ISOLATION AND SENSITIVITY OF BACTERIA ISOLATE FROM VAGINAL DISCHARGE TO ANTIBIOTICS


ISOLATION AND SENSITIVITY OF BACTERIA ISOLATE FROM VAGINAL DISCHARGE TO ANTIBIOTICS  

ABSTRACT

Normal vaginal flora contains a wide range of microorganisms. Bacterial vaginosis BV is the main reason of vaginal discharge. Many gram positive and gram negative rods i.e. E.coli, Klebsiella, Proteus, Acinetobacter and Pseudomonas spp. are major contributors in bacterial vaginosis. Aim: The present study was conducted to elucidate the frequency of various bacteria in high vaginal swabs and sensitivity pattern of bacteria to antibiotics that are currently used Material and Methods used are a  total of 6 High vaginal swabs (HVS) which were collected from patients presenting with symptoms of vaginal discharge. Swabs were inoculated on blood and Chocolate agar. After overnight incubation plates were examined for growth, colonial morphology, final confirmation was done on the basis of biochemical testing. Antibiotic sensitivity testing was done by (modified Kirby-Bauer’s) disc diffusion method using amikacin(30μg), ampicillin(10μg), amoxicillin(10μg),) clavulanic acid, imipenem(10μg),  ciprofloxacin(10μg),  and cefixime(5μg). After overnight incubation plates were examined to read the susceptibility zone. Results showed  that Highly sensitive antibiotics against bacteria were imipenem (27mm), and Ciprofloxacin (28mm)  whereas least affective antibiotics against gram negative rods were penicillins, amikacin due to indiscriminate use of antibiotics. In conclusion, high prevalence of gynecological infections demands that the patients who have vaginosis must be investigated regularly and carefully through culture and identification of causative bacteria. Emergence of antibiotic resistance must be controlled in order to avoid improper use, frequent abuse, insufficient dosages, trouble-free availability of antibiotics and treatment schedule must be designed subsequent to proper laboratory investigations.

TABLE OF CONTENTS

Title page     ____ ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____i

Approval page  ___  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  ii

Dedication ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  iii

Acknowlwdgement ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___ iv

Table of Contents  ____   ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  _v

List of table   ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  _viii

Abstract   ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___ix

CHAPTER ONE

1.1  Introduction   ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  __  1

1.2  Background of the study    ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  __ 1

1.3  Antibiotic sensitivity  ___  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  __  2

1.4  aim  ____  ____  ____  ____  ________  ____  ____  ____  ____  ____  ____  ____  ____  ___  __  4

1.5 Objectives of the study    ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___ 

CHAPTER TWO 

2.1  Literature review   ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  5

2.2  Origin of antibiotics resistance  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____ 5

2.3  Development of Antibiotic Resistance____  ____  ____  ____  ____  ____  ____  ____  ____  ___   7

2.4  Consequence of Antibiotic Resistance____  ____  ____  ____  ____  ____  ____  ____  ____  ___  8

2.5  Regulatory issues to Antibiotic Resistance    ____  ____  ____  ____  ____  ____  ____  ____  ___9

2.6  Mechanism of Antimicrobial Resistance  ____  ____  ____  ____  ____  ____  ____  ___  ___  _  10

CHAPTER THREE

3.1  Materials and methods  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  14

3.2  Preparation of chocolate  agar  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  15

3.3  Preparation of blood  agar  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  _  15

3.4  Sub culture in agar plate  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  15

3.5  Agar slant sub culture in Bijou bottle   ____  ____  ____  ____  ____  ____  ____  ____  ____  __15

3.6  Gram staining  ____  ____  ____  ____  ____  ____  ____  ____  ____   ____  ____  ____  ___  _  16

3.7  Biochemical tests  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  16

3.7.1  Catalase test  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ______  16

3.7.2  Oxidase test  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ______  17

3.7.3  Methyl red test   ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___ 17

3.7.4  Indole test    ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  __  17

3.7.5  Vogwsproskauer test  ____  ____  ____  ____  ____  ____  ____   ____  ____  ____  ___  ___   17

3.7.6  Citrate test  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  __ 18

3.8  Antibiotic culture sensitivity test  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  __  18

CHAPTER FOUR

4.1  Results  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  _ 20

CHAPTER FIVE

5.1  Discussion  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  ____  24

CONCLUTION  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ____  ___  ___  ___  _25

REFERENCE

CHAPTER ONE

1.1 INTRODUCTION 

1.2 Background of the Study

Antimicrobial resistance is a global concern, particularly pressing in developing nations where  infectious diseases, poverty and malnutrition are endemic. Infections caused by resistant bacteria have been shown to be more frequently associated with increased morbidity and mortality than those caused by susceptible pathogens. In areas of concentrated use, such as hospitals, antimicrobial resistance  lead to hospital stays, increased health care costs and in extreme cases untreatable infections. The lack of clinical microbiology laboratories to identify the specific etiologic agents and their antimicrobial susceptibility testing has increased empirical therapy which in turn leads to emergence of AMR. Moreover, self-antibiotic prescription, lack of access to local antibiogram data and poor awareness of prescriber about AMR were the leading local factors for AMR development in Ethiopia (Aberaet al., 2014). 

Studies have shown that besides the temporal changes in profile of infecting microorganisms and pattern of resistance over time, antimicrobial resistance profile of bacteria varies among population because of difference in geography, local antimicrobial prescribing practices and prevalence of resistant bacterial strains. Such differences are never stable and may change rapidly especially in places where misuse of antibiotics are common particularly in developing countries. A systematic review in Ethiopia has also indicated a trend towards an increasing resistance rates among pathogens such as Escherichia coli, Proteus, Klebsiella, Pseudomonas,Citrobacter and Acenotobacter to commonly prescribed antibiotics, including Ampicillin, Amoxicillin, Amikasin, Imipenem, Cefixime and Ciprofloxacin (Moges et al.,2014). Thus, up to date information on microbial resistance is needed at local level to guide the rational use of the existing antimicrobials. 

The adult human vagina is a complex biota containing a profusion of microorganisms.  These can be either unicellular or multicellular and are present everywhere in nature. They include  bacteria, fungi,  archaea,  protists,  some  microscopic  plants  such  as green  algae  and  animals  such  as  planktons  and  palanarian. On account of their nature, viruses may or may not be included. Bacteria   and   yeast   form   normal   flora   of   this ecosystem, which is normally found on the skin and every opening of the body such as mouth, ears, rectum and vagina. Even  a  neonate  carries  specific  flora  of  his/  her  mother  and soon develops own floral  community. This flora persists till death of the individual. An adult human carries normal flora consisting of more   than 200 bacterial species.  Normally these are harmless and are involved in benefiting their hosts. Yet some are parasitic in nature, living at the expense of their host, and some are even pathogenic.These pathogenic microbes, after getting a chance, invade their   hosts   and   lead   to   opportunistic   infection. These diseases caused by normalflora are termed endogenous diseases (Khan et at.,2002).

Resistance of bacteria to antimicrobial agents is an imminent threat to patient management all over the world. This issue has plagued policy makers and clinicians everywhere but there seems to be no simple way of circumventing the problem. Rapidly rising antibiotic resistance is a challenge to comprehensive patient care in all branches of medical science. The interaction between various clinical bacteria and the antimicrobial agents is a complex issue involving the prokaryotic adaptive mechanisms and genetic changes.  This complex interaction must be studied in depth in order to achieve a sustainable and effective solution to the looming threat of antibiotic resistance. Earlier, the problem of antibiotic resistance was primarily a concern for not so comical infections. But now, even community acquired infections are caused by organisms with high levels of antibiotic resistance. As a report had demonstrated, such multi-drug resistant community acquired infections can be a cause of significant. 

Earlier, such drug resistant organisms were said to infect mainly patients with identifiable risk factors or profound immune suppression. But now, reports are showing such infections in seemingly normal healthy persons. Also, such drug-resistant infections may complicate the newly emerging infectious diseases. For example, influenza epidemics are sometimes reported to be complicated by superadded infection with drug-resistant bacteria (Hageman et al., 2004). The issue of drug resistance in clinical bacteria is such a vital threat that the UN held a special assembly in 2016 to address only this issue. In that assembly, the issue was said to be of as much importance as climate change and it was deemed to require a global response (Farr, 1994) and   non-pregnant women  attending  the  University  of  Maiduguri Teaching Hospital (UMTH), Maiduguri, Nigeria”. 

1.3 Antibiotic Sensitivity

Antibiotic   sensitivity   is   a   term   used   to   describe   the susceptibility of bacteria to antibiotics. Antibiotic sensitivity testing (AST) is  usually  carried  out  to  determine  which antibiotic  will  be  most  successful  in  treating  a  bacterial infection  in  vivo. Testing  for  antibiotic  sensitivity  is  often done  by  the Kirby-Bauer  method  while  other    methods include  the  Stokes  method, E-test (also  based  on  antibiotic diffusion)   and      Agar   and   Broth dilution   methods   (for Minimum  Inhibitory  Concentration determination). Muller Hinton   agar   is   most   frequently   used   in   this   antibiotic susceptibility test. Our study was aimed at the isolation, identification and antibiotic sensitivity testing of URINARY TRACT INFECTION (UTI) causing bacteria

1.4 AIM

To isolate, identify and check antibiotic sensitivity of bacteria implicated in URINARY TRACT INFECTION (UTI)s.

1.5 OBJECTIVES OF THE STUDY

To isolate bacteria from the urinary tract of women of child-bearing age using vaginal swab.

To identify the isolated bacteria.

To determine the antibiotic sensitivity of bacteria isolated.

.

ISOLATION AND SENSITIVITY OF BACTERIA ISOLATE FROM VAGINAL DISCHARGE TO ANTIBIOTICS



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