Isolation And Sensitivity Of Bacteria Isolate From Vaginal Discharge To Antibiotics

Project and Seminar Material for Microbiology

Isolation And Sensitivity Of Bacteria Isolate From Vaginal Discharge To Antibiotics


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.

Chapter One


1.1 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 micro organisms 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 micro organisms. 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”.

1.2 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

1.3 Aim

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

1.5 Objectives of the Study

  1. To isolate bacteria from the urinary tract of women of child-bearing age using vaginal swab.
  2. To identify the isolated bacteria.
  3. To determine the antibiotic sensitivity of bacteria isolated.

Chapter Five

Discussion and Conclusion

5.1 Discussion

Vaginal flora contains a range of microorganisms normally. Lactobacillus spp. play fundamental role in maintaining acidic vaginal pH and prevent the overgrowth of potentially harmful and opportunistic bacteria. Vaginal infections are a great threat for women’s health related to common gynecological problem. This study demonstrates the prevalence of vaginal pathogens among the women study group. Vaginal infections are increasing due to vagina colonization by pathogenic bacteria other than the protective bacteria. The results of this study are similar to the study conducted by Lakshmi et al.,(2001). They reported the prevalence of vaginal infections in India.(Larsen, 2001)In our study E.coli was the most frequent bacteria followed by Klebsiella , Pseudomonas Citrobacter , Proteus and Acinetobacter respectively, similar to a study conducted by Dutta et a.,.(1995). McDonald et al.,(1994) also found E. coli to be the important bacteria associated with bacterial vaginosis. The most useful antibiotics against gram negative rods in our study were Imipenem (27mm) and Ciprofloxacin (28). Antibiotics like Imipenem are extremely effective but expensive. Tariq et al.,(2017)reported similar findings. Whereas the antimicrobials with least affectivity against gram negative rods were Penicillins (Ampicillin, amoxicillin-clavulanic acid), Amikacin due, probably, to indiscriminate use of antibiotics.

5.2 Conclusion

High prevalence of gynecological infections demands that the patients who suffer from the symptoms of gynecology must be investigated carefully. The elevated prevalence of vaginal bacteria in the recent study stress that high vaginal swab culture provides a good laboratory means for the identification of causative bacteria.It is therefore recommended for roUrinary Tract Infection(UTi)ne use. There is a need for stakeholders to be aware of antibiotic resistance in order to avoid improper use and frequent abuse of available antibiotics. Treatment schedule must be designed subsequent to proper laboratory investigations

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