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ISSN: 2320 – 7051

Int. J. Pure App. Biosci. 2 (1): 213-222 (2014)

Review Article

International Journal of Pure & Applied Bioscience

Pseudomonas in Biodegradation
Shivendra Sharma* and Hardik Pathak
JECRC University Jaipur, Rajasthan
*Corresponding Author E-mail: [email protected]
______________________________________________________________________________

ABSTRACT
Due to careless waste disposal, illegal waste dumping and accidental spills large number of hazardous
compound are released into the environment. The most common chemicals involved in environmental
contamination are petroleum hydrocarbons, polynuclear aromatic hydrocarbons (PAHs).these products
are hazardous to the surroundings as well as to life forms. The most common technique to remove
pollutants from the natural environment and convert the pollutants to a less harmful product using the
microbiological community of the contaminated environment is bioremediation. Pseudomonas sp. is the
most frequently found bacteria in nature which is used in biodegradation process. Degradation of
petroleum hazards by Pseudomonas sp. sp. is the best carrier based inoculums. Pseudomonas sp.
degrades the various hydrocarbons by the different catabolic pathways. The final products can be carbon
dioxide, water and simpler compounds which do not affect the environment. Pseudomonas sp. used to
biodegrade of Naphthalene, Pyrine and Phenanthrene.

Keywords: Petroleum hydrocarbons, PAHs, Pollutants, Biodegradation, Pseudomonas.

____________________________________________________________________________________

INTRODUCTION
The development of human society all over history has lead to rising disruption of the natural equilibrium
and the rate of different types of pollution. The planet depends on oil, and the use of oil as fuel has lead to
intensive economic development worldwide. The great need for this power source has led to the gradual
exhaustion of normal oil capital. on the other hand, mankind will witness the results of oil utilization for
centuries after its termination. Ecological pollution with petroleum and petrochemical products has been
recognized as a significant and serious problem1.
Worldwide industrial and agricultural developments have released a large number of natural and synthetic
hazardous compounds into the environment due to careless waste disposal, illegal waste dumping and
accidental spills. There are numerous sites in the world that require cleanup of soils and sludge. In the
United States, it has been estimated that contaminated site treatment costs may approach 1.7 trillion
dollars over the next 30 years2.
Oil spills in the environment cause long-term damage to aquatic flora, soil ecosystems, human health and
natural resources. Petroleum oil spills tend to be associated with offshore oil rigs and tankers in marine
related accidents. In contrast, land oil spills often go unnoticed by everyone except environmentalist, yet
land oil spills contribute to the pollution of water supply and soil. Typical sources of land oil spill include
accidents as well as oil from vehicles on the road. Characterization of spilled oil and its derivatives is very
important in order to predict the behavior of oil and its long-term effects on the environment, and in order
to select the proper cleaning methods. The potential danger which petroleum hydrocarbons pose to
humans and the environment makes testing and characterization of the biodegradation and
biotransformation processes of hydrocarbons in contaminated soil necessary in order to develop
bioremediation techniques for cleaning such soils to levels that ensures its safe disposal or reuse3.
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Polycyclic Aromatic Hydrocarbons (Pahs)
The most common chemicals involved in environmental contamination are petroleum hydrocarbons,
polynuclear aromatic hydrocarbons ( Benzo (a) pyrene), solvents, pesticides, lead, chlorinated
hydrocarbons (CFH), heavy metals (Cr, Cd Pb, MTBE, Zn and As) and Gasoline. Soil contamination with
polycyclic aromatic hydrocarbons (PAHs) is a vital problem worldwide. Soil organic carbon (SOC) is one
of the important factors that can influence the concentrations of PAHs in soils. Polycyclic aromatic
hydrocarbons (PAHs) are widely distributed and relocated in the environment because of the incomplete
combustion of organic matter.
Polycyclic aromatic hydrocarbons (PAHs) are a group of compounds containing carbon and hydrogen and
are composed of two or more fused aromatic rings in linear, angular and cluster arrangements. PAHs are
the pollutants present in air, soil and sediments. These compounds enter into the environment from many
ways. PAHs and their derivatives are results of incomplete combustion of organic materials which arise
from natural combustion like; forest fires and volcanic eruptions. PAHs are widely found in high
concentrations at many industrial sites, particularly those associated with petroleum, gas production and
wood preserving industries4. PAHs are a group of several hundred individual organic compounds, which
contain two or more aromatics rings and generally occur as complex mixtures rather than single
compounds. PAHs are classified by their melting and boiling point, vapor pressure, and water solubility,
depending on their structure5.
Bioremediation
Bioremediation is a complex process, with biological degradation taking place in the cells of
microorganisms which absorb pollutants, where if they have specific enzymes, the degradation of
pollutants and their corresponding metabolites will take place. Hydrocarbons from oil are used as a source
of nutrients and energy for microorganism growth, and at the same time microorganisms decompose them
to naphthenic acids, alcohols, phenols, hydroperoxides, carbonylcompounds, esters, and eventually to
carbon dioxide and water6.
Bioremediation is best technique to completely remove PAHs from the environment or convert them to
less harmful compounds. Bioremediation of suspended naphthalene or 2-methylnaphthalene as a single
substrate and their mixture was studied using the bacterium Pseudomonas sp. putida. Polycyclic aromatic
hydrocarbons (PAHs) are one of the major groups of these contaminants7. PAHs constitute a diverse class
of organic compounds consisting of two or more aromatic rings with various structural configurations8.
Being a derivative of benzene, PAHs are thermodynamically stable. In addition, these chemicals tend to
adhere to particle surfaces, such as soils, because of their low water solubility and strong hydrophobicity,
and this result in greater persistency under natural conditions. This persistency combined with their
potential carcinogenicity makes PAHs problematic environmental contaminants9,10.
Bioremediation is the use of living organisms to degrade or detoxify hazardous wastes into harmless
substances such as carbon dioxide, water and cell biomass. It uses relatively low-cost, low-technology
techniques, which generally have a high public acceptance. Bioremediation is considered a non-
destructive, cost- and treatment effective and sometimes logistically favorable cleanup technology, which
attempts to accelerate the naturally occurring biodegradation of contaminants through the optimization of
limiting conditions. Bioremediation is an option that offers the possibility to destroy or render harmless
various contaminants using natural biological activity. As such, it uses relatively low-cost, low
technology techniques, which generally have a high public acceptance and can often be carried out on
site1. It will not always be suitable, however, as the range of contaminants on which it is effective is
limited, the time scales involved are relatively long, and the residual contaminant levels achievable may
not always be appropriate11,12.
Bioremediation can be divided into two types: first is natural attenuation, which can be applied when the
natural conditions are suitable for the performance of bioremediation without human intervention, and
second is engineered bioremediation, which is used when is necessary to add substances that stimulate
microorganisms. The first one is more attractive because of its low cost, minimum of maintenance and
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minimal environmental impact. Still, this technology is applicable only in cases when the natural level of
biodegradation is higher than the degree of pollution migration. Nevertheless, this technology is more
often used as a supplement to the other technologies, or after finished engineered bioremediation in order
to prevent migration of pollution from the treated area. Engineered bioremediation is faster than natural
attenuation because it includes microbial degradation stimulation, by controlling the concentrations of
pollution, oxygen, nutrients, moisture, pH, temperature, etc13,14. Engineered bioremediation is applied
when it is essential to carry out cleaning in a short time or when the pollution is very rapidly expanding.
Its application reduces the costs due to the shorter treatment of land and lower number of sampling and
analysis, and it is important for political and psychological needs when the community is exposed to
pollution.
Pseudomonas sp. Involved in Biodegradation:
Some microorganisms found are capable of the transforming and degrading the pollutants, which can also
contaminate the environment. Some species of microorganisms: bacteria, yeasts, and fungi obtain both
energy and tissue-building material from petroleum. The fuel eating bacteria known as Pseudomonas sp.
have evolved a taste for hydrocarbons are the major component of fossil fuels. Degradation of oils by
Pseudomonas sp. is the best carrier based inoculums. These bacteria are found in different environments
such as soil, water, and plant and animal tissue. Various different species of this bacterium are
opportunistic pathogens that affect humans, animals, and plants.
Biodegradation is a natural process by which microbes break down oil into other substances. The final
products can be carbon dioxide, water, and simpler compounds which do not affect the environment.
Pseudomonas sp. sp. ubiquitous in soil and water are considered as scientific and technological
importance. They comprise a taxon of metabolically versatile organisms which is capable of utilizing a
wide range of simple and complex organic compounds. They are known to be involved in biodegradation
of natural or man-made toxic chemical compounds15. Pseudomonas sp. is a prolific producer of a
number of extra cellular enzymes (like lipase). Bioremediation is the optimization of biodegradation. Two
forms of technology can accomplish this acceleration: One is fertilizing (adding nutrients) and the other is
Seeding (adding microbes). These additions are necessary to overcome certain environmental factors that
may prevent biodegradation. Petroleum is a complex mixture of thousands of compounds.
These can be divided into four major groups: the alkanes, the aromatics, the resins, and the asphaltenes. In
general, the alkane fraction is the most biodegradable rather than the polar fraction (i.e., the resins and
asphaltenes) is resistant to biological degradation. The aromatic compounds, especially the polycyclic
aromatic hydrocarbons (PAHs) are of intermediate biodegradability, but these are of most concern owing
to their toxicity and tendency to bioaccumulat16. The bioavailability of weekly soluble hydrophobic
compounds for microbial conversion is usually low and thus limits their degradation rate in aqueous
medium. The use of surfactants has been found to degradation of crude oil17,18.
Contamination of groundwater is also a potential problem. The other impact noticed was on surface
water, mostly nearby the streams, which receive a lot of untreated effluent containing oil and grease as
well as non-biodegradable detergents. Emtiazi et al., studied the biodegradation of petroleum oil by a
Pseudomonas sp. isolated from a petroleum-contaminated soil was instable. They were showed that the
isolates were immobilized on Perlite, they were more stable for oil degradation. The generation time was
for degradation of petroleum oil, dodecane and octadecane was 20, 22, and 25 h. 19.
Catabolic Pathways of PAHs degradation:-(Naphthalene and Pyrine)
Jones et al., recognized the biodegraded petroleum-derived aromatic hydrocarbons in marine sediments.
They investigated the immense biodegradation of alkyl aromatics in marine sediments which eventuate
prior to detectable biodegradation of n-alkane profile of the crude oil and the microorganisms namely:
Arthrobacter, Burkholderia, Mycobacterium, Pseudomonas sp. , Sphingomonas, and Rhodococcus were
observed to be involved for alkyl aromatic degradation20.
Zhang et al., investigated a bacterial isolate, designated as DQ8, was capable of degrading diesel, crude
oil, n-alkanes and polycyclic aromatic hydrocarbons (PAHs) in petroleum. Strain DQ8 was assigned to
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the genus Pseudomonas sp. aeruginosa based on biochemical and genetic data. The metabolites
identified from n-docosane as substrate suggested that P. aeruginosa DQ8 could oxidize n-alkanes via a
terminal oxidation pathway21.
Malik et.al., investigated the metabolic capability of 15 bacterial isolates isolated from oil contaminated
site by using enrichment culture technique which were able to degrade aromatic and polyaromatic
fractions. The results showed that the aromatics compounds (benzene, toluene and xylene) were
vaporized in the 4th day of incubation, while the efficiency on polyaromatic fractions (anthracene,
phenanthrene and pyrene) was 46.17 to 55.3% after 24 days of incubation. The ability of degrading long
chain n-alkanes and crude oil at high concentrations makes the consortium potentially useful for
bioremediation and microbial enhanced oil recovery22. Ashok et.al., isolated four bacterial strains which
were able to degrade naphthalene, anthracene or mixture of both from the soil of oil refinery. Out of four
isolates two of them were identified as belonging to the genus Micrococcus, and other two were identified
as Pseudomonas sp. and Alcaligenes respectively. These species degrade at the rate of 89%, 67.5% and
92.1°/o of high molecular weight plasmid DNA23.
Naphthalene
Naphthalene is the primitive and most soluble PAH in nature. It is found naturally in fossil fuels like; coal
and oil. Burning of fuels, tobacco or wood produces naphthalene as a product. It has a strong odour smells
like coal tar or mothballs. It is used to make products like; moth balls, dyes, leather goods, and as a
household fumigant. Its melting point is 80.2ºC and the molecular weight is 128.2. It degrades more
rapidly as compared to other PAHs found in nature. Bacterial strains that are able to degrade aromatic
hydrocarbons have been isolated mainly from the soil. These generally belong to gram-negative and are
from the genus Pseudomonas sp.
Davies and Evans were the first ones who investigated biochemical sequence and enzymatic reactions
which leads to the degradation of naphthalene. Naphthalene dioxygenase enzyme is required for studying
naphthalene activity however; it is unstable in nature. Naphthalene oxygenase has also been isolated from
cells of Corynebacterium renale which was able to use naphthalene as a main source of carbon and
energy24. Degradation of naphthalene is best studied due to its simplicity. It starts through the
multicomponent enzyme called naphthalene dioxygenase which attacks on the aromatic ring to form cis-
(1R, 2S)-dihydroxy-1,2-dihydronaphthalene (cis-naphthalene dihydrodiol). The cis-naphthalene
dihydrodiol formed by naphthalene dioxygenase and it is subsequently dehydrogenated to 1,2-
dihydroxynaphthalene by an enzyme cis-dihydrodiol dehydrogenase. 1,2-dihydroxynaphthalene is
metabolized to salicylate via 2-hydroxy-2H-chromene-2-carboxylic acid, cis-o-hydroxybenzalpyruvate
and 2-hydroxy-benzaldehyde. However, 1,2-dihydroxynaphthalene is non enzymatically oxidized to 1,2-
naphthaquinone. Salicylate convert to catechol via decarboxylated, and it is further metabolized by ring
fission in meta and ortho-pathways.
Plasmid NAH7 in Pseudomonas putida G7 used to encode bacterial degradation of naphthalene. NAH7
has two operons which contain the structural genes for naphthalene degradation. One operon contains the
gene for the upper catabolic pathway and encodes the enzymes necessary for the conversion of
naphthalene to salicylate. The second operon contains the gene for the lower catabolic pathway and
encodes the enzymes necessary for the metabolism of salicylate through the catechol meta-cleavage
pathway to pyruvate and acetaldehyde. Parales et al., reported aspartate 205 in the catalytic domain of
naphthalene dioxygenase which is a necessary residue in the major pathways of electron transfer to
mononuclear iron at the active site. Several naphthalene degrading bacteria were isolated from oil
contaminated soil in a crude oil extraction and desalination center in Omidieh, Ahvaz, Iran. Bacterial
strains were isolated and identified by biochemical and morphological tests. This investigation exhibited
the most of the isolated bacteria were belong to Staphylococcus sp, Corynebacterium sp, Pseudomonas
sp. Bacillus sp, and Micrococcus sp. These species were able to degrade naphthalene. The efficiency of
naphthalene as the only source of carbon and energy was evaluated by High performance liquid
chromatography (HPLC) analysis. HPLC analysis showed that the Bacillus sp. and Pseudomonas sp. are
capable to degrade naphthalene 86% and 80%, respectively and Corynebacterium sp and Staphylococcus
sp, degrade77% and 69% after one week incubation25.
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Fig.1:The
1:The proposed pathway for naphthalene biodegradation by bacteria

Survey et al., studied the soil near several gas stations in Karachi, Pakistan. He has isolated and identified
60 bacterial strains including Staphylococcus (11.5٪) Corynebacterium (5٪), Bacillus (10٪),
Pseudomonas sp. (8.3٪), Escherichia (33.3٪) and Klebsiella (10٪). These bacteria were capable of
degradation of the hydrocarbons. Coral et al successfully isolated 50 bacterial strains that all belong to
Pseudomonas sp. and were capable of degrading naphthalene. Since native bacteria of contaminated
areas are in contact with aromatic compounds, these bacteria should be able to degrade the materials
surrounding them. 38
Walczak et al and Bestett et al., have studied the degradability of naphthalene by native bacteria. In this
study, the most dominant naphthalene degrading bacteria isolated from the Maroon II oil field belonged to
Pseudomonas sp. and Bacillus.. Therefore, the results of this study showed that these bacteria are native
in this area26,27.
Pyrene
Pyrene is a polycyclic aromatic hydrocarbon, composed of four fused fused benzene rings and the chemical
formula is C16H10. It forms during incomplete combustion of organic compounds. It is a byproduct of
gasification processes and other incomplete combustion processes. This colourless solid is the smallest
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peri-fused PAH (one where the rings are fused through more than one face). Its melting point is 145-
148ºC. Many bacteria have been isolated which are capable of degrading pyrene. Mycobacterium (Gram-
positive species) use it as a sole carbon and energy source to perform their metabolic activity.
Mycobacterium spp. is known to have high cell surface hydrophobicity and adhere to the emulsified
solvent droplets. Other pyrene degrading strains isolated include Rhodococcus sp., Bacillus cereus,
Burkholderia cepacia, Cycloclasticus sp. P1, Pseudomonas sp. fluorescens, Pseudomonas sp. stutzeri,
Sphingomonas sp. VKM B-2434, Sphingomonas paucimobilis, and Stenotrophomonas maltophilia.
Heitkamp et al., found the three products of ring oxidation, pyrene-cis-4,5-dihydrodiol, pyrene-trans-4,5-
dihydrodiol, and pyrenol, and four products of ring fission, 4-hydroxyperinaphthenone, 4-phenanthroic
acid, phthalic acid, and cinnamic acid by multiple analyses, including UV, infrared, mass spectrometry,
NMR, and GC. The formation of pyrene-cis-4,5-dihydrodiol by dioxygenase and pyrene-trans-4,5-
dihydrodiol by monooxygenase suggested multiple initial oxidative attacks on pyrene. Pyrene-1,2-diol
derived from the dioxygenation at pyrene 1,2-C positions is metabolized to 4-hydroxyperinaphthenone
via cis-2-hydroxy-3-(perinaphthenone-9-yl)-propenic acid and 2-hydroxy-2H-1-oxa-pyrene-2-carboxylic
acid28.
Kafilzadeh et. al., isolated bacterial strain from soil of the landfills in Shiraz and evaluation of their
growth kinetic and investigated the pyrene degradation. The isolated bacterial strains were
Mycobacterium sp., Corynebacterium sp., Nocardia sp., Pseudomonas sp. Rhodococcus sp. and
Micrococcus sp. which were potentially capable to degrade pyrene hydrocarbon. After 10 days of
incubation, the pyrene biodegradation value evaluated by high performance liquid chromatography
(HPLC) was 89.1%, 79.4%, 75.3%, 68.2%, 62.3% and 56.8% for each strain respectively. Therefore these
bacteria could be used to clean the soils which are polluted with pyrene29. Moody et.al., reported that the
degradation of anthracene and phenanthrene by Mycobacterium sp. strain PYR-1. After the incubation of
14 days 92% and 90 % of degradation was obtained of anthracene and phenanthrene, respectively.
Metabolites of anthracene and phenanthrene by the UV-visible light absorption, high-pressure liquid
chromatography (HPLC) retention times and mass spectrometry analysis were also identified30. Schneider
et.al., studied the degradation of three polycyclic aromatic hydrocarbons (PAH), pyrene (PYR), benz [a]
anthracene (BAA), and benzo [a] pyrene (BaP), by isolated the Mycobacterium sp. strain RJGII-135 from
abandoned coal gasification site soil by analog enrichment techniques. By high-resolution mass spectral
and fluorescence metabolites of these PAHs were identified. The ability of this bacterium to degrade these
PAH is well supported and used in remediation of sites containing mixtures of these PAH31.
Bisht et.al., isolated naphthalene and anthracene degrading bacteria from non-contaminated soil of
rhizosphere of Populus deltoids. Four isolates were determined by HPLC analysis i.e. Kurthia sp.,
Micrococcus varians, Deinococcus radiodurans and Bacillus circulans utilizing the chrysene, benzene,
toluene and xylene. Among these isolates Kurthia sp and B. circulans showed positive chemotactic
response for naphthalene and anthracene. After 6 days of incubation it was found that B. circulans
SBA12 and Kurthia SBA4 degraded 87.5% and 86.6% of anthracene while, Kurthia sp. SBA4, B.
circulans SBA12, and M. varians SBA8 degraded 85.3 %, 95.8 % and 86.8 % of naphthalene
respectively32. Pathak et al. investigated 4T engine oil biodegradation potential of Pseudomonas sp.
serratia strains, were isolated from contaminated soil from Sitapura industrial area Jaipur. MSM broth
was used in enrichment technique supplemented with 1% v/v hydrocarbon substrate (4T engine oil)33.
Juhasz et.al., analyzed that the degradation of low molecular weight PAHs by isolating numerous genera
of bacteria, fungi and algae have been done while the degradation by high molecular weight PAHs
compounds are generally recalcitrant to microbial attack, although some fungal and bacterial isolates were
identified which were able to degrade four ring PAHs as sole carbon and energy source. This review
concern about the presence of benzo [a] pyrene in the environment and the ability of bacteria, fungi and
algae to degrade the toxic, carcinogenic and mutagenic four fused benzene ring compound, benzo [a]
pyrene34.
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Pyrene is a tetracyclic aromatic hydrocarbon with a symmetrical structure which is one of the top 129
pollutants as ranked by the U.S Environmental Protection Agency. Scientists collected sample from three
different stations in landfills areas in the city of Shiraz, Iran. The bacterial strains were isolated and then
identified by standard bacteriological tests. Isolated bacteria including Mycobacterium sp., Pseudomonas
sp. Rhodococcus sp. and Micrococcus sp. were potentially capable of degrading pyrene hydrocarbon.
They showed high growth rate during increasing the optical density (OD600). Its biodegradation value
evaluated by high performance liquid chromatography (HPLC) was 89.1%, 79.4%, 75.3%, 68.2%, 62.3%
and 56.8% for each strain respectively 10 days after incubation. The highest pyrene degradation rate was
found in Mycobacterium sp. and Corynebacterium sp. with 89.1% and 79.4% values; therefore these
bacteria could be used to clean the soils which are polluted with pyrene degradation occurred in
Mycobacterium and Corynebacterium. This indicates the high potency of these bacteria in contact with
most PAHs. Thus, these two bacteria were introduced as the indicator degrader strain in the landfills’
area35.
Fig. 2:Proposed catabolic pathways of pyrene by bacteria

Compound designations: 1, pyrene; 2, pyrene-cis-1,2-dihydrodiol; 3, pyrene-1,2-diol; 4, 2-hydroxy-3-

(perinaphthenone-9-yl)-propenic acid; 5, 2-hydroxy-2H-1-oxa-pyrene-2-carboxylic acid; 6, 4-
hydroxyperinaphthenone; 7, 1,2-dimethoxypyrene; 8, pyrene-trans-4,5-dihydrodiol; 9, pyrene-cis-4,5-dihydrodiol;
10, pyrene-4,5-diol; 11, phenanthrene-4,5-dicarboxylic acid; 12, 4-carboxyphenanthrene-5-ol; 13, 4-carboxy-5-
hydroxy-phenanthrene-9,10-dihydrodiol; 14, 4-carboxyphenanthrene-5,9,10-triol; 15, 2,6,6’-tricarboxy-2’-
hydroxybiphenyl; 16, 2,2’-dicarboxy-6,6’-dihydroxybiphenyl; 17, phthalic acid; 18, 4-phenantroic acid; 19, 3,4-
dihydroxy-3,4-dihydro-phenanthrene-4-carboxylic acid; 20, phenanthrene-3,4-diol; 21, 4-phenanthroic acid methyl
ester, 22, 4-hydroxyphenanthrene; 23, 7,8-benzocoumarin; 24, 2-hydroxy-2-(phenanthrene-5-one-4-enyl)-acetic
acid; 25, 5-hydroxy-5H-4-oxa-pyrene-5-carboxylic acid; 26, pyrene-4,5-dione; 27, 4-oxa-pyrene-5-one.
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Bishnoi et al., studied pyrene degradation by Pseudomonas Putida and Pseudomonas paucimobilis were
selected for further study. After 42 days, Pseudomonas Putida degraded 59.8% of pyrene and
Pseudomonas Paucimobilis degraded 52% of pyrene. In the current study, this bacterium also was
isolated as the pyrene-using strain and its degradation value was 68.2%36. In another study in normal
conditions by Shafiee et al., after 10 days 60% of pyrene was consumed by soil bacteria. Studies on
compounds with low molecular weight, e.g., the average of phenanthrene and anthracene degradation by
soil bacteria, was reported as 48.44% and 30.19%, respectively, after 24 h 37.

CONCLUSION
Bioremediation offers an alternative method for detoxification of contaminant. It is a natural process
which relies on bacteria, fungi and plants to alter contaminants. These organisms carry out their
normal life processes using these contaminants as their source of nutrients. Metabolic processes of these
organisms are capable of using chemical contaminants as energy source, rendering the contaminants
harmless or less toxic in most cases. This study shall provide a better solution for bioremediation of
spilled petroleum hydrocarbons in soil and water ecosystems. It will provide an idea on distribution of
microorganisms in the environment which have the ability to degrade the hydrocarbons and investigation
of the response of microorganisms towards different petroleum oils.
Acknowledgement
I Pray To Almighty for all Known, Unknown Hands And Learned Soul That Enlightened My Path And
Helped Me In Meeting The Final Of Endeavour With Great Exultation. I Am Deeply Inducted To Dr.
Hardik Pathak HOD Biotechnology, JECRC UNIVERSITY, For Her Indispensable And Unstained
Cooperation As My Research Guide.

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