Antibiotic susceptibility tests are presented as the zone of inhibition using the disc-diffusion method, and categorized as resistant, intermediate or susceptible. DNA samples from antibiotic-resistant bacteria were analysed for the presence or absence of resistance genes using polymerase chain reaction (PCR). Laboratory analyses were conducted by trained staff at the International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b). The aim of the study was to identify the antibiotic-susceptibility profiles and resistance genes of bacteria (Escherichia coli) obtained from humans, poultry and the environment. Bacterial isolates previously identified with resistance to third-generation cephalosporins or carbapenems were included in the analysis. Bacterial samples originated from rural households and poultry farms (broiler chickens) in Mirzapur, Tangail district; and urban food markets in Dhaka city, Bangladesh. Environmental samples included surface water, water supply, wastewater, soil, animal faeces (poultry and cattle) and solid waste. The survey was part of a wider research project, Spatial and Temporal Dynamics of Antimicrobial Resistance Transmission from the Outdoor Environment to Humans in Urban and Rural Bangladesh. The research was funded by NERC/BBSRC/MRC on behalf of the Antimicrobial Resistance Cross-Council Initiative award NE/N019555/1. Full details about this dataset can be found at https://doi.org/10.5285/dda6dd55-f955-4dd5-bc03-b07cc8548a3d