Introduction and Aim: Antibiotic resistance is one of the major problems in the medical world, and the sewage treatment plants are among the primary habitats to harbor antibiotic resistance bacteria (ARB) especially multi-drug resistance (MDR). So the aim of this study was to identify the prevalence of multi-drug resistant bacteria in the sewage-water treatment plants (STPs). Materials and Methods: Samples were collected from sewage treatment plants (after its secondary treatment) from 6 different regions in south India. Serial dilution was performed to screen the Gram-negative bacterial isolates and identified through VITEK identification system. The multi-drug resistance was identified using disk-diffusion and transconjugation studies were performed. Results: The total of 101 Gram-negative bacteria was isolated and identified as Escherichia coli, Pseudomonas aeruginosa, Serratia odorifera, Serratia liquefaciens, Pseudomonas luteola, Aeromonas spp., Proteus spp. Based on the susceptibility pattern, 55/101 were found to be multi-drug resistance and out of 101 isolates, 14% were found to be resistance to meropenem (carbapenem). Conjugation studies showed that the donor MDR E. coli was able to conjugate the resistance to recipient E. coli AB1157. Conclusion: Our study concluded that the transconjugation might be one of the mechanisms by which acquired resistance can be transferred between the bacterial populations including clinical pathogens. © 2017 Indian Association of Biomedical Scientists. All rights reserved.