ABSTRACT

The antibacterial activity of  bile on bacteria isolated from  faeces, urine and blood  was determined in this study precisely Escherichia coli and Salmonella spp   . Bile from three animal origin(cow ,goat and chicken) was extracted using alcohol and water and their antibacterial activity tested using the agar well diffusion method  at different concentrations . The crude extracts exerted the highest inhibition zone diameter within the range of 20mm-49mm and 38mm-48mm against Salmonella spp and Escherichia coli respectively, followed by the alcoholic extract with inhibition zone diameter in the range of  6mm-35mm and 7mm-33mm against Salmonella spp and Escherichia coli respectively. While the aqueous   extract exhibited the least activity   in the range of 5mm-25mm and 7mm- 25mm against Salmonella spp and  Escherichia coli respectively.   The MIC of the bile extracts which was determined using the tube macrodilution method and the MBC showed  that  bile have  both bactericidal and bacteriostatic activity on the test organisms. The findings of this work has shown that bile contains some substances that can exert inhibitory effect on bacteria hence can be employed  in the treatment of diseases caused  by these groups of bacteria.

TABLE OF CONTENT

Title page i

Declaration ii

Certification iii

Dedication iv

Acknowledgment v

Table of content vi

List of tables vii

Abstract viii

CHAPTER ONE

 INTRODUCTION 1

1.1       Background of the Study              1

1.2       Aim of the Study 4

1.3       Specific Objectives 4

1.4       Significance 4

CHAPTER TWO

LITERATURE REVIEW 5

2.1      Cholanology 5

2.2      Chemical Structure of Bile And Evolution 5

2.3      Bile and microorganisms: Previous Studies 6

2.4      Overview of Enterobacteriaceae Cell Envelope 8

2.5      Mechanism of Action of Bile on Cells 10

CHAPTER THREE

MATERIALS AND METHODS 14

3.1        Materials 14

3.1.1     Equipments 14

3.1.2     Media, Reagents and Solvents Used 15

3.2        Methods 16

3.2.1     Bile Sample Collection 16

3.2.2     Bile Extraction 16

3.2.3     Preparation of Extract Concentration 16

3.2.3.1  Preparation of Aqueous Bile Solution 17

3.2.3.2  Preparation Alcoholic Bile Solution 17

3.2.3.3  Preparattion of the Crude Extract 17

3.2.4     Media Preparation 18

3.2.5     Isolation of the Test Organisms 18

3.2.6     Identification 18

3.2.6.1  Grams Reaction 18

3.2.6.2  Citrate Utilization Test 19

3.2.6.3  Indole Test 19

3.2.6.4  Oxidase Test 20

3.2.6.5  Methyl Red-Voges Proskauer Test. 20

3.3   Bile Activity Assay 21

3.3.1      Preparation of Standard Innoculums of Test Organisms 21

3.3.2      Antibacterial Susceptibilty Testing 21

3.3.3      Well Diffusion Method Assay 21

3.3.4      Determination Of Minimum Inhibitory Concentration (MIC) Using the         

             Macrodilution Method 22

3.3.5      Determination of Minimum Bactericidal Concentration (MBC) 22

CHAPTER FOUR

RESULTS 23

4.1       Morphological And Biochemical Properties of the Isolates 23

4.2       Antibacterial Activity of Bile on the Test Organisms Using 

 Well Diffusion Method Assay 25

4.3       Antibacterial Activity of Bile on the Test Organism Using 

 Tube Macrodilution Assay 29

CHAPTER FIVE

DISCUSSION 37

5.1      Conclusion 40

5.2      Recommendations 40

          REFERENCE        

          APPENDIX

                                        LIST OF TABLES

 1:     Morphological and biochemical properties of the 25

         isolates    

 2:     Antibacterial activity of aqueous bile solution 

         from cow, goat and chicken against the isolates 27

3:      Antibacterial activity of alcoholic exracts of bile

         from cow, goat and chicken against the isolates 28

 4:    Antibacterial activity of crude extracts of bile

        from cow,goat and chicken against the isolates 29

 5a :   Mnimum inhibitory concentration (MIC) and minimum   

         bactericidal concentration (MBC) of the cow

         bile against salmonella spp   32

 5b:      Minimum inhibitory concentration (MIC) and minimum 

            bactericidal concentration (MBC) of the goat

            bile against salmonella spp 33

 5c :     Minimum inhibitory concentration (MIC)  and minimum

            bactericidal concentration (MBC) of the chicken

            bile against salmonella spp 34

 6a:      Minimum inhibitory concentration (MIC) and minimum

            bactericidal concentration (MBC) of the cow 

            bile extracts against escherichia coli 35

 6b:      Minimum inhibitory concentration (MIC) and minimum 

            bactericidal concentration (MBC) of the goat 

            bile against escherichia coli 36

 6c:      Minimum inhibitory concentration (MIC) and minimum 

            bactericidal concentration (MBC) of the chicken 

            bile against  escherichia coli 37