MICROBIOLOGICAL AND PHYSICOCHEMICAL ASSESSMENTS OF DIESEL CONTAMINATED SOIL.

ABSTRACT

Microbiological and physicochemical assessments of diesel contaminated soil were done. Four soil samples were collected from diesel engine generator sites. One sample was collected from diesel oil free environment. The soil samples were analyzed for total heterotrophic bacteria (THB), total heterotrophic fungi (THF), hydrocarbon utilizing bacteria (HUB) and hydrocarbon utilizing fungi (HUF). The result showed that total heterotrophic bacterial count ranged from 1.52 x 107-2.63x107 cfu/g while total heterotrophic fungal count ranged from 3.0x106 -7.4x106cfu/g. The hydrocarbon utilizing bacteria was in the range of 5.6x105-1.66x106 cfu/g while hydrocarbon utilizing fungi ranged from 1.5x105 -5.3x105cfu/g. The bacteria isolated were Bacillus sp., Micrococcus sp., Pseudomonas sp., Staphlococcus sp., Streptococcus sp., Corynebacterium sp., Flavobacterium sp., Arthrobacter sp., Acinetobacter sp., Klebsiella sp., and Alicagenes sp. Micrococcus sp., Pseudomonas sp., and Bacillus sp., showed heavy growths as all the organisms grow in the diesel media at different rates. The fungi isolated were Aspergillus sp.,Fusarium sp., Geotricum sp., Rhizopus sp., and Cladosporium sp. Aspergillus sp. Showed the highest occurrence of growth among the fungi isolated. The following physicochemical properties of pH, conductivity, Ca2+, Mg2+, oil and grease, PO4, SO4, NO3, TPH and BTEX were done during the course of this study. Diesel pollution increased the concentrations of NO3, SO4, TPH, Ca2+ and Mg2+ of the contaminated soil. There were no significant differences in the BTEX concentrations, conductivity and pH of the samples analyzed. The result showed that the pollution of soil with diesel oil causes great effects on soil physicochemical 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 ix

List of figures                                                                                    x

Abstract xi

CHAPTER ONE

1.0 Introduction 1

1.1 Aims and objectives    2

CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Biology of soil 3

2.2 Soil profile 4

2.2.1 Structure of Soil Profile 5  

2.3       Soil Structure 6

2.4 Chemical properties of soil 7

2.4.1    Cation exchange 10

2.4.2. Soil pH    8

2.4.3. Heavy metals 9

2.5 Soil microbial community 9

2.6 Components of soil microbiota 10

2.7 Diesel 11

2.8 Chemical properties of diesel soil 11

2.9 Major sources of diesel contamination of soil 12

2.10 Diesel Degradation by Microbes 12

2.11 Health and Environmental Risks Regarding contamination of soil by

Diesel fuel 13

2.12 Bioremediation 14

2.13 Advantages of bioremediation 15

2.14 Disadvantages  of bioremediation 16

CHAPTER THREE

3.0 MATERIALS AND METHOD

3.1 Study area 17

3.2 Sample collection 17

3.3 Preparation of media 17

3.4 Preparation of sample and isolation 18

3.6 Microbial characterization and Identification 19

3.6.1 Identification of Bacterial Isolates 19

3.6.2 Gram Staining 19

3.6.3 Motility Test 20

3.6.4 Indole test 20

3.6.5 Catalase Test 20

3.6.6 Oxidase Test 21

3.6.7 Methyl red-Voges Proskauer Test 21

3.6.8 Identification of fungal Isolates 22

CHAPTER FOUR

4.0 RESULTS 43

CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATIONS

5.1 Discussion 29

5.2 Conclusion 54

5.3 Recommendation 33

Reference 55

Appendix 40

LIST OF TABLES

Table          Title                                                                                                   Page

 1:              Microbial count (cfu/g) 25

 2:              Bacterial isolates and their percentage occurrence 26

3:               Fungal isolates and their percentage occurrences 27

4:               The value of the parameters of the contaminated and                         28

                  control soil samples

LIST OF FIGURES

Figure       Title                                                                                      page

1:              TPH Chromatograph for sample A                                             29

2:              TPH Chromatograph for sample B                                             30  

3:              TPH Chromatograph for sample C                                             31

4:              TPH Chromatograph for sample D                                             32

5:              TPH Chromatograph for sample E                                             33

CHAPTER ONE

1.0 INTRODUCTION

There is rapid exponential increase in the world population especially in the last 100 years. This rapid increase in the world population has resulted in the increase in the demand for hydrocarbon products such as diesel, petrol, kerosene, pesticides, agricultural chemicals, and industrial chemicals (Chakrabarty et al., 1998). This increase in hydrocarbon products suffers the environment, as the environment pays the price for the high usage of these products. Although many of the chemicals are utilized or destroyed, a high percentage is released into the air, water and soil representing a potential environmental hazard (Alexander, 1995; Stephen et al., 2011). Processing and distribution of petroleum hydrocarbons as well as the use of petroleum products lead to contamination of soil (Ayotamuno et al., 2006). These processing, distribution, usage and disposal of petroleum (directly or indirectly) lead to heavy contamination of soil leading to oil contamination of soil.

Oil contamination is one of the most dangerous pollution factors known today. It causes a lot of threat to our environment. They cause changes in soil properties due to the contamination with petroleum derived substances and they can be very hard to control in most serious cases. Hydrocarbon spill or contamination of soil can lead to water and oxygen deficits as well as shortage of available forms of nitrogen and phosphorus (Wyszkowska and Kucharski, 2000). Hydrocarbon spill is a serious threat to the ecology. According to Dorn et al., (1998), hydrocarbon contains substances that are toxic to the flora and fauna found in the ecosystem.

Diesel oil, which is one of the major products of crude oil, constitutes a major source of pollution in our environment. Diesel contains low molecular weight compounds and high proportion of saturated hydrocarbon that are usually more toxic than long chained hydrocarbons. With the combined dependence on diesel oil by some vehicle, and generators, greater quantities are being transported over long distances. Therefore diesel engine oil can enter into the soil through wrecks of oil tankers carrying diesel oil, cleaning of diesel tanks by merchants, warship carrying diesel oil and motor mechanics (Hill et al., 1980). Diesel pollution is on the rise in Nigeria, as well as other developing countries (Stephen et al., 2011). The resultant effects of this increase in diesel oil usage include contamination of soil and water bodies. This contamination can also occur when such generators are serviced and the oil discarded carelessly in the environment. This often results in distortion of soil physical, biological and chemical properties. This soil’s physical properties also include the microorganisms that form the normal fauna of the soil. This research work was undertaken to assess the effect of diesel oil on soil microorganisms and the physiochemical properties of the contaminated the soil. Many microorganisms have the ability to utilize hydrocarbons (diesel, fuel and kerosene) as sole sources of carbon and energy for metabolic activities but the microbial utilization of these hydrocarbons depends on the chemical nature of the compound within the petroleum mixture and on environmental determinants and biodegradability. Since hydrocarbons are natural products, it is not surprising to find organisms that are capable of degrading these energy-rich substrates.

1.1 AIMS AND OBJECTIVES

i To isolate, identify and characterize diesel oil degrading bacteria from diesel oil contaminated soil.

ii To isolate, identify and characterize diesel oil degrading fungi from diesel oil contaminated soil.

iii To check and characterize the physiochemical properties of diesel oil contaminated soil

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