ABSTRACT
Samples were collected from diesel engine generator sites, mechanic workshop and filling station. One sample was collected from diesel oil free environment. The soil samples were analyzed for Total Heterotrophic Bacteria (THB), Fungal Count (TC), Hydrocarbon Utilizing Bacteria (HUB) and Hydrocarbon Utilizing Fungi (HUF). Total Heterotrophic Bacteria Count ranged from 1.73x107– 4.94 x 107while Total Fungi Count ranged from 1.28 x 107 – 2.1 x 107.The bacteria isolated were Bacillus spp., Pseudomonas spp., Micrococcus spp., Escherichia coli, Klebsiella spp., and Serratia spp. Bacillus spp., Micrococcus spp., and pseudomonas spp., showed heavy growths as all the organisms grow in the diesel media at different rates. The fungi isolated were Aspergillus spp., Fusarium spp., and Rhodotorula spp. Aspergillus showed the highest occurrence of growth among the fungi isolated. Analysis of Variance (ANOVA) of the mean count was done. From the chart, the uncontaminated soil (control) had the highest occurrence in Total Heterotrophic Bacteria count among others. The following physico-chemical properties of pH, conductivity, oil and grease, Mg2+, PO4, SO4, NO3, TPH, and TOC were done during the course of this study. Diesel pollution increased the concentrations of NO3, SO4, TPH, Mg2+, PO4, pH, TPH and TOC of the contaminated soil compared to the control. The result showed that the pollution of soil with diesel oil causes great effects on soil physico-chemical characteristics and thus affects microbial nature and population of such soil.
TABLE OF CONTENTS
Title page i
Certification ii
Dedication iii
Acknowledgment iv
Table of contents v
List of tables viii
Abstract ix
CHAPTER ONE
1.0 INTRODUCTION 1
1.1 Aims and Objectives of the Study 2
CHAPTER TWO
2.0 LITERATURE
REVIEW 3
2.1 Historical
Review 3
2.2 Types of
Diesel 3
2.3 Chemical
Properties of Diesel Contaminated Soil 4
2.4 Effects of
Diesel on Soil Macronutrients 4
2.5 Effects of
Diesel on Soil pH 5
2.6 Diesel and
Heavy Metals Accumulation in Soil 5
2.7 Effect of
Diesel on Soil Microbial Activity 6
2.8 Effects of
Diesel Contaminated Soil on Growing Plants 6
2.8.1 Economic
Losses 7
2.8.2 Air and water
contamination 8
2.8.3 Increase in
Soil Salinity 8
2.9 Environmental
Processes on Soil Contaminated with Diesel 8
2.10.1 Uses of
Microorganisms to Remediation Diesel Contaminated soil 10
2.10.2 Bioremediation
of Diesel Contaminated Soil 12
2.10.3 Microorganisms
found in diesel contaminated soil. 13
2.11 Major Sources of Diesel Contaminated Soil 13
CHAPTER THREE
3.0
Materials and Method 15
3.1 Materials 15
3.2 Methods 15
3.3 Sample
Collection 15
3.4 Processing of
Samples 15
3.5 Enumeration
of Total Heterotrophic Bacteria and Fungi 16
3.6 Inoculation
of Media 16
3.7 Characterization
and Identification 17
3.8 Characterization
of Bacterial Isolates 17
3.8.1 Colony
features: 18
3.8.2 Microscopic
characteristic 18
3.9 Gram Staining 18
3.10 Biochemical
Test 19
3.10.1 Catalase Test 19
3.10.2 Coagulase test 19
3.10.3 Methyl Red Test 20
3.10.4 Voges-Proskauer
Test 20
3.10.5 Indole Test 20
3.10.6 Motility Test 21
3.10.7 Identification
of Fungal Isolates 21
3.11 Identification
of Bacterial Isolates 21
3.12 Determination
of Physiochemical Parameters 22
3.12.1 Determination
of Organic Carbon and Organic Matter 22
3.12.2 Determination
of Exchangeable Cation 23
3.13 Determination
of Frequency of Occurrence 23
3.14 Statistical Analysis 23
CHAPTER FOUR
4.0 RESULTS: 24
CHAPTER FIVE
5.0 Discussion,
Conclusion, and Recommendation 31
5.1 Discussion 31
5.2 Conclusion 33
5.3 Recommendation 33
REFERENCES
APPENDIX
LIST
OF TABLES
Table Title Page
1: Mean count of
microorganisms isolated from the soil samples (cfu/g) 26
2: Biochemical characterization of bacteria
isolates 27
3: Bacteria isolates and their percentage
occurrence
28
4: Fugal isolates and their
percentage occurrence 29
5: The value of the
physicochemical parameters of the contaminated and 30
control soil samples
CHAPTER
ONE
1.0 INTRODUCTION
Soil contamination with oil spill is the
major global concern today. This contamination causes serious human health
hazards and organic pollution of ground water, which limit it use, causes
economic loss, environmental problems and also decrease in the agricultural
productivity of the soil. (Thapa et al.,
2012) Environmental contamination with petroleum hydrocarbons is an inevitable
action that strikes many geographical regions to variable extent depending on
the local environmental law. Petroleum products continue to be used as the
principle source of energy, however despite its importance usage, petroleum
hydrocarbon also acts as a globally environmental pollutant, (Smeta et al.,
2012) since the petroleum hydrocarbon are used widely, oil spills are
inevitable even in virtually habitats areas like Antarctica (Smeta, et al.,
2012). Hydrocarbons are bio persistent, bio accumulative and can cause
deleterious effects to aquatic fauna and flora as well as to humans (Bebson et al., 2007). However, not all
hydrocarbon contamination is anthropogenic. Approximately five million joules
of crude oil and refined oil enter the environment each year as a result of
anthropogenic sources such as oil spill.
Extensive changes in marine, as well as
terrestrial ecosystem resulting from the grounding of prestige spill, (2002)
have recently increased the attention of environmentalist, chemists,
biotechnologists and engineers (Smlta et
al., 2012). Diesel spills are by far the largest of all products from crude
oil, because it is used in many sectors, which include, transport, and non-
transport/commercial use (SAPIA, 2007). Diesel is widely used for engines of
cars, generators, industrial trucks and most agricultural machineries (Walker et al., 1978; SAPIA, 2007). The
consumption of diesel has continued to grow worldwide. The increased usage of
diesel has lead to an increase in accidental spillages of diesel, and
consequently environmental pollution, including soil contamination. Soil contamination
by diesel occurs through; leakages from storage containers, refueling of
machineries, wrecks of oil tankers and through improper disposal by mechanics
when cleaning diesel tankers (Alkio et
al., 2005). Diesel spill have been implicated in the destruction of
forests, aquatic and terrestrial ecosystems (Alkio et al., 2005).
Diesel oil is a common product of crude
oil distillation with a very complex composition. It consists mainly of low
molecular weight alkanes and polycyclic aromatic hydrocarbon (Bona et al., 2011). The fate of polycyclic
aromatic hydrocarbon (PAHs) in nature may be of great human health importance,
since PAHs have been considered toxic for plants and carcinogenic to people
(Alkio et al., 2005). In case of an
uncontrolled industrial leakage, diesel oil and it constitute might act as a
persistent water and soil pollutant. According to Nogueria et al., 2011, petroleum compounds can decrease the availability of
water, oxygen and nutrients in the soil which in consequence, may decline the
rate of seed germination.
1.1 Aims and Objectives of the Study
1.
To isolate, identify and
characterize diesel degrading bacteria from diesel contaminated soil.
2.
To isolate, identify and
characterize diesel degrading fungi from diesel contaminated soil.
3.
To check and characterize the
physico-chemical properties of diesel contaminated soil.
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