6 0,292 0,340 0,8 4412,8 895,0 1122,4 46,0 14,0 393,8 382
B3 8,7 22,2 72,7 0,324 0,308 1,0 4372,2 899,3 1109,2 68,1 16,7 230,3 334

Figure 6. Variation of COD and BOD5 contained in leachates from basins B1, B2 et B3 during the three campaigns.
Figure 7. Variation of pH, conductivity, turbidity and SS contained in leachates from basins B1, B2 et B3 during the three campaigns.
Figure 8. Variation of color, Abs at 254nm, phenol and surfactant contained in leachates from basins B1, B2 et B3 during the
three campaigns.
Figure 9. Variation of phosphorus, NH4+ and NO3- contained in leachates from basins B1, B2 et B3 during the three campaigns.
234 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

In our case, the BOD5/COD ratio ranges from 0,14 to 0,3, indicating that the
leachates in the three basins are intermediate leachates with mean biodegradability.
However, it is possible to envisage a biological treatment for these effluents (Renou et
al., 2008).

Variation of Turbidity, SS and Color

Table 6. Physicochemical characterization of leachates from different landfills

Landfills
Parameter
(Morocco)

(Morocco)

(Morocco)

(Morocco)

(Algeria)5
ElKerma
El jadida

(China)6
Beijing
Agadir
Rabat

Fez
1

4
pH 8,4 8,84 7,11 4,3 8,19 8
COD (mg/L) 14 900 920 53 999,6 72 000 19 333 8528
BOD5 (mg/L) 8700 55,33 20 000 44 000 3301 5669
NH4+ (mg/L) 3740 114 2,41 0,10 2726 1154
NO3- (mg/L) - 2,37 11,3 62 0,92 2,21
Total Phosphorus (mg/L) 49,73 - - 188,4 0,60 5,2
Turbidity (NTU) 1619 - - - 732 -
Conductivity (mS/cm) 22,8 27,18 - 14,7 120,3 -
1
Bakraouy et al. (2016)
2
Chofqi et al. (2004)
3
Ezzoubi et al. (2010)
4
Jirou et al. (2014)
5
Bennama et al. (2010)
6
Zhu et al. (2013)

Generally, in liquids with a pH between 5 and 9, the suspended particles carry a

negative charge on the surface of the liquid and are of a size ranging between 0,01 and 1
μm (Duan et al. 2003, Verma et al., 2010).
It is known that the presence of the suspended matter causes an increase in the
turbidity of any liquid. For the three basins, SS ranges from 0,2 to 2,54 g/L, while
turbidity ranges from 67 to 186 NTU. These values are lower than those reported by
Mahmud et al. (2012).
Concerning color, it follows the same variation undertaken by the COD. Indeed, the
color intensifies during the winter season and becomes weaker during the summer
season. This can be explained by the fact that the color is governed by the organic matter
contained in the leachate (Aziz et al., 2007, Assou et al., 2014).

Variation of Conductivity
The conductivity values obtained ranged from 38 to 13,5 mS/cm, with an average of
22 mS/cm. It approaches the values recorded by Bakraouy et al. (2016) which is 22,8
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 235

mS/cm, Muller et al. (2015) which is 30 mS/cm, as well as that of Chofqi et al. (2007)
which is 26,5 mS/cm. we notice that these values are quite high, showing that the
leachates of the three basins are highly mineralized. Moreover, the conductivity reaches
low values during the winter season, which can be explained by the dilution of the
leachate by stormwater.
While during the summer season, the conductivity values are high, which can be
related to the bacterial activity that is favored by the increase in temperature. This
increase stimulates enzymatic reactions, thereby releasing high amounts of mineral
elements such as chlorides. Khattabi et al. (2002) demonstrated that conductivity is
closely related to the content of chloride ions.

Variation of Phenol
Phenol usually comes from resins, paints, plastics, sand molding foundries. In
addition, phenol is the most important benzene derivative, after styrene. Ozonation,
chlorination, coagulation-flocculation, or adsorption on activated carbon (Aravindhan et
al., 2009) have been developed for its elimination due to its toxicity. It is considered to be
an inhibitor of biomass in biological treatments.
We note that the concentration of phenol admits a maximum value of 1258 mg/L and
a minimum value of 945,8 mg/L. These values are higher than those reported by Silva et
al. (2013), which reaches 162 mg/L, as well as that recorded by Bannama et al. (2010),
which reaches 37 mg/L.
It should be noted that the concentration of phenol increases over time, since
anaerobic storage promotes the degradation of phenolic derivatives while producing
phenol.

Variation of Surfactants
For the three basins, the analysis of surfactants revealed moderate values ranging
from 18 to 77,6 mg/L.
According to Wang et al. (2004), surfactants come from various domestic, urban and
industrial activities (production of detergents and fertilizers, pharmaceutical and
petrochemical industries, etc.). They are major inhibitors of the development of biomass
promoter of biodegradability. Borghi et al. (2011) explained that when the chain length of
the alkyl group increases, the hydrophobicity of the molecule increases also, inducing
high toxicity. On the other hand, a high number of ethylene oxide units reduces the
hydrophobicity of the molecule and induces a decrease in toxicity.

Variation of Total Phosphorus

Phosphorus usually comes from detergents (used by households as well as industrial
activities), fertilizers, etc. When its concentration in surface waters is high, phosphorus-
dependent organisms (such as plants and algae) grow excessively to the detriment of
236 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

other organisms, causing the imbalance of the aquatic ecosystem. This phenomenon is
called: Eutrophication. The elimination of phosphorus is therefore one of the major
environmental issues.
The values obtained during the sampling campaigns ranged from 9,3 to 34,4 mg/L.
These values exceed those reported by Silva et al. (2013), with an average of 19,1 mg/L,
and by Zhu et al. (2013), with an average of 5,2 mg/ L, but still much lower than that of
the Agadir landfill, which reached 188,4 mg/L (Jirou et al., 2014).

Variation of Absorbance at 254nm

The absorbance at 254 nm (Abs 254 nm) characterizes the conjugated compounds
(aromatic nuclei) and is generally used for the determination of organic matter (Chin et
al., 1994). It is considered as an additional parameter indicating the quality of most
wastewater (Rivas et al., 2003). This parameter also informs us about the aromaticity and
the hydrophobicity of the molecules contained in a sample by means of the SUVA index
which is given by the ratio of the absorbance at 254 nm/COD (Liu et al., 2015).
In the case of the three basins, this absorbance admits a maximum value of 0,434 and
a minimum value of 0,237 (at a dilution factor of 100 times).

Variation of Ammonium Ions

Ammonium ions (NH4+) originate essentially from the biodegradation of dead
organic matter. It is generally rapidly reabsorbed by aquatic organisms, but it may be
present in high concentrations in polluted and low oxygenated waters,
Biological treatments are the most used for the removal of ammonium ions. Indeed,
this is achieved by the processes of nitrification and denitrification, at the end of which
the nitrogen is converted into nitrates and then nitrogen gas.
Nitrification corresponds to the oxidation of ammonium ions to nitrites and nitrates in
aerobic conditions.
It is carried out in two stages:

Nitritation reaction (The microorganisms involved are of the genus Nitrosomonas)

3
NH4+ + O2 → NO−
2 + H2 O + 2H
+
2

Nitratation reaction (The microorganisms involved are of the genus Nitrobacter)

1
NO− −
2 + O2 → NO3
2
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 237

Moreover, the elimination of these two elements (nitrates and nitrites) is commonly
called denitrification, it is carried out under anoxic conditions. This reduction in nitrogen
gas is governed by the reaction below:

NO− −
3 → NO2 → NO → N2 O → N2

High concentrations of ammonium ions are typically characteristic of aged or

stabilized leachate, while lower concentrations are characteristic of young leachates
(Christensen and Kjeldsen, 1989, Kjeldsen et al., 2002).
Concentrations found during our study range from 9,33 mg/L to 1421,5 mg/L. It
should be noted that ammonium ions concentration is higher in the winter season, which
is mainly due to the cessation of the nitrification process, which is strongly related to
temperature. Indeed, the increase in temperature leads to better nitrification, while the
decrease in temperature to values below 10°C, leads to a drop in nitrification rate,
estimated at half by Rim and Al. (1997).

Variation of Nitrates
Nitrates result from the oxidation of nitrogen, they represent the most common
nitrogenous form in natural waters. According to Rodier et al. (2009), nitrates represent a
major part of mineral nitrogen, which together with organic nitrogen constitute the total
nitrogen. Toxicity of nitrates has been widely studied by Karine et al. (2003).
Indeed, nitrates alter the quality of water as well as human health. On the one hand,
the reduction of nitrates to nitrites favors the formation of methemoglobin which causes
the oxidation of iron II to iron III, which makes it incapable of transporting oxygen. On
the other hand, nitrates contribute to the endogenous synthesis of N-nitroso compounds
which are strongly linked to the risk of cancerous diseases.
Nitrate levels in the three basins ranged from 172 to 592 mg/L. These values remain
below the maximum value determined by Souabi et al. (2011) for leachates from the city
of mohammedia in Morocco, which reaches 845 mg/L.
In a research conducted by Tahiri et al. (2014), the characterization of the leachate
from the city of Meknes reveals a concentration of nitrates about 751,1 mg/L, which
exceeds that revealed by the present study. On the other hand, the nitrate concentration is
lower in the leachate of the city of El jadida, 2,4 mg/L (Chofqi et al., 2007), of Fez
citywhich reaches 11,3 mg/L (Ezzoubi et al., 2010) and of Oran city which reaches 0,92
mg/L (Bennama et al., 2010).
Throughout Figure 3, we notice that the nitrate concentration is higher in the summer
season, due to the nitrification of ammonium ions and their conversion to nitrates. The
latter are accumulated in the leachate, since the denitrification is carried out under
optimum temperature conditions ranging from 35 to 50°C.
238 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Characteristics of Leachate Used in this Study

Table 7 reports the physicochemical characteristics of the leachate used in this study.
The leachate studied has a blackish color, a fecal odor and an average turbidity of
180 NTU. Organic load expressed in terms of COD reaches 21 g/L, which shows that this
leachate is young and biodegradable. This concentration is higher than that reported by
Chofqi et al. (2007), which was 990 mg/L for the leachate from the El Jadida landfill.
Furthermore, we note high concentrations of nitrogen pollutants and moderate phenol
content (96,2 mg/L) and surfactants (47,1 mg/L). These concentrations are much lower
than those detected by Bakraouy et al. (2016) for the Rabat landfill, being 241,8 mg/L for
phenol and 58 mg/L for surfactants.
The metal charge contained in the leachate from the Ouled Berjal landfill is typical of
a domestic landfill since the concentration of metallic elements is similar to that detected
in other household landfills. According to Christensen et al. (2001), heavy metals
composition of a household landfill is as follows:

 The concentration of Zinc varies from 30 to 106 μg/L;

 The concentration of chromium varies from 20 to 1500 μg/L;
 The concentration of copper varies from 5 to 1500 μg/L;

Chromium comes from cardboard paper and wood that is collected with household
waste. Its content is 190 μg/L, it is much lower than the concentration detected in the
leachate of the Mohammedia landfill (Chtioui et al. 2005).

Table 7. Characteristics of leachate used in this study

Parameter Value
Turbidity (NTU) 180
COD (mg/L) 21120
Phenol (mg/L) 96,2
NH4+ (mg/L) 536,9
NO3- (mg/L) 456,0
NO2- (mg/L) 760,0
Total phosphorus (mg/L) 31,5
Surfactant (mg/L) 47,1
Abs 254 nm (FD = 200) 0,258
Color (FD = 20) 0,569
Hg (µg/L) 6,5
Cr (µg/L) 190
Cd (µg/L) 25
Zn (µg/L) 85
Pb (µg/L) 85
Cu (µg/L) 45
*
FD: Dilution factor
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 239

LEACHATE TREATMENT USING DIFFERENT FLOCCULANTS

MIXED WITH FERRIC CHLORIDE

This part concerns the study of the coagulation flocculation of young leachate. These
tests were carried out by the addition of ferric chloride mixed with three flocculants,
namely: chitosan, Superfloc SD2065 and Himoloc.
The dose of the coagulant was set at 6g/L (determined from preliminary tests), while
flocculant doses ranged from 3,3 to 20 mL/L. The evaluation of the quality of the
treatment was determined by monitoring removal efficiencies of: Turbidity, COD,
phenol, color, Abs at 254 nm and the volume of sludge collected after 24 hours.
Figure 9 illustrates the variation of these parameters as a function of different
concentrations of the flocculants.

Elimination of Abs at 254nm

Referring to Figure 10 (a), Abs at 254 nm decreases progressively with the addition
of the different flocculants until reaching a minimum value, and then increases following
an excess of flocculant.
The removal efficiencies of Abs at 254 nm obtained by the addition of the three
flocculants vary from 75 to 85%. Superfloc SD2065 flocculant and chitosan achieved a
removal of around 83%. In the case of Himoloc flocculant, the highest removal was 81%
at a concentration of 20 mL/L.
The results we obtained remain superior to those obtained by Ntampou et al. (2006),
with a removal of 75% of Abs at 254nm, adding 365 mg/L of coagulant.
Tzoupanos et al. (2008) conducted coagulation flocculation tests via a new coagulant
named PSI. Removal efficiencies of Abs at 254 nm obtained varied from 15 to 70%. The
best removal was 75%, obtained by the addition of 300 mg/L of PACSi 1/15 coagulant.
These authors show that Alumina sulphate is the least suitable coagulant for the
elimination of Abs at 254 nm.

Elimination of Phenol

Phenol is composed of a benzene aromatic ring (aromatic hydrocarbon) and a

hydroxyl function. It is the simplest molecule in the phenol family. The choice of this
molecule results from its frequency in waste water from various industries.
240 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Figure 10. (Continued)

 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 241

Figure 10. Effect of different flocculants on the elimination of pollution in leachate.

Figure 10 (b) shows the effectiveness of ferric chloride mixed with the three
flocculants in terms of phenol removal. The maximum removals were achieved using
Superfloc SD2065 and chitosan, and are respectively 99 and 88%. The optimal dose of
both flocculants is 13 mL/L. For Himoloc flocculant, phenol was reduced by 81% by
adding 3 mL/L.
242 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Achour and Guesbaya (2005) conducted CF testing for the removal of phenolic
organic compounds and humic substances. Low phenol removal was achieved ranging
from 9,26 to 32%. The authors concluded that phenolic structures are little affected by
Coagulation flocculation. Krou (2010) showed that charcoal adsorption is an effective
method for phenol removal. The disadvantage of this technique is the high cost generated
by the regeneration of coal. Indeed, the pollutant remains concentrated on the surface of
the solid which itself becomes a waste that must be transported and treated within
specialized sites.

Elimination of COD

We note that COD removal increases with the increase in the concentration of the
three flocculant. However, beyond the optimal concentration, COD elimination
decreases. This is due to the excessive adsorption of polymer on the colloidal surfaces,
which induces the production of stabilized colloids. These colloids are positively charged
and cause electrostatic repulsion of suspended particles (Abu Hassan et al., 2009).
The most effective flocculant is Superfloc SD2065, with a maximum removal of
around 86% at a concentration of 13 mL/L. In addition, a dose of 10 mL/L of Himoloc
flocculant leaded to an elimination of 82% of COD initially contained in the leachate.
COD was reduced by 73% by adding 17mL/L of chitosan. Several researchers use ferric
chloride as coagulant for the treatment of landfill leachate. This is the case of Tatsi et al.
(2003), who studied the treatment of stabilized leachate from the Thessaloniki landfill in
Greece. Without pH adjustment, the addition of 1,5 g/L of FeCl3 + eliminated 80% of
COD, while a dose of 1,5 g/L of Al3+ leaded to an elimination around 38%. These authors
also tested the effect of several flocculants: Anionic (A321), cationic (K1370 and K506)
and neutral (N200) on the treatment of young and partially stabilized leachates. They note
that in the case of partially stabilized leachates, the addition of these flocculants has no
effect on improving the reduction of organic matter. In addition, for young leachates, the
elimination of organic pollution was improved by the simultaneous effect of pH
adjustment (to a value of 10) and the addition of flocculants. As a result, 80% of COD
was removed by the addition of flocculant A321 and ferric chloride with a concentration
of 1,5 g/L (Tatsi et al., 2003).

Volume of Sludge

According to Assou et al. (2015), the volume of sludge produced strongly depends on
physicochemical characteristics of the leachate as well as the effectiveness of
Coagulation flocculation treatment.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 243

Based on Figure 10 (d), we note that the mixture of ferric chloride with Superfloc
SD2065 flocculant induces the formation of less sludge (after 24 hours of settling),
compared to the other two flocculants. Indeed, the minimum volume of sludge is 43% for
the mixture FeCl3+Superfloc SD2065, 46% for FeCl3+Himoloc and 50% for
FeCl3+chitosan.
At low concentrations, FeCl3+chitosan mixture produces less sludge. According to
Renault et al. (2009), chitosan significantly increases the density of sludge and facilitates
drying. In addition, chitosan is a biodegradable biopolymer, which allows sludge
degradation by microorganisms.
Removal efficiencies obtained slightly exceed those achieved by Maranon et al.
(2008), which reached 32 and 30% respectively at 30 min and 24h decantation.
Assou et al. (2015) noted that the volume of sludge produced increases dramatically
as the concentration of flocculants increases. According to these authors, this may be due
to the cationic nature of the flocculants used in the study. These flocculants have a high
molecular weight, which promotes bond formation between small flocs that have the
ability to capture colloidal particles and promote their settling.
These same authors add that organic polymers generally produce less sludge than
inorganic salts. Consequently, sludges resulting from coagulation via ferric chloride
mixed with a polyelectrolyte are compact.

Elimination of Turbidity

Figure 10 (e) shows that turbidity removal is little affected by the change in
concentration of different flocculants.
Indeed, removal efficiencies are of the order of:

 43% for chitosan;

 95% for Superfloc SD2065 flocculant and Himoloc flocculant.

However, it is clear that chitosan is less effective in eliminating turbidity compared to

the other two flocculants.
It should be noted that for chitosan, the best removals are reached at low
concentrations. According to Ahmad et al. (2006), this could be explained by its high
charge density which lead to high adsorption of the polymer and less quantity required
(Ariffin et al., 2005).
Assou et al. (2014), examined the effect of ferric chloride mixed with three different
flocculants, namely: Astral, Superfloc and Alginate. For turbidity, FeCl3 and Superfloc
mixture showed the best removal efficiency (66%).
244 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

The optimization of the operating conditions of the Coagulation flocculation of

industrial wastewater was the subject of the study conducted by Martin et al. (2011).
Indeed, the experimental design methodology was used to determine the optimal doses of
coagulant (Aluminum Polychloride) and Flocculant (Actipol A-401), leading to a
maximum reduction in COD, turbidity and TOC. As a result, the turbidity was reduced
by 96% following the addition of 1 mL/L of the coagulant and 6,13 mL/L of the
flocculant.

Elimination of Color

Figure 10 (f) shows that color removal is little influenced by the flocculant
concentration. The maximum values reached:

 99,2% for chitosan at a concentration of 20 mL/L;

 99,4% for the Superfloc flocculant at a concentration of 13,3 mL/L;
 99,2% for the Himoloc flocculant at a concentration of 10 mL/L.

The elimination efficiencies we obtained exceeded those achieved by Aziz et al.

(2007). Indeed, these authors realized an elimination of 97% of the color following the
addition of 2,5 g /L of ferric chloride at a pH of 6.
In a similar study, Maranon et al. (2008) concluded that the optimal dose of ferric
chloride useful for the treatment of young leachate is in the order of 0,5 g/L. This dose
eliminates 28,1% of COD, 78,4% of color and 90,2% of turbidity. These researchers also
tested the effect of ferric chloride (at 0.3 g Fe3+/L) mixed with five flocculants (A-401, A-
352, C-472, C-421 and Polifloc A-1849), keeping the initial pH of the leachate. They
concluded that the addition of flocculants did not improve treatment efficiency in terms
of COD, color and turbidity. Nevertheless, the authors noticed an improvement of the
settling, due to the formation of flocs which decant quickly.

Figure 11. Picture of coagulation flocculation tests using Himoloc flocculant.

 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 245

Conclusion

Comparing the results obtained following the addition of the various flocculants, we
note that the mixture of ferric chloride with Himoloc is better suited to the treatment of
leachate from ouled Berjal landfill with a minimum volume of 10 mL/L (Figure 11).

EFFECT OF ORDER OF INTRODUCTION OF THE REAGENTS ON THE

EFFICIENCY OF COAGULATION FLOCCULATION TREATMENT

The purpose of this section is to reveal the effect of order of introduction of the
reagents on the efficiency of treatment of landfill leachate by coagulation flocculation.
In this context, leachate was tested for coagulation flocculation by ferric chloride and
three different flocculants, namely: Chitosan, Superfloc SD2065 and Himoloc.
The optimal doses were determined following preliminary tests, and are as follows:

 8 g/L for ferric chloride;

 20 mL/L for chitosan;
 10 mL/L for Superfloc SD2065;
 13 mL/L for Himoloc.

We varied the order of introduction by adding:

 Ferric chloride first followed by flocculant.

 Flocculant first followed by coagulant;
 Coagulant and flocculant at the same time.

The experimental approach was carried out in three stages:

 The first reagent is added to the sample and stirred at 150 rpm for 10 minutes;
 The second reagent is added and stirred at the same speed for 10 min;
 The sample is then subjected to slow stirring at 30 rpm for 30 min;
 The supernatant is removed after 1 hour of decantation.

The effect of the order of introduction of the reagents was evaluated in terms of
elimination of turbidity, COD, color, Abs at 254 nm, phenol, surfactant, ammonium ions,
nitrites, nitrates and total phosphorus.
246 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Elimination of Turbidity

The results reported in Figure 12 (a) show that the introduction of coagulant followed
by flocculant provides the best turbidity removal efficiencies.
Turbidity was reduced by 90% by chitosan and Himoloc, and 87% by Superfloc
SD2065.
In a study conducted by El Bada et al. (2010), doses of lime, calcite and soda,
ranging from 0 to 10 g/L were added to leachate from the Azemmour landfill to eliminate
turbidity, COD and metals. In terms of turbidity, lime was the most effective in
eliminating 80%.
Abu Hassan et al. (2009) studied coagulation flocculation of wastewater from textile
industry using chitosan. The results showed that the reduction in turbidity and COD were
respectively 94,9 and 72,5% following the addition of 30 mg/L of chitosan at
a pH of 4.

Elimination of COD

COD removal is maximum when the coagulant and the flocculant are added
simultaneously. COD was reduced by 86% by mixing the ferric chloride with the
Himoloc flocculant, and 82% by the other two flocculants.
These efficiencies exceed those obtained by other researchers (Zeng et al., 2008, El
bada et al., 2010, Ez-zoubi et al., 2010).

Figure 12. (Continued)

 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 247

Figure 12. (Continued)

248 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Figure 12. Effect of the order of introduction of the reagents on removal of pollutants in the leachate.

Precipitation by lime, NaOH and calcite has been studied by El bada et al. (2010).
They showed that the COD decreases with the increase in the concentration of the three
coagulants. Its elimination reached 31, 24 and 16 respectively for lime, NaOH and
calcite.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 249

Ez-zoubi et al. (2010) obtained a maximum reduction of 35% in terms of COD. The
authors showed that COD removed corresponds to the hard COD and its removal is due
to the adsorption on ferric chlorides (Wang et al., 2002).

Elimination of Color

From Figure 12 (c), there is little difference between the different flocculants as well
as between the different orders of introduction. Indeed, color removal exceeded 97%.
Rasool et al. (2016) used experimental design methodology for the
optimization treatment of landfill leachate using basil seed powder as a biocoagulant.
Optimal conditions for the three factors are a contact time of 15 min, a pH of 7 and a
Alum:Basil ratio of 1:1. COD and color were eliminated by 64,4% and 77,8%
respectively
Syafalni et al. (2012) evaluated the treatment efficiency of landfill leachate
with lateritic soil as coagulant. An optimal dose of 14 g/L at a pH of 2 makes it
possible to reduce the COD by 65,7%, color by 81,8% and the ammoniacal nitrogen by
41,2%.
Zahrim et al. (2010) performed coagulation flocculation of highly concentrated dye
solutions (Acid Black 210). The authors compared the effect of five flocculants, mixed
with aluminum sulphate used as a coagulant. The best removal efficiency of color (37%)
was obtained by adding polyDADMAC flocculant.

Elimination of Phosphorus

Phosphorus exists in different forms such as mineral salts or organic compounds.

These are either solubilized or fixed on suspended solids (Rodier et al., 1996).
Orthophosphates react rapidly with ferric chloride by their negative charges to form
amorphous precipitates.
We conclude that there is little difference between the three flocculants, because the
total phosphorus removals are 85%. However, these efficiencies are obtained by different
orders of introduction. For chitosan, the maximum removal was obtained by the
simultaneous introduction of the two reagents, whereas for Superfloc 2065 and Himoloc,
the maximum removal was obtained by introducing the coagulant first.
Leachate from the controlled landfill of Fez city in Morocco has been treated by
coagulation flocculation using ferric chloride. A concentration of 100 mg/L of this
coagulant reduced the orthophosphate content by 12,1%. Ez-zoubi et al. (2010)
concluded that the low abatement is due to the low concentration of orthophosphates in
raw leachate.
250 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Moreover, in the study of Jirou et al. (2014), the Agadir landfill leachate was
subjected to continuous aeration for 60 days. Total phosphorus was reduced by 92,1% at
the end of the test, while in the control sample the removal was only 51,2%.
Khattabi et al. (2002) studied elimination of the total phosphorus contained in
leachate from Etueffont landfill by natural lagooning. The authors achieved a reduction
of 70% in summer and 14% in winter. The low rate reached in the winter season was
explained by the decrease in algal assimilation of phosphorus as well as the re-solution of
this element, which is favored by a low pH (of the order of 4).

Volume of Sludge

Volume of sludge is one of the most important parameters in coagulation flocculation

process. Indeed, it is taken into consideration when choosing reagents as well as their
optimal doses. In general, it is necessary to find a compromise between this parameter
and the efficiency of the treatment, considering the additional costs generated by the
treatment of sludge which themselves become a source of nuisance.
For the three flocculants, the minimum volume was obtained by the simultaneous
introduction of the reagents: 330 mL/L for chitosan, 300 mL/L for Superfloc SD2065 and
280 mL/L for Himoloc.
Previous studies on the use of chitosan for the treatment of water from a dairy plant
(Chi et al., 2006), and for the treatment of kaolinite suspensions (Divakaran et al., 2001),
indicated that sludge produced is non-toxic and can be used to stimulate plant growth.
Khalil et al. (2015) recorded the volume of sludge produced by the coagulation
flocculation of leachate from Fez landfill after 60min of settling. Therefore, the minimum
volume of sludge was obtained by ferric chloride (245 mL/L), followed by iron sulfate
(264 mL/L), aluminum sulphate (568 mL/L) and finally lime (647 mL/L).

Elimination of Abs at 254nm

Coagulation flocculation is very effective in eliminating Abs at 254 nm which

characterizes conjugated compounds (aromatic nuclei) (Chin et al., 1994).
By introducing the coagulant first, Abs at 254 nm was reduced by 81% for chitosan
and 84% for Superfloc SD2065. Moreover, 88%, was eliminated by the simultaneous
introduction of ferric chloride and flocculant Himoloc.
Singh et al. (2012) evaluated the effectiveness of coagulation by ferric chloride and
magnetic ion exchange (MIEX), for the treatment of leachate from three different
landfills. A maximum reduction of 94 and 48% of the Abs at 254 nm was obtained
respectively by the two treatment techniques.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 251

Elimination of Phenol and Surfactant

Phenol and detergents are major inhibitors of the development of biomass promoting
biodegradability. Their elimination is therefore of great necessity.
From Figure 12, the introduction of chitosan and Superfloc SD2065 flocculant first
improves detergent removal, with removals of 64 and 57%, respectively. The lowest
removal (23%) was obtained by Himoloc.
Unlike phenol which elimination is favored by the introduction of the coagulant first,
specifically for chitosan (64%) and Himoloc.

Elimination of Nitrogen Compounds

Figure 13 reports the effect of the order of introduction of the reagents on the
removal of nitrogen compounds.
According to Rodier et al. (1996), mineral nitrogen composed of ammonia, nitrates
and nitrites, constitutes the major part of the total nitrogen. Elimination of nitrogen
compounds is of great importance in the field of water treatment, given their negative
effect on the environment as well as on human health.
According to Umapriya and Shrihari (2010), elimination of ammoniacal nitrogen is
probably done by the adsorption of ammonium ions on the coagulant. Thanks to their
positive charge, the ammonium ions act as complementary species to improve the effect
of Al3+ and Fe3+ cations on the coagulation of the suspended matter, thus acting bridging
elements as follows:

Al3+ ---- MES ---- NH4+ ---- MES ---- Al3+

Fe3+ ---- MES ---- NH4+ ---- MES ---- Fe3+
Fe3+ ---- MES ---- NH4+ ---- MES ---- Al3+

We note that the maximum removals in terms of ammonium ions are quite low. They
are of the order of 14, 46 and 39% respectively for chitosan, Superfloc SD2065 and
Himoloc.
Orders of introduction leading to these results differ from one flocculant to another.
Indeed, for chitosan, the most suitable is to introduce the flocculant first. While for the
other two flocculants, it is advisable to introduce the coagulant first.
For nitrites, elimination efficiencies reached:

 64% for chitosan and 83% for Himoloc by introducing the flocculant first;
 74% for Superfloc SD2065 by introducing reagents at the same time.
252 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

Figure 13. Effect of order of introduction of the reagents on elimination of

nitrogen compounds.

In addition, the elimination of nitrates is favored by adding the coagulant first, for
Superfloc SD2065 (99%) and Himoloc (62%). While adding ferric chloride first followed
by chitosan reduced the nitrate content by 55%.
 Treatment of Leachate from the Ouled Berjal Landfill in Morocco … 253

Ez-zoubi et al. (2010) studied the effect of coagulation flocculation on the

elimination of nitrogen compounds contained in the landfill leachate of Fez city.
Removals obtained reached 44, 13,3 and 31% respectively for ammoniacal nitrogen,
nitrates and nitrites.
Bashir et al. (2010) studied the treatment of landfill leachate through an ion exchange
resin. 94,2% of the ammonium ions were removed by adding a dose of 24,6 cm3 of resin,
with a mixing speed of 147 rev/min for a contact time of 6 min.
Adlan et al. (2011) studied the combination of coagulation and dissolved air flotation
for the treatment of landfill leachate. The optimization of the flow rate, the coagulant
dosage, the pH and the injection time was carried out by means of experimental plans.
Under optimal conditions, turbidity, COD, color and NH3-N decreased by 50%, 75%,
93% and 41%, respectively.
At the end of this study, we conclude that the order of introduction of the reagents
has a significant effect on the efficacy of coagulation flocculation treatment.
This effect can be summarized as follows:

 Elimination of turbidity, ammonium ions, Abs at 254 nm, phenol, total

phosphorus, nitrates is favored by the introduction of the coagulant first;
 A minimum volume of sludge and a high elimination of COD are achieved by the
simultaneous introduction of reagents;
 Elimination of detergents is favored by the introduction of flocculant first.

CONCLUSION

In this chapter, we studied the treatment of leachate from ouled Berjal discharge by
flocculation coagulation.
For young leachate from Ouled Berjal landfill, we evaluated the effectiveness
of ferric chloride as coagulant mixed with various flocculants, such as:
Chitosan, Himoloc DR3000 and Superfloc SD2065. The best results were
obtained by mixing the coagulant with Himoloc DR3000 polymer, with removal
efficiencies of 90, 86, 99, 83 and 99,4% respectively for turbidity, COD, phenol, Abs at
254nm and color. Optimum doses are 6g/L for the coagulant and 13,3 mL/L for the
flocculant.
We also examined the effect of order of introduction of reagents on the
effectiveness of coagulation floculation. Indeed, we concluded that the elimination of
turbidity, ammonium ions, Abs at 254 nm, phenol, total phosphorus and
nitrates is favored by the introduction of the coagulant first. Minimal volume
of sludge and high COD removal are achieved by simultaneous introduction
254 Hajar Bakraouy, Salah Souabi, Khalid Digua et al.

of the reagents, while removal of detergents is promoted by introduction of the flocculant

first.

ACKNOWLEDGMENTS

This study was conducted as part of the research and development program initiated
by the State Secretariat for Sustainable Development.

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In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 13

OMW PRETREATMENT BY ASSISTED

SEDIMENTATION METHODS:
COAGULATION/FLOCCULATION

Gassan Hodaifa* and Cristina Agabo García

Molecular Biology and Biochemical Engineering Department,
Chemical Engineering Area, University of Pablo de Olaivde,
Seville, Spain

ABSTRACT

Natural and assisted particles sedimentation are crucial operations during the
treatment of wastewater generated in many industrial processes, above all in the
wastewater treatment field. This operation is usually used as an effluent pre-treatment;
normally essential for reducing suspended solids, organic load, turbidity and colour so as
to achieve the correct clarification of wastewater. This chapter introduce the coagulation
and flocculation concepts as well as the influence of operational conditions on assisted
sedimentation.
At industrial level, the factual use of both processes has been proved on different
wastewater. In this sense, this chapter outline researches about combining or comparing
assisted sedimentation (coagulation, electrocoagulation, neutralization, etc.) with other
operations such as oxidation processes in order to evaluate the solids removal of the
complete designed wastewater treatment focusing on OMW treatment. In addition, in this
work new trends about multifunctional and novel eco-friendly bio-degradable flocculants
and coagulants were reviewed and suggested for OMW pretreatment.

*
Corresponding Author Email: [email protected].
260 Gassan Hodaifa and Cristina Agabo García

Keywords: OMW, colloids, neutralization, flocculation, coagulation

1. INTRODUCTION

In the last century, two-phases production process (TP-PP) has become to the most
efficient and eco-friendly olive oil production extraction method. As the most used in
Spanish mills, TP-PP applies a centrifugation step after common operations: olive
washing, crushing and malaxing. This operation involve the use of horizontal centrifuge
‘decanter’ with two exists: one for crude olive oil which contain impurities and the other
for a wet solid residue named in Spain ‘alperujo.’
In this context, olive oil mill wastewater (OMW) is a result of the mixture of
different wastewaters generated during olive oil production process. In other words, it is
the sum of wastewater from washing olives (WOW), vegetation water, wastewater from
olive oil washing (WOOW) and others wastewaters such as additional water added to
malaxing system, cleaning devices wastewaters, etc. (Dermeche et al., 2013; Garcia-Ivars
et al., 2015).
On average, olives washing machine needs 1 m3 of drinking water per each
tonne of olives. This wastewater contains basically natural organic matter (NOM) coming
from sources of natural waters and the impurities such as dust, little stones,
microorganisms or even herbicides and pesticides (Guardia-Rubio et al., 2006 and 2007).
Avoiding water addition in malaxing system, TP-PP generates a 0,33-0,35 m3 of
wastewater per tonne of olives and bringing tannins, pectins, lignines, long fatty acids,
reduced carbohydrates, proteins and bacterial and plants growth inhibitors
phenolic compounds (Dermeche et al., 2013; Bouknana et al., 2014; Gerasopoulos et al.,
2015).
Regarding OMW treatment, natural or assisted particles sedimentation is a
fundamental step to be used as effluent pretreatment. This basic operation contribute to
the treatment of wastewater by reducing its suspended solids, organic load, turbidity and
colour. Since the presence of different kinds of suspended solids in olive oil mills
wastewater (OMW), colloids are charged stabilized particles that avoid aggregation
leading to non-sedimentable particles.
Coagulation and flocculation techniques consist of using chemical agents that reduce
the charge of the particles and promote micro-particles formation. It is commonly named
coagulation and flocculation indistinctly. However, on the one hand, coagulation
comprises the electrostatic destabilization of the colloids that form microflocs. But on the
other hand, flocculation is the flocs growth by chemical bonds formation resulting in
sedimentable cluster of particles (Grant et al., 2001; Martínez-Nieto et al., 2011; Rytwo
et al., 2013).
 OMW Pretreatment by Assisted Sedimentation Methods 261

2. FACTORS INVOLVED IN FLOCCULATION AS A PRETREATMENT

Flocculation/sedimentation are effective operations, which try to remove the majority

of particles and natural organic matter. Normally, particles that are difficult to be
separated by gravity or natural sedimentation. In this sense, flocculation produces
aggregations of particles forming flocs. This fact occurs due to the combination of
different mechanisms:

1. Charge neutralization.
2. Entrapment mainly by Van der Waals forces.
3. Adsorption forces.
4. Complexation with coagulant metal ions into insoluble particulate aggregates.

The charge neutralization mechanism is proved to be more efficient when pH is

optimized. However, due to the different composition of effluents the rest of removal
mechanisms will be different because the wide range of structures (Matilainen et al.,
2010; Martínez-Nieto et al., 2011).
In coagulation or flocculation, mixing energy promotes formation of larger structures
(Matilainen et al., 2010). In addition, these structures play a secondary role in treatment
by adsorbing other soluble wastewater pollutants such as pathogens (Talaie, 2008).
In general, flocculation could be divided into initial-flocculation stage, growth stage
and the steady stage. Flocs keep growing until reaching a steady-state size through a
balance between flocs formation and flocs breakage (Jarvis et al., 2005). After reaching
steady-state, flocs size is governed by the shear conditions in the flocculation unit
(decanter). According to Jarvis et al., (2005) it can be expressed the rate of floc growth,
Rfloc, as following in Eq. (1):

𝑅𝑓𝑙𝑜𝑐 = ∝· 𝑅𝑐𝑜𝑙 − 𝑅𝑏𝑟 (1)

where ‘Rcol’ is the rate of particles collision and ‘Rbr’ is the rate of flocs breakage. ‘α’ is
the collision efficiency factor that means the number of effective collisions which result
in attachment among the total of collisions. This factor is not constant, while particles
sizes are increasing, the number of particles in the system is reduced leading to a decrease
in efficiency of collision (α·Rcol). When both parts of the equation have the same value,
the net rate of flocs growth is zero. At this point, the steady-state is reached and the size
of the flocs have got its maximum level. It is important to take account that floc breakage
may not be readily re-formed in some irreversible breakages and the ‘α’ parameter would
be equal to 0 (Gregory and Dupont, 2001).
Floc stability analysis in suspension is possible by determining not only the strength
but also the number of the bonds holding the floc together. The floc strength can be
262 Gassan Hodaifa and Cristina Agabo García

approached by macro- and microscopic views. Macroscopic techniques for determining

floc strength use different devices (e.g., Impeller, ultrasonic, multigrid mixer) to create
hydrodynamic stress conditions to cause floc breakage. It can be measured the energy
required for floc breakage, the ratio of the floc size or floc erosion in order to determine
the correlation between the shear rate and the resulting floc size. However, microscopic
techniques include micromechanical method and micromanipulation in order to isolate
individual flocs and determine the force required to pull or compress a floc until breakage
(Gregory, 2003).
Analysing flocculation efficiency is also necessary to select the optimal flocculant in
a sedimentation system. In the case of wastewater, it is necessary to characterize the
water before and after flocculation tests determining various water quality parameters
such as: pH, alkalinity, turbidity, conductivity, temperature, cations/metals and anions
content, UV254, COD, total carbon (TC), total organic carbon (TOC), inorganic carbon
(IC), total nitrogen (TN) and phosphate, etc. (Sinha et al., 2004).
Particles size distribution is also important to be studied. Small flocs removal in
conventional wastewater treatment is the most difficult task because sedimentation
process can only remove particles in specific size ranges efficiently. Nevertheless, the
elimination of small particles (smaller than 3 µm, such as pathogens) is crucial to avoid
their penetration in filter layer after flocculation steps which cause fouling in the filter
(Wang et al., 2018).
Additionally, it is important to characterize the metal coagulant used, as well as, to
register the time of sedimentation. Characterization of available coagulants consist of
determining some chemistry features such as acidity, solubility, active metal content,
anion content, and speciation (Sinha et al., 2004). Normally, coagulation conditions (pH,
coagulant dosage) are optimized for turbidity and TOC removal (Swietlik et al., 2004;
Matilainen et al., 2010). In brief, both natural and assited sedimentation-flocculation
depend on two factors:

a) Collision rate, that depends on the number and the size of the colloidal particles,
as well as, mixing conditions (timing and velocity), of course surface, and size of
the decanter (Grant et al., 2004; Matilainen et al., 2010). In this sense, the
behavior of floc size increasing thorough the time in each aggregate is related to
the rest of particles aggregations by an infinite number of non-lineal differential
equations.
b) Particles interaction. Attractive and repulsive forces between particles are due to
surface charges. This also involves the physico-chemical characteristic of
effluent particles such as size, functionality, charge and hydrophobicity.

Normally, organic matter with high molecular mass needs less coagulant dosage due
to that the mechanism of removal is, basically, neutralization of charges. In addition,
 OMW Pretreatment by Assisted Sedimentation Methods 263

organic matter with high molecular mass is mostly hydrophobic (e.g.,

aromatic compounds), which is more easy to remove than hydrophilic organic
compounds with low molecular mass (Swietlik et al., 2004; Sharp et al., 2006; Korshin et
al., 2009).

2.1. Mixing Conditions

Agitation is crucial for a complete coagulation/flocculation process. Wastewater

flocculation is normally carried out in three stages. Firstly, a strong but brief wastewater-
flocculant mixing is necessary to achieve uniform dispersion of the flocculant, promoting
particles collisions and micro-flocs formation. Secondly, the agitation of the mixture
must be reduced in order to keep the integrity of flocs in suspension during mixing to
favouring their growth and avoiding their disintegration. Finally, the mixture must left to
settle (Sinha et al., 2004; Swietlik et al., 2004).
Chen et al., (2017) tested the effect of energy input on flocculation and flotation
performance of fine scheelite using sodium oleate. The more energy input applied (>2.04
kJ/m3) the more flocs with these characteristics: small, branchy and globule-like;
however they suffered an inevitably redispersion. On the other hand, low energy input
promotes larger flocs with more branches but with weak resistance to breakage. Wang et
al., 2018 programed two mixing steps. In the first one, the agitation was 400 rpm during 1
min and in the second one; they tested different agitation velocities 60, 120 and 180 rpm.
Results concluded that a moderate shear rate (120 rpm) optimized the shape of flocs for a
good flocculation as well as removal of particle with a size < 3 µm whereas the highest
mixing rate contributed to rapidly steady stage reaching and major breakage of flocs.

2.2. pH Effect

pH conditions of wastewater have a great influence in particle interaction because

they affect to surface charge of colloids, organic matter and dissolved-phase of coagulant
species and flocs (Sahu and Chaudhari, 2013; Yan et al., 2017). In addition, all-metal
cations are hydrated, to some extent and each coagulant has minimum pH condition on
which it keeps its solubility. For example, iron has two oxidation states, Fe2+ and Fe3+ or
ferrous and ferric forms, respectively. Iron species in aqueous solution is subject to
equilibrium. The iron hydroxides formed, especially the Fe2+ form, have very low
solubility. The presence of iron species in solution is consequently influenced by the pH
value. In natural waters, the pH value is not too low to avoid hydroxides from forming,
and under oxidizing conditions, virtually all the iron is precipitated as Fe(OH)3. Figure 1
shows the iron species distribution as a function of pH solution value. Dissociation
264 Gassan Hodaifa and Cristina Agabo García

products of ferric chloride (normally used as coagulant) are cationic so interaction with
negative colloids is easy under correct pH value. Fe+3 is arounded by six molecules of
water which will be progessively replaced by hydroxyl ions due to polarization and loss
of protons because of pH values (Sahu and Chaudhari, 2013).

Figure 1. Chemical equilibrium diagram of iron species presence in function of pH value at initial iron
concentration = 10 M. The diagram was obained by using Medusa-Hydra chemical equilibrium
software.

Salt concentrartions have also a crucial influence due to the supply of divalent cations
and destabilizing anions (bicarbonate, chloride, and sulfate). As an example, Ferric
chloride, ferric sulphate and alum are acidic, leading to lower pH condition after
application (Yan et al., 2008). Moreover, alkalinity and pH are related. In water with
lower alkalinity, coagulant addition may consume all the available alkalinity, depressing
the pH to values favourable for coagulation. In contrast, at higher alkalinity conditions it
is necessary the addition of great amount of coagulant in order to decrease pH values.

2.3. Temperature Influence

Floc size, strength and re-construction of flocs after shear break-up during
flocculation process are influenced by temperature due to changes in coagulant
chemistry. Temperature affects the solubility of the metal hydroxide precipitate and the
rate of formation of the metal hydrolysis products. In addition, temperature conditions
also affect the viscosity of clarified water after flocculation.
Gregory et al., (2004) compared aluminium sulphate, ferric sulphate and
polyaluminium chloride products (PACL). It was concluded that warmer temperatures
 OMW Pretreatment by Assisted Sedimentation Methods 265

(>15ºC) generally produce bigger flocs that not only are easily to be broken but also
tough to be reformed. On the other hand, lower temperatures favoured lower size flocs
which suffer less number of breakages and recover their structure better. Moreover, the
variation of size and strength of flocs depends on the type of flocculant used. As an
example, the results of this study showed that aluminium based coagulants produce flocs
more sensible to temperature variation than the ones formed using ferric sulphate. In
addition, flocculation size also depends on the dosage of flocculant. At lower aluminium
dosage (1.4 mg/L), the size of flocs had more variation than at higher dosage (3.4 mg/L)
in the case of using PACL. In many cases, as lower temperatures favour
flocculation/coagulation process, it is better wait for the cooling of warm water before
starting the coagulation step. In full-scale plants, cold temperatures tend to reduce
kinetics of hydrolysis reactions and particle flocculation formation due to a decrease of
alum and PACL solubilities and an increase in water viscosity. For this reason, higher
coagulant dosages and additional flocculation time are required at a low temperature
(Frank et al., 2000).

2.4. Classification and Coagulant Dosage

In general, it can be distinguished between coagulant (inorganic) and flocculant

(organic). Conventionally, coagulants are mineral inorganic salts formed by the mixture
of a strong acid e.g., HCl or H2SO4 and a weak base e.g., Al(OH)3 or Fe(OH)3
(Tzoupanos and Zouboulis, 2011). The most common coagulant salts are ferric sulphate
(Fe2(SO4)3), aluminium sulphate (Al2(SO4)3 or Al2(SO4)3 · 14H2O) and ferric chloride
(FeCl3) or (FeCl3 · 6H2O). In contact with water, salts are dissociated to their ions and
later hydrolysed in soluble structures positively charged which attach into the surface of
negative colloids (Sinha et al., 2004; Talaiekhozani et al., 2017).
Using inorganic coagulant it can be obtained three kinds of species in the effluent:
mononuclear, medium polynuclear and precipitated/colloidal. For medium/monomer
species formation there is a major COD removal by complexation, adsorption, charge
neutralization or co-precipitation (Yan et al., 2008). However, in the case of higher
polymer species (flocculant) is more common the removal by bridging flocculation; in
other words, when flocs are formed because of the adsorption of more than one particle
into the surface of the flocculants (Hogg, 2013).
Metallic coagulants have been used for several decades to precipitate soluble metals
as hydroxides. However, the main drawback resides on the generation of large quantities
of chemical sludge that must be eliminated after coagulation due to their hazardous
character. The amount of sludge produced is also important in order to obtain a major
clarified phase. Talaiekhozani et al., (2017) showed that potassium ferrate (K2FeO4)
produces less amount of sludge than others ferrous and aluminium salts and therefore,
266 Gassan Hodaifa and Cristina Agabo García

less amounts of problems with final disposal of sludge. This fact is considered as one of
the advantage of using potassium ferrate among inorganic salts.
On the other hand, among polymers, it can be found inorganic polymeric coagulants
and synthetic and natural organic polymeric materials that are used as flocculants (dos
Santos et al., 2018). Polymers can be cationic, anionic or non-ionic (neutrally charged).
By and large, polymers show low diffusion rates raising the viscosity of solution. For this
reason, it is compulsory, a great mechanical dispersion of the polymer into the water, as it
mentioned before, by applying a strong first step of mixing in a level that polymers can
not be degraded (Sahu and Chaudhari, 2013).
Since the beginning of the last decade, inorganic polymerized flocculants (IPFs) have
been increasingly used in water treatment not only in Europe and Japan but also in North
America owing to availability and reduced price. IPFs are formed by the addition of
anions to conventional coagulant salts acquiring new coagulant properties. IPFs are
considered to be improved in particulation, organic removal, lower alkalinity
consumption and lesser sludge production (Sinha et al., 2004; Tzoupanos and Zouboulis,
2011). As an example, Van Benschoten and Edzwald (1990) compared the alum with and
prepared PACL. Results claimed that PACL showed a shape based on small spheres (<25
mm) and/or chain-like structures, whereas alum appeared as a fluffy and porous flocs
(ranging from 25 to 100 mm). The advantages of the structures formed by PACL
included: lesser turbidity in clarified water and less sensibility to temperature changes.
These benefits could arise from differences in their hydrolysis, nucleation, growth, and
aggregation leading to a new structure formation which interact more efficiently with
more fractions of organic matter (Van Benschoten and Edzwald, 1990). Others such as
polymeric ferric sulphate (PFS) are being used not only because it is superior capability
in removing turbidity, algae, colour and natural organic matter (NOM) but also avoiding
corrosion (Shi et al., 2004).
It is well known the advantage of synthetic polyelectrolytes compared to natural
ones. In a study conducted by Aguilar et al., (2003) some polymers were used as a
coagulant aids with ferric sulphate in liquid effluent from a slaughterhouse. Results
showed that, using anionic polyacrylamide or polyvinyl alcohol polymers increase the
total particulates removal from 80 to 99 and 93%, respectively (Aguilar et al., 2003).
However, organic polyelectrolytes generally present toxicity due to its monomer presence
in clarified water. In fact, some countries have already forbidden or restricted the use of
some polyelectrolytes for drinking water applications (WHO, 2000). For this reason,
there is a growing interest in replacing synthetic polyelectrolytes with natural and
sustainable alternatives, such as combination of PAM with activated starch (Lapointe and
Barbeau, 2017).
Natural organic coagulants presents a minimal health risk to living organisms, and
are highly biodegradable compared to inorganic coagulants. In this sense, exist a large list
of natural coagulant–flocculants that come from vegetable, animal, or microorganism
 OMW Pretreatment by Assisted Sedimentation Methods 267

sources used in the treatment of natural water and wastewater, such as Nirmali seed,
maize mesquite, species of Opuntia, chitosan and xanthan (Choumane et al., 2017). As an
example, herbal coagulants (mustard seed extract, guar gum, Moringa oleifera) have
proven effective for treatment of drinking water and wastewater (Sanghi et al., 2006). In
addition, they could be more efficient that inorganic coagulants. In other study conducted
by dos-Santos et al., (2018) three tannin-based coagulants (Tanfloc, Acquapol C1 and
S5T) were compared to aluminium sulphate in order to treat starch effluent. Conclusions
showed that the highest colour and turbidity removal occurred for natural coagulants
Acquapol S5T and Tanfloc SL. at 320 mg/L dos-Santos et al., (2018). Sinha et al., (2004)
proposed a strategy to select the most effective coagulant by three steps:

 Effluent characterization with the aim to determine its hidrofobicity, level of

organic matter and content of ions. The content of ions is related to alkalinity and
therefore to pH. Higher alkalinity waters normally have higher pH. This tests
give information about the ease of the water to be treated and which coagulants
must be selected depending on its capacity of consumption of alcalinity.
 Coagulant characterization acidity, solubility, active metal content, speciation or
anion content (such as sulfate and chloride ions). These anion can be tested as a
colloidal neutralizer using a synthetic colloidal dispersion (e.g., humic/fulvic acid
and particulate matter) in order to determine the optimal dosage of coagulant. It
is also useful to determine the alkalinity consumption of coagulants in the case of
treating water. For example, in the case of inorganic metallic coagulants (alum
and ferric chloride), their acidic charcter consume more alkalinity than PACLs
(Sahu, and Chaudhari, 2013). In spite of these additional tests, companies
normally give the related information.
 To develop flocculation tests with the aim to optimize the operating conditions at
pilot plant level (coagulant dosage and pH). Sinha et al., (2004) proposed that
aluminuin based alternative coagulants will increasingly be used in water
treatment instead of aluminum sulphate beacuse of its expensiveness,
availability, and advantages such as non-acidic, higher active metal content or
less sludge production. It was also obtained higher water quality observing that
50 mg/L of PCL-4 was necessary to achieve 50% COD removal and turbidity < 1
NTU instead of 75 mg/L of alum. In addition, they concluded that optimizing pH
of coagulation can be achieved with a lesser amount of coagulant. In this case,
the coagulation at lower pH would be very effective using alum based inorganic
and organic coagulant. However, if the coagulation must be at higher pH (around
6.5–7.0), then PACl-4 would be the most appropriate choice.

It is important to indicate that coagulant dosage can be determined by testing

different concentrations of flocculant. However, there is a specific concentration at which
268 Gassan Hodaifa and Cristina Agabo García

treatment efficiency is optimized. At higher concentration of coagulant, separation

efficiency can not increase but it starts colloids re-stabilization (Jaouani et al., 2005).

3. COMBINATION OF FLOCCULATION WITH OTHER

OPERATIONS IN OMW TREATMENT

Some authors focused their researches on combining assisted sedimentation with

other operations in order to reduce cost and increase the solids removal of wastewater
treatments. In this sense, Murcia et al., (2018) have tested flocculation (cationic organic
flocculant) as a pretreatment before a heterogeneous oxidation process (using TiO2 as a
catalyst) in a dairy wastewater. Following the steps established by Sinha et al., (2004) the
previous characterization of the wastewater is important. As the main compounds of this
wastewater have electronegative charges (amino acids, fat acids, sulphates, carbonates,
microorganisms), it was used different cationic flocculants. Results showed a significant
decrease of turbidity values (from 2100 to 95 NTU) using the highest cationic charged
flocculant.
In the case of using olive mill wastewater, as a source of wastewater, different studies
were carried out. OMW has high organic load, high inorganic concentration, and acidic
pH. Organic load can be positively and negatively charged particles. Suspended and
colloidal matter typically consists of pectins, proteins, oils, tannins, phenols and
carboxylic acids which can have both positive and negative charges (Sarika et al., 2005).
One of the most important molecules in OMW are phenolic compounds. These molecules
in contact with water dissociates into negative anions (negatively charged phenoxide
ions) and leaving hydrogen atoms to form H3O+ (Nassar et al., 2014). Because of that, at
acidic conditions an electrostatic attraction will exist between the cationic flocculants and
the phenoxide molecules. However, at basic conditions, positively charged sodium
phenoxdies prefer adsorption onto anionic flocculants. Hence, the characterization of
OMW in content of phenols and pH conditions would be recommendable before selecting
the flocculant. On the other hand, acidic pH and the polyphenols’ complexing abilities
increase the solubility of heavy metals making inorganic coagulant excellent alternatives
in OMW treatment (Gernjak et al., 2004).
Considering all this facts, researchers normally use charged flocculants in order to
treat them. Michael et al., (2014) studied a pilot-scale treatment of OMW by flocculation
following by solar Fenton. They developed a coagulation step by using two assisted
sedimentation agents i) 6.67 g/L of FeSO4·7H2O (inorganic coagulant) and ii) 0.287 g/L
of FLOCAN 23 (anionic polyelectrolyte flocculant). It was reached 44% of COD
removal and 94% of total suspended solid (TSS) removal. In addition, final pH values
keep similar than original. In this point, it is important to remark that some coagulants
 OMW Pretreatment by Assisted Sedimentation Methods 269

modify pH after treatments such as FeCl3 that reduce pH values to 2-3 favouring AOPs
after flocculation (Papaphilippou et al., 2013). Ginos et al., (2006) also have used a
combination of two agents: i) 5000 mg/L of Fe(II) or 30,000 mg/L of lime (inorganic
coagulants) and ii) 287 mg/L FO-4700-SH (cationic flocculant). The conclusions calimed
that 230 mg/L cationic polyelectrolyte is more effective alone than in combination,
reducing TSS 97.4%, total phenolic compounds (TPCS) 49.7% and COD 17.1%.
However, the amount of sludge produced during the combined use of inorganic coagulant
and poly-electrolyte was lower than that during separation with the poly-electrolyte
alone. Fe(II) and polyelectrolyte did not change pH conditions but the use of lime made
the sample alkaline. But combined process using lime always resulted in greater TPCs
removal than treatment with Fe(II) and therefore it is reduced the toxicity of samples.
Considering the restriction in using organic polycations, before mentioned, it is
important to decide which process is better according to the searched criteria: more
quality, flocculant more eco-friendly, less sludge amount formed, final pH conditions,
etc. In addition, velocity, as well as, volume sludge formation during flocculantion is also
important. In this sense, Martínez-Nieto et al., (2011) studied kinetic equation of
flocculation operation after OMW treatment by Fenton. In this research, it was registered
volumes and velocities of the formed sludge using different concentrations of several
coagulants/flocculants. In the same study Nalco-77171 reached the maximum volumes of
formed sludge and clarified water, 13,5% v/v and 86,5% v/v, respectively. Furthermore,
the main quality parameters of wastewater were determined obtaining %DQOremoval =
18.3% and %Ironremoval = 99.6%. Hodaifa et al., (2015) proposed a treatement using
flocculation as a pretreatment before a foto-Fenton operation. Results showed that the
flocculant with the major capability of sludge formation (Nalco-9913, 3% v/v) was not
the flocculant with the major capability of reducing organic load (QG-2001, DQOremoval =
20.4%).
In spite of high efficiencies in using charged organic polymeric flocculants in OMW,
others authors have tested other kinds of coagulant/flocculant with similar results
chitosan, starch, alum and ferric chloride (Meyssami and Kasaeian, 2005). Even
neutralization as an assited sedimentation operation was tested as pretreatments. Mert et
al., (2010) proposed two pretreatments before Fenton operation: i) Chemical pre-
treatment based on acid cracking and coagulation–flocculation which achieve 67%
CODremoval and ii) Fenton process with >80% COD removal.
Different authors evaluate organic removal capacity of electrocoagulation method. In
this case, the reaction of charged ionics species from wastewater and
coagulants/flocculants improve remotion of organic load. In this sense, Hande and
Arslan-Alaton (2014) used acid cracking (at pH 2.0 and T = 70 °C) and filtration as
pretreatments. Pretreated wastewater was subsequently treated by different techniques: i)
Coagulation with FeCl3, ii) Ca(OH)2 precipitation, iii) electrocoagulation using stainless
steel electrodes and iv) Fenton process as chemical treatment. Among them,
270 Gassan Hodaifa and Cristina Agabo García

electrocoagulation and Fenton reaction appeared as more effective treatments, especially

in removing total phenolic compounds.

4. NEW TRENDS

New trends in investigations lead to search multifunctional and novel eco-friendly

biodegradable flocculants and coagulants (Yan et al., 2009). This is the case of Ferrate
(VI) which was previously used as a coagulant but it was recently discovered other useful
functions. The main objectives of using Ferrate (VI) include:

 Water desinfection.
 Degradation of organic and inorganic contaminants in advanced oxidation
process (AOPs).
 Elimination of suspended particles and heavy metals in water.

Ferric chloride salt also has multiple functions: acting as a catalyst in AOPs and as an
inorganic coagulant during sedimentation. However, this chemical mineral present
corrosive effect in pipes and pumps. On the other hand, Ferrate (VI) is a strong oxidizer,
which in acidic condition, has the highest oxidation potential even beyond ozone
contributing to AOPs and therefore, disinfection. In addition, it is not corrosive and has a
high solubility in water (approximately 15 g/L) where is converted into trivalent iron or
ferric hydroxide (Fe(OH)3) which is a strong coagulant. Hence, ferrate (VI) is suggested
as a novel chemical able to eliminate heavy metals, suspended particles and organic
matter, without producing hazardous by-products. In any case, more researchs at large-
scale must be conducted in order to study this treatment deeply (Chen et al., 2015;
Talaiekhozani et al., 2017). Furthermore, over the last years, there is a tendency of
synthetizing composite coagulants or using natural biodegradable flocculants. Preparation
of composite coagulants consist of adding organic or inorganic modifiers to the
respective coagulants in order to improve coagulation-flocculation efficiency by adding a
high cationic charge. As an example Al–Fe composite unify advantages of both Al and
Fe polymers, to overcome the problems associated with the individual coagulants. In this
sense, other metals have been incorporated to aluminium ferric polymers such as Zn and
Si (Zhu et al., 2012; Wei et al., 2016). Jia et al., (2017) used composite coagulant
polymeric aluminum ferric sulfate (PAFS) in order to treat urban wastewater where
optimal conditions were [PAFS] = 45 mg/L and pH = 8.5. The removal %COD and
%turbidity were 82.8% and 98.2%, respectively. However, the presence of sulphates or
sodium ions (which could be sometimes found in OMW) could interfere into coagulation
(Jia et al., 2017). On the other hand, some researchers have used natural and eco-friendly
flocculating agents such as ones based on cellulose or derived from plants (Khiari et al.,
 OMW Pretreatment by Assisted Sedimentation Methods 271

2010; Zhu et al., 2010; Nourani et al., 2016; Kono, 2017; Ajao et al., 2018) to reduce the
toxicity and the treatment costs. However, no studies were carried out in OMW.
Considering these new trends, new studies of applicability of these novel flocculants
must be conducted in order to improve flocculation pretreatment of OMW.

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In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 14

CHITOSAN BASED FLOCCULANTS FOR THE

REMOVAL OF HEAVY METAL IONS

Sayaka Fujita1,* and Nobuo Sakairi2

1
Division of Applied Chemistry and Biochemistry,
National Institute of Technology, Tomakomai College,
Tomakomai, Hokkaido, Japan
2
Faculty of Environmental Earth Science, Hokkaido University,
Sapporo, Hokkaido, Japan

ABSTRACT

This chapter focuses on the preparation and characterization of the chitosan based
flocculant for removal of heavy metal ion prepared from chitosan by N-acylation with
ethylenediaminetetraacetic acid monoanhydride (EDTAM). In this study, a series of
chitosan flocculants with various degree of substitution were prepared by changing the
molar ratio of EDTAM to chitosan in the preparation. The structural characterization of
the chitosan flocculants by FTIR and 13C NMR spectroscopy were performed. The newly
introduced functional group provided properties such as being a strong chelating reagent
and an amphoteric polyelectrolyte. It was found that chitosan flocculants had good water
solubility in both acidic and basic regions and precipitated in a narrow pH region, which
is close to its isoelectric point. The chitosan flocculants has an ability to remove heavy
metal ion from aqueous solution by controlling pH or initial concentration of the
flocculant, and removed Cu(II) almost completely. The flocculation mechanism was
investigated using Cu(II) and found that flocculation occurred by charge neutralization
was sensitive both pH conditions and amount of chelated Cu(II) on chitosan flocculants.
The complexation stoichiometry of Cu(II) and the EDTA residues of chitosan flocculant

*
Corresponding Author Email:[email protected].
278 Sayaka Fujita and Nobuo Sakairi

determined by UV-vis titration. As a result, the chitosan flocculants can easily remove
Cu(II) from aqueous solution by only flocculation/precipitation process, which indicated
that the chitosan flocculants could be applicable for remediation and treatment system of
wastewater.

Keywords: chitosan, flocculation, chelating, EDTA, removal of heavy metal ion

INTRODUCTION

Among the large number of environmental pollutants, heavy metals are major water
and soil pollutants. Metals are introduced into aquatic systems by the weathering of soils
and rocks, volcanic eruptions, and a variety of human activities, such as mining,
processing, or using metal-containing materials or products. Most heavy metals are
highly toxic or carcinogenic, and tend to readily accumulate in living organisms through
the food chain. Various methods are available for the removal of heavy metal ions,
including chemical precipitation [1], ion exchange [2], adsorption [3], oxidation [4],
electrochemical treatment [5], and membrane separation [6].
As a water treatment method, flocculation is an efficient and cost-effective
technology for the removal of suspended insoluble organic and inorganic substances from
industrial and domestic wastewater. There are two major types of flocculants: inorganic
additives such as polyaluminum chloride and polyferric sulfate, and organic
macromolecules such as polyacrylamide [7]. These three commonly used flocculants
cause aggregation of dispersed particles and hydrophobic colloids into larger flocs that
can be easily settled and separated [8, 9]. They are used mainly for removal of insoluble
organic and inorganic compounds from contaminated water. As it is considered difficult
to accumulate metal ions into large flocs, flocculation has been regarded as a less
effective method for removal of metal ions.
Chitosan is a linear polysaccharide of -(1,4)-linked 2-amino-2-deoxy-D-
glucopyranose, a semi-synthetic polysaccharide made from such abundant and renewable
biomass as shells of crabs, shrimps, and insects. Behaving as a cationic polyelectrolyte in
acidic aqueous solution, chitosan and its grafted polymers have been investigated as
flocculants to reduce turbidity in wastewater. Chitosan has also the amino groups which
reactive with various transition metal ions. However, the ability of chelating with metal
ions is weaken by protonation of the amino groups, and chitosan has limitation of using
as a flocculant for removal of heavy metal ion.
The purpose of this study is to explore the properties of the chitosan based flocculant
prepared form chitosan with ethylenediaminetetraacetic acid (EDTA) (Figure 1) such as
structural characterization, removal of Cu(II), complexation stoichiometry in order to
clarify flocculation mechanism.
 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 279

EXPERIMENTAL

Materials

Chitosan (deacetylation degree of 95%, average molecular weight of 5-10  105

g/mol) was pur-chased from Hokkaido Soda Co., Ltd. (Hokkaido, Japan). Ethylened-
iaminetetraacetic acid (EDTA) was obtained from Dojindo Laboratories Co., Ltd.
(Kumamoto, Japan). Cu(NO3)2∙3H2O of analytical grade and other regent of preparative
grade were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), and all
chemicals were used without further purification. Water was purified by treatment with
ion-exchanger followed by reverse osmosis with a Millipore Millex-3 apparatus.

Figure 1. Scheme for chitosan based flocculant (EDTA grafted chitosan) synthesis through N-acylation
of chitosan with EDTA monoanhydride.

Apparatus

Fourier transform infrared spectra (FTIR) were recorded on a FT-210 spectrometer

(Horiba Ltd., Japan) using a potassium bromide pellet at a resolution of 2 cm-1. The
samples were scanned from 4000 to 500 cm-1 using an average of 16 scans. 1H NMR and
13
C spectra were recorded on a JNM-ECA600 spectrometer (Jeol Ltd., Japan) at 600.17
and 150.91 MHz, respectively. Turbidity was measured with a V-560 UV-visible
spectrometer (Jasco Co., Japan), using a quart cell with an optical path length of 1 cm at
600 nm. Zeta potential was con-ducted on a Delsa Nano HC particle analyzer (Beckman
Coulter Inc., USA) with a laser light with wave-length of 658 nm at 25°C, using the same
Beckman apparatus with the standard flow cell. The zeta-potential value was calculated
using Smoluchowski equation [10]. Concentration of Cu(II) in aqueous solution was
measured by A-2000 atomic absorption spectrometer (Hitachi Ltd., Japan) which
equipped with a flame atomizer (air/acetylene) and a hollow cathode lamp of Cu
(wavelength of 324.7 nm).
280 Sayaka Fujita and Nobuo Sakairi

Preparation of Chitosan Flocculants

A series of six chitosan based flocculants were prepared from chitosan with EDTA.
EDTA monoanhydride was prepared by a procedure reported by Capretta et al. [11].
Chitosan (2.50 g, 15.6 mmol of D-glucosamine units) was dissolved in 2% aqueous acetic
acid-methanol (1000 ml, 1:1(v/v)). EDTA monoanhydride (0.53 g, 1.95 mmol, 0.25
molar equivalents to the glucosamine unit) was added to the solution, and then the
mixture was stirred vigorously at room temperature for 24 h. The pH of the reaction
mixture was adjusted to 8 by 2% aqueous NaHCO3, and the same amount of EDTAM
was added to the mixture. The mixture was stirred at room temperature for 12 h, and
precipitated by acidification of the mixture to pH 2-3 with 2 mol/l HCl. The resulting
precipitate was separated by filtration, washed with methanol, and then dissolved in 2%
aqueous NaHCO3. The solution was subjected to ultrafiltration using a membrane
(Advantec UK-10; Toyo Roshi Co., Ltd., Japan) with molecular weight cutoff 20,000
against deionized water, and lyophilized to give chitosan flocculant; ED-ch (Na form) 1
(2.59 g, D.S. 7.5%) as white hygroscopic amorphous powder. Using a similar method to
prepare the ED-ch, the other ED-ch 2–6 were prepared. The molar ratio of the chitosan
and EDTA for the preparation of these ED-ch are listed in Table 1.

Solubility Tests

The solubility was evaluated by turbidity measurement based on the method re-
ported by Kubota et al. [12]. Fine powder of ED-ch or chitosan (20 mg) was prepared by
dissolving in 0.1 mol/l HCl (20 ml), and adjusted the pH by addition of small amount of 3
mol/l HCl or 3 mol/l NaOH solutions. The optical transmittance of each sample was
recorded on UV-visible spectrometer at 600 nm. The zeta potential value of each sample
was also measured at different pH.

Removal of Cu(II) from Aqueous Solution

ED-ch (1.00 g) was dissolved in deionized water (50 ml) to prepare 20g/l of ED-ch
stock solution. The ED-ch stock solution was diluted with deionized water to provide
various concentrations of ED-ch solution. A Cu(II) solution (1000 ppm) prepared from
Cu(NO3)2 and 0.1 mol/l HNO3 was diluted with deionized water to give various
concentration of Cu(II) solution. One ml of the ED-ch solution was added to 19 ml of the
Cu (II) stock solution in a 50 ml centrifuge tube, and pH of the mixture was adjusted by
the addition of small amounts of 3 mol/l HNO3 or 3 mol/l CH3COONa. The mixture was
shaken at 120 rpm at room temperature for defined times. The resulting precipitate was
 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 281

settled by centrifugation at 4,000 rpm for 15 min and the supernatant was collected
immediately. The residual Cu(II) concentration in the supernatant was determined by
atomic absorption spectrometer.
The removal of Cu(II) with ED-ch was calculated according to the following
equation:

Removal (%) = (C0-Csup)/C0  100

where C0 is the initial Cu(II) concentration and Csup is the residual Cu(II) concentration of
the supernatant. The experiments were performed three times.
For comparison, the same experiments were also performed using 20 g/l of chitosan
solution in aqueous acetic acid, which was prepared by dissolving in 1wt% acetic acid
aqueous.

Table 1. Preparation of properties of chitosan flocculants

Sample Molar ratio of the starting material D.S. (%)a Water solubilityb
(EDTAM:chitosan)
1 0.25:1 7.5 Insoluble
2 0.5:1 21 Insoluble
3 1:1 39 Soluble
4 1:1 45 Soluble
5 1.5:1 58 Soluble
6 3:1 79 Soluble
a
calculated from integral of H-1protons.
b
sample (5 mg) was dissolved in H2O (5 ml). Transmittance at 600 nm of the sample 95% higher than deionized
water referred as “soluble”.

UV-vis Spectroscopic Titration

Complexation stoichiometry was determined by UV-vis titration. Stock solutions of

ED-ch 6 (3.49  10-3 mol/l) and Cu(II) (2.36  10-3 mol/l) were prepared at pH 5.5 as
follows. ED-ch (75 mg) was dissolved in an appropriate quantity of deionized water, the
pH of the solution was adjusted to 5.5 by the addition of small amounts of 3 mol/l HNO3
or 3 mol/l CH3COONa, and the solution was diluted to 50 ml with deionized water.
Cu(NO3)2 (58 mg) was dissolved in an appropriate quantity of deionized water, the pH of
the solution was adjusted to 5.5, and then the solution was diluted to 50 ml. Analytical
samples were prepared by mixing the Cu(II) stock solution (1 ml) with 2 ml of
appropriately diluted ED-ch stock solution. All analytical samples had the same Cu(II)
concentration, and the molar ratio between EDTA residues and Cu(II) was varied from 0
282 Sayaka Fujita and Nobuo Sakairi

to 2.0. After stirring each freshly prepared analytical sample for 30 min at room
temperature, the UV-vis spectrum was recorded between 200 and 900 nm.

Zeta Potential Measurements

ED-ch 4 (0.25 g) was dissolved in an appropriate quantity of deionized water, the pH

of the solution was adjusted by the addition of small amounts of 3 mol/l HNO3 or 3 mol/l
CH3COONa, and then the solution was diluted to 50 ml with deionized water, giving ED-
ch stock solutions (5.0 g/l). Cu(II) stock solutions (175 mg/l) were prepared and adjusted
the pH of the solution, and then diluting the solution to 50 ml with deionized water.
Analytical samples, with varying molar ratios (0 – 3.0) between EDTA residues and
Cu(II) were prepared by mixing the ED-ch stock solution (0.5 ml) with 6.5 mL of
appropriately diluted Cu(II) stock solution. All analytical samples had the same ED-ch
concentration (0.35 mg/l, 1.1 × 10-3 mmol/l). After stirring the freshly prepared analytical
solutions for 30 min at room temperature, the zeta potential of each sample was
measured.

RESULTS AND DISCUSSION

Preparation of Chitosan Flocculants

In this study, as shown in Figure 1, the preparation of chitosan flocculants was

performed using EDTAM as an acylation regent. Since EDTA grafted chitosan so far
reported was synthesized by using EDTA dianhydride in acetic acid/methanol, the
product was insoluble due to crosslining reactions partially occurred between chitosan
chains [13, 14]. We have been reported the synthesis of EDTA grafted chitosan by use of
EDTA monoanhydride (EDTAM) instead of its dianhydride, in order to prevent the
crosslining reaction to obtain the corresponding water soluble material [15]. A series of
six flocculants (ED-ch) 1-6 were prepared by varying the molar ratio of EDTAM to
chitosan (Table 1). N-acylation of chitosan with EDTAM was carried out under
continually homogenous conditions. Chitosan was treated with EDTAM in aqueous
acetic acid-methanol, and then the reaction was continued under slightly basic media of
pH 8 in aqueous medium where O-acylation of chitosan would be restricted by
hydrolysis. Precipitate obtained by acidification of the reaction mixture with aqueous HCl
was collected by filtration. The carboxyl groups of EDTA residue were converted to
sodium carboxylate groups by dissolving the product in aqueous NaHCO3, and
subsequently the solution was subjected to ultrafiltration. Finally, lyophilization was
performed to give ED-ch as white cotton-like hygroscopic amorphous.
 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 283

Structural Characterization of Chitosan Flocculants

Figure 2 shows the FTIR spectra of chitosan and ED-ch 1-6. These spectra were
normalized by the absorption of C-O-C in the glucopyranose at 1071 cm-1 [16]. With the
exception of the absorption at 1071 cm-1, these spectra display common adsorption bands.
The broad absorption at 3435 cm-1 corresponds to NH and O-H stretching vibration, and
absorption band between 2959 and 2888 cm-1 are attributable to the C-H stretching
vibration. The characteristic absorption bands were observed at 1653, 1592, and 1324 cm-
1
due to the C=O stretching vibration (amide I), N-H bending vibration (amide II), and C-
N stretching vibration (amide III) from the secondary amide, respectively [17]. The
absorption bands of -(1,4) glycoside bridge at 1151 and 914 cm-1. The intensity of
the bands at 1653 and 1592 cm-1 of ED-ch were obviously increased as compared to those
of chitosan, and the new absorption bands at 1636 and 1405 cm-1 due to C-O
asymmetrical and symmetrical stretching vibration of the carboxylate group was detected
in the spectrum of ED-ch [18, 19]. Furthermore, absorption band of ester (C=O stretch)
was not observed at 1720-1770 cm-1 [20, 21]. These observations suggested highly
selective formation of the amino group of chitosan by the reaction with EDTAM. In the
spectra of ED-ch 1 and 2, the appreciable alternation was not observed comparing with
that of chitosan. The spectra of 3-6 showed that intensity of the amide I and II bands at
1653 and 1592 cm-1 increased with an increase in the molar ratio of EDTAM to chitosan.

Figure 2. FTIR spectra of (a) chitosan, ED-ch (b) 1, (c) 2, (d) 3, (e) 5 and (f) 6. These spectra were
normalized by the peak intensity of the carbonyl bands at 1071 cm-1.

Figure 3 shows the 13C NMR spectra of chitosan in DCl/D2O and ED-ch 3 in
NaOD/D2O. In the spectrum of chitosan, the resonances at 98.3, 77.8, 75.6, 70.9, 61.1
284 Sayaka Fujita and Nobuo Sakairi

and 56.8 ppm were assigned to the carbonyl carbon, C1, C4, C5, C3, C6 and C2 [22]. In
the spectrum of ED-ch, the new resonances appearing at 61.6 and 55.0 ppm due to two
kinds of methylene carbon in EDTA moiety [23]. Moreover, the new resonances at 182.5
and 182.1 ppm were assignable to carbonyl carbons of carboxylate groups in the EDTA
moiety, and the signal at 177.5 ppm corresponds to carbonyl carbon of amide group.
These results also supported the assigned structure of ED-ch.
The degree of substitution (D.S.) of the EDTA residue in the products was calculated
the area ratio between the anomeric protons of substituted (H-1’) and unsubstituted (H-1)
glucosamine residues in 1H NMR spectrum (spectra not shown). The results of
calculation are shown in Table 1. The D.S. of ED-ch was able to change by the molar
ratio of starting chitosan and EDTAM used.

Figure 3. 13 C NMR spectra of chitosan in DCl/D2O (top) and ED-ch 3 (bottom) in NaOD/D2O at rt.

Solubility of Chitosan Flocculants

Solubility is one of the most basic characteristics in utilizing as flocculant. We

investigated water solubility by turbidity measurement, and the sample was determined
soluble when the transmittance was higher than 95%, compared to that of deionized water
(Table 1). The transmittance of ED-ch 1 and 2 (D.S. 7.5 and 21%) were lower than 95%
and these were poorly soluble. ED-ch 3-6 with over D.S. 39% was found to be dissolved
readily in deionized water without adding any acid in contrast to original chitosan, and
the transmittance of sample was higher than 95%. This can be caused by that inter and
 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 285

intramolecular hydrogen bonds of chitosan were disrupted by introduction of EDTA as

bulky group, and the carboxyl groups were also enhanced affinity in water.
The dependence on solubility of ED-ch 3 was investigated by turbidity measurement
in aqueous solution in the pH range of 1-8 (Figure 4). ED-ch 3 was soluble in the wide
range of pH region, including both acidic and basic region. The zeta potential of ED-ch 3
was positive value at acidic conditions, while it changed to negative value at neutral or
basic conditions, which suggested that ED-ch behaved as a cationic or an anionic
polyelectrolyte. These results are explainable by consideration that the amino and
carboxyl groups can be protonated or deprotonated depending on pH of the solution. The
transmittance became unstable in the narrow pH region of 3.3-4.5 which approaches the
isoelectric point (IEP) of ED-ch 3. The transmittance decreased in this pH range, the
transmittance gradually increased to maximum of approximately 95% after leaving this
sample to stand for 10 min. As shown in Figure 4a, b, fine particles presented in the
suspension gradually became larger aggregates and settled out in the clear.
The solubility and IEP of ED-ch with various D.S. are summarized in Table 2. The
IEP were approximately determined from interpolation as dotted line in the zeta potential
data. The insoluble pH region narrowed and moved to the lower pH region with
increasing D.S.. The IEP also shifted to lower pH with increasing D.S.. It is reasonable
for ED-ch with higher D.S. to precipitate in the lower pH region because it has fewer
amounts of amino groups.

Figure 4. The optimal transmittance (T%) and the zeta potential of aqueous solutions of ED-ch 3 and
chitosan under various pH. Inset: The photographs of ED-ch with 0.1% aqueous solution at pH 4.0 (a)
immediately after adjusting pH and (b) after leaving to stand for 10 min.
286 Sayaka Fujita and Nobuo Sakairi

Table 2. Solubility of ED-ch with various D.S. at various pH levels

a
opened bar means “soluble”; closed bar means “insoluble”; hatched bar means “the transmittance was
unstable”.
b
determined from the zeta potential.

Effect of pH on the Removal of Cu(II)

The removal experiments depending on pH were undertaken by use of Cu(II) as a

model pollutant of heavy metal ion. After addition of ED-ch, it was observed visually
formation of blue-colored flocculent precipitates at pH 3.0-6.0, whereas the precipitate
formation was not observed under other pH conditions. Furthermore, the blue precipitates
could be readily separated by centrifugation at 4000 rpm.
Figure 5 shows the pH dependent removal of Cu(II) by ED-ch 3 for contacting time
of 24 h and that of chitosan is shown in Figure 5 for comparison. The removal Cu(II) by
ED-ch 3 was remarkably different from that of chitosan. The removal Cu(II) increased
with increasing pH up to 4.5 from 3.0 and the maximum removal of 100% was obtained
at pH 4.5, which was higher than that of chitosan at pH 6.0. These results showed that
ED-ch has the excellent flocculation ability of toward to Cu(II). The optimal pH of ED-
ch flocculation suggest that the electrostatic attraction between NH3+ and COO- in ED-ch
is reduced the repulsion of the polymer at around isoelectric point [24], which would
cause a high efficient flocculation ability.
By changing the contact time, the removal of Cu(II) was examined in Cu(II)
solutions of the initial concentration in range of 10-50 mg/l under the optimum pH of 4.5
(Figure 6). Surprisingly, the removal of Cu(II) with ED-ch was extremely quick and
highly efficient, and the removal ratio reached approximately 100%. It took only 10 min
to be equilibrium in Cu(II) solution of 50 mg/l. The solid EDTA-chitosan needed 4 h to
attain equilibrium conditions using Co(II) and Ni(II) aqueous solution (100 mg/l) [25].
The extremely short contact time necessary for the water-soluble ED-ch is explainable by
 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 287

consideration of homogeneity of the reaction. When both solutions of ED-ch and Cu(II)
are mixed, the chelation of Cu(II) with EDTA occur quickly in the homogeneous
conditions.

Figure 5. The effect of pH on the removal of Cu(II) contacting with chitosan and ED-ch 3 by
flocculation/precipitation. Initial concentration of Cu(II) 10 mg/l, dose of polymer ligand 1 g/l, contact
time 24 h, shaking rate 120 rpm, rt.

Figure 6. The effect of contact time on the removal of Cu(II) by ED-ch 3 at pH 4.5. Insrt: The
magnified figure at range of contact time 0-30 min. Initial concentration of Cu(II) 10-50 mg/l, dose of
polymer ligand 1 g/l, shaking rate 120 rpm, rt.
288 Sayaka Fujita and Nobuo Sakairi

Complexation Stoichiometry

The ED-ch exhibited a high removal efficiency for Cu(II) by flocculation within a
narrow pH range, which is close to the IEP of ED-ch. It was also confirmed that most
Cu(II) could be removed at the pH value if excess amounts of EDTA residues were added
relative to Cu(II) (EDTA/Cu(II) molar ratio = 8.6). It is well known that EDTA forms
chelate complexes with Cu(II) in a 1:1 molar ratio in a range of pH values. As EDTA and
Cu(II) form 1:1 chelate complexes with an extremely high stability constant (log K =
18.7) [26], the excess amounts of ED-ch necessary for flocculation were unexpected. To
determine the stoichiometry between EDTA residues in ED-ch and Cu(II), we carried out
UV-vis titrations [27, 28] of Cu(II) with ED-ch at pH 5.5. As precipitate formation can
influence the absorbance of these samples, the experiment was carried out using ED-ch 6,
which is soluble over a wide range of pH.
Figure 7 shows the UV-vis spectra of ED-ch-Cu(II) chelates at pH 5.5. The spectra
show an adsorption maximum at 730 nm for the complex formed between Cu(II) and
different concentrations of ED-ch. The absorbance increased with increasing
concentrations of ED-ch. To determine the complex stoichiometry, the CEDTA/CCu was
plotted against absorbance of complex at 730 nm (Figure 7 right). Absorbance of the ED-
ch-Cu(II) complex increased in a nearly linear manner with increasing CEDTA/CCu, when
CEDTA/CCu was less than 1.0, and then reached plateau when CEDTA/CCu was more than 1.0.
This plateau of the plot (that is, absorbance was not changed with increasing CEDTA/CCu)
indicated that the chelation reaction had reached saturation. The intersection at the
turning point in the plot occurred at CEDTA/CCu = 1.0, suggesting that the EDTA residues
of ED-ch form chelation complex with Cu(II) in 1 : 1 molar ratio. This result was agreed
the stoichiometry of original EDTA with Cu(II).

Figure 7. UV-vis spectra of Cu(II) (7.87  10-4 mol/l) at pH 5.5 with ED-ch 6 (left) (from a to u: 0, 9.96
 10-5, 1.99  10-4, 2.99  10-4, 4.00  10-3, 4.98  10-4, 5.97  10-4, 6.97  10-4, 7.97  10-4, 8.96  10-4,
9.96  10-4, 1.10  10-3, 1.19  10-3, 1.29  10-3, 1.39  10-3, 1.49  10-3, 1.59  10-3, 2.69  10-3, 1.79 
10-3, 1.89  10-3, and 1.99  10-3 mol/l). Plot of absorbance at 730 nm against CEDTA/Ccu (right).
 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 289

Effect of Complexation Stoichiometry on the Removal of Cu(II)

As described in the previous section, the EDTA residues in ED-ch form chelate
complexes with Cu(II) in a 1:1 molar ratio at pH 5.5. Therefore, we expected that
discovery of new flocculation method other than pH control would increase remarkably
the removal efficiency of ED-ch. The removal ability under higher Cu(II) concentration
was investigated to clarify the effect of complexation stoichiometry on flocculation.
Experiments on the removal of Cu(II) by flocculation with ED-ch 4 were carried out by
changing the EDTA/Cu(II) molar ratio at pH 5.5, which is not closed to the IEP of ED-ch
4 (IEP = 3.0), and the results are shown in Figure 8. The theoretical removal efficiencies,
shown by dashed lines in the figure, were calculated by assuming 1:1 complexation
between the EDTA residues in ED-ch and Cu(II) and complete separation of ED-ch–
Cu(II) complexes from aqueous solution. The zeta potential of the mixture solution of
ED-ch and Cu(II) are also shown in figure.

Figure 8. Effect of EDTA/Cu(II) molar ratio on removal of Cu(II) by ED-ch 4 (left) and zeta potential
of mixture solution of ED-ch (right) at pH 5.5. Initial concentration of Cu(II): 10 mg/l, shaking rate:
120 rpm, rt.

The removal efficiency initially increased with increasing the molar ratio of
EDTA/Cu and the maximum removal was almost 100% at the molar ratio of 1.26-1.35.
Subsequently the removal efficiency gradually decreased with increasing the molar ratio.
The zeta potential increased with increasing the molar ratio of EDTA/Cu(II), and reached
approximately zero when the molar ratio was 1.2. This condition is the same as the
optimum conditions for high removal efficiency. These results indicated that the
flocculation process is controlled by the charge neutralization mechanism. Subsequently,
when the removal efficiency decreased with increasing molar ratios (above the optimum
molar ratio), the zeta potential also decreased to negative values, preventing flocculation
[29, 30, 31]. These charges should increase repulsion among particles, and thereby hinder
flocs growing into larger ones.
290 Sayaka Fujita and Nobuo Sakairi

We thought about factors that could affect charge neutralization to achieve effective
flocculation of ED-ch. EDTA is known to have four dissociation constants, and exists as
a zero-valent anion (H4Y) to a tetravalent anion (Y4-) at various pH values [32]. As EDTA
exists as a tetravalent anion (Y4-) under neutral and basic conditions, the EDTA residues
in ED-ch are expected to exist as trivalent anions, i.e., with three carboxylate ions. When
an EDTA residue forms a chelate complex with Cu(II), EDTA residue exist as
monovalent anion because two carboxylate ions are used by chelate formation. Therefore,
the coordination with Cu(II) should decrease the negative charges of the EDTA
residues in ED-ch.

CONCLUSION

A novel chitosan flocculant for removal of heavy metal ion was easily obtained by N-
acylation of chitosan with EDTA which is a strong chelating agent. Having property as
an amphoteric polyelectrolyte, EDTA grafted chitosan (ED-ch) had good water solubility
in both acidic and basic regions and precipitated in a narrow pH region, which is close to
its isoelectric point. The ED-ch could remove heavy metal ion from aqueous solutions,
and Cu(II) was almost completely removed by flocculation/precipitation process. The
removal of Cu(II) could be controlled by the pH of solutions as well as initial
concentration of the flocculant. Zeta-potential measurement of the Cu(II) solution in the
presence of ED-ch proved that the flocculation could be explained by the charge
neutralization. ED-ch can easily get rid of heavy metal ions from aqueous solution, and
are expected to be applicable for remediation and treatment system of wastewater.

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Co(II) and Ni(II) by EDTA- and/or DTPA-modified chitosan: Kinetic and
equilibrium modeling, Chem. Eng. J., 161, 73-82.
[26] R. W. S. & C. N. R. (1956). A Rapid Electrochemical Method for the
Determination of Metal Chelate Stability Constants, J. Am. Chem. Soc., 78, 5513-
5518.
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interaction with selected transition and post-transition metal ions, Coord. Chem.
Rev., 245, 11–16.
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of an amphoteric chelating polymer flocculant with Cu(II), Pb(II), Cd(II), and
Ni(II), Spectrochim. Acta., Part A, 118, 765–775.
[29] E. A. L., M. T. O., J. L. J. & A. O. (2014). Coagulation–flocculation mechanisms in
wastewater treatment plants through zeta potential measurements, J. Hazard.
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 Chitosan Based Flocculants for the Removal of Heavy Metal Ions 293

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In: Flocculation: Processes and Applications ISBN: 978-1-53614-339-3
Editor: Eleonora Vollan © 2019 Nova Science Publishers, Inc.

Chapter 15

DETERMINATION OF THE KINETIC COEFFICIENT

OF AGGREGATION AND THE KINETIC COEFFICIENT
OF RUPTURE IN THE TURBIDITY REMOVAL PROCESS

Joseane Debora Peruço Theodoro1,

Paulo Sergio Theodoro2
and Rosangela Bergamasco3
1
Department of Environmental Academic,
Federal Technological University of Paraná,
Londrina, Paraná, Brazil
2
Department of Chemical Engineering, State University
of West Paraná, Toledo, Paraná, Brazil
3
Department of Chemical Engineering, State
University of Maringá, Maringá, Paraná, Brazil

ABSTRACT

Obtaining drinking water using a coagulation / flocculation process is intended to

remove impurities that are suspended or dissolved in contaminated water through the use
of natural and chemical coagulating agents. In order to achieve satisfactory removal of
contaminants from water, it is necessary to obtain efficiency in water treatments. In the
treatment of water either by sedimentation or flotation followed by filtration is preceded
by the coagulation process of the suspended particles of the aqueous medium and by the
formation of flocs. It is of fundamental importance that during the process of particle
removal the ideal value of the agitation rate of the aqueous medium, the agitation time
and the sedimentation or flotation time of the flocs formed is determined, in order to
obtain the optimization of the treatment process. In this way, the present study aimed to
296 J. D. Peruço Theodoro, P. S. Theodoro and R. Bergamasco

apply the Bratby method in the characterization of the turbidity removal process, through
the determination of the kinetic aggregation coefficient (KA) of the flocs and the kinetic
coefficient of rupture (KB) of the flocs. The experimental data were obtained by turbidity
determination (uT) of turbid water, consisting of a liquid solution containing kaolin and
humic acid. The coagulating agent used was iron chloride. The results were evaluated
based on flocculation kinetics by means of the aggregation constants (KA) and rupture
(KB). It was observed that floc aggregation resulted in the removal of 80% of turbidity,
with sedimentation time in 80 minutes and mean velocity gradient (GT) at 100 rpm,
resulting in a ratio of the kinetic coefficients of fracture and aggregation flock (KB/KA)
equals 0.005.

Keywords: ferric chloride, kinetic study, Bratby method

1. INTRODUCTION

According to Oliveira (2008) during flocculation, the agitation imposed on the liquid
medium promotes two effects simultaneously: aggregation and rupture (Figure 1).
Basically, the aggregation is the result of the encounters of the destabilized particles,
being that the agitation promotes a higher rate of encounters, forming agglomerates or
flocs; in the rupture occurs the erosion of the flocs by shearing forces, which are usually
accentuated with intense agitation or a great flocculation time, causing the partial or total
degradation of the flocs in a few seconds.

Source: FILHO (2010).

Figure 1. Aggregation and rupture of flocs in the flocculation step.

The use of mathematical modeling related to flocculation kinetics aims at estimating

the flocculation kinetics considering the aggregation and rupture phenomena (DI
BERNARDO, 2005).
According to Oliveira (2008) the Bratby Method equation proposed by Argaman and
Kaufman (1970) to describe the kinetics of flocculation in a static reactor, assuming the
 Determination of the Kinetic Coefficient of Aggregation … 297

number of primary particles equal to the remaining turbidity, is similar to Equation (1)
given by :

𝑑𝑛
𝑑𝑡
= −𝐾𝐴 𝑛𝑇 𝐺𝐹 + 𝐾𝐵 𝑛0 (𝐺𝐹 )2 (1)

In which:
n0 = number of primary particles per unit volume at time t = 0;
nT = number of primary particles per unit volume at time t;
dn/dt = variation of number of primary particles per unit volume with according to
time;
KA = coefficient of aggregation;
KB = coefficient of rupture; and
GF = mean velocity gradient during flocculation.

The term − KA.nT.GF indicates the disappearance of primary particles to give

rise to the flocs and the term +KB.n0.(GF)2 indicates the appearance of particles by erosion
of the flocs. The variation of the number of primary particles in relation to time
is given by the sum of these two simultaneous effects. Integrating Equation (1) and
assuming the number of primary particles equal to the remaining turbidity, we
get Equation. (2):

𝑁0 𝐾 𝐾 −1
𝑁1
= [𝐾𝐵 𝐺𝐹 + (1 − 𝐾𝐵 𝐺𝐹 ) 𝑒 −𝐾𝐴 𝐺𝐹 𝑇𝐹 ] (2)
𝐴 𝐴

In which:
N0= initial turbidity of the supernatant;
N1=final turbidity of the supernatant; and
TF=flocculation time.
Rearranging Equation (2), we have Equation (3):

𝐾
1 (1− 𝐵𝐺𝐹 )
𝐾𝐴
𝐾𝐴 = 𝑙𝑛 [ ] (3)
𝐺𝐹 𝑇𝐹 1 𝐾
( 𝑁 − 𝐵𝐺𝐹 )
0 𝐾𝐴
𝑁

Assuming that there is no more aggregation or disintegration of primary particles of

the flocs after a relatively long sedimentation period, Equation (3) can be equaled to zero,
resulting (Bratby, 1977) in Equation (4):
298 J. D. Peruço Theodoro, P. S. Theodoro and R. Bergamasco

𝐾𝐵 1
= 𝑁 (4)
𝐾𝐴 𝐺𝐹 0
𝑁

2. METHODOLOGY

Preliminary tests were performed to evaluate the removal of the parameter turbidity
present in the water in the study, in which the composition consists of the presence of
humic acid at the initial concentration of 4 mg/L and caulin at a concentration of 5 g/L of
humic acid as the stock solution, then it was diluted to obtain solutions of turbidity values
150 uT, for the ferric chloride inorganic coagulating agent. To adjust the pH, 0.1 M
sodium hydroxide solution and 1M hydrochloric acid were used. It was used a 1 g/L
concentration solution for the coagulant ferric chloride.
Coagulation / flocculation / sedimentation assays were performed, aliquots being
removed at various times (1 to 60 minutes) and turbidity parameters were determined.
The turbidity was determined in a HACH DR / 2010 spectrophotometer at wavelength
860 nm, according to a procedure determined by Standard Methods (APHA, 2012).
The coagulation / flocculation assays were performed in Jar Test brand MILAN
Scientific Equipment. The pH parameter was determined in pHmeter TECNAL model
pHmeter TEC-2. The speeds used in the coagulation / flocculation assays to provide fast
mixing was set at 100 rpm and a fast mixing time of 1 min, while that the speed to
provide the slow mixing at 10 rpm and slow mixing time of 10 minutes, being the
sedimentation time = 60 minutes during the development of the experiments
(MADRONA, 2010).
The kinetic study of the aggregation and rupture of the particles by the Bratby
method was determined for maximum time of removal of 80 minutes (collections of 1
min, 20 min, 40 min, 60 min and 80 min), for fast mixing at 100 rpm and fast mixing
time of 1 minute, slow mixing at 10 rpm and slow mixing time of 10 minutes, settling
time = 80 minutes during the development of the experiments.
Figure 2 shows the flowchart of the kinetic study performed for the coagulation /
flocculation process.

Figure 2. Flowchart of the methodology used for the kinetic study of the coagulation / flocculation
process.
 Determination of the Kinetic Coefficient of Aggregation … 299

3. RESULTS

The results for the kinetic study of aggregation and rupture (KB/KA) (Equation 4) in
the coagulation / flocculation / sedimentation process are shown in Table 1, which
indicates the turbidity parameter removal data obtained in the experiment and turbidity
initial dose is 150 uT.
Figure 3 represents a graphic of N0/N (Equation 4) which stands for the ratio of initial
turbidity to final turbidity versus time.
After the linear adjustment (Figure 3), it was verified that the ratio of turbidity
parameter removal increased as a function of time in the coagulation / flocculation
process with the use of the chemical coagulant ferric chloride.

Table 1. Turbidity parameter removal data

t (minute) Turbidity (uT) N0/N KB/KA

1 70 2.14 0.04
20 42 3.57 0.02
40 15 10 0.01
60 10 15 0.006
80 8 18.75 0.005

Figure 3. Ratio of turbidity parameter as a function of time.

Figure 4 shows the graphic of the coefficient of rupture with the coefficient of
aggregation (KB/KA) (Equation 4) as a function of time. According to Moreira (2003) the
size of the flocs, once formed or aggregated, any disturbance of the system, however
small, may cause its rupture.
300 J. D. Peruço Theodoro, P. S. Theodoro and R. Bergamasco

Figure 4. Relationship of the coefficient of rupture and aggregation (KB/KA) as a function of time.

Thus, according to Figure 4, the KB/KA ratio is inversely proportional to time. Figure
5 shows a graphic of KB/KA as a function of turbidity parameter.

Figure 5. KB/KA as a function of turbidity parameter.

According to Figure 5, the KB/KA ratio is directly proportional to the turbidity

parameter.
Figure 6 shows a graph of N0/N as a function of KB/KA.

Figure 6. Relation of initial turbidity and final turbidity (N0/N) as a function of the ratio of the
coefficient of rupture and coefficient of aggregation (KB/KA).
 Determination of the Kinetic Coefficient of Aggregation … 301

In Figure 6, the removal ratio of the turbidity parameter was found to decrease with
the ratio of the coefficient of rupture to the coefficient of aggregation in the water
treatment process with the use of ferric chloride as a coagulant.
According to Moreira (2003) that the chemical coagulant aluminum sulphate
Al2(SO4)3.18H2O was used for the pH of coagulation / flocculation to values between 6.0
and 6.5 with reason to allow the prevailing coagulation mechanism by scanning, it was
concluded that the higher the coefficient of aggregation tended to be, the lower the
relationship between the flocculation times in the batch reactor and the continuous flow
reactor.
Many water treatment plants (ETAs) that work in hydraulic overload generally
choose to work with iron salts in preference to chemical coagulants based on aluminum
salts. This is happening, because the iron allows to obtain higher values of aggregation
constants and lower values of rupture constants when compared with aluminum salts, the
quality of the decanted water can be significantly improved for a flocculation system
operating with the same time of hydraulic detention (MOREIRA, 2003).
Bache and Gregory (2010) affirm that the size of the floc can be pointed as the factor
that exerts greater influence on the density and the force of formation of flocs. According
to Bache et al., (1997) and Jarvis et al., (2005) the strength of the floc is directly related
to the structure of the floc and, therefore, dependent on the formation process of the flocs.
The flocs do not grow infinitely. The growth reaches a steady state for a given shear
condition. Generally the growth of the flocs is related to the break, so the rate of
aggregation is the balance between the formation and rupture of the aggregates.

CONCLUSION

The results evaluated based on the kinetics of flocculation by means of the

aggregation constants (KA) and rupture (KB) by the Bratby method showed that the floc
aggregation resulted in the ratio directly proportional to the turbidity parameter and
inversely proportional to the time.

REFERENCES

APHA – American Public Health Association. Standard Methods for the Examination of
Water and Wastewater. 22 ed. Washington, 2012.
Argaman, Y. and Kaufman, W. J. (1970) Turbulence and Flocculation. J. Sanit. Eng. Div.
ASCE 96, 223.
302 J. D. Peruço Theodoro, P. S. Theodoro and R. Bergamasco

Bache, D. H.; Johnson, C.; Mcgillian, J. F.; Rasool, E. A conceptual view of floc
structure in the sweep floc domain. Water Science and Technology. v.36, p. 49-56,
1997.
Bache, D. H. and Gregory, R. Flocs and separation processes in drinking water treatment:
a review. Journal of Water Supply: Researche and Technology-AQUA. v.59, n.1,
2010.
Bratby, J. et al. Design of flocculation systems from batch test data. Water SA. v.3, n. 4,
p. 173-182, 1977.
Di Bernardo, L. et al. Use of mathematical modeling for the design of mechanized
flocculation chambers in series in water treatment plants. Sanitary and
Environmental Engineering, v. 10, n. 1, p. 82-90, 2005.
Filho, S. S. F., Physical-Chemical Processes I - Coagulation. Escola Politécnica da USP
Department of Hydraulic and Sanitary Engineering- (2010).
Madrona G. S. Extraction / Purification of the Moringa oleifera Lam Seed Active
Compound and its use in the Treatment of Water for Human Consumption. Thesis of
Doctorate in Chemical Engineering, UEM, Maringá -PR, 2010.
Moreira, H. A.- The Flocculation Process in the Treatment of Water Supply:
Transposition of Jury Test Results for Real-Scale Flocculation Systems -22º
Brazilian Congress of Sanitary and Environmental Engineering Joinville - Santa
Catarina - I-082, 14-19 of September 2003.
Oliveira, D. S. Evaluation of the turbidity removal efficiency due to variations in the
length of helical tubular flocculators / Danieli Soares de Oliveira. Dissertation
(master’s degree) - Federal University of Espírito Santo, Technological Center p.
116, 2008.
Jarvis, P.; Jefferson, B.; Gregory, J. and Parsons, S.A. A Review of floc strength and
breakage. Water Research. v.39, p. 3121-3137, 2005.
 INDEX

algal cells, 116, 117, 123, 124, 126, 131, 132, 212,
A
214
algal organic matter, vii, x, 107, 108, 110, 111, 131,
acetic acid, 25, 108, 280, 281, 282
134, 135, 137, 139, 140, 141, 142, 216
acid, viii, xii, xv, 13, 14, 15, 16, 18, 20, 21, 22, 23,
alkalinity, 172, 262, 264, 266, 267, 275
25, 26, 27, 36, 37, 39, 43, 48, 50, 72, 74, 76, 77,
aluminium, vii, x, 5, 6, 32, 116, 118, 120, 121, 122,
108, 111, 122, 134, 136, 158, 187, 190, 200, 201,
124, 134, 137, 143, 264, 265, 267, 270, 271
219, 229, 258, 265, 267, 269, 272, 277, 278, 279,
aluminum sulfate, vii, ix, 47, 48, 50, 56, 69, 145
280, 281, 282, 284, 296, 298
amino acids, 110, 112, 113, 120, 126, 141, 268
acidic, x, xv, 8, 22, 29, 30, 72, 107, 115, 116, 117,
ammonium ions, xiv, 218, 225, 229, 236, 237, 245,
120, 122, 125, 126, 158, 203, 208, 226, 229, 264,
251, 253
267, 268, 270, 277, 278, 285, 290
amorphous precipitate, 249
activated carbon, 15, 109, 131, 134, 141, 178, 214,
anaerobic digestion, 66
215, 235, 256
aquatic systems, 203, 278
acylation, viii, xv, 277, 279, 282, 290
aqueous solutions, 4, 38, 41, 43, 113, 285, 290, 291,
adsorption, 3, 6, 7, 21, 26, 27, 38, 83, 84, 85, 86, 87,
292
88, 89, 109, 118, 120, 121, 122, 125, 131, 134,
aromatic compounds, 4, 263
136, 137, 140, 141, 157, 170, 177, 178, 182, 208,
aromatic rings, 35
209, 215, 221, 235, 242, 243, 249, 251, 254, 255,
assisted sedimentation, viii, xiv, 259, 268
256, 257, 261, 265, 268, 278, 283, 288, 291, 292
atmosphere, 4, 54, 99
advanced oxidation process, 170, 176, 270
agglomeration, 2, 50, 83, 85, 220, 223
aggregation, vi, viii, 2, 82, 83, 84, 85, 86, 87, 88, 89, B
97, 98, 103, 105, 121, 138, 220, 260, 266, 278,
295, 296, 297, 298, 299, 300, 301 bacteria, 8, 9, 10, 11, 17, 18, 20, 22, 27, 50, 56, 82,
aggregation process, 84, 87, 138 156
agriculture, 30, 48, 66, 102, 171 bacterium, 11, 13, 20, 21, 28, 34
aldehydes, 110, 112, 113 bicarbonate, 229, 264
algae, viii, ix, 1, 10, 18, 22, 23, 81, 82, 108, 109, biochemistry, 136
111, 116, 117, 122, 124, 126, 130, 131, 135, 136, biocompatibility, 3, 10, 23
137, 138, 139, 156, 198, 200, 208, 210, 214, 215, bioconversion, 136
216, 223, 235, 266 biodegradability, viii, 1, 2, 3, 4, 8, 10, 17, 30, 35,
172, 219, 222, 229, 234, 235, 251, 273
304 Index

biodegradation, 38, 219, 229, 236, 275 chelating, xv, 27, 170, 178, 277, 278, 290, 292
biofuel, 116, 126 chemical, vii, viii, ix, x, xi, 1, 2, 3, 4, 7, 8, 10, 11, 12,
biogas, 66, 99, 101, 102, 219 13, 14, 15, 16, 21, 22, 23, 24, 26, 28, 29, 30, 32,
biological activity, 11, 43 33, 42, 43, 47, 48, 66, 69, 81, 83, 85, 87, 97, 102,
biological processes, 99 104, 116, 143, 144, 145, 147, 150, 152, 155, 156,
bioluminescence, 40 157, 166, 169, 170, 171, 174, 176, 180, 181, 186,
biomass, 11, 21, 24, 32, 34, 36, 49, 102, 103, 116, 198, 199, 215, 222, 223, 225, 226, 229, 256, 260,
123, 126, 136, 235, 251, 278 262, 264, 265, 269, 270, 272, 274, 275, 278, 295,
biomaterials, 2, 40 299, 301
biomedical applications, 37 chemical bonds, 260
biomolecules, 6, 45 chemical characteristics, ix, 69
biopolyelectrolytes, vi, xi, 169, 170, 171, 172, 173, chemical interaction, 85
174, 176 chemical properties, 3, 33
biopolymers, ix, 2, 4, 8, 10, 22, 24, 25, 26, 32, 35, chemical reactions, ix, 47, 48
38, 41, 43, 47, 48, 49, 51, 52, 56, 57, 58, 59, 61, chemical reactivity, 3
62, 64, 67, 86, 113, 122, 124, 131, 223, 226, 243, chitosan, vi, viii, xii, xiv, xv, 2, 6, 8, 12, 19, 23, 24,
292 25, 27, 28, 30, 31, 33, 35, 37, 38, 39, 42, 43, 44,
bioremediation, 22, 40 116, 117, 124, 125, 129, 135, 169, 171, 172, 173,
biosafety, viii, 1, 2, 30, 31 174, 175, 176, 178, 209, 210, 211, 213, 214, 215,
biosensors, 41 218, 222, 226, 227, 239, 241, 242, 243, 244, 245,
biosynthesis, 10, 44, 137 246, 249, 250, 251, 252, 253, 254, 255, 258, 267,
biotechnological applications, 12, 20, 31 269, 277, 278, 279, 280, 281, 282, 283, 284, 285,
biotechnology, 23, 65, 177 286, 287, 290, 291, 292, 293
blue-green algae, 198, 200 chlorination, 108, 133, 136, 166, 215, 235
Bratby method, viii, 296, 298, 301 chlorine, 48, 82, 138, 166, 275
Brazil, vii, viii, ix, xii, 1, 44, 69, 70, 71, 76, 78, 79, chromatography, 122, 272
81, 91, 92, 143, 145, 152, 155, 157, 165, 166, chromium, 27, 144, 152, 177, 238
179, 180, 186, 187, 188, 295 climate, 54, 99, 223
coagulant, v, vii, viii, ix, xi, xii, xiii, xiv, 7, 13, 16,
31, 35, 38, 41, 48, 49, 51, 53, 54, 57, 58, 59, 60,
C
61, 62, 64, 65, 69, 70, 72, 74, 75, 78, 86, 116,
117, 119, 121, 123, 124, 125, 126, 128, 129, 130,
carbon, 11, 15, 28, 34, 43, 44, 49, 109, 131, 132,
131, 133, 136, 143, 144, 145, 146, 147, 149, 150,
134, 136, 141, 170, 171, 172, 177, 178, 200, 214,
152,155, 157, 158, 159, 161, 163, 164, 165, 166,
215, 226, 235, 256, 262, 284
167, 171, 172, 174, 176, 178, 179, 181, 185, 193,
carbon tetrachloride, 43
195, 197, 199, 201, 202, 207, 208, 209, 210, 211,
carboxyl, 6, 14, 21, 26, 126, 282, 285
212, 215, 218, 220, 222, 223, 226, 239, 242, 244,
carboxylic acids, 112, 268
245, 246, 249, 250, 251, 252, 253, 255, 257, 258,
carboxylic groups, 27
261, 262, 263, 264, 265, 266, 267, 268, 269, 270,
carboxymethyl cellulose, 26, 32
271, 274, 275, 298, 299, 301
cell surface, 116, 117, 122, 127
coagulation mechanism, 108, 117, 118, 136, 176,
cellular organic matter, 109, 113, 128, 132, 138, 139,
215, 301
141
coagulation process, 8, 70, 180, 209, 213, 265, 295
cellulose, 2, 3, 6, 10, 11, 12, 15, 17, 23, 26, 29, 32,
coagulation-flocculation, vii, viii, xi, xiii, 15, 16, 25,
33, 35, 37, 38, 40, 43, 44, 177, 270, 273, 275, 290
29, 33, 36, 42, 64, 71, 73, 74, 76, 77, 78, 145,
charge density, 7, 8, 87, 115, 124, 171, 172, 173,
149, 155, 169, 170, 171, 172, 173, 174, 176, 186,
174, 176, 223, 243
197, 198, 199, 201, 202, 203, 204, 206, 207, 208,
charge neutralization, xv, 5, 6, 7, 26, 28, 108, 118,
209, 212, 213, 215, 235, 256, 263, 270, 271, 273,
122, 123, 176, 203, 208, 261, 265, 277, 289, 290
274
 Index 305

colloids, 6, 8, 29, 34, 37, 38, 83, 84, 96, 133, 141, dosage, xiii, 6, 8, 9, 22, 24, 30, 139, 174, 197, 199,
215, 220, 221, 222, 242, 260, 263, 265, 268, 278 201, 202, 203, 204, 205, 206, 207, 209, 210, 212,
composition, vii, viii, x, xii, 8, 16, 20, 21, 22, 26, 30, 216, 253, 262, 265, 267
39, 49, 53, 89, 107, 110, 128, 129, 130, 135, 136, drinking water, x, xiii, 48, 49, 62, 63, 64, 66, 67, 79,
137, 144, 179, 219, 224, 238, 261, 298 107, 108, 110, 126, 133, 134, 138, 140, 166, 197,
compounds, vii, viii, x, xiii, 4, 29, 44, 49, 82, 107, 198, 199, 200, 208, 213, 214, 215, 216, 260, 266,
108, 109, 110, 112, 115, 118, 120, 122, 124, 125, 267, 273, 295, 302
126, 131, 136, 197, 198, 219, 226, 229, 236, 237,
242, 249, 250, 251, 252, 253, 260, 263, 268, 269,
E
270, 272, 275, 278
compression, 28, 84
effluent, viii, x, xii, xiv, 11, 17, 20, 28, 29, 30, 32,
conductivity, x, xii, xiii, 143, 145, 149, 150, 171,
33, 38, 41, 44, 92, 95, 97, 98, 99, 101, 118, 143,
172, 180, 181, 182, 187, 188, 190, 193, 194, 225,
144, 145, 146, 147, 149, 150, 152, 153, 167, 170,
231, 234, 235, 262
179, 180, 181, 182, 185, 186, 188, 191, 259, 260,
consumption, xiii, 53, 64, 70, 95, 101, 144, 170, 174,
262, 265, 266, 267, 273, 274, 275
186, 187, 190, 191, 193, 194, 217, 218, 266, 267
electric conductivity, xii, 150, 181, 187
consumption patterns, xiii, 217, 218
electrical conductivity, x, xii, 143, 149, 150, 171,
contaminated water, 278, 295
172, 179, 181, 182, 188, 193, 194
contamination, 2, 27, 53, 170, 219, 223
electrocoagulation, vi, viii, xii, xiv, 170, 177, 187,
coordination, 27, 140, 290
188, 190, 191, 193, 194, 195, 259, 269
copolymer, 16, 26, 28, 37, 38
electrostatic interaction, 6, 26, 27, 85, 117, 120, 121,
copolymerization, 12, 13, 23
125, 126, 127, 171, 174, 254
cost, viii, ix, xii, 1, 2, 5, 10, 12, 13, 14, 15, 16, 17,
energy, ix, xiii, 12, 47, 66, 82, 84, 85, 87, 88, 89, 90,
21, 25, 30, 34, 47, 70, 99, 102, 109, 131, 177,
91, 93, 94, 96, 97, 99, 101, 102, 103, 130, 134,
187, 188, 191, 194, 199, 222, 242, 268, 278
177, 187, 190, 191, 194, 219, 261, 262, 263, 271,
cost effectiveness, 199
274
cotton, 74, 159, 160, 274, 282
energy consumption, xiii, 101, 187, 190, 194
covalent bond, 121
environmental conditions, 4, 110
cultivation, 22, 110, 123, 136
environmental impact, viii, x, xii, xiii, 48, 49, 50,
culture, 8, 11, 20, 28, 66, 111, 114, 116, 136
143, 144, 179, 217
culture media, 8
environmental issues, 236
culture medium, 20, 28, 116
erosion, 82, 262, 296, 297
cyanobacteria, xiii, 18, 27, 64, 109, 110, 111, 133,
ethylenediaminetetraacetic acid, 279
135, 137, 138, 197, 198, 208, 209, 210, 211, 212,
exopolysaccharides, 20, 22, 32, 35, 137
213, 214, 215, 216
experimental design, 191, 222, 244, 249
extracellular microcystins, 199, 201, 202, 212, 215
D extracellular organic matter, 109, 123, 135, 138
extraction, xi, 3, 7, 14, 30, 32, 33, 36, 37, 39, 43, 49,
degradation, 4, 11, 56, 70, 124, 229, 235, 243, 296 53, 54, 62, 70, 71, 72, 152, 159, 166, 169, 225,
denitrification, 99, 236, 237, 256 260, 274
derivatives, 10, 11, 12, 21, 28, 34, 42, 86, 171, 235 extracts, vii, ix, 47, 49, 57, 62, 64
detergents, xiv, 218, 235, 251, 253, 254
diatoms, 109, 111, 135, 136
F
disinfection, 82, 108, 134, 136, 198, 270, 275
dispersion, 12, 58, 95, 96, 97, 171, 174, 263, 266,
fats and oil, xii, 169, 170, 171, 172, 173, 174, 175
267
fermentation, viii, xiii, 1, 2, 6, 21, 30, 180, 217, 229
ferric chloride, vii, viii, ix, x, xiii, xiv, 5, 47, 48, 50,
62, 116, 117, 133, 143, 144, 145, 149, 171, 197,
306 Index

199, 200, 201, 202, 203, 204, 205, 206, 208, 211,
H
212, 213, 215, 218, 219, 221, 239, 241, 242, 243,
244, 245, 246, 249, 250, 252, 253, 264, 265, 267,
health, xi, xiii, 2, 5, 27, 32, 39, 70, 99, 103, 133, 156,
269, 270, 274, 296, 298, 299, 301
157, 160, 161, 163, 165, 171, 197, 198, 213, 214,
fertilizers, 29, 102, 235
215, 219, 222, 237, 251, 266
filters, xi, 73, 74, 155, 159, 198
health effects, 215
filtration, vii, ix, xi, xii, xiii, 7, 69, 70, 71, 73, 74, 75,
health problems, 2, 27
76, 77, 78, 81, 86, 109, 122, 128, 131, 133, 137,
health risks, 70
140, 155, 157, 159, 161, 162, 163, 164, 165, 170,
heavy metals, 15, 20, 22, 27, 35, 40, 156, 170, 176,
179, 180, 181, 182, 184, 185, 208, 217, 254, 269,
226, 238, 268, 270, 278, 291
280, 282, 295
human, xiii, 5, 27, 33, 52, 53, 64, 70, 71, 171, 186,
floc properties, x, 13, 108, 128, 129, 131, 134, 141
197, 198, 214, 219, 222, 237, 251, 278
flocculant, v, vii, viii, ix, xiv, xv, 1, 2, 6, 7, 8, 9, 11,
human health, xiii, 5, 27, 171, 197, 198, 219, 222,
13, 15, 16, 17, 18, 20, 21, 22, 23, 24, 25, 26, 27,
237, 251
28, 29, 30, 33, 34, 35, 37, 38, 39, 40, 41, 42, 43,
humic substances, 120, 122, 123, 126, 127, 131, 137,
44, 57, 58, 62, 65, 104, 172, 174, 176, 178, 214,
139, 142, 229, 242
216, 218, 220, 222, 226, 227, 239, 241, 242, 243,
hydrogen, 6, 7, 10, 11, 22, 86, 117, 120, 121, 122,
244, 245, 246, 249, 250, 251, 253, 254, 258, 262,
125, 127, 130, 229, 268, 272, 285
263, 265, 267, 268, 269, 274, 275, 277, 278, 279,
hydrogen bond(s), 6, 7, 10, 11, 22, 86, 117, 120, 121,
280, 284, 290, 291, 292, 293
122, 125, 127, 130, 285
flotation, ix, 81, 87, 98, 104, 109, 128, 134, 135,
hydrogen peroxide, 272
139, 214, 220, 226, 253, 263, 271, 273, 295
hydrolysis, 32, 84, 87, 118, 120, 203, 220, 221, 264,
food, viii, xi, 1, 2, 3, 6, 11, 14, 29, 34, 36, 37, 38, 39,
265, 266, 282
40, 41, 44, 45, 56, 66, 169, 171, 278
hydrophilicity, 3, 15, 111, 212
food chain, 278
hydrophobicity, 86, 111, 131, 235, 236, 262, 292
food industry, 14, 29, 44, 66, 171
hydroxide, 48, 72, 116, 118, 121, 158, 190, 201, 264,
formation, xiv, 2, 5, 9, 23, 25, 26, 27, 30, 39, 82, 83,
270, 298
85, 86, 87, 88, 95, 96, 108, 118, 122, 126, 133,
hydroxyl, 3, 6, 11, 12, 21, 26, 27, 120, 239, 264
134, 136, 138, 140, 198, 218, 223, 229, 237, 243,
hydroxyl groups, 3, 11, 12, 21, 26, 120
244, 260, 261, 263, 264, 265, 266, 269, 272, 283,
286, 288, 290, 295, 301
fouling, 108, 122, 135, 136, 139, 176, 213, 216, 262 I
freshwater, 18, 122, 139, 200
fungi, viii, 1, 9, 10, 17, 18, 21, 23, 53, 56, 57, 226 impurities, x, 57, 70, 95, 107, 128, 129, 131, 132,
156, 157, 260, 295
inorganic coagulants, xiv, 122, 144, 180, 218, 222,
G 266, 269
intermolecular interactions, 4, 130, 131
genus, 51, 64, 236
intracellular, xiii, 109, 136, 197, 199, 201, 202, 203,
glucose, 10, 20, 23, 111
204, 205, 206, 207, 208, 209, 210, 211, 212, 213
glycoproteins, 17, 20
ions, xiv, 4, 12, 15, 20, 22, 24, 27, 28, 38, 41, 42, 83,
glycosaminoglycans, 24
84, 125, 134, 178, 203, 218, 225, 229, 235, 236,
gravity, 49, 96, 157, 171, 261
237, 245, 251, 253, 261, 264, 265, 267, 268, 270,
green alga, 109, 111, 117, 126, 135, 138, 198, 200
273, 278, 290, 291
greenhouse gas emissions, 99
iron, vii, ix, 27, 47, 48, 49, 50, 70, 73, 82, 84, 137,
guar gum, 29, 39, 86, 173, 267
156, 159, 203, 221, 222, 237, 250, 256, 263, 264,
270, 296, 301
 Index 307

neutral, xi, xii, xiii, 3, 7, 16, 22, 24, 26, 59, 72, 74,
K
112, 116, 120, 125, 155, 162, 180, 187, 193, 203,
208, 222, 226, 242, 285, 290
kinetic study, 296, 298, 299
neutralization, xv, 5, 6, 7, 26, 28, 83, 85, 108, 118,
kinetics, vii, x, 6, 34, 81, 86, 88, 265, 291, 296, 301
122, 157, 174, 176, 193, 203, 208, 221, 259, 260,
261, 262, 265, 269, 277, 289, 290
L nitrates, 218, 225, 236, 237, 245, 251, 252, 253
nitrification, 99, 100, 236, 237, 256
landfill leachate, 16, 31, 219, 222, 242, 245, 249, nitrogen, 29, 43, 44, 49, 97, 99, 100, 101, 102, 103,
250, 253, 254, 255, 256, 257, 258 104, 171, 172, 180, 226, 236, 237, 238, 249, 251,
lignin, 10, 24, 26, 28, 33, 37, 173 252, 253, 255, 262
nitrogen compounds, 44, 251, 252, 253
M nitrogen dioxide, 43
nitrogen gas, 236, 237
meat processing wastewater, vi, 169, 172, 173 nucleic acid, 18, 20, 21, 110, 120, 126, 134
mesquite gum, viii, xii, 169, 171, 173, 176 nutrients, xiii, 22, 180, 197, 198, 257
metabolites, 20, 133, 137, 199, 208, 212
metal ion, viii, xv, 12, 20, 27, 35, 38, 41, 42, 261, O
277, 278, 286, 290, 291, 292
metal salts, 28, 70, 134, 141, 214, 221, 222 oil, xii, 5, 11, 18, 20, 21, 29, 30, 32, 36, 42, 85, 86,
metals, 4, 5, 15, 20, 21, 22, 27, 35, 40, 48, 49, 50, 62, 170, 174, 175, 260, 272, 273
156, 170, 176, 221, 223, 226, 238, 246, 262, 265, oil production, 260
268, 270, 278, 291 organic coagulant(s), vii, ix, xi, 69, 74, 144, 147,
methodology, 4, 39, 53, 72, 145, 160, 190, 222, 244, 149, 155, 157, 158, 164, 165, 185, 195, 221, 222,
249, 255, 256, 274, 298 266, 267
methylene blue, 26, 97, 178 organic compounds, x, 49, 107, 108, 109, 110, 120,
microcystin, xiii, 133, 198, 199, 202, 203, 204, 205, 126, 131, 136, 229, 242, 249, 263
206, 208, 209, 210, 212, 213, 214, 215, 216 organic matter, vii, viii, x, xii, 33, 43, 48, 49, 57, 82,
microcystis aeruginosa, 198 100, 101, 107, 108, 109, 110, 111, 112, 113, 118,
microorganisms, ix, 2, 9, 10, 11, 17, 30, 49, 50, 59, 120, 121, 122, 123, 128, 131, 132, 133, 134, 135,
66, 81, 82, 101, 132, 156, 226, 229, 236, 243, 136, 137, 138, 139, 140, 141, 142, 157, 170, 179,
260, 268, 275 184, 203, 208, 214, 216, 219, 223, 229, 234, 236,
molecular weight, x, 4, 5, 7, 8, 20, 22, 24, 86, 87, 96, 242, 258, 260, 261, 262, 263, 266, 267, 270, 273,
107, 111, 113, 131, 157, 226, 243, 273, 279, 280 275
molecules, 3, 12, 24, 25, 30, 84, 85, 87, 88, 108, 113, organic polymers, 33, 243
120, 226, 236, 264, 268 organic solvents, 11, 291
Moringa oleifera, ix, xi, xii, 32, 47, 51, 64, 125, 155, oxidation, viii, xiii, xv, 12, 110, 124, 135, 136, 137,
156, 157, 158, 160, 161, 163, 164, 165, 166, 179, 139, 142, 170, 176, 177, 188, 191, 216, 217, 236,
181, 222, 256, 267, 302 237, 259, 263, 268, 270, 271, 272, 274, 275, 278,
291
N
P
nanocomposites, 38, 40, 274
nanomaterials, 38, 170 pectin, 10, 14, 15, 27, 32, 33, 34, 35, 37, 39, 40, 42,
nanoparticles, 21, 37, 274 43, 44
natural polymers, 6, 11, 31, 32, 117, 125 peptides, x, 107, 110, 111, 113, 114, 115, 118, 119,
natural resources, ix, 47, 144 120, 121, 122, 123, 126, 131, 134, 138, 141, 142
natural sciences, 64
308 Index

pH, ix, x, xi, xii, xiii, xiv, xv, 3, 5, 6, 7, 8, 13, 16, 20, polysaccharide(s), v, vii, viii, 1, 2, 3, 4, 6, 7, 8, 9, 10,
22, 23, 27, 35, 48, 58, 59, 60, 61, 62, 69, 71, 72, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,
73, 74, 75, 76, 77, 78, 107, 115, 116, 117, 118, 24, 25, 26, 27, 28, 29, 30, 33, 34, 36, 37, 38, 39,
119, 120, 121, 122, 123, 124, 126, 128, 130, 131, 40, 41, 42, 45, 110, 111, 113, 115, 117, 120, 122,
132, 135, 139, 143, 145, 146, 147, 155, 158, 159, 124, 126, 129, 133, 136, 137, 278
160, 161, 162, 163, 164, 165, 169, 172, 173, 174, potassium, 136, 137, 142, 215, 265, 279
175, 179, 181, 182, 183, 187, 188, 189, 190, 191, potato, 12, 14, 22, 28, 29, 34, 40, 43
192, 193, 194, 197, 199, 200, 201, 202, 203, 204, potato starch, 22, 28, 29, 34, 40
205, 206, 207, 208, 210, 211, 212, 213, 218, 220, precipitation, xv, 7, 9, 116, 117, 118, 170, 220, 221,
221, 222, 223, 225, 226, 227, 229, 230, 231, 234, 265, 269, 278, 287, 290
242, 244, 246, 249, 250, 253, 261, 262, 263, 264, preparation, iv, vii, viii, xv, 1, 23, 35, 49, 57, 58,
267, 268, 269, 270, 277, 280, 281, 282, 285, 286, 158, 181, 277, 280, 282, 291
287, 288, 289, 290, 291, 292, 293, 298, 301 proteins, x, 12, 17, 20, 86, 107, 110, 111, 113, 114,
pharmaceutical, 3, 6, 11, 15, 17, 23, 45, 235 115, 118, 119, 120, 121, 122, 123, 126, 127, 129,
pharmacology, 2, 17 131, 134, 137, 138, 141, 142, 260, 268
phenol, xiv, 218, 219, 225, 232, 235, 238, 239, 241, pulp, 11, 14, 28, 32, 36, 38, 41, 42, 43, 72, 291
242, 245, 251, 253 purification, xiv, 3, 10, 30, 48, 64, 138, 188, 218,
phenolic compounds, 226, 260, 268, 269, 270, 272 273, 274, 279
phosphorus, xiv, 29, 30, 49, 101, 172, 180, 218, 225,
226, 230, 233, 235, 238, 245, 249, 250, 253
Q
photocatalysis, 170, 177, 272, 274
physical properties, 22, 134, 214
quality control, 105
physicochemical characteristics, 11, 175, 226, 238,
quaternary ammonium, 13
242
physicochemical properties, 3, 4, 8, 14
phytoplankton, x, 107, 109, 110, 112, 113, 114, 122, R
136, 139
plants, v, vii, viii, ix, x, xiii, 1, 2, 3, 7, 10, 11, 12, 14, raw materials, viii, 1, 5, 11, 56
16, 17, 30, 47, 48, 49, 50, 51, 52, 53, 54, 55, 58, reactants, 5, 88, 95
59, 62, 63, 65, 66, 67, 70, 81, 82, 89, 91, 94, 95, reaction temperature, 5
97, 98, 99, 101, 103, 107, 108, 109, 174, 178, reaction time, 5
197, 198, 235, 260, 265, 270, 292, 301, 302 reagents, xiv, 48, 85, 88, 218, 245, 248, 249, 250,
pollutant, xiv, 25, 29, 144, 188, 218, 223, 242, 286 251, 252, 253
pollution, viii, xii, 5, 27, 35, 99, 103, 144, 187, 219, recovery, 2, 25, 28, 29, 38, 66, 99, 101, 102, 171,
241, 242 219
polyacrylamide, 5, 11, 13, 15, 16, 23, 26, 29, 32, 37, removal of heavy metal ion, viii, xv, 277, 278, 290,
38, 41, 44, 226, 227, 257, 266, 273, 274, 278, 292 291
polyamides, 86 repulsion, 7, 9, 49, 82, 84, 208, 220, 242, 286, 289
polyelectrolyte complex, 23, 39
polyhydroxyalkanoates, 31 S
polymer, vii, ix, xiv, 3, 4, 6, 7, 8, 12, 23, 24, 25, 26,
33, 40, 69, 71, 72, 79, 86, 87, 88, 89, 117, 121, salts, 5, 28, 50, 70, 82, 84, 89, 116, 117, 129, 134,
124, 131, 171, 218, 222, 223, 242, 243, 253, 265, 135, 141, 182, 188, 194, 214, 221, 222, 243, 249,
266, 273, 274, 275, 286, 287, 292 265, 266, 275, 301
polymeric materials, 125, 176, 266 science, 33, 45, 104, 138, 214
polymers, viii, xiv, 1, 3, 6, 11, 25, 26, 31, 32, 33, 63, sedimentation, vii, viii, ix, x, xi, xii, xiv, 16, 18, 26,
70, 83, 86, 87, 89, 96, 104, 113, 116, 117, 121, 32, 47, 49, 50, 66, 69, 70, 71, 73, 74, 75, 76, 77,
124, 125, 170, 218, 223, 243, 258, 266, 270, 273, 78, 86, 87, 89, 109, 126, 128, 139, 140, 143, 145,
278 146, 149, 155, 157, 161, 162, 163, 164, 171, 179,
 Index 309

181, 185, 208, 216, 259, 260, 261, 262, 268, 269, stoichiometry, xv, 277, 278, 281, 288, 289
270, 272, 275, 295, 297, 298, 299 sulfate, vii, ix, 5, 27, 44, 47, 48, 50, 56, 62, 69, 84,
seed, 16, 28, 32, 41, 157, 158, 163, 165, 166, 181, 138, 145, 200, 250, 264, 267, 270, 273, 274, 275,
249, 267, 274 278, 292
separation, ix, x, xi, 10, 35, 38, 81, 82, 86, 87, 96, synthetic polymers, 3, 86, 258
97, 102, 103, 104, 108, 109, 128, 133, 135, 136,
138, 139, 140, 141, 157, 169, 171, 173, 174, 178,
T
213, 215, 216, 226, 257, 268, 269, 278, 289, 291,
292, 302
tannin, xii, 144, 145, 146, 147, 149, 150, 151, 152,
sewage, 16, 17, 20, 21, 38, 44, 50, 63, 97, 100, 101,
179, 180, 181, 184, 185, 267
176
technology, 4, 11, 25, 104, 108, 130, 138, 166, 170,
shear, xiv, 83, 84, 85, 128, 129, 130, 131, 134, 141,
214, 215, 278
199, 208, 212, 218, 223, 261, 262, 263, 264, 275,
temperature, xiv, 3, 5, 7, 8, 9, 20, 48, 55, 56, 99, 110,
301
138, 172, 200, 201, 218, 223, 229, 235, 237, 262,
shear rate, 128, 129, 130, 131, 134, 141, 262, 263,
264, 265, 266, 280, 282
275
testing, 66, 145, 178, 242, 267
sludge, vii, ix, xiv, 5, 13, 14, 18, 21, 22, 29, 34, 37,
toxic effect, 198
47, 48, 49, 50, 63, 70, 99, 100, 101, 102, 103,
toxic metals, 49
104, 133, 144, 177, 218, 220, 222, 223, 239, 242,
toxic substances, x, 143
243, 250, 253, 257, 265, 266, 267, 269, 274
toxicity, 5, 20, 27, 28, 70, 138, 220, 222, 235, 266,
sodium, 19, 27, 40, 44, 50, 72, 86, 122, 134, 144,
269, 271
158, 190, 195, 201, 211, 225, 263, 268, 270, 271,
toxin, 199, 203, 208, 209
273, 282, 298
transition metal, 278, 292
sodium hydroxide, 72, 158, 190, 201, 298
transition metal ions, 278, 292
solid phase, 157, 291
turbidity, vi, viii, ix, xi, xii, xiii, xiv, 16, 17, 28, 29,
solid state, 22, 292
30, 31, 35, 38, 42, 47, 58, 59, 60, 61, 62, 63, 64,
solid waste, xiii, 144, 217, 218, 223
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 82, 91,
solubility, xv, 11, 24, 43, 118, 221, 223, 262, 263,
120, 126, 128, 153, 155, 156, 157, 160, 163, 164,
264, 267, 268, 270, 277, 280, 284, 285, 290
165, 166, 170, 171, 172, 173, 175, 179, 181, 183,
solution, xi, xii, xv, 4, 5, 7, 16, 25, 28, 35, 48, 58, 72,
184, 185, 194, 197, 199, 200, 201, 202, 203, 204,
74, 78, 83, 85, 87, 88, 120, 125, 149, 155, 157,
205, 206, 207, 208, 210, 212, 213, 218, 220, 225,
158, 159, 165, 181, 184, 187, 194, 200, 208, 219,
230, 231, 234, 238, 239, 243, 244, 245, 246, 253,
225, 227, 250, 257, 263, 266, 273, 275, 277, 278,
259, 260, 262, 266, 267, 268, 270, 272, 278, 279,
279, 280, 281, 282, 285, 286, 289, 290, 296, 298
280, 284, 285, 295, 296, 297, 298, 299, 300, 301,
solvents, 11, 12, 24, 291
302
species, 21, 28, 49, 50, 51, 52, 53, 54, 59, 64, 89,
109, 110, 111, 112, 113, 114, 115, 120, 121, 124,
131, 135, 136, 212, 221, 251, 263, 264, 265, 267, V
269
specific adsorption, 83, 84 velocity, 87, 89, 90, 91, 94, 95, 96, 97, 128, 133,
spectroscopy, xv, 273, 277 140, 157, 262, 269, 296, 297
stability, ix, 8, 15, 17, 20, 23, 47, 49, 82, 122, 174, viscosity, 14, 96, 264, 265, 266
261, 273, 288, 292 volatile organic compounds, 229
stabilization, 9, 118, 121, 222, 268
starch, 2, 3, 12, 13, 16, 17, 22, 27, 28, 29, 30, 32, 33, W
34, 35, 37, 39, 40, 42, 43, 111, 116, 124, 125,
129, 266, 267, 269, 273, 290 waste management, 48, 218, 219
starch granules, 39 wastewater treatment, viii, ix, xi, xiv, xv, 2, 6, 13,
stock, 58, 200, 280, 281, 282, 298 17, 18, 21, 24, 30, 32, 33, 35, 36, 39, 40, 42, 43,
310 Index

44, 65, 66, 79, 81, 82, 89, 97, 99, 101, 102, 103, water-soluble polymers, 104
104, 118, 125, 133, 169, 170, 171, 172, 174, 178, waterways, 103
180, 227, 259, 262, 268, 271, 275, 291, 292
water purification, 138, 188
X
water quality, viii, x, xiii, 71, 82, 105, 107, 108, 110,
138, 142, 170, 172, 197, 198, 262, 267
xanthan gum, 27, 29
water shortages, 219
water treatment, viii, x, xii, xiii, 23, 26, 32, 43, 44,
48, 49, 63, 64, 65, 66, 69, 70, 73, 78, 79, 82, 91, Z
102, 104, 107, 108, 110, 111, 115, 120, 123, 126,
133, 134, 137, 138, 140, 141, 142, 144, 156, 162, zeta potential, xi, 6, 61, 62, 115, 169, 170, 171, 172,
165, 166, 167, 187, 188, 193, 195, 198, 208, 212, 173, 174, 175, 176, 178, 220, 222, 279, 280, 282,
213, 214, 215, 216, 222, 251, 254, 255, 258, 266, 285, 286, 289, 292
267, 272, 273, 274, 275, 278, 295, 301, 302