Isolation And Identification Of Crude Oil Degrading Bacteria From Soil Polluted With Automobile Lubricants

Title page

Approval page

Dedication

Acknowledgement

Abstract

Table of content

List of tables

List of figures

CHAPTER ONE – INTRODUCTION AND LITERATURE REVIEW

1. 1.        Introduction
   2.         Literature review
      1.     Component of crude oil
      2.      Crude oil degrading bacteria
         1.       Bacteria
         2.       Fungi and Yeasts
         3. Algae
      3. Biodegradation and Bioremediation
      4.  Physicochemical versus biological methods for degrading petroleum hydrocarbon pollutants
      5.   Microbial degradation of petroleum hydrocarbon
      6.   Factors affecting petroleum hydrocarbon degradation
         1.   Temperature

1. 1. 1. 1.    Nutrient
         2.    Moisture

CHAPTER TWO – MATERIALS AND METHODS

2.0.        Materials

2.1.        Sample collection

2.2.        Reagents and media

2.4.        Methods

2.5.        Media preparation

2.5.1.      Preparation of Bushnell Haas broth ( Mineral salt broth)

2.5.2.      Preparation of Bushnell Haas agr ( mineral salt agar)

2.5.3.      Preparation of nutrient agar

2.6.         Isolation of crude oil degrading bacteria

2.6.1.      Inoculation of sample into Bushnell Haas broth

2.6.2.       Serial dilution

2.6.3.      Inoculation into Bushnell agar

2.6.4.      Viable plate count

2.6.5.       Pure culture

2.6.6.       Preparation of agar slant

2.6.7.       Inoculation into agar slant

2.7.          Identification of isolates

2.7.1.      Morphology examination and identification

2.7.1.1.   Macroscopy

2.7.1.2.   Microscopy

2.7.1.3.    Gram staining

2.7.2.       Biochemical tests

2.7.2.1    Catalase test

2.7.2.2.   Sugar fermentation test

2.7.2.3.   Nitrate reduction test

2.7.2.4.   Indole test

2.7.2.5.   Starch hydrolysis test

2.7.2.6.   Simmon citrate test

2.7.2.7.   Methyl red test

2.7.2.8.  Voges Prokaurer test

2.7.2.9.   Motility test

2.8.         Determination of rate of utilization of crude oil by the different isolates

CHAPTER THREE- RESULTS

3.1          Colony morphology of isolates

3.2          Microscopy of isolates

3.3.         Biochemical tests of selected isolates

3.4.         Optical densities of selected isolates

3.5.         Graph of optical densities of isolates

CHAPTER FOUR- DISCUSSION AND CONCLUSION

4.1.         Discussion

4.2.         Conclusion

REFERENCES

                                                      LIST OF TABLES

Table 1.           Colony morphology of isolates after inoculation into Bushnell Haas agar-23

Table 2.            Morphology of isolates

Table 3.            Morphology and biochemical tests results of selected isolates

Table 4.             Optical densities of isolates

                                                                   LIST OF FIGURES

Figure 3.1:              Time-course of medium optical density change during growth of IS1 isolate (Micrococcus spp} on mineral salt medium with specific concentration of lubricating oil (crude oil product).

Figure 3.2:                 Time-course of  medium optical density change during growth of IS2 isolate (Streptomyces spp}  on mineral salt medium with specific concentration of lubricating oil (crude oil product