The aim of this study was to isolate the antimicrobial-resistant enterobacteria from cattle feces and to identify the extendedspectrum beta-lactamase (ESBL) and carbapenemase gene among them. A total of 70 cattle fecal swab samples were collected from Durg, Chhattisgarh, India, and cultured in nutrient broth. Bacterial isolation recovered 120 gram-negative bacteria. Antimicrobial susceptibility testing showed 90 (75%) coliform isolates (66 Escherichia coli, 20 Klebsiella sp., and two each of Enterobacter sp. and Citrobacter sp.) resistant to amoxicillin, amoxicillin-clavulanic acid, cefixime, ceftazidime, imipenem, or meropenem. Bacterial isolates showed the highest resistance rate for ceftazidime (61.6%), followed by cefixime and amoxicillin-clavulanic acid (60%). The ESBL phenotype, assessed by combination disk method, was shown by 36 (40%) of the resistant isolates. PCR amplification revealed 20% and 31.6% positivity of bla TEM and bla SHV genes, respectively. bla TEM and bla SHV coexpression was recorded among eight (12.1%) E. coli and two (10%) Klebsiella sp. isolates. Restriction endonuclease analysis indicated clonal bla TEM and clonal and nonclonal bla SHV genes. Nucleotide sequencing confirmed the bla TEM and bla SHV genotypes. None of the isolates showed amplification of the carbapenemase gene. Phenotypic and genetic detection and characterization of ESBLs among Enterobacteriaceae bacteria illustrate the importance of planning and execution of effective control programs for emerging antimicrobial-resistant pathogens.
Significance and impact of the study: The emergence of carbapenemases and extended-spectrum b-lactamases (ESBLs) in bacteria has now become a global problem leading to failure of advanced antimicrobial therapy. We report for the first time the bla NDM-1 gene in non-fermenting Gram-negative bacterial isolates, and the bla KPC-2 gene in an Escherichia coli isolate from goat meat in India. The presence of carbapenemase genes in bacteria from goat meat develops serious concerns about public health and food safety. Analysis of genetic determinants of b-lactam resistance in bacteria would be helpful to formulate a suitable control strategy.
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