Yearly discharge of antibiotic-resistant bacteria (ARB) from combined sewer overflow (CSO) was estimated. The volume of CSO was estimated from operating data of the pumping station. In the target sewer catchment, 23% of the total of the volume of combined sewage was discharged untreated as CSO. Combined sewage contained 3-log larger E. coli than secondary treatment effluent although the abundance of antibiotic-resistant E. coli was not significantly different. In the target-combined sewer catchment, a yearly total of 4.8 × 10 16 CFU of E. coli was discharged from 6.1 × 10 6 m 3 of CSO, while 1.3 × 10 12 CFU of E. coli from 2.1 × 10 7 m 3 of effluent from the wastewater treatment plant (WWTP). This E. coli discharge was equivalent to 7.9 × 10 9 CFU/m 3 from CSO, and 6.2 × 10 4 CFU/m 3 from WWTP effluent. Consequently, a yearly total discharge of antibiotic-resistant E. coli from CSO was 3.7-log larger than the WWTP effluent. The small-flow CSO events, which had hourly flow rate smaller than five times of the average dryweather flow, accounted for 43% of the total CSO volume, but 79% of the total discharge of antibiotic-resistant E. coli due to a small dilution factor with stormwater and frequent discharge. Reduction of small-flow CSO events would be important for effective reduction of ARB discharge from CSO.

This study investigated the impact of each treatment stage of the activated sludge process on the fate of antibiotic resistant bacteria (ARB) in wastewater treatment plants (WWTPs). Wastewater and sludge samples were collected monthly at each stage of a commercial-scale WWTP. After 20–25 strains of indicator Escherichia coli were isolated from each sample on Chromocult Coliform Agar, antibiotic resistance of the isolates to amoxicillin (AMX), ciprofloxacin (CIP), norfloxacin (NFX), kanamycin (KM), sulfamethoxazole/trimethoprim (ST) and tetracycline (TC) were tested with the Kirby–Bauer disk diffusion method. As a result, activated sludge in the aeration tank and return sludge had higher abundance of antibiotic resistant E. coli than influent wastewater and secondary treatment effluent. AMX resistant E. coli was enriched in return sludge at the secondary clarifier. Higher temperature was also likely to cause an increase of AMX resistant E. coli in sludge. The antibiotic resistance profile of E. coli in secondary treatment effluent was more dependent on activated sludge than influent wastewater. These results suggested that activated sludge in WWTP possibly serves as a reservoir of ARB, and that behavior of ARB in WWTP differs by antibiotic classes.