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.
.