The behavior and removal of roxithromycin (ROX), oxytetracycline (OTC), chlortetracycline (CTC), and enrofloxacin (ENR) were investigated during the steady state of sludge anaerobic digestion (AD) in semi-continuous mode (37°C). Sludge was spiked at realistic concentrations (50μg/L of each antibiotic) and then used to feed the bioreactor for 80days. Antibiotics were extracted from the substrate and digested sludge samples by accelerated solvent extraction (ASE). Accurate determination of antibiotics was obtained by the standard addition method (SAM) associated with the liquid chromatography-tandem mass spectrometry (LC-MS/MS). The presence of antibiotics at a concentration of 2.5μg/g TS had no inhibitory effects on methane (CH4) production, total and volatile solids (TS and VS) removal as well as chemical oxygen demand (COD) removal. During the steady-state, antibiotics were removed significantly by 50, 100, and 59% respectively for the ROX, OTC, and CTC. Furthermore, ENR removal was not statistically significant and was estimated at 36%. This study highlighted that AD process could partially remove parent compounds, but ROX, CTC, and ENR persisted in the digested sludge. Hence, AD could be considered as a sludge treatment for mitigating, but not suppressing, the release of antibiotics through sludge application.
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