229. Molecular Typing by RAPD, Characterization and Antibiotic Resistance Profile of ESBL Producing and Non-ESBL Producing Klebsiella Species Isolated From Diarrheal Stool and Environmental Samples

Abstract

BackgroundExtended-spectrum β-lactamase is a major public health problem in hospitals and community that mediate resistance to Penicillin, Cephalosporins, and Monobactams. Data regarding the detection of TEM, CTX-M, and SHV genes by molecular techniques and typing by RAPD in ESBL producing bacteria can be useful in epidemiology and risk factors associated with infections.MethodsTotal of 140 samples were collected. Well water (n = 50), Fish effluents (n = 40), and Diarrheal stool samples (n = 80). Antibiotic susceptibility test was done using the Kirby–Bauer disc diffusion method. Phenotypic detection of ESBL enzyme was done by Double disk diffusion test. PCR analysis was carried out for β-lactamase genes TEM, SHV, and CTX-M. Molecular Typing was done by RAPD.Results38 (57.57%) Klebsiella spp. isolated from Fish Effluents,11 (57.89%) from Well water and 15 (18.98%) from Diarrheal stool samples. ESBL producers were 4 (26.66%) from stool and 12 (31.57%) from fish effluents. Stool isolates showed high resistance to Ampicillin (86.7%), Cefuroxime (83.3%), Cefepime (76.7%), and Ceftazidime (70%). Fish effluents were more resistant to Cefeperazone sulbactum (95.9%), Ampicillin (81.6%) while well water isolates showed high resistance to Ampicillin (94.7%) and Erythromycin (73.7%). Molecular identification showed the presence of more than 2 genes among the isolates. Prevalence of gene bla- TEM was highest, followed by bla-CTX-M and bla-SHV. Genetic relatedness are expressed as percentage similarity and presented as dendogram.ConclusionThe study shows high prevalence of ESBL among Klebsiella isolates mainly rom Fish effluents and diarrheal stool samples. It shows 24% ESBL positive rate. Antibiotic-resistant bacteria from fish effluents highlights the associated human health risk when they enter food chain and become passive carriers. Practice of routine ESBL testing with conventional antibiotic susceptibility testing would be useful for combatting multi drug resistance.Present study shows high prevalence of TEM gene among Klebsiella species. RAPD-PCR will help in descriminating isolates and reflecting the genotypes circulating in the settings either hospital or in community.Disclosures All authors: No reported disclosures.