ABSTRACT

Microbiological analysis was conducted on fresh and fermented palm wine in Umudike, Abia state. Fresh and fermented palm wine samples were collected from a palm wine tapper immediately after tapping and from different palm wine bars respectively, using tightly capped sterile containers and was taken to a microbiology laboratory for analysis. Spread plate and streak plate method was used for the isolation of microorganisms. The microorganisms isolated were identified based on their cultural, morphological and bio-chemical characteristics. The microorganisms include Bacillus spp, Streptococcus spp, Lactobacillus spp, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Acetobacter spp, Candida spp, and Saccharomyces spp. The mean average microbial counts of the sample were 2.7 × 104cfu/ml, 3.1 × 104cfu/ml, 0.3 × 104cfu/ml, and 3.2× 104cfu/ml for heterotrophic count, fungal isolates, coliform bacteria count and isolates on De Man, Regosa and Sharpe agar respectively. Saccharomyces spp occurred in all the samples (100.0% occurrence) while Bacillus spp, Klebsiella spp and Escherichia coli had the least occurrence (20.0%). In the assessment of the survival of isolates in relation to time (0-96hrs) Klebsiella pneumoniae, Staphylococcus aureus were eliminated from the sample after 24hours while Streptococcus spp, Lactobacillus spp, Escherichia coli and Candida spp were eliminated after 48hours. Bacillus spp, Acetobacter spp and Saccharomyces spp survived beyond 72 hours of fermentation. The presence of Bacillus spp, Staphylococcus aureus, Klebsiella pneumoniae, and Candida spp indicated unhygienic handling of the beverage. It is recommended that appropriate agencies should ensure that proper personal and environmental hygiene is enforced to prevent outbreak of disease.

TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgements iv

Table of Contents v

List of Tables vii

Abstract viii

CHAPTER ONE 1

INTRODUCTION 1

Background of the study 1

General objective 2

Specific objectives 3

CHAPTER TWO 4

LITERATURE REVIEW 4

2.1 Production of palm wine 4

2.2 Biochemical constituents of palm wine 5

2.2.1 Sugar identified and their concentration in palm wine. 6

2.2.2 pH and organic acid concentration in palm wine 7

2.2.3 Ethanol concentration in palm wine 8

2.2.4 Minerals and trace elements present in palm wine 9

2.2.5 Odorant of palm wine 9

2.3 Microbial communities in palm wine 10

2.4 Fermentation of palm wine 12

2.5 Uses of palm wine 13

CHAPTER THREE 14

MATERIALS AND METHODS 14

3.1 Study Area 14

3.2 Sample collection 14

3.3 Preparation of media 14

3.4 Serial dilution 15

3.5 Isolation of Microorganisms 15

3.6 Characterization and identification of isolates 15

3.6.1 Fungi 15

3.6.2 Bacteria 16

3.6.2.1 Gram staining 16

3.6.2.2 Spore staining 16

3.6.2.3 Catalase test 17

3.6.2.4 Coagulase test 17

3.6.2.5 Oxidase test 17

3.6.2.6 Citrate test 17

3.6.2.7 Indole test 18

3.6.2.8 Carbohydrate fermentation test 18

CHAPTER FOUR 19

Results 19

CHAPTER FIVE 25

DISCUSSION, CONCLUSION AND SUMMARY 25

5.1 Discussion 25

5.2 Conclusion 26

5.3 Recommendation 26

REFERENCES 27