Total of 261 samples of fish and environmental samples (i.e. wash water, swabs of hand/ gloves of workers, fish contact surfaces i.e. knives, cutting boards and working tables) were collected from two Pangasius processing factories (PPF1 and PPF2). A total of seventy-one (71) isolates of Escherichia coli were selected to study the prevalence of antibiotics resistance using disk agar diffusion method. Overall, it was determined that 61% (22/36) of PPF1 isolates were resistant except to colistin while 68.57% (24/35) of PPF2 isolates were resistant except kanamycin. High resistance was against ampicillin in both PPF1 and PPF2 isolates (47.22% and 42.86%), followed by cefotaxime (33.33% and 40%) respectively. Varying resistance response to all other tested antibiotics such as streptomycin, meropenem, tetracycline, sulfamethoxazole/trimethoprim and nalidixic acid was also observed among the E. coli isolates from both factories. About 50% of the multidrug resistant (3-9 antibiotics) among PPF1 were observed whereas there were 45.83% multi-drug resistant (3-7 antibiotics) among PPF2 isolates. The result from this study reflected that there was a prevalence of multi-drug resistance of E. coli isolated during the processing of Pangasius at the studied factories. Therefore, there is a need for an effective risk management assessment models and management plans from stakeholders involved in the Pangasius value chain (i.e. farmers, processors and government) to ensure the food safety of production chain.