1.

0 INTRODUCTION

1.1 BACKGROUND OF THE STUDY

The term fish is applied to a variety of vertebrates of several evolutionary lines. It describes a life-

form rather than a taxonomic group. As members of the phylum Chordate, fish share certain features

with other vertebrates. J. Fish. Biol.(2021). These features are gill slits at some point in the life cycle,

a notochord, or skeletal supporting rod, a dorsal hollow nerve cord, and a tail. Living fishes represent

some five classes, which are as distinct from one another as are the four classes of familiar air-

breathing animals—amphibians, reptiles, birds, and mammals. Dev. Dynam.(2022). For example, the

jawless fishes (Agnatha) have gills in pouches and lack limb girdles. Extant agnathans are the

lampreys and the hagfishes. As the name implies, the skeletons of fishes of the class Chondrichthyes

(from chondr, “cartilage,” and ichthyes, “fish”) are made entirely of cartilage. Chandra Segaranet al.

(2022). Modern fish of this class lack a swim bladder, and their scales and teeth are made up of the

same placoid material. Sharks, skates, and rays are examples of cartilaginous fishes. The bony fishes

are by far the largest class. Examples range from the tiny seahorse to the 450-kg (1,000-pound) blue

marlin, from the flattened soles and flounders to the boxy puffers and ocean sunfishes. Unlike the

scales of the cartilaginous fishes, those of bony fishes, when present, grow throughout life and are

made up of thin overlapping plates of bone. Bony fishes also have an operculum that covers the gill

slits. W.B. Wosiacki (2019).

The study of fishes, the science of ichthyology, is of broad importance. Fishes are of interest to

humans for many reasons, the most important being their relationship with and dependence on the

environment. A more obvious reason for interest in fishes is their role as a moderate but important

part of the world’s food supply. This resource, once thought unlimited, is now realized to be finite and

in delicate balance with the biological, chemical, and physical factors of the aquatic environment. S.

Hegde, G. Kumar et al (2022).

A fish is a cold blooded aquatic gill breathing oviparous vertebrae whose body is covered by dermal

scales. Fish is highly proteinous, has high digestibility and is a rich source of lysine and

1
sulphurcontaining amino acids.

Fish plays a hugely important role in global food security. It provides more than 3.1 billion people

with at least 20% of their animal protein but above all it is an important source of fatty acids and

micronutrients (FAO 2021).

Fish however is susceptible to damage as soon as it is harvested hence the urgent need for

preservative measures. (Akintola et al., 2020) reported a 50% annual loss of fish in Nigeria to post

harvest spoilage irrespective of the method of preservation employed. High ambient temperatures and

humid tropical conditions speed up spoilage rates in fresh water fishes (Saliu, 2018).

1.2 Justification of the Study

Smoked fish consumed mostly in tropical region comes from local smokers and such required

attention hence fish form a major part man and animal protein.

Fish smoking is expensive energy source;therefore, there is need to replace it with other energy

sources in the smoking of fish, since they are less expensive energy sources.

Smokedcatfish are subjected to effects of smoke and heat therefore needs proper handling.

1.3.1 The Broad Objective of the Study

The Broad objective of the study is to investigate the effects of smoke on quality of in the

tropicalCatfish.

1.3.2Specific objectives of the study were to determine;

i. the proximate composition of smoked dried pond bred Catfish.

ii. the sensory attributes of smoked dried pond bred Catfish.

iii. the microbiological stability of smoked dried pond bred Catfish.

2
2.0 LITERATURE REVIEW

2.1 African Catfish (Clariasgariepinus )

Catfish (order Siluriformes) are a diverse group of fish They are named for their

prominent barbels, slender, whisker-like tactile organs near the mouth, which give the image of cat-

like whiskers.Catfish are very diverse, ranking second or third in diversity among orders

of vertebrates, with almost 3,000 known species. Lundberg andFriel (2013).About one in every ten

species of fish, and one in every 20 vertebrates.

Catfish are found primarily in freshwater environments of all kinds, with species on every continent

except Antarctica. Some species from the families Ariidae and Plotosidae are also found in marine

environments. They feature some of the smallest known vertebrates, including the candiru (toothpick

fish), the only vertebrate parasite to attack humans, as well as Mekong giant catfish, the largest

reported freshwater fish.

Catfish include bony-plated types and also smooth, naked types, but they do not have scales. Not all

catfish families have prominent barbels; what defines a fish as being in the order Siluriformes are

certain features of the skull and swim bladder. Owe, (2011).Unlike most fish, catfish do not have

scales. They usually have an adipose fin (fleshy, rayless posterior ffin), and often spine-like rays

(spines) present at the front of the dorsal and pectoral fins.The dorsal fin of most catfish species

technically has two spines, the first one very short.Sutton, (2017). As noted, the catfish body may be

naked or covered with bony plates. Normally, there are up to four pairs of barbels on the head, with

two on the chin, one nasal, and one maxillary, but the nasal and chin barbels may be absent. Dong,

and Yang (2017) With the exception of the Diplomystidae (among extant families), the maxilla is

toothless.Some catfish are known to communicate with one other. There are some species that use a

method known as stridulation to produce communicative sounds, such as marine catfish that have the

base of the pectoral fin rubbing against the pectoral girdle to produce sounds. Others make use of the

springferderapparat (elastic spring apparatus or modified swim bladder mechanism) to create

vibrations in the swim bladder to produce sounds. Friel, (1997).Some catfish species can inflict

3
painful stings with their spines, such as the case with the stone and tadpole catfishes and the madtom.

The strong, hollow, bonifiedleading ray on their dorsal and particularly the pectoral fins can emit a

stinging protein, which comes from poison glands in the epidermal tissue covering the spines. In

members of the family Plotosidae, and of the genusHeteropneustes, this protein is so strong it may

hospitalize humans unfortunate enough to receive a sting. Stings from Plotosuslineatus can result

indeath. Asagba andEriyamremu, (2018) in many catfish, these fins may be held open and locked in

place. Most are passive stingers, but Heteropneustesfossilis of India is aggressive, with records of

attacks on humans.Siluriformes are monophyletic in their development of the upper jaw, which is able

to move and support the catfishbarbels.

TAXONOMY OF AFRICAN CATFISH

Classification of African Catfish

Kingdom : Animalia
Phylum : Chordata
Class : Actinopterygii
Order : Siluriormes
Family : Clariidae
Genus : Clarias
Species : C. gariepinus
Scientific Name : Clariasgariepinus
Taxonomy classification of catfish Heliyon, (2023).

4
2.2 NUTRITIONAL COMPOSITION OF CATFISH

2.2.3. Nutritional composition of catfish

Fish is a rich source of chemical composition such as proteins, fat (poly-unsaturated fatty acids),

vitamin (including, vitamin A, vitamin B2, vitamin B6 and others), minerals (iron, calcium, iodine,

potassium and other minerals), and carbohydrates. In many part of the world people use fish as the

main component in their diet, as a good source of nutrition. Praveen Kumar G 2017. Fish constitute

more protein source than other animals protein source.

Generally fish is made up of percent water, percent protein, percent fat and percent minerals and

percent carbohydrate.Olayemi F (2017).Fats from fatty fish species contain poly unsaturated fatty

acids (PUFAs) namely, EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) (omega 3 fatty

acids) which are essential for proper growth of children and prevent the cardiovascular diseases such

as coronary heart disease. Ochiai., Ozawa. (2017).

The nutritional value of fish used as a source of food is obtained from its chemical composition,

which varies widely from species to species and within the same species. Farid., Latifa. (2019) the

feeding habit, sex and seasonal variations are the main factors which can affect the chemical

composition in fish. Knowledge on chemical composition of fish is vital to develop processing

technology for fish and fish products (both in commercial and industrial level), in order to compare its

value with other foods as a source of protein, in fortification for product development and nutritional

enrichment, to ensure that they are within the range of the dietary requirements, commercial

specifications and maximum utilization; minimize loss of nutritional contents.

2.3 Fish Smoking

Smoking is one of the oldest fish preservation methods. It consists in the application of wood smoke

on food. Aremu and Ibrahim. (2013). Smoking can inhibit the formation of toxins in products (Huong,

2014), reduce the growth of bacteria, due to lower water activity by smoking in combination with

salting and drying which creates a physical surface barrier Amaefule, Ekwu. (2017)

5
Smoking is a culinary technique that involves flavoring and preserving catfish by exposing it to

smoke from burning wood or other materials. This process imparts a distinctive smoky flavor to the

fish while also extending its shelf life. A recent review by Smith et al. (2023) highlighted the growing

popularity of catfish smoking among food enthusiasts. The study emphasized the importance of using

high-quality wood chips, controlling smoking temperatures, and experimenting with various brines or

rubs to achieve the desired flavor profile. However, further research is needed to explore the potential

health effects associated with smoked catfish consumption and to ensure proper food safety measures

are followed throughout the smoking process.

Traditionally, fish is smoked in pits or on raised smoking “tables” where the control of heat is difficult

and at times impossible. (Kumolu-Johnson et al. (2010). Nowadays, shifting for high sensory quality

product is the main purpose of smoking. The smoked products have higher moisture and lower salt

content than in the past (Kolodziejskaet al.,(2002).

Depending on the way smoke gets into products, the quality of smoked fish is affected by raw

material, salting method, brining concentration, condition processing, and composition of smoke,

smoking method, smoke agents and storage conditions (Huong, 2014).

2.4 Methods of Smoking Fish

2.4.1 Hot smoking

Fish is smoked until cooked in order to obtain a product with extended shelf-life, since alternative

preservation methods such as refrigeration are absent in remote fishing villages where most fish

processing takes place (Aremu et al., 2013). Hot smoking is known as the traditional smoking method

of smoking fish in the tropics.

Hot smoking entails the application of much more heat, through the burning of large quantity of fuel–

wood. The products from hot smoking are tastier and have longer shelf lives (Aremu et al., 2013;

Arasonet al., 2014)). Hot smoking occurs within the range of 52 to 80ºC (126- 176ºF). Within this

temperature range, catfish are fully cooked, moist and flavorful. High temperatures beyond the

specified can lead to excessive shrinkage. It also reduces yield, as well as other essential nutrients

6
present in the fish (Myrvold, 2011). Safe hot smoked fish requires at least 3.5% water phase salt

(WPS) and must have achieved an internal product temperature of at least 145 °F (62.8 °C) for at least

30 minutes. This prevents the production of toxins by Clostridium botulinum (Huong, 2014).

Additionally, water activity (aw) of hot smoked fish products must be less than 0.85 to make products

stable at room temperature (Arason et al., 2014).

2.4.2 Cold smoking

Cold smoking is practiced in advanced countries where alternative means of preserving the fish such

as refrigeration is available. Cold smoking is primarily to improve the flavor of the fish and retain its

nutritive value (Eyo, 2001). Cold smoking is a smoking method where the temperature is maintained

between 20 to 30°C (68- 86°F). In this temperature range, fish take on a smoked flavor, but remain

relatively moist (Myrvold, 2011; Huong, 2014). Arason et al., (2014) suggests that the relative

humidity during cold smoking should be remained in the range of 75- 85%. Vacuum packed and

chilled storage should be followed by cold smoking because product is not completely preserved

(Rørvik, 2000).

2.4.3 Smoke Drying

2.5 Preservation Effect of Smokeon Catfish

Smoke contributes to fish preservation by acting as an effective antioxidant, bacteriostatic and

bactericidal agent as well as by providing a protective film on the surface of smoked fish. Smoking

kills food poisoning and spoilage bacteria or renders them harmless by altering the chemistry of the

environment these spoilage organisms need to grow (Hilderbrand, 2001). Smoked foods are generally

better preserved and protected against insect’s infestation than untreated products (Crynen, 2007).

Smoking preserves fish by drying, cooking and depositing natural wood–smoke chemicals like tars,

phenols and microorganisms on the flesh of the fish. The phenolic fraction of wood smoke is said to

possess the highest inhibiting ability on bacteria (Aremu, et al., 2013). The smoke determines the

colour which is one of the qualities that attracts consumers. It imparts aroma, taste and colour on

processed fish (Kumolu-Johnson, 2010). Compounds such as carbonyls and acids; as well as phenolic

7
fractions contribute significantly in the overall flavor of smoked fish. The colour of smoked fish is

relatively not independent on the method as well as the type of wood or agricultural waste used in

smoking the fishDien, H.A., Montolalu, R.I., &Berhimpon, S. (2019). The colour ranges from black,

dark brown, golden brown or light brown to dirty white. Although preservation methods for fish and

products are often used in order to make the fish safer and to prolong its shelf– life, the determination

of some proximate parameters such as moisture, ash, lipid and protein contents are necessary so as to

ascertain that they meet the requirements of food regulations and commercial specifications (Aremu,

et al., 2013).

2.6 Effect of Smoke on Nutritive Value of Fish

Fish is smoked until cooked in order to obtain a product with extended shelf-life, since alternative

preservation methods such as refrigeration are absent in remote fishing villages where most fish

processing takes place. Amaefule KU, Ekwu AO. (2017).

Hot smoking is the traditional methods of fish smoking in the tropics. Hot smoking entails the

application of much more heat, through the burning of large quantity of fuel-wood. (Aremu, et al.,

2013).This process has effect on the nutritive value of fish, and is also responsible for significant

modifications of the organoleptic properties of fishmeal (Kjallstrand and Petersson, 2001).

Some studies have reported reductions in the quality of protein as a result of smoking. The loss in

available lysine may vary from 6-33% at 25oC to 53-56 % at 40C during hot smoking and a 25% loss

of available lysine on the surface and a 12% loss at the center of hot smoked fish (Akinwumi, 2014).

The lysine loss is partly attributed to high temperature effect as a Millard reaction. This reaction

involves the amino acids with sugars.

These changes are necessary to give the characteristic golden- brown colour of smoked fish. Losses

in other amino acids such as arginine, histidine and a decline in net protein utilization (NPU) have

been observed in smoked fish. A number of chemical reactions may take place during heat treatment

including decomposition, dehydration of serine and threonine, loss of sulphur from cysteine oxidation

of cystane and methionine, cyclization of glutamic and aspartic acids and threonine.

8
However, the positive effect of heat intensity on the shelf-life of smoked product has been reported by

several researchers. CO2 is reported to have an important effect on microbial aldehydes all of which

have powerful bactericidal action and prevent the growth of other growth, exerting a selective

inhibitory action.

Akinwumi, (2014) in his study on the Effects of Smoking and Freezing on the Nutritive Value of

African Mud Catfish, (Clariasgariepinus) observed that, the proximate values of the smoked fish

samples in all the parameters studied were statistically significant to those in the fresh fish samples

(control). There was significant increase in the protein level in smoked catfish, when compared with

the frozen catfish; therefore, he suggests that protein was not lost during smoking. This finding is in

agreement with the observations of Puwastien et al., (1999). Similarly, Fapohunda and Ogunkoya

(2006) reported that smoke drying methods increased the protein, ash and fat contents of C.

gariepinus.

(Cardinal et al., 2019) observed decrease of moisture, carbohydrate, lipid, potassium and vitamin C

contents and increase of protein, ash, crude fiber, and phosphorus and iron contents in smoked C.

gariepinus. The authors further noted that the increase in the ash content in the smoked fish was due to

fthe loss of humidity and that the significant reduction in the moisture content when the fish was

smoked as a result of the loss in moisture during hot smoking. The components in the wood/charcoal

via pyrolysis are broken down in the process of burning to form smoke, thus it gives the fish a unique

aroma and, improves its taste and color because of the presence of a range of phenolic compounds,

nitrites and formaldehyde present in the smoke (Akinwumi, 2014).

The control of the organoleptic characteristics can be of real interest to processors who want to adapt

their products to consumers demand. The method of smoke generation and the smoking process used

have a considerable influence on the sensory characteristics of smoked fish, particularly on smoke

flavour perception.

9
2.7 Health Implication of Smoke

Wood combustion is typically inefficient and a multitude of partially oxidized organic chemicals are

generated in smoke. Smoke contains a large number of chemicals, many of which have been

associated with adverse health impacts. Examples include polycyclic aromatic hydrocarbons (PAHs),

dioxins, and formaldehyde, nitrogen and sulfur oxides (relevant for formation of e.g. nitrosamines).

Hokkanen, M.,et al (2018).

Furthermore, heavy metals are also found in combustion gases. The types and amount depend on the

kind of wood used, the temperature, the method used for developing the smoke, the water contents in

the wood and possible other parameters Aheto et al. (2017). There has been considerable evidence

which suggested that smoked foods may contain traces of compounds with mutagenic or carcinogenic

potential (Muyelaet al., 2012). The smoking process contaminates fish with polycyclic aromatic

hydrocarbons (PAHs), especially benzo[a]pyrene (Bomfehet al., 2019). and nitrosamines, which are

known carcinogens.

Consuming smoked food increases the risk of gastrointestinal cancer. Hence, safety of food is a

growing concern worldwide and PAHs residues if present in food above the maximum residual limit

(MRL) pose a serious threat to the public health (Muyelaet al., 2012). Therefore, there are strong

pressures on chemical safety for smoked products from the EU institutions. Thus the Codex

Alimentarius Commission on contaminants in food, at its 29th session from 16 to 20 April 2007

established a reflection on reducing levels of Polycyclic Aromatic Hydrocarbons (PAHs) in food dried

and smoked. In addition, the EU Regulation 1881/2006/EC on the content of PAH in smoked products

has been amended (European Union (EU), 2011), and in September 2014, these levels were lowered

from 5 to 2 μg/kg for benzo[a]pyrene and from 30 to 12 μg/kg for PAH4 (Bourgeois, et al., 2014)

The decision of the Codex Alimentarius Commission is very important for the activity of the smoked
fish industry because it will lead to the questioning of certain practices and the necessity of an
absolute control of processes to produce smoked fish presenting both excellent organoleptic and all
guarantees in terms of food safety.

10
2.7.1 Biological effects of smoke on catfish

Generally, Cold smoking will typically reduce the level of microorganism by 90 to 99%. But after the

cold smoking there are no such steps to eliminate or reduce the level of microorganisms. Typical

temperature used for cold smoking is 22-28˚C. However, this temperature is not sufficient to

eliminate the risk from Listeria monocytogens, a gram positive, facultative anaerobic, psychotropic

bacteria causing deadly septicemia, meningitis, spontaneous abortion, and foetal death in adult human

beings. Specific high risk categories like persons with altered immune system, pregnant ladies, old

aged persons etc. will be more susceptible to listeriosis followed by accidental inclusion.

Comparatively high temperature used in hot-smoking process and long-time of exposure to that

temperature (60-70˚C for 2-3 h) can inactivate the L. monocytogenseffectively, provided the raw

material is not extra-ordinarily contaminated with the bacteria prior to processing. At the same time

listericidal process should be validated to ensure that the treatments are effective and can be applied

continuously. But the hot smoked products are susceptible to post-process contaminations from many

of the micro-organisms due to improper handling and storage of the products. Sufficient heat

treatment, proper hygienic handling and cold chain maintenance during distribution can reduce the

risk of biological hazards in smoked fish and fishery products.

Another important biological hazard associated with storage of smoked fish is Clostridium botulinum.

The toxin produced by C. botulinum can lead to botulism, serious illness and death to the consumer.

Even a few micrograms of intoxication can lead to ill health with symptoms like weakness, vertigo,

double vision, difficulty in speaking, Swallowing and breathing, abdominal swelling, constipation,

paralysis and death. The symptoms will start within 18-36 h after consumption of the infected

product. By achieving proper salt concentration in processed fish, proper refrigeration during storage

and reduced oxygen packaging like Modified Atmosphere Packaging (MAP) and vacuum packaging

of the products can prevent the occurrence of C. botulinum in smoked fish and fishery products,

especially type E and non-proteolytic types B and F.

Salt along with smoke effectively prevents the toxin formation from type E, B and F.In cold smoked

fish and fishery products, which undergo mild heat processing, the presence of spoilage organisms

11
prevents the growth of C. botulinum and toxin production. Whereas in hot-smoked products, high

temperature application causes damages to spores of C. botulinum thus prevents the toxin formation.

Same process also prevents the prevalence of spoilage organisms and thus extends the shelf life of the

product. Thus, the time- temperature combination for smoking, along with salt concentration plays

critical roles in safety and quality aspects of the smoked fish and fishery products.

2.8 Chemical Effects of Smoke on Nutritional Quality of Catfish

2.8.1 Polycyclic aromatic hydrocarbons (PAHs)

PAHs are large class of organic compounds containing two or more fused aromatic rings made up of

carbon and hydrogen atoms. Incomplete combustion(pyrolysis), during smoking can lead to formation

and release of PAHs into the smoked product. Some of them are carcinogenic and mutagenic

substances causing serious health issues to the consumers, Hitzel, A., et al (2014). Processing

procedures such as smoking, drying, roasting, baking, frying and barbecuing/grilling can lead to

formation of PAHs in food items. Many reports indicate that individual PAHs in smoked fish can go

up to a level of 200µg/Kg. Among the 33 PAHs evaluated by the scientific committee on Food (SCF,

2002) of EU, 15 were found to be having mutagenicity/Geno toxicity in somatic cells of experimental

animal in-vivo. They are benzo[a]anthracene, benzo[b]-, benzo[j]-and benzo[k]fluoranthene,

benzo[ghi]perylene, benzo[a]pyrene, chrysene, cyclopenta[cd]pyrene, dibenz[a,h]anthracene,

dibenzo[a,e]-, dibenzo[a,h]-, dibenzo[a,i]-, dibenzo[a,l]pyrene, indeno[1,2,3-cd]pyrene and 5-

methylchryseneBomfeh, K.et al(2019).

The carcinogenic and genotoxic potentials of PAH are largest among the high molecular weight

PAH, i.e. compounds with 4 rings or more. Among them benzo[a]pyrene is regarded as potentially

genotoxic and carcinogenic to humans. They can cause long-term adverse health effects following

dietary intake of PAH. The PAH contamination in smoked products can be significantly reduced by

using indirect smoking process instead of direct smoking of the fish. In indirect smoking, the smoke

generated in an external smoking kiln, under controlled conditions, is used for smoking process. The

smoke produced can be even, washed before coming into contact with the food material processed. In

12
addition to that, use of lean fish for smoking, and cooking at lower temperature for longer time can

also reduce the PAH contamination significantly. If the smoke condensate is used for smoking, usage

of smoke condensate from reputed reliable resources approved by competent authority can effectively

reduce the occurrence of PAH contamination in the final product. The formation of PAH in smoked

fish can be minimised by following Code of Practice for the Reduction of Contamination of Food

with Polycyclic Hydrocarbons (PAH) from Smoking and Direct Drying Processes (CAC/RCP 68-

2009) given by Codex Alimentarius Commission. EU No.835/2011 specifies that maximum level of

benzopyrene, and PAH4 (benzo[a]pyrene + chrysene+ benz[a]anthracene+benzo[b]fluoranthene)

should be 2µg/Kg wet weight and 12µg/Kg in meat of smoked fish and fishery products.

13
Table 1: Estimated daily intakes (EDI) and margin of exposure (MOE) for polycyclic aromatic
hydrocarbons (PAH) through consumption of smoked or grilled fish.
Smoked dried
Parameters Smoked fish fish Reference
Benzo(a)pyrene (µg/kg) 21.8 ± 21.2 78.5 ± 53.8

PAH4 (µg/kg) 119.3 ± 107.5 484.2 ± 305.6 IkoAfé

et al. (2021
BaPEstimated daily intake BaP: 2.3-809.9 BaP: 2.5-1,974.8
(ng/kg bw/day
BaPMargin of exposure BaP: 30,978-86 BaP: 27,718-35
.
Abbreviations: PAH4, sum of benzo[a]pyrene, chrysene, benzo[b]fluoranthene, and
benz[a]anthracene; bw, body weight.
2.8.2 Heterocyclic amines (HCAs): Smoking catfish at high temperatures can also lead to the

formation of HCAs, which are formed from the reaction of amino acids and creatinine in the fish with

the heat. HCAs are potential carcinogens and may contribute to the risk of certain cancers when

consumed.Asamoah, E. K. (2021).

2.8.3 Acrylamide: The Maillard reaction during smoking can lead to the formation of acryl amide, a

chemical that forms in starchy foods cooked at high temperatures. Acryl amide has been associated

with potential neurotoxicity and carcinogenicity.

Loss of Nutrients: Smoking can cause a loss of essential nutrients in the catfish, such as omega-3

fatty acids and vitamins, which are important for human health.

2.9 Microbial Flora 0f Smoked Catfish

Microbial flora of fish depends on the microbial content off the water in which they live as the slime

that covers the surface of fish has been found to contain great variety of bacteria genera. D. K.&

Adjei, J. K. (2013).). Many dangers therefore exist if fish harvested from polluted water is eaten raw,

and because of the high microbial load of freshly harvested fish it is susceptible to rapid spoilage.

Hence preservation of fresh fish becomes very important. This can be achieved by freezing, drying

through smoking and sun-drying, canning, etc.

14
 Hence smoked fishes are less prone to microbial spoilage than fresh fish. However spoilage still

occurs as a result of growth of microbes due to partial dehydration during smoking (Abolagba OJ,

Uwagbai EC(2011). Contamination of fish and other fishery products by microbes has been a serious

threat to human health.There are four main factors responsible for fish spoilage once it is out of its

natural habitat (water) and these include: Autolysis which usually precedes bacterial spoilage and

involves the breakdown of protein and lipids to amino acids and fats by muscle enzymes. The activity

of microorganism is another factor which uses the amino acid produced by autolysis for proliferation

Okonta, A.A., and J.K. Ekelemu, (2019). Others are chemical deterioration and insect attack which

cause considerable deterioration. However, spoilage of fresh and highly preserved fish products is

mostly caused by microbial action. Foods of high sugar/salt contents are therefore most likely to be

spoiled by any kind of microbe, Adebayo-Tayo BC, et al (2018). It has been reported that serious

disease outbreak had occurred in both man and animals after consuming some dried fish feed and

food. This could be as a result of disease causing microorganism like Escherichia coli, Salmonella

typhi, Vibrio cholera which results from poor handling/cross-contamination and improper processing

practice of ready to eat “smoked fish” products.

15
3.0 MATERIALS AND METHODS

3.1Sample procurement

Nine fishes of vary weight of freshly caught catfish(Clariasgariepinus) was obtained from De-

Famous Integrated Farms & Business fishpond in Gboko-west Benue State, while rice husk was

gotten from Rice mill, palm frond from De-Famous Farms and sawdust was gotten from sawing mill

Gboko local government Benue State, Nigeria.

3.2Sample

All the catfish were killed, eviscerated and thoroughly rinsed with clean saltwater Bonoco , & Kurt

Kaya. (2018) and let to thaw for 30 min. All the pre-processed catfish was equally divided into three

portions of each after thawed.

3.3Smoking method

All portion of the pre-processed catfish were smoked in an open air kiln (drum kiln) constructed by

CEO, De-Famous integrated farms & Business. The drying was done at moderate temperature for

72hrs. The fish was turned at every 2 hrto prevent hardening. It was then let to cool(Olayemiet

al.2011). Packaged and stored in transparent air-tight polyethene bags as samplefor further use.

16
 2 hours and above

A chart showing the process of smoke drying of fish

Source: Ajue, 2020

17
Figure 1: De-Famous traditional drum kiln (Ogunbamboetal., 2018)

18
3.4Proximate Analysis of Smoked, Dried Pond Bred Catfish

The proximate analyses (moisture content, protein content, ash content, and fat content and crude

fiber) of the samples were analyzed according the official methods of analysis described by the

Association of Official Analytical Chemists (2021), while carbohydrate was calculated by differences.

3.4.1 Moisture Content

Moisture content was determined according to AOAC (2021)protocol. Five grams (5g) of each

sample were placed into apreviously washed, dried and weighed moisture cans, dried inthe oven at

105°C for 4hours and placed indesiccators to cool. Thereafter the process was repeated at a chosen

time intervalother than 4hr until a constant weight was obtained. Theweight of the moisture lost was

calculated as a percentage of weight of sample analyzed as expressed below

% Moisture content = W2-W3/W2-W1×100

Where W1=weight of the empty moisture can, W2=weightof moisture can + sample before drying

and W3=Weight ofmoisture can + sample dried to constant weight.

3.4.2. Crude Fiber Content

Five grams (5g) of the sample was boiled for 30min with150ml of a solution containing 1.25g H2SO4

per 100ml underreflux. The solution was filtered through two-fold muslincloth on a fluted funnel and

washed with boiling water untilit is no longer acidic. The residue was returned to the flaskand boiled

for 30min with 150ml of solution containing1.25g of carbonate free NaOH per 100ml. After washing,

thesample was allowed to drain dry before being transferred in aweighed crucible where it was dried

in the oven at 105°C to aconstant weight which was recorded. The sample was finallyincinerated in a

muffle furnace and the weight of the ash wastaken which was used to determine the fiber thus:

% Crude fiber=100 (W2-W3)

Where: W2=Weight of crucible + sample after oven dryingand W3=Weight of crucible + sample after

incineration AOAC (2022).

19
3.4.3. Fat Content

Soxhlet fat extraction method was used. The boilingflask were dried in the oven and cooled in the

dessicator. Twogram (2g) of each sample was placed in the marked thimblesand weighed. The boiling

flask was filled with 300ml ofpetroleum ether at 60°C. The Soxhlet apparatus wasassembled and

allowed to reflux for 5-6h thereafter thethimble was removed, petroleum ether collected and the

flaskdried at 105°C for 1hour. It was cooled and weighed. Fatcontent was calculated as follows:

% oil=W2-W1/W3 ×100/1

Where: W1=Weight of empty flask, W2=Weight of flask +extract and W3=Weight of sample used.

3.4.4. Protein

The Kjeldahl method was used. 0.2 grams (0.2g) of thesamples were mixed with 15ml of

concentratedtetraoxosulphate (VI) acid in a Kjeldahl digestion flask. Atablet of selenium catalysts was

added and the mixture washeated inside a fume cupboard. The digest was transferredinto a 100ml

volumetric flask, made up with equal volume of45% sodium hydroxide (NaOH) solution and poured

into aKjeldahl distilled apparatus. The mixture was distilled andthe distillate was collected into a 4%

boric acid solutioncontaining 2 drops mixed indicator (mixture of methylred and bromacresol green).

A total of 15ml distillate wastitrated against 0.02N tetraoxosulphate (VI) acid solutionuntil the color

changed from green to a deep red or pink endpoint. Total nitrogen was calculated and multiplied with

3.4.5. Sensory evaluation

Samples produced were presented to 10 semi-trained panelists using 9 hedonic scale were asked to

assess the coded samples in order of like and dislike based on colour, flavour, texture, mouthfeel, taste

and overall acceptability as described by ( Renitaet al,2021). Scoring was done on the 9-point

Hedonic scale where 9 = like extremely and 1 = dislike extremely.

3.5 Microbial Analysis

One gram (1g) representative sample was obtainedaseptically from the muscle of the smoked

catfishsamples. The samples were grounded and serial dilutions(10-1 10-3log cfu/g). of the

homogenized samples were madeusing sterile distilled water. All chemicals used were ofanalytical

20
grade. Each analysis was carried out in triplicates. Allmicrobial analysis was done following the

methodsprescribed by (A.O.A.C., 2020).

3.5.1 Total Plate Count (TBC)

This was done using the pour plate method of (A.O.A.C.2021). One milliliter of the serially diluted

samples wastaken in duplicates and plate count agar was poured at400C on the plates. The samples

and the medium wereproperly mixed, allowed to set and incubated at 350C and370C for 24h. The

number of colonies on the plates wascounted. Coliform counts value is expressed in logarithm colony

forming unit per gram (log cfu/g).

Coli form count

The presumptive coliform count was determined by plating 1 ml of the serially diluted composite

flour samples on MacConkey’s agar and incubated at 37ºC for 24 hr according to Hartman (1985)

method. The method of AOAC (2005) was used to determine the sample was inoculated and

incubated at 37ºC for 24 hr. After 24 hr of incubation the coli form counts are expressed in logarithm

colony forming unit per gramme (log cfu/g).

21
Table 1: Proximate composition of smoked catfish using Rice husk, Palm front, and sawdust (%)

Samples moisture protein fiber fat ash CHO

A 19.72±0.0354ᶜ 59.13±0.0212ᶜ 0.02±0.00ᵇ 14.84±0.021ᵃ 4.165±0.0212ᵃ 2.12±0.0141ᵇ

B 26.85±0.0141 57.07±0.0849ᵇ 0.015±0.00ᵇ 11.45±0.028ᶜ 3.37±0.0070 ᵇ 1.24±0.0283ᶜ

C 27.78±0.141ᵃ 53.16±0.0212ᶜ 0.04±0.00ᶜ 13.36±0.021ᵇ 3.190±0.0141ᶜ 2.47±0.0141ᵃ

Mean value in the column with different superscript are significantly (p< 0.05)Each value represents
the mean ± standard deviation of duplicate determinations; Carbohydrate percent calculated as the
(100 – total other components);SD = Standard deviation; CV = Coefficient of variation.

Key:

Sample A: Rice husk

Sample B: Palm frond

Sample C: Sawdust

22
Table 2: Sensory attributes of the smoke catfish using different sources

Sample Texture mouthful Color taste flavor Overall acceptability

A 7.4ᵃ 7.6ᵃ 7.5ᵃ 7.7ᵃ 7.7ᵃ 7.58ᵃ

B 7.1ᵃ 7.4ᵃ 7.Iᵃ 7.4ᵃ 7.2ᵃ 7.24ᵇ

C 7.1ᵃ 7.0ᵃ 7.2ᵃ 7.7ᵃ 7.1ᵃ 7.22ᶜ

Mean value in the column with different superscript are significantly (p< 0.05). Each value represents
The mean duplicate determinations
Key:
Sample A: Rice husk

Sample B: Palm frond

Sample C: Sawdust

23
Table 3: microbial load on smoked catfish using Rice husk, palm fronds and sawdust as a heat
sources

Sample Total plate count (× 10⁵log cfu/g) Coli form count (× 10⁵log cfu/g )

A 5.405±0.0919ᵃᵇ 4.28±0.0141ᵇ

B 5.55±0.0566ᵃ 4.415±0.0354ᵃ

C 5.27±0.0283ᵇ 4.11±0.0424ᶜ

Mean value in the column with different superscript are significantly (p< 0.05). Each value represents
The mean duplicate determinations
Key:
Sample A: Rice husk

Sample B: Palm frond

Sample C: Sawdust

24
4.0 RESULTS AND DISCUSSION

The smoked fish quality depends on variousfactors, which can be grouped as shown in Figure 1.

To provide constant quality of the finished productall parameters should be considered and

necessaryadjustments made in the processing technologies.

4.1 Raw material

The raw material quality is the main factor affectingsmoked fish quality. Raw material quality has

intrinsicfactors like size, fat content and skin properties, that

4.2 Proximate analysis

There is need for regular monitoring of the levels of PAHs in smoked fish due to the mutagenic and

fcarcinogenic potential of these compounds. Smoked fish has being a common food and the smoking

processes such as combustion temperature, duration of smoking, fuel sources as well as environmental

contaminants differentiates organoleptic properties that are much appreciated by consumers.

Considering the result above, the developed smoking kiln could be used effectively to achieve smoked

fish products free from microbial and dust contaminants from the smoking environments and the fish

handlers during the smoking process. However more microbial analysis should be carried out to

ascertain the durability and the best storage condition and material for the smoked fish. Also more

work should be done to identify the organisms found and recorded in the result of the total microbial

count to confirm that they are not producers of microbial toxins like Staphylococcus toxin which

could constitute harm upon consumption or use in animal.

Raw material quality and its storage conditions are the important parameters for the smoked fish

quality. Better raw fish quality can be provided, when reaching low storage temperature as fast as

possible after catching. The selection of proper pretreatment methods and technology can provide and

ensure constant quality. Smoking conditions have a significant influence on the shelf life of product,

and make changes in volatile compounds, and provide better sensory parameters and texture.

Therefore, systematic research of factors and their combinations is necessary in order to achieve

stable quality of smoked.

25
26
4.3 Result and Discussion

Proximate composition of the samples

Table 1 shows the proximate composition of the samples. Statistical differences (p<0.05) were

observed in all the samples. The moisture content of the samples ranged from 20.70 to 74.98% with

sample A having the highest value, while sample D had the lowest value. However, samples B, C and

D showed a lower moisture content compared to sample A. This might be due to the loss of water

during drying, smoking and the combination of smoking-drying. A study on the influence of the

traditional smoking and/or drying on the quality of catfish by Ali et al. (2011) showed that the

percentage moisture content was low in smoked-dried catfish compared to other samples. The

moisture content results agreed with the work of Cardinal et al. (2001) and Abolagba and Osifo

(2004). They reported that fish weight loss is due to the dehydration during smoking. The protein

content of samples A, B, C and D ranged from 21.71 to 63.00%, with sample D having the highest

value, while sample A has the least value. The increase observed in the protein content of the

processed catfish could be attributed to the loss of water which lead to an increase in the dry matter

content per unit of weight of the sample (Ikeme and Gugnani, 1988; Ime-Ibanga and Fakunle, 2008).

The crude fiber of the catfish ranged from 0.00 – 0.04 % with sample A having the least, while

sample B has the highest value.

The fat content of the sample A to D ranged from 0.50 to 8.57%, with sample B having the highest

value, while sample A has the lowest value. The ash content of the catfish ranged from 1.13 to 4.89%

with sample A having the least value, while sample B has the highest value. There is a correlation

between percentage crude fiber and percentage total ash. This is an indication that total amount of

minerals available in sample C is greater than other samples including the control.

Sensory attributes of the samples Table 2. showed the results obtained for the sensory attributes of the

processed catfish. Mean values obtained for appearance ranged from 7.4 for sample B to 8.6 for

sample D, while that of colour ranged from 7.1 for sample B to 8.5for sample D. The result showed

that there is no significant difference (p<0.05) between the samples in terms of appearance and

colour. All the panelists did not show total dislike for any of the samples appearance and colour. The

27
mean scores obtained for the flavour ranged from 7.2 for sample B to 8.6 for sample D, while that of

taste ranged from 6.2 for sample B to 8.2 for sample D. Texture mean scores ranged from 7.3 for

sample B to 8.3 for sample D. Flavour plays an important factor in consumers preferences and

products, hence sample D would be most preferred than other two samples. Overall acceptability of

the catfish products ranged from 7.1 to 8.6, which indicated that all the catfish products were accepted

by the panelists based on a mean scores obtained, which is greater than 5.0. Sample D, smoked-dried

catfish was the most preferred and accepted one by the panelists as shown in Table 2.

Microbiological quality of the samples

Table 3 shows the microbiological quality of the samples. Statistical differences (p<0.05) were

observed in all the samples Sample D had the lowest total plate count, while sample B had the highest

total plate count. This may be due to the combined processing method used. The aggregate

Salmonella-Shigella count ranged from 0.50 x 10-5 CFU/g to 0.92 x 10-5 CFU/g with sample A

having the highest value, while sample C had the least value.

28
5.0 CONCLUSION AND RECOMMENDATION

5.1 Conclusion

From result above, the selection of proper pretreatmentmethods and technology can provide
andensure constant quality. Smoking conditions have asignificant influence on the shelf life of
product, andmake changes in volatile compounds, and providebetter sensory parameters and texture.

According result, smoked fish has being a common food and the smoking processes such as
combustion temperature, duration of smoking, fuel sources as well as environmental contaminants
differentiates organoleptic properties that are much appreciated by consumers.

Considering the result above, the developed smoking kiln could be used effectively to achieve smoked
fish free from microbial and dust contaminants from the smoking environments and the fish handlers
during the smoking process.

Raw material quality and its storage conditions are the important parameters for the smoked fish
quality.

Smoked catfish may be contaminated by various toxic compounds including carcinogenic

compounds. Most of the chemical hazards reported in this project are processing contaminants. Some
of them can be formed when high temperature is reached inside the product (heterocyclic amines and
nitrosamines) and others during pyrolysis of the fuel during processing such as (PAHs).

5.2 Recommendation

Smoking conditions have a significant influence on the shelf life of product, and make changes in
volatile compounds, and provide better sensory parameters and texture. Therefore, systematic
research of factors and their combinations is necessary in order to achieve stable quality of smoked

There is need for regular monitoring of the levels of PAHs in smoked fish due to the mutagenic and
carcinogenic potential of these compounds.

However more microbial analysis should be carried out to ascertain the durability and the best
storage condition and material for the smoked fish.

Also more work should be done to identify the organisms found and recorded in the result of the total
microbial count to confirm that they are not producers of microbial toxins

29
In traditionally smoked fish, most of the chemical hazards mentioned in this review exceed the
maximal limits established by ISO. Several actions should be addressed to decrease them in smoked
fish as they are highly consumed products.

30
 References

Abdel-Shafy, H. I., & Mansour, M. S. M. (2016). A review on polycyclic aromatic hydrocarbons:

Source, environmental impact, effect on human health and remediation. Egyptian Journal of
Petroleum, 25, 107–123.

Asamoah, E. K., Nunoo, F. K. E., Addo, S., Nyarko, J. O., &Hyldig, G. (2021). Polycyclic aromatic
hydrocarbons (PAHs) in fish smoked using traditional and improved kilns: Levels and human
health risk implications through dietary exposure in Ghana. Food Control, 121, 107576.

Campbell, L. M., Osano, O., Hecky, R. E., & Dixon, D. G. (2003). Mercury in fish from three rift
valley lakes (Turkana, Naivasha and Baringo), Kenya, East Africa. Environmental Pollution,
125, 281–286.

Codex Alimentarius Commission (CAC), (2009). Code of practice for the reduction of contamination
of food with polycyclic aromatic hydrocarbons (PAH) from smoking and direct drying
processes. CAC/RCP, 68.

Desta, Z., Borgstrøm, R., Rosseland, B. O., &Dadebo, E. (2007). Lower than expected mercury
concentration in piscivorous African sharptooth catfish Clariasgariepinus (Burchell). Science
of the Total Environment, 376(1–3), 134–142.

European Food Safety Authority (EFSA). (2008). Polycyclic aromatic hydrocarbons in food.
Scientific opinion of the panel on contaminants in the food chain adopted on 9 June 2008.
EFSA Journal, 724, 1–114.

Essumang, D. K., Dodoo, D. K., & Adjei, J. K. (2013). Effect of smoke generation sources and smoke
curing duration on the levels of polycyclic aromatic hydrocarbon (PAH) in different suites of
fish. Food and Chemical Toxicology, 58, 86–94.

Essumang, D. K., Dodoo, D. K., & Adjei, J. K. (2014). Effective reduction of PAH contamination in
smoke cured fish products using charcoal filters in a modified traditional kiln. Food Control,
35(1), 85–93.

Gao, P., da Silva, E., Hou, L., Denslow, N. D., Xiang, P., & Ma, L. Q. (2018). Human exposure to
polycyclic aromatic hydrocarbons: Metabolomics perspective. Environment International,
119, 466–477.

Gebremedhin, K., &Berhanu, T. (2015). Determination of some selected heavy metals in fish and
water samples from Hawassa and Ziway Lakes. Science Journal of Analytical Chemistry., 3,
10–16.

31
Gómez-Estaca, J., Gómez-Guillén, M. C., Montero, P., Sopelana, P., &Guillén, M. D. (2011).
Oxidative stability, volatile components and polycyclic aromatic hydrocarbons of cold-
smoked sardine (Sardinapilchardus) and dolphin fish (Coryphaenahippurus). LWT - Food
Science and Technology, 44, 1517–1524.

Hokkanen, M., Luhtasela, U., Kostamo, P., Ritvanen, T., Peltonen, K., &Jestoi, M. (2018). Critical
effects of smoking parameters on the levels of polycyclic aromatic hydrocarbons in
traditionally smoked fish and meat products in Finland. Journal of Chemistry, 2018, 1–14.

Igwe, J.C., Odo, E.O., Okereke, S.E., Asuqou, E.E., Nnorom, I.C., Okpareke, O.C. (2012). Levels of
polycyclic aromatic hydrocarbon (PAHs) in some fish samples from Mushin area of Lagos,
Nigeria: Effects of smoking, Terrestrial Aquat. Environment Toxicology 6 (1), 2012) 30–35.

Ingenbleek, L., Veyrand, B., Adegboye, A., Hossou, S. E., Koné, A. Z., Oyedele, A. D., et al. (2019).
Polycyclic aromatic hydrocarbons in foods from the first regional total diet study in Sub-
Saharan Africa: contamination profile and occurrence data. Food Control, 103, 133–144.

Kassegne, A. B., Okonkwo, J. O., Esho, T. B., Mekonnen, K. N., Malehase, T., & Asfaw, S. L.
(2018). Distribution of total mercury in surface sediments and African catfish
(Clariasgariepinus) from Akaki River catchment and Aba Samuel Reservoir, downstream to
the mega-city Addis Ababa, Ethiopia. Emerging Contaminants, 4, 32–39.

Mahugija, J. A. M., &Njale, E. (2018). Effects of washing on the polycyclic aromatic hydrocarbons
(PAHs) contents in smoked fish. Food Control, 93, 139–14.

Phelps, D. W., Fletcher, A. A., Rodriguez-Nunez, I., Balik-Meisner, M. R., Tokarz, D. A., Reif, D.
M., Germolec, D. R., & Yoder, J. A. (2020). In vivo assessment of respiratory burst inhibition
by xenobiotic exposure using larval zebrafish. Journal of Immunotoxicology, 17, 94–104.

SCF (Scientific Committee on Food) (2002). Opinion of the Scientific Committee on Food on the
risks to human health of polycyclic aromatic hydrocarbons in food expressed on 4th
December 2002. Brussels: European Commission, European Commission, Health and
Consumer Protection Directorate-General, 2002 SCF/CS/CNTM/PAH/29 Final.

Silva, B. O., Adetunde, O. T., Oluseyi, T. O., Olayinka, K. O., &Alo, B. I. (2011). Effects of the
methods of smoking on the levels of polycyclic aromatic hydrocarbons (PAHs)in some
locally consumed fishes in Nigeria. African Journal of Food Science, 5(7), 384–391.

Squadrone, S., Chiaravalle, E., Gavinelli, S., Monaco, G., Rizzi, M., &Abete, M. C. (2015). Analysis
of mercury and methylmercury concentrations, and selenium: Mercury molar ratios for a
toxicological assessment of sperm whales (Physeter macrocephalus) in the most recent

32
 stranding event along the Adriatic coast (Southern Italy, Mediterranean Sea). Chemosphere,
138, 633–641

Straaten, P. V. (2000). Human exposure to mercury due to small scale gold mining in northern
Tanzania. Science of the Total Environment, 259, 45–5

Tongo, I., Ogbeide, O., &Ezemonye, L. (2017). Human health risk assessment of polycyclic aromatic
hydrocarbons (PAHs) in smoked fish species from markets in Southern Nigeria. Toxicology
Reports, 4, 55–61.

Yusuf, K. A., Ezechukwu, L. N., Fakoya, K. A., Akintola, L. S., Agboola, J. I., &Omoleye, T. O.
(2015). Influence of fish smoking methods on polycyclic aromatic hydrocarbons content and
possible risks to human health. Africa Journal Food Science, 9(3), 126–135.

Ubwa, S. T., Ubwa, A., & J., Tarzaa, L, Tyohemba R. L., Ahile, U. J. (2015). Effects of Traditional
Smoking Methods on the Concentrations of Polynuclear Aromatic Hydrocarbons (PAHs) in
Some Species of Smoked Fish Traded in Benue State Nigeria.

Journal of Food Research, 4, 2.World Health Organization International Agency for Research on
Cancer. (2010). some non-heterocyclic polycyclic aromatic hydrocarbons and some related
exposures. IARC Monography Evaluation Carcinogenesis Risks Human, 92, 1–853.
Available from: http://publications.iarc.fr/110.

Squadrone, S., Brizio, P., Abete, M.C. et al. Polycyclic Aromatic Hydrocarbons and Mercury in
Smoked Catfish: A Case Study. Water Air Soil Pollut 232, 252 (2021).
https://doi.org/10.1007/s11270-021-05204-1

Oluwaniyi OO, Adeyanju IA, Osibanjo O. (2016). The effects of smoking methods on the proximate
composition, lipid oxidation and mineral contents of catfish (Clariasgariepinus). Food Science
& Nutrition, 4(6), 854-860.

Amaefule KU, Ekwu AO. (2017). Effects of smoking on the quality and shelf stability of African
catfish (Clariasgariepinus) during ambient storage. Food Science & Nutrition, 5(4), 832-840.

Nkafamiya II, Bemji MN, Hassan LG. (2018). Comparative effect of sun and solar drying on the
proximate composition of catfish (Clariasgariepinus). Journal of Nutrition & Food Sciences,
8(1), 1-5.

33