PLASMID PROFILING OF MULTIDRUG RESISTANCE CLINICAL ISOLATES OF (Pseudomonas aeruginosa)

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ABSTRACT

The emergence of multiple antibiotic resistance in bacteria and the indiscriminate use of antibiotics contribute to the dissemination of resistant pathogens in the environment which may cause problems in therapy and is a serious public health issue. This study was conducted to determine the incidence of Pseudomonas aeruginosa in clinical samples and their antibiotics resistant pattern.

The organisms were isolated using standard microbiological techniques and the antibiotic susceptibility was determined using disc diffusion method. All the 20 Pseudomonas aeruginosa isolates in this study were subjected to antibiotic sensitivity assay by disc diffusion method using twelve (12) antibiotic agents which include Cefuroxime sodium, Ceftazidime, Ciprofloxacin, Meropenem, Zinacef, Cefuroxime, Cefepime, Ampicillin, Levofloxacin, Norfloxacin, Moxifloxacin, Mipenem. The plasmid profiling was also carried out and visualized under UV light trans-illuminator.

All the isolates were susceptible to one of the antibiotics which is Cefepime and resistant to the remaining eleven (11) antibiotics (Ciprofloxacin, Cefuroxime sodium, Ceftazidime, Meropenem, Zinacef, Cefuroxime, Ampicillin, Levofloxacin, Norfloxacin, Moxifloxacin, Mipenem). This indicates that the isolates was susceptible to 8% of the 12 antibiotics used and resistant to 92 % of the 12 antibiotics.

This study shows that the resistance is not plasmid mediated in all of the clinical isolates but maybe due to other intrinsic factor, as antibiotic resistance can either be due to the presence of plasmid or as a result of some cellular factors.

TABLE OF CONTENTS

CERTIFICATION - - - - - - - - - ii

DEDICATION - - - - - - - - - iii

ACKNOWLEDGEMENT - - - - - - - - iv

TABLE OF CONTENTS - - - - - - - - v

LIST OF FIGURE - - - - - - - - - viii

LIST OF PLATES - - - - - - - - - ix

LIST OF TABLES - - - - - - - - - x

ABSTRACT - - - - - - - - - xi

CHAPTER ONE

1.0 INTRODUCTION - - - - - - - - 1

1.1 Background - - - - - - - - - 1

1.2 Growth Characteristics - - - - - - - 2

1.3 Aims and Objectives - - - - - - - - 7

1.3.1 Aim - - - - - - - - - - 7

1.3.2 Objectives - - - - - - - - - 7

CHAPTER TWO

2.0 LITERATURE REVIEW - - - - - - - 8

2.1 General Characteristics of Pseudomonas aeruginosa- - - - 9

2.2 Environmental Range - - - - - - - - 13

2.3 Infections associated with Pseudomonas aeruginosa - - - - 14

2.3.1 Nosocomial infections - - - - - - - 16

2.4 Laboratory Identification - - - - - - - 17

2.5 Antimicrobial Resistance - - - - - - - 18

2.6 Multidrug Resistance - - - - - - - - 19

2.6.1 Mechanism of multidrug resistance in Pseudomonas aeruginosa - - 20

2.7 Plasmids - - - - - - - - - - 20

2.7.1 Plasmids and bacterial resistance - - - - - - 22

CHAPTER THREE

3.0 MATERIALS AND METHODS - - - - - - 24

3.1 Study Design and Population - - - - - - - 24

3.2 Biochemical Identification of isolates - - - - - 24

3.3 Antibiotic Susceptibility Test - - - - - - 25

3.4 Plasmid Extraction - - - - - - - - 27

3.4.1 Cell Harvest and Lysis - - - - - - - 28

3.4.2 Neutralization - - - - - - - - - 28

3.4.3 Column Activation - - - - - - - - 28

3.4.4 Column Loading - - - - - - - - 28

3.4.5 Column Washing - - - - - - - - 28

3.4.6 Elution - - - - - - - - - 29

3.5 Gel Electrophoresis - - - - - - - - 29

3.6 Statistical Analysis - - - - - - - - 29

CHAPTER FOUR

4.0 RESULTS - - - - - - - - 30

4.1 Identification of isolates - - - - - - - 30

4.2 Population distribution - - - - - - - 30

4.3 Antibiotic susceptibility pattern of Pseudomonas aeruginosa - - 38

4.4 Plasmid profiling - - - - - - - - 42

CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATIONS - - 44

5.1 Discussion - - - - - - - - - 44

5.2 Conclusion - - - - - - - - - 45

5.3 Recommendations - - - - - - - 46

REFERENCES - - - - - - - - 47

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PLASMID PROFILING OF MULTIDRUG RESISTANCE CLINICAL ISOLATES OF (Pseudomonas aeruginosa)

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