This study collected ten treated wastewater samples from Vinh Long General Hospital to determine their physicochemical characteristics and antibiotic properties. All treated wastewater samples collected during the monitoring periods complied with national regulations. In addition, these samples did not contain bacteria such as Salmonella, Shigella, and Vibrio cholerae. The investigation yielded a total of 25 Escherichia coli isolates. The E. coli isolates exhibied highest antibiotic resistance rate to ampicillin (100%), followed by ciprofloxacin, amoxicillin/clavulanic acid, and cefazolin (96%, 92%, and 92%, respectively). The resistance rate to fosfomycin was 88%, whereas 80% of the isolates were resistant to sulfamethoxazole-trimethoprim. The resistance rate to gentamicin was 72%, whereas that to imipenem and tetracycline was 52%. In addition, 44% isolates were resistant to chloramphenicol, and 32% of isolates were colistin-resistant. Among analyzed isolates, three were resistant to 10 of 11 tested antibiotics but only displayed intermediate resistance to imipenems (carbapenems). Surprisingly, 23 out of 25 isolates showed a positive ESBL phenotype. Eleven of them had both the blaTEM and blaCTX-M-1 group structural genes, while twelve only had the blaCTX-M-1 group gene. Furthermore, none of the isolated E. coli isolates exhibited the blaSHV gene. The minimum inhibitory concentration (MIC) of colistin exceeded 4 μg/mL in 8 out of 25 (32%) isolates. Seven of eight isolates (87.5%) carried the mcr-1 gene, while one (12.5%) carried the mcr-8 gene. None of the other mcr (mcr-2 to mcr-9) genes were found.