Sensitivity assessment of denitrifying bacteria against typical antibiotics in groundwater

The effects of antibiotics on nitrate denitrification in groundwater have acquired growing concern. Denitrification is a microbially mediated process. The effects of antibiotics on denitrification were mainly reflected in denitrifying bacteria. However, little is known about the relationship between antibiotics and denitrifying bacteria. Based on this, both direct antimicrobial susceptibility testing and microbial batch-culture experiments were conducted to assess the influences of typical antibiotics on denitrifying groundwater bacteria, mainly Pseudomonas (46.17%). Denitrifying bacteria, screened from a long-term groundwater denitrification environment, were tested for sensitivity to five typical antibiotics in groundwater: sulfamethoxazole (SMX), erythromycin (ERY), enrofloxacin (ENR), clindamycin (CLI), and tetracycline (TCY). The results showed that the sensitivity of denitrifying bacteria to antibiotics is mainly related to the type and concentration of antibiotics. For antibiotic types, the order of sensitivity by quantitative assessment is ENR > TCY > SMX > ERY > CLI. Fluoroquinolones (FQs) represented by ENR were selected to explore their concentration effects. The influences on denitrifying bacteria were divided into the high concentration effect (500 μg L-1 to 100 mg L-1) and the low concentration effect (100 ng L-1 to 10 μg L-1) with about 100 μg L-1 as a boundary. Exposure to high concentrations had significant inhibitory effects on bacterial growth and exhibited dose dependency, especially for ciprofloxacin (CIP). The low concentration effect was independent of concentration, which may be stimulation or inhibition. The stimulation mainly occurred due to ENR-exposure. For inhibitory effects, Lomefloxacin (LOM) was more effective than other FQs. Especially for inhibition at ng-level exposure, LOM and norfloxacin (NOR) exposures led to the highest and lowest inhibition rates, respectively.