Detection and characterization of extended-spectrum beta-lactamases (bla(CTX-M-1) and bla(SHV)) producing Escherichia coli, Salmonella spp. and Klebsiella pneumoniae isolated from humans in Mizoram

Aim: The present study was conducted to isolate and characterize the extended spectrum beta-lactamases (ESBLs) producing enteric bacteria in human beings in Mizoram, India. Materials and Methods: Fecal samples were collected from human beings with or without the history of diarrhea from different hospitals of Mizoram. Samples were processed for isolation and identification of Escherichia coli, Salmonella and Klebsiella pneumoniae. All the isolates were subjected to antibiotic sensitivity assays. Phenotypically, ESBLs production ability was determined by double discs synergy test (DDST) method. ESBLs producing isolates were subjected to polymerase chain reaction (PCR) for detection of ESBLs genes. Plasmids were cured by acridine orange. Transfer of resistance from a donor to recipient strains was done by in vitro horizontal method. Results: A total of 414 enteric bacteria were isolated from 180 fecal samples (113 were from diarrheic patients and 67 were from non-diarrheic patients), of which 333 (80.44%), 52 (12.56%), and 29 (7.00%) were E. coli, K. pneumoniae and Salmonella spp., respectively. Double discs synergy test (DDST) exhibited 72 (21.62%) E. coli, 12 (23.08%) K. pneumoniae and 4 (13.79%) Salmonella spp. were ESBLs producers. Altogether, 24 (13.04%) isolates were found to be positive for at least one resistance genes under this study. A total of 36 (8.70%) E. coli, 4 (0.97%) K. pneumoniae and 2 (0.48%) Salmonella spp. were found to be positive for bla(CTX-M-1) gene by PCR. Similarly, 5 (1.21%) E. coli and 4 (0.97%) K. pneumoniae isolates were found to be positive for bla(SHV) gene. A total of 3 (0.72%) K. pneumoniae isolates were recorded as positive for both bla(CTX-M-1) and bla(SHV) genes. All the isolates were carrying plasmids ranging between 0.9 kb and similar to 30 kb. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method. Conclusion: ESBLs producing enteric bacteria are circulating in human population in North Eastern Region of India. Indiscriminate use of antibiotics should be avoided to control the menace of multidrug resistance bacteria in the environment, animals, and human beings.