Populations of antibiotic-resistant coliform bacteria change rapidly in a wastewater effluent dominated stream

Incomplete elimination of bacteria and pharmaceutical drugs during wastewater treatment results in the entry of antibiotics and antibiotic-resistant bacteria into receiving streams with effluent inputs. In Mud Creek in Fayetteville, AR, ofloxacin, trimethoprim, and sulfamethoxazole have been detected in water and sediment, and tetracycline has been detected in sediment downstream of treated effluent input. These antibiotics have been measured repeatedly, but at low concentrations (<1μg/L) in the stream. To determine if effluent input results in detectable and stable changes in antibiotic resistances downstream of effluent input, antibiotic resistance in Escherichia coli and total coliform bacteria in Mud Creek stream water and sediment were determined using a culture-based method. Isolated E. coli colonies were characterized for multiple antibiotic resistance (MAR) patterns on solid media and to evaluate E. coli isolate richness by amplification of a partial uidA gene followed by denaturant gradient gel electrophoresis (DGGE). Despite temporal variability, proportions of antibiotic-resistant E. coli were generally high in effluent and 640m downstream. The MAR pattern ampicillin-trimethoprim-sulfamethoxazole was associated with a DGGE profile that was detected in effluent and downstream E. coli isolates, but not upstream. Percent resistance among coliform bacteria to trimethoprim and sulfamethoxazole was higher 640m downstream compared to upstream sediment and water (with one exception). Resistance to ofloxacin was too low to analyze statistically and tetracycline resistance was fairly constant across sites. Resistances changed from 640m to 2000m downstream, although dissolved nutrient concentrations within that stream stretch resembled effluent. Antibiotic resistant bacteria are entering the stream, but resistances change within a short distance of effluent inputs, more quickly than indicated based on chemical water properties. Results illustrate the difficulty in tracking the input and fate of antibiotic resistance and in relating the presence of low antibiotic concentrations to selection or persistence of antibiotic resistances.