BIOSTIMULATION OF BACTERIA DEGRADATION OF CRUDE OIL POLLUTED SOIL USING PLANTAIN PEELS

Biostimulation of bacteria degradation of crude oil polluted soil using plantain peels were studied in University of Abuja permanent site. Top soil was collected randomly from different points, sieved and weighed out 3kg into each plastic bucket, and were polluted with 300ml, 250ml and 200ml on each. The plastic buckets were labeled in single form with cotton wool plugged at the base to retain the water. (A,B,C,D,E,F,G,H,I,J) with the control as J without crude oil. After 30 days, the sun-dried plantain peels (PP) was added (200g-300ml,150g-250ml and 120g-200ml) and allowed to stand then Soil sample were analyzed. The results revealed that plantain peels had a crude oil utilizing bacteria (CUB) count 7.5 x 109±0.1 cfug-1 (200g PP) followed by 5.8 x 109± 1.4 (150g PP) and 5.0 x 109± 0.7 (120g PP) for plantain peels. Four micro-organisms were isolated including: Pseudomonas aeruginosa, Bacillus species, Staphylococcus aureus and Actinomycetes species. The result of this study indicates that plantain option in the degradation of total petroleum hydrocarbon in the so plantain peels possess bio-utiliziing potentials and thus suggest that the waste should be considered as an option in the degradation of total petroleum hydrocarbon in the soil.

Keyword: Crude oil, Bacteria, Plantain peels, Mineral salt medium and N-Hexane

TABLE OF CONTENTS

TITLE PAGE..............................................................................................................i

DEDICATION...........................................................................................................ii

DECLARATION......................................................................................................iii

CERTIFICATION.....................................................................................................iv

ACKNOWLEDGEMENTS.......................................................................................v

ABSTRACT..............................................................................................................vi

TABLE OF CONTENTS........................................................................................vii

LIST OF TABLES.................................................................................................viii

CHAPTER ONE

INTRODUCTION

1.1 Justification of study ……………………………………………………...3

1.1. Aim and Objectives of the study.......................................................................4

CHAPTER TWO

2.1. Background of study.........................................................................................5

2.1.1. History of crude oil in Nigeria........................................................................5

2.2.

Biostimulation...................................................................................................7

2.3.

Hydrocarbon....................................................................................................11

2.4. Relation of micro-organisms to petroleum.....................................................13

2.5. Fossil fuel formation/ petroleum formation....................................................14

2.6. Petroleum biodegradation...............................................................................15

2.7. Composition of crude oil................................................................................16

2.8. Uses of crude oil.............................................................................................17

2.9. Physicochemical properties of plantain..........................................................17

CHAPTER THREE

3.0. Materials and methods....................................................................................19

3.1. Sample collection............................................................................................19

3.2. Determination of pH and Temperature of the normal Soil and contaminated

Soil with Crude oil..........................................................................................19

3.3. Enumeration of total heterotrophic bacteria (THB)........................................20

3.4. Bacteria counts and isolation..........................................................................21

3.5. Identification of organisms.............................................................................22

3.6. Gram stain.......................................................................................................22

3.7. Catalase test....................................................................................................23

3.8. Indole test........................................................................................................23

3.9. Oxidase test......................................................................................................23

3.10. Motility test....................................................................................................24

3.11. Spore test........................................................................................................24

3.12. Fermentation test............................................................................................24

3.13. Isolation of crude oil degrading bacteria.......................................................25

3.14. Contents of mineral salt medium...................................................................25

3.15. Application of plantain peels.........................................................................26

3.16. Residual crude oil after contamination..........................................................27

CHAPTER FOUR

4.0.

Results.............................................................................................................29

4.1. Effect of crude oil on soil bacterial population...............................................29

4.2. Physico-chemical characteristics of contaminated soil...................................39

4.3. Isolates from crude oil polluted soil................................................................41

4.4. Biochemicaland Morphological characteristics of the organisms isolated.....45

CHAPTER FIVE

5.0. Discussion, Conclusion and Recommendation……….............................46

5.1.

Conclusion......................................................................................................50

5.2.

Recommendation............................................................................................51

5.3.

References.......................................................................................................52

5.4.

Appendix.........................................................................................................58