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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