ABSTRACT
The microbiological and physicochemical characteristics of soil contaminated with petroleum products were carried out. A total of 3 sampling sites ( Filling station, mechanic workshop and areas around power generating plant) were investigated. The media used were Nutrient agar, Sabouraud dextrose agar, Mineral salt agar for Isolation of total heterotrophic bacteria and fungi and hydrocarbon-utilizing bacteria and fungi, respectively. The pour plate method is used for the isolation of microorganisms. The total heterotrophic bacterial count ranged from 1.4×107±0.36 - 1.7×108±0.06 cfu/g, while the total heterotrophic fungal count ranged from 6.0×105±0.63 - 9.0×106±4.0 cfu/g. The hydrocarbon-utilizing bacterial count ranged from 3.0×105±0.81 - 2.3×106±0.17 cfu/g, while hydrocarbon-utilizing fungal count ranged from 1.0×106±0.2 - 1.4×106±0.2 cfu/g. The microorganisms isolated were Bacillus species, Escherichia coli, Staphylococcus species, Klebsiella species, Micrococcus species, Penicillium species, Aspergillus species, Rhizopus species and Mucor species. The result of the physicochemical parameters ranged as follows: Oil and grease, 520mg/kg to 615mg/kg; total organic carbon, 22.10% to25.65%; pH, 3.8 to 4.3; Conductivity, 129µS/cm to 134µS/cm; Phosphate, 1.92 mg/kg to 2.26 mg/kg; Sulphate, 12.70mg/kg to 16.30mg/kg; Nitrate, 1.72mg/kg to 2.19mg/kg; Total Iron, 39.5mg/kg to 53.4mg/kg; Copper, 1.92mg/kg to 2.26mg/kg; Nickel, 0.081mg/kg to 0.102mg/kg; Cadmium, 0.049mg/kg to 0.085mg/kg, Barium, 0.034mg/kg to 0.046mg/kg; Manganese, 7.08mg/kg to 9.47mg/kg; Sodium, 13.7mg/kg to 18.4mg/kg; Magnesium, 1.86mg/kg to 3.72mg/kg; Potassium, 4.11mg/kg to 5.21mg/kg; and Calcium, 129mg/kg to 169mg/kg. It has been observed that the used petroleum had impacted negatively on the soil within the vicinity of the polluted sites and could affect vegetation around the area.
TABLE OF CONTENTS
Title
page i
Certification ii
Dedication iii
Acknowledgement iv
Table
of Contents v
List
of Tables x
Abstract ix
CHAPTER
ONE: INTRODUCTION 1
1.1 Aims and objectives 3
CHAPTER
TWO: LITERATURE REVIEW 4
2.1 The Biology of Soil 4
2.2 Petroleum and its Derivatives 5
2.3 Environmental contamination of soil 6
2.4 Sources of Petroleum contamination in soil 6
2.5 Effect of petroleum contamination 6
2.6 Effect of soil contamination on ecosystem 7
2.7 Clean up of soil 7
2.7.1 Bioremediation of
sites contaminated by petroleum derivatives 10
2.8 The physical characteristics of soil 11
1.8.1 Soil texture 11
1.8.2 Soil structure 12
1.8.3 Soil tilth 13
1.8.4 Soil water 13
1.8.5 Soil organic matter 14
1.9 Chemical properties of soil 14
1.9.1 Cat ion exchange 14
1.9.2 Soil pH 15
1.9.3 Soil salinity 16
1.9.4 Heavy metals 16
CHAPTER THREE: MATERIALS
AND METHODS 19
3.1 Study area 19
3.2 sample collection 19
3.3 Microbiological
methods 19
3.3.1 Media 19
3.4 Enumeration of Total Heterotrophic Bacteria
and Fungi 20
3.5 Enumeration of hydrocarbon-utilizing Bacteria
and Fungi 20
3.6 Characterization and identification of
Hydrocarbon utilizing isolates 21
3.6.1 Macroscopic
Examination 22
3.6.2 Microscopic
Examination 22
3.6.3 Gram staining 22
3.6.4 Lacto phenol cotton
blue 22
3.6.5 Spore Staining 23
3.6.6 Motility Test 23
3.7 Biochemical Tests 24
3.7.1 Catalase test 24
3.7.2 Citrate test 24
3.7.3 Oxidase test 24
3.7.4 Indole test 24
3.7.5 Urease test 25
3.7.6 Coagulase test 25
3.7.7 Methyl red- Voges
proskquer test 25
3.7.8 Triple Sugar Iron
Agar (TSIA) Test 25
3.7.9 Starch Hydrolysis
Test 26
3.7.10 Sugar Fermentation
Test 26
3.8 Determination of physicochemical parameters 27
3.8.1 pH Determination 27
3.8.2 Electrical
Conductivity 27
3.8.3 Determination of
sulphates 27
3.8.4 Determination of
nitrates 28
3.8.5 Determination of
phosphates 28
3.8.6 Heavy metal
Analysis 29
3.8.7 Determination of
oil and grease and Total Petroleum Hydrocarbon(TPH) 29
3.8.8 Determination of
Total Organic Carbon 29
3.8.9 Determination of
Exchangeable Cat ions 30
CHAPTER FOUR: RESULTS 31
CHAPTER FIVE: DISCUSSION,
CONCLUSION, RECOMENDATION 41
5.1 Discussion 41
5.2 Conclusion 44
5.3 Recommendation 44
References
Appendix
LIST OF TABLES
Table Title Page
1 Total Microbial counts of Soil
Samples 33
2 Percentage occurrence of Bacterial
Isolates 34
3 Isolation and Identification of
Bacterial Isolates 35-36
4 Percentage Occurrence of Fungal
Isolates 37
5 Isolation and Identification of
Fungal Isolates 38
6 Analysis of Physicochemical
Parameters 39-40
CHAPTER ONE
1.0. INTRODUCTION
Man’s technological and scientific advances
have caused environmental changes that are
impossible
to evaluate and fully comprehend. Pollution of the environment is one of the
major
effects
of man’s technological advancement. Pollution results when a change in the
environment
harmfully
affects the quality of human life, animals, microorganisms and plants ((Okoh,
2006).
Accidental
and deliberate crude oil spills are significant sources of environmental
pollution,
posing
serious environmental problem such as damage to vegetation, soil-borne
microorganisms
and
soil fertility. (Nweke and Okpokwasili, 2004).
Pollution
of the environment by petroleum products is an inevitable consequence of oil
production, transportation and distribution activities. Large amounts of
petroleum products handled on land every year create the possibility for land
contamination.
In
addition, large volumes of crude oil and/or refined petroleum products are
transported across the world's oceans from producing areas to consumer
countries (Atlas, 1981). Consequently, a substantial fraction of this oil is
released into the sea either by accident or during normal tanker operations. It
has been estimated that approximately 0.1 % of transported crude oil (about 35
million metric tons) enters the sea annually from tankers alone (Energy
Information Administration, 1992).
Other
pollution sources include ballast water discharges, natural seepage, blow out
of wells, leakages from pipelines and storage tanks, industrial wastes and
runoffs and sometimes sabotage (Atlas, 1981).
The
toxicity of crude oil or petroleum products varies widely, depending on their
composition, concentration, environmental factors and on the biological state
of the organisms at the time of the contamination. The discharge of used oil
from vehicles, motorcycles, filling stations and generator house are major
source of oil pollution in the environment.
Used
oil is any oil that has been refined from crude oil or any synthetic oil that
has been used and as a result of such use, is contaminated by physical and
chemical impurities. Used oil is typically contaminated or mixed with dirt,
fine particle, water or chemicals, all of which affect the performance of the
oil and eventually render it unstable. Used oil does not include product
derived from vegetables or animal fats or petroleum distillates used as
solvents. Used oil is generated from
many different sources. The most common sources are manufacturing companies,
electric generator, mining /smelter operations, automotive maintenance
facilities and air conditioning repair facilities (Kobayashi and Rittman,
1992).
Biodegradation of hydrocarbons by natural population of microorganisms represents one of
the primary mechanisms of eliminating petroleum pollution from the environment
(Leahy and Colwell, 1990). The ability to degrade and/or utilize hydrocarbon
substrates is exhibited by a wide range of bacteria and fungi (Atlas, 1981).
The ability to isolate high numbers of certain oil-degrading microorganisms
from oil-polluted environment is commonly taken as evidence that these
microorganisms are the active degraders of the pollutants in the environment
(Okerentugba and Ezeronye, 2003).
Petroleum
products such as engine oil, petrol, diesel and kerosene are used daily in
various forms in mechanic workshops, generator house and filling station. These
products tend to harden or change the texture of the soil, which may have
effects on the microbiological and physicochemical properties of the
contaminated soil. Therefore, due to the increasing number of mechanic
workshops, filling stations, generator house and their indiscriminate disposal
of used oil into the environment, there is need to consider options for their
removal from the environment due to the environmental hazards associated with
petroleum products.
1.1. AIMS AND OBJECTIVES
1. To assess the microbiological and
physiochemical characteristics of soils contaminated with used petroleum products.
2. To determine the effect of used petroleum
on the microbiological and physiochemical
characteristics of soil.
3. To
evaluate the potential of microorganisms isolated from oil contaminated soil.
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