ANTIBIOTIC RESISTANCE PATTERN OF BACTERIAL ISOLATES FROM WASTE DUMP SITE

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ABSTRACT

Efficient bio-medical waste management is a major challenge in Africa, and Nigeria in particular. Uncontrolled and excessive use of antibiotics by human may cause an increase in the prevalence and distribution of resistance genes in the environmental samples such as biomedical wastes. Antibiotics resistance is a global problem currently threatening the treatment of infections in plants, animals and humans. This study investigated the antibiotics resistant pattern of bacterial isolates from waste dump sites in Federal Medical Centre, Aba Road, Umuahia, Abia State. The isolates present in the samples were isolated and identified using cultural and biochemical techniques. The isolates were subjected to antimicrobial susceptibility to determine the resistant pattern of the isolates to some antimicrobial agents. Exactly fifty (50) samples from different dump sites from Federal Medical Centre, Umuahia were analyzed. A total of 41 bacterial species were isolated from the 50 samples. Bacillus spp occupied 36.9% of the total number of the isolates, Escherichia coli occupied 17.1%, Streptococcus spp occupied 22.0%, and Klebsiellaspp occupied 7.3% of the total number of isolates. The isolates showed multidrug resistance to Chloramphenicol, Septrin, Augmentin, Amoxycillin and Streptomycin. From the result, Bacillus spp showed the highest resistance to almost all the antibiotics tested. All the Bacillus spp isolates (100%) were resistant to Augmentin,93.3% of Bacillus spp showed resistance to Amoxycillin, 53.3% were resistant to Streptomycin, 86.7% were resistant to Chloramphenicol, 13.3% were resistant to Gentamicin while 6.7% were resistant to Pefloxacin. All the Bacillus sppisolates were sensitive to Ciprofloxacin. Among the isolates, Klebsiellaspp showed the least resistant to the antibiotics tested, only Augmentin, Amoxycillin and Chloramphenicol were resistance recorded. The high resistant to antibiotics as observed in this study is very alarming and thus highlights the global problem of antibiotics resistance to the public health.




TABLE OF CONTENTS

Title page                                                                                                                                i

Dedication                                                                                                                              ii

Acknowledgements                                                                                                                iii

Certification                                                                                                                           iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          viii

List of Figures                                                                                                                         ix

Abstract                                                                                                                                  x

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Statement of Problem                                                                                                 3

1.2       Justification                                                                                                                3

1.3       Aim and Objectives                                                                                                    4

CHAPTER TWO

2.0       Literature Review                                                                                                       5

2.1       Antibiotic Resistance                                                                                                  6

2.1.1    Emergence of Resistance                                                                                           7

2.2       Problems of Antibiotic Resistance                                                                             9

2.3       Genetic Basis of Antibiotic Resistance                                                                      10

2.3.1    Inherent Resistance                                                                                                    10

2.3.2    Mutational resistance                                                                                                  10

2.4       Mechanisms for Emergence Resistance                                                                     11

2.4.1    Plasmid Mediated Resistance                                                                                     11

2.4.2    Reduced Membrane Permeability                                                                              12

2.4.3    Modification of Target Resistance Site                                                                      12

2.4.4    Rapid Extrusion or Effluxpumps                                                                                13

2.4.5    Chromosome Mediated-Resistance                                                                            13

2.5       Mechanism of Resistance Transfer                                                                            14

2.5.1    Horizontal transfer of antibiotic resistance genes                                                      14

2.6       Specific Mechanisms of Resistance                                                                           17

2.6.1    Staphylococcus aureus                                                                                               17

2.6.2    Escherichia coli                                                                                                          18

2.6.3    Pseudomonas aeruginosa                                                                                           18

2.7       Waste                                                                                                                          20

CHAPTER THREE

3.0       Materials and Method                                                                                                 23       

3.1       Collection of Samples                                                                                                23

3.2       Media Preparation                                                                                                      23       

3.3       Isolation Characterization and Identification of Bacterial Isolates                                    23

3.4       Biochemical Test                                                                                                        24

3.4.1    Gram Staining                                                                                                            24

3.4.2    Motility Test                                                                                                               24

3.4.3    Catalase Test                                                                                                               25

3.4.4    Coagulase Test (Slide test)                                                                                         25

3.4.5    Citrate Utilization Test                                                                                               25

3.4.6    Indole Test                                                                                                                  26

3.4.7    Urease Test                                                                                                                 26

3.4.8    Triple Sugar Iron Test (TSI)                                                                                       27

3.4.9    Endospore formation stain                                                                                          27

3.5       Antibiotics susceptibility test                                                                                     27

CHAPTER FOUR

4.0       Result                                                                                                                          29

CHAPTER FIVE

5.0       Discussions                                                                                                                 38

5.1       Conclusion                                                                                                                  39

5.2       Recommendations                                                                                                      40

References


 

LIST OF TABLES

Table                                          Title                                  Page

4.1:     Prevalence of bacteria isolates from different dumpsite at Federal Medical   30

Centre, Aba Road, Umuahia, Abia State, Nigeria.

4.2:     Antibiotic susceptibility pattern of the isolates from Federal Medical Centre,  Aba road, Umuahia, Abia State Nigeria.   31

4.3:      Percentage resistance of the isolates on different antimicrobial agents.             35

 

 



 

                                            LIST OF FIGURES

Figure               Title                                                  Page

4.1:      Average percentage resistance of the isolates to different antimicrobial agents.          36

4.2:       Multidrug resistance of the isolates from different dumpsites at Federal Medical      37

             Centre, Aba road, Umuahia, Abia State Nigeria.

 

 

 

 

CHAPTER ONE


1.0      INTRODUCTION

Efficient bio-medical waste management is a major challenge in Africa, and Nigeria in particular (Fongwa, 2002). Uncontrolled and excessive use of antibiotics by human may cause an increase in the prevalence and distribution of resistance genes in the environmental samples such as bio-medical waste (Iversen et al., 2002).

In the early 1970s, physicians were finally forced to abandon the belief that, given the vast array of effective antimicrobial agents, virtually all-bacterial infections were treatable. This optimism was shaken by the emergence of resistance to multiple antibiotics among such pathogens as Staphylococcus aureus, Pseudomonas aeruginosa and a host of others (Lowy, 2003).

Multidrug resistance is now the norm among these pathogens. Staphylococcus aureus is perhaps the pathogen of greatest concern because of its intrinsic virulence, its ability to cause a diverse array of life threatening infections, and its capacity to adapt to different environmental conditions (Lowy, 2003). Pseudomonas aeruginosa is a highly relevant opportunistic pathogen. One of the most worrisome characteristics of Pseudomonas aeruginosa is its low antibiotic susceptibility. Escherichia coli is responsible for more than 80% of urinary tract infections (UTI) (Wikipedia, 2007), and is noted for producing an enzyme (β- lactamase) which degrades penicillin during clinical trials, and currently resists a vast number of other antibiotics (Sherman, 2006).

Antibiotic resistance is the ability of a microorganism to resist the effect of an antibiotic; these organisms thwart antibiotics by interfering with their mechanism of action. Resistance to antibiotics has spread fast to a number of commonly used antibiotics such that “in the 1990s, we have come to a point for certain infections that we do not have agents available”. (Lewis, 1995).

Antimicrobial drug resistance in bacterial pathogens is of National and International concern (WHO, 2001; CDC, 2001). Although the misuse of antimicrobial agents is accepted as a major driving force behind the spread of resistance, the nature of this relationship is complex. Resistance to antibiotics is sometimes acquired by a change in the genetic make-up of a bacterium, which can occur by either a genetic mutation or by transfer of antibiotic resistance genes between bacteria in the environment (Williams, 2002).

Infectious diseases are becoming more difficult and more expensive to treat, because our current antibiotics are becoming less useful against antibiotic resistant pathogenic bacteria. When antibiotics fail to work, the consequences include extra consultation with the doctor, hospitalization or extended hospital stays, a need for more expensive antibiotics to replace the older ineffective ones, loss of workdays, and sometimes, death (APUA, 1999).

The increased use of antibiotics in health care, as well as in agriculture and animal husbandry, is in turn contributing to the growing problem of antibiotic resistant bacteria. The antibiotic does not technically cause resistance, but allows a situation where an already existing variant can flourish (APUA, 1999). Products such as disinfectants, sterilants, and heavy metals used in industries and in household products along with antibiotics, are creating selective pressure in the environment that lead to mutations in microorganisms (Baquero et al., 1998).

In environment with multiple stresses, such as antibiotics and heavy metals, it would be ecologically favorable, in terms of survival, for a bacterium to acquire resistance to both stresses. If the resistance is carried on plasmid, those bacteria with clusters of resistance genes on plasmids are likely to simultaneously pass on those genes to other bacteria, with a better chance of survival (Lawrence, 2000).


1.1       Statement of Problem

Antibiotic resistance is a global problem currently threatening the treatment of infections in plants, animals and humans. Bacteria with elevated antibiotic resistance levels present a potential public health risk because infections with these organisms do not respond to one or more of the drugs commonly used to treat them. According to CDC, (2000), drug resistant pathogens are a growing menace to all people, regardless of age, gender, or socioeconomic background.

There is also, the risk of transferring antibiotic resistance to bacterial populations in the environment because antimicrobial resistance genes and their genetic vectors, once evolved in bacteria of any kind anywhere, can spread indirectly through the world’s interconnecting commensal, environmental, and pathogenic bacteria anywhere else (O’Brien, 2002).


1.2       Justification

The proliferation of dumpsites and open drainages within hospital and the lack of efficient waste collection and treatment processes is of public health concern because microorganisms and heavy metals among others are the major pollutants of dumpsites and wastewater.

Therefore, there is need to study the antibiotic resistance pattern of bacterial isolates from hospital dump sites.

In view of the increasing resistance among various pathogens to common antibiotics, including broad-spectrum antibiotics, there is need to research into preventable causes of antibiotic resistance in order to lower the incidence of drug resistance.

Antimicrobial resistance is driving up health care cost, increasing the severity of disease, and increasing the death rates from certain infections (NIAID, 2006). A number of clinically important microbes have developed resistance to available antimicrobials including Staphylococcus aureus and Pseudomonas aeruginosa which cause skin, bone, lung and bloodstream infections and Escherichia coli which causes urinary tract infections (Wikipedia, 2007).


1.3       Aim and Objectives

The aim of this study is to ascertain the antibiotic resistance pattern of bacterial isolates from waste dump site in Federal Medical Centre Umuahia.

The objectives of this work are therefore:

1. To isolate and identify bacteria pathogens from hospital dump site.

2. To determine antibiotic resistance pattern of the bacterial isolates.

3. To deduce the public health importance of the antibiotic resistant bacterial isolates.

 

 

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