Impact of oxytetracycline on anaerobic wastewater treatment and mitigation using enhanced hydrolysis pretreatment.

In this study, two parallel-operated up-flow anaerobic sludge bed reactors, one used to treat synthetic wastewater spiked with oxytetracycline and the other used to treat the same wastewater after enhanced hydrolysis, were used to evaluate the impact of oxytetracycline on anaerobic digestion and resistance development and the efficacy of enhanced hydrolysis pretreatment on the elimination of adverse effects. The reactors were operated under a constant organic-loading rate (10 g/L/d) with increasing oxytetracycline doses (0 mg/L to 200 mg/L) over a period of 15 months. For the reactor without pretreatment, the chemical oxygen demand removal reached up to 89.5%%at oxytetracycline doses ranging from 0 mg/L to 100 mg/L, which collapsed at higher oxytetracycline doses. Miseq sequencing showed that a diverse hydrolysis/fermentation/acetogenesis bacterial community was maintained as the oxytetracycline dose was increased from 0 mg/L to 100 mg/L, while extreme dominance of Macellibacteroides (65.70%%- 71.56%) was found to occur at higher oxytetracycline doses. The total abundance of antibiotic resistance genes increased from 1.3 × 10-1 copies per cell to 2.6 × 10-1 copies per cell with increasing oxytetracycline dose from 0 mg/L to 5 mg/L, remained unchanged at oxytetracycline doses ranging from 25 mg/L to 100 mg/L, and then increased to 4.8 × 10-1 copies per cell and 1.3 copies per cell at oxytetracycline doses of 150 mg/L and 200 mg/L, respectively. Multidrug resistance developed in response to oxytetracycline treatment at 200 mg/L. Poor chemical oxygen demand removal and a marked enrichment in antibiotic resistance genes was validated using a full-scale up-flow anaerobic sludge bed system fed with an influent oxytetracycline concentration of approximately 200 mg/L. For the reactor treating wastewater pretreated with enhanced hydrolysis (85 °C for 6 h), the chemical oxygen demand removal rate and antibiotic resistance genes level over the whole oxytetracycline dose range were found to be similar to those achieved with zero oxytetracycline treatment. These results demonstrated that the control of conventional pollutants and ARGs could be achieved simultaneously in the UASB reactor by employing enhanced hydrolysis pretreatment.