ANTIBACTERIAL ACTIVITY OF BILE ON BACTERIA ISOLATED FROM FAECES, URINE AND BLOOD.

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