Molecular Characterization of Clinically Relevant Extended-Spectrum beta-Lactamases bla(CTX-M-15)-Producing Enterobacteriaceae Isolated from Free-Range Chicken from Households in Bangladesh

The study explored the potential colonization and characterization of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in the gut of free-range poultry from the rural households in Bangladesh. From 48 households located in several rural regions (eastern, western, and southern) of Bangladesh, 180 poultry fecal samples were collected to isolate ESBL-producing Enterobacteriaceae. ESBL producers were characterized by susceptibility testing, conjugation experiment, conventional polymerase chain reactions (PCRs), and multilocus sequence typing (MLST) followed by sequencing. Total 23% (42/180) poultry were ESBL positive consisting of Escherichia coli (n = 41) and Klebsiella pneumoniae (n = 1). ESBL producers were resistant to Cefotaxime (CTX; 100%), Cefepime (100%), Amoxicillin-clavulanic acid (36%), Ciprofloxacin (31%), and Trimethoprim-sulfamethoxazole (24%), and 12% isolates were multidrug resistant. All ESBL producers were carrying bla(CTX-M-15)-like genotype.Isolates were also carrying genes for quinolone resistance [qnrS1, aac(6 ')-Ib-cr], silver resistance (silE), and mercury resistance (merA). Isolates were negative for 025b-ST131 clone, mcr-1, and bla(OXA-48) gene. The repetitive element PCR revealed 15 different clones of E. coli and some of these clones were found to be common in 3 sampling locations. MLST analysis of E. coli revealed 9 different sequence types (STs); ST4, ST156, ST542, ST1140, ST1290, ST4628, ST5114, ST9768, and ST11317. ESBL producers were carrying transferable plasmids and 4 different plasmid replicon types; IncI1 (29%), IncY (7%), IncFIB (7%), and IncF1A (5%). The findings from the study confirmed that free-range poultry are potential ESBL carriers with coresistance to other antibiotic classes, metals, and biocides. This study confirms that free-range poultry in Bangladesh living close to humans without any direct antibiotic exposure could carry ESBL bacteria. Free-range poultry could be reservoir as well as a potential spreader of pathogenic E. coli and antibiotic- or biocide-resistant genes.