PCR-Based Detection of Extended-Spectrum β-Lactamases (bla CTX-M-1 and bla TEM ) in Escherichia coli, Salmonella spp. and Klebsiella pneumoniae Isolated from Pigs in North Eastern India (Mizoram)

Abstract

Cephalosporins are major antimicrobials used to treat serious infections. However, their effectiveness is being compromised by the emergence of extended-spectrum β-lactamases (ESBLs). A total of 138 enteric bacteria were isolated from 53 faecal samples of pigs collected from different districts of Mizoram, of which 102 (73.91%) were Escherichia coli, 26 (18.84%) were Salmonella spp. and 10 (7.25%) were Klebsiella pneumoniae. Phenotypic confirmatory test (Double Discs Synergy Test) showed that 8 (5.80%) E. coli isolates were ESBLs producer. PCR analysis confirmed that out of the eight isolate, 7 (5.07%) harboured bla CTX-M-1 gene and/or bla TEM gene. Of the eight positive isolates, 7 (5.07%) and 3 (2.17%) were found to be positive for bla CTX-M-1 gene and bla TEM gene, respectively, of which 3 (2.17%) isolates were positive for both the genes. Only 4 (2.90%) E. coli isolates carried bla CTX-M-1 gene alone. Agarose gel electrophoresis showed that all the isolates were carrying plasmids ranging between 0.9 and~30kb. Out of the seven isolates positive for bla CTX-M-1 and/or bla TEM , 2 (1.84%) isolates were confirmed for bla CTX-M-1 gene in their plasmid. Only one E. coli isolate was found to be positive for both the genes in its plasmid. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method.