ANTIMICROBIAL PROFILE OF PATHOGENIC BACTERIA ISOLATED FROM MUNICIPAL WASTE WATER IN UMUAHIA, ABIA STATE

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ABSTRACT

Antimicrobial profile of pathogenic bacteria isolated from municipal waste water system in Umuahia, Abia State. Two (2) samples each from four (4) sites (Bawas Street, Umudike junction, Ndoki Street and Old timber junction) were used. Seven (7) bacterial genera; Enterobacter, Escherichia, Klebsiella, Pseudomonas, Salmonella, Shigella and Staphylococcus were isolated. All the isolates had a 100% prevalence. The invitro antibiotic susceptibility test was done using the Kirby bauer disc diffusion method. Both Gram negative and Gram positive multiple disc were used which consist of ceftazidime (30ug), cefuroxime (30ug), gentamicin (10ug), ciprofloxacin (5ug), ofloxacin (5ug), augmentin (30ug), nitrofurantoin (300ug), ampicillin (10ug) and cefixime (5ug). At Bawas street, the percentage resistance and susceptible of pathogenic organisms to antibiotics were; ceftazidime 35.7% and 57.1%,cefuroxime 35.7% and 57.1%, gentamicin 28.6% and 64.3%, ciprofloxacin 28.6% and 57.1%, ofloxacin 14.3% and 64.3%, ampicillin 14.3% and 85.7%, nitrofurantoin 42.9% and 57.1%, augmentin 14.3% and 85.7% and cefixime 64.3% and 28.6% respectively. At Umudike junction, the percentage resistance and susceptible of pathogenic organisms to antibiotics were; ceftazidime 50% and 28.6%, cefuroxime 42.9% and 35.7%, gentamicin 50% and 42.9%, ciprofloxacin 42.9% and 57.1%, ofloxacin 35.7% and 50%, ampicillin 14.3% and 78.6%, nitrofurantoin 42.9% and 50%, augmentin 14.3% and 85.7% and cefixime 71.4% and 28.6% respectively. At Ndoki street, the percentage resistance and susceptible of pathogenic organisms were; ceftazidime 64.3% and 14.3%, cefuroxime 64.3% and 28.6%, gentamicin 71.4% and 14.3%, ciprofloxacin 50% and 42.9%, ofloxacin 57.1% and 35.7%, ampicillin 28.6% and 42.9%, nitrofurantoin 57.1% and 21.4%, augmentin 14.3% and 85.7% and cefixime 78.6% and 7.1% respectively. At Old timber junction, the percentage resistance and susceptible of organisms to antibiotics were ceftazidime 85.7% and 0%, cefuroxime 71.4% and 28.6%, gentamicin 85.7% and 0%, ciprofloxacin 71.4% and 28.6%, ofloxacin 78.6% and 14.3%, ampicillin 57.1% and 42.9%, nitrofurantoin 92.9% and 0%, augmentin 21.4% and 57.1% and cefixime 92.9% and 0% respectively. The overall prevalence of multiple drug  resistance (MDR) in this study was more in Old timber junction 73% and followed by Ndoki street 54% while in Umudike junction 40% and Bawas street 31% were found to be less. From the above results it was observed that the presence of antibiotic resistance organisms in this waste water should not be overlooked. Since this organisms may be vital to the safety and well-being of patients who are hospitalized and individuals susceptible to infection. Therefore, individuals should avoid the use of waste water or water contaminated by waste water for irrigation of farm, washing of plants and animals food and others, proper waste water treatment plant should be established and improved sanitary measure should be practice.

 

 






TABLE OF CONTENT

Title page                                                                                                                                i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                  iv

Table of Contents                                                                                                                    v

List of tables                                                                                                                           viii

Abstract                                                                                                                                   ix

Chapter One                                                                                                                            1

Introduction                                                                                                                            1

1.1 Aim and Objective                                                                                                            3

Chapter Two                                                                                                                           4

Literature Review                                                                                                                   4

2.1 Waste                                                                                                                                4

2.1.1 Waste water                                                                                                                    4

2.1.2 Municipal waste water                                                                                                   6

2.2 Antimicrobial agents                                                                                                          7

2 2.3 Antimicrobial and bacteria in the environment                                                             8

2.3.1 Antimicrobial in the environment                                                                                  8

2.3.2 Bacteria in the environment                                                                                          9

2.4 Antimicrobial resistance                                                                                                   14

2.4.1 Antimicrobial resistance in medical                                                                              14

2.4.2 Antimicrobial resistance in the environment                                                                 16

2.5 Identification of resistance and resistant bacteria in the environment                                    19

2.6 Sources of antibiotics and resistance into municipal waste water system                         20

2.7 Input of resistant bacteria into municipal waste water system                                         21

2.8 Understanding the interaction of bacteria and antimicrobials in the Environment                                                                                                                    27

2.9 Antimicrobial activity                                                                                                       29

Chapter Three                                                                                                                         30

3.0 Materials and Methods                                                                                                     30

3.1. Materials                                                                                                                          30

3.1.1 Study Area                                                                                                                     30

3.1.2. Sample Collection                                                                                                        30

3.1.3 Sterilization                                                                                                                   30

3.1.4 Media preparation                                                                                                         31

3.1.5 Isolation of microorganisms                                                                                          31

3.1.6 Morphological and biochemical characteristics            of bacterial isolates                                    32

3.1.7 Gram staining technique                                                                                               32

3.1.8 Biochemical Tests                                                                                                          33

3.1.8.1 Oxidase test                                                                                                                33

3.1.8.2 Indole test                                                                                                                   33

3.1.8.3 Coagulase test                                                                                                             33

3.1.8.4 Citrate utilization test:                                                                                                34

3.1.8.5 Catalase test                                                                                                                34

3.1.8.6 Motility test                                                                                                                34

3.1.8.7 Urease test                                                                                                                  35

3.1.9 Antimicrobial Susceptibility Test                                                                                  35

Chapter Four                                                                                                                           36

Result                                                                                                                                      36

Chapter Five                                                                                                                           49

Discussion                                                                                                                               49

Conclusion                                                                                                                              50

Recommendation                                                                                                                   51

References

 

 

 

 

 

 

 

LIST OF TABLES

 

Table                         Title                                      Page

 

Table 4.1         The occurrence of different bacterial isolates at different sampling  sites                 41       

 

Table 4.2         The morphological and biochemical characteristics of the bacterial isolates              43

 

Table 4.3         The antimicrobial profile of Gram positive and Gram negative bacterial isolates and their percentage          45

 

 



 

 

CHAPTER ONE

INTRODUCTION


Hospitals and clinics are major reservoirs for large numbers of pathogenic bacteria comprised of resident and comity introduced strains (Periasamy and Sundaram, 2013). High usage of antibiotics to treat infections in patients serves as a selective pressure for resistance development and there are concerns with transmission and their long term survival in the environment (Alamet al., 2013). Dissemination of antibiotic resistant bacteria (ARB) from hospitals can occur via various routes such as hospital wastewater, discharged patients and health care workers (Alam et al., 2013; Novo and Manaia, 2010). Antibiotics in wastewater can arise from excretion in urine and faces, direct disposal of expired drugs, and accidental spilling; these events could serve as additional selective pressure on bacteria while in wastewater. An elevated level of antibiotics and other pharmaceuticals in the environments are considered favorable for the selection of antibiotic resistance and most probably important hotspots for horizontal gene transfer (HGT) of resistance genes, and therefore conducive sites for resistance evolution. The possible persistence and further dissemination of ARB in natural aquatic environments could ultimately lead to an increase in the pool of antimicrobial resistance determinants. The transfer of resistance into current and emerging pathogens are major concerns that are being entertained with regards to the continuous introduction of ARB and their resistance genes into the environment (Baquero, 2008; Kemper, 2008; Kummerer, 2009). 

Antibiotics are used extensively to prevent or to treat microbial infections in human and veterinary medicine. Apart from their use in aquaculture, they are also employment to promote more rapid growth of livestock. Most of the compounds used in medicine are only partially metabolic by patients and are then discharged into the hospital sewage system or directly into municipal waste water if used at home. Along with excreta, they flow with municipal waste water to the sewage treatment plant (STP). They may pass through the sewage system and end up in the environment, mainly in the water compartment. Antibacterial substances used for livestock enter the environment when manure is applied to fields. These antibiotics may either end up in soil or sediment or in ground water. Antimicrobial agents are also used to treat infections in intensive fish farming where they are added directly to the water, resulting in high local concentrations in the water compartment and adjoining sediments. Some antibiotics such as streptomycins are used in fruit growing, others in bee-keeping. Disinfectants are widely used in the food and glue industries, medicine and livestock rearing. In addition to antimicrobials and disinfectants, resistant bacteria themselves are excreted by humans and animals and are emitted into sewage or manure and other environmental compartment (Richardson and Bowron, 1985; Kummerer, 2003; Goletet al., 2001; Zuccato et al., 2000). The unwanted effects of microbial growth have long been controlled through use of antimicrobials. It has also long been recognized that susceptibility to such chemicals varies markedly between different groups of organisms and within these groups. The different mechanisms of action and the methods used to evaluate susceptibility are crucial for the result of susceptibility testing and the evaluation of resistance. Resistance is a description of the relative in susceptibility of microorganism to a particular treatment under a particular set of conditions. Therefore, care must be taken in interpreting the literature and the reader is advised to refer for details to the literature. For antibacterial, resistance is usually quantified as the minimum concentration required asserting a definable effect (e.g. growth inhibition) on a population of cells. Wherever there is a change in susceptibility that renders an agent ineffective against a certain organism, this organism is referred to as resistant. Many organisms have always been in sensitive to and are thereby intrinsically resistant to a particular agent by nature of their physiology or biochemistry. Susceptible organisms can become insensitive by mutation or by incorporation of the genetic information which encodes the resistance. This paper summarizes findings on antibiotic resistance in different environmental compartments. It is structured as follows: first, a brief background is given on resistance and antimicrobials in the environment. Next, different sources for the input of resistant bacteria and antibiotics into the environment such as hospital effluent and municipal sewage or waste water including sewage treatment plants are discussed. Findings on other compartments of the aquatic environment such as surface water, ground water, sea water and sediments are briefly summarized. Issues in our understanding of the interaction of bacteria and antimicrobials in the environment are outlined before research needs are addressed (Gilbert and McBain, 2003).


1.1 AIM AND OBJECTIVE

ü  To isolate and identify pathogenic bacteria from municipal waste water system 

ü  To determine the antimicrobial profile of the pathogenic bacteria

 

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