Anaerobic digestion is one of the best ways to re-use animal manure and agricultural residues, through the production of combustible biogas and digestate. However, the use of antibiotics for preventing and treating animal diseases and, consequently, their residual concentrations in manure, could introduce them into anaerobic digesters. If the digestate is applied as a soil fertilizer, antibiotic residues and/or their corresponding antibiotic resistance genes (ARGs) could reach soil ecosystems. This work investigated three common soil emerging contaminants, i.e., sulfamethoxazole (SMX), ciprofloxacin (CIP), enrofloxacin (ENR), their ARGs sul1, sul2, qnrS, qepA, aac-(6′)-Ib-cr and the mobile genetic element intI1, for one year in a full scale anaerobic plant. Six samplings were performed in line with the 45-day hydraulic retention time (HRT) of the anaerobic plant, by collecting input and output samples. The overall results show both antibiotics and ARGs decreased during the anaerobic digestion process. In particular, SMX was degraded by up to 100%, ENR up to 84% and CIP up to 92%, depending on the sampling time. In a similar way, all ARGs declined significantly (up to 80%) in the digestate samples. This work shows how anaerobic digestion can be a promising practice for lowering antibiotic residues and ARGs in soil.

Natural ecosystems are frequently exposed to complex mixtures of different chemicals. However, the environmental risk assessment is mainly based on data from individual substances. In this study, the individual and combined effects on the terrestrial earthworm E. fetida exposed to the anionic surfactant sodium lauryl ether sulphate (SLES) and the pesticides chlorpyrifos (CPF) and imidacloprid (IMI) were investigated, by using the avoidance behaviour as endpoint. Earthworms were exposed to a soil artificially contaminated with five sub-lethal concentrations of each contaminant, both as single substances and in combination of binary and ternary mixtures. Overall results showed that IMI provoked the highest avoidance effect on earthworms, with a concentration value that induced an avoidance rate of 50% of treated organisms (AC50) of 1.30 mg/kg, followed by CPF (AC50 75.26 mg/kg) and SLES (AC50 139.67 mg/kg). The application of the Combination Index (CI) method, indicated that a deviation from the additive response occurred for most of the tested chemical mixtures, leading to synergistic or antagonistic avoidance responses. Synergistic effects were produced by the exposure to the two lowest concentrations of the CPF+IMI mixture, and by the highest concentrations of SLES+CPF and SLES+CPF+IMI mixtures. On the contrary, antagonistic effects were observed at the lowest concentrations of the binary mixtures containing the SLES and at almost all the tested concentrations of the SLES+CPF+IMI mixture (with the exception of the highest tested concentration). These results show that the avoidance test is suitable to assess the detrimental effects exerted on earthworms by chemical mixtures in soil ecosystems and the use of behavioural endpoints can increase the ecological significance of environmental risk assessment procedures.

River contamination is due to a chemical mixture of point and diffuse pollution, which can compromise water quality. Polycyclic Aromatic Hydrocarbons (PAHs) and emerging compounds such as pharmaceuticals and antibiotics are frequently found in rivers flowing through big cities. This work evaluated the presence of fifteen priority PAHs, eight pharmaceuticals including the antibiotics ciprofloxacin (CIP) and sulfamethoxazole (SMX), together with their main antibiotic resistant genes (ARGs) and the structure of the natural bacterioplankton community, in an urbanized stretch of the river Danube. SMX and diclofenac were the most abundant chemicals found (up to 20 ng/L). ARGs were also found to be detected as ubiquitous contaminants. A principal component analysis of the overall microbiological and chemical data revealed which contaminants were correlated with the presence of certain bacterial groups. The highest concentrations of naphthalene were associated with Deltaproteobacteria and intI1 gene. Overall, the most contaminated site was inside the city and located immediately downstream of a wastewater treatment plant. However, both the sampling points before the river reached the city and in its southern suburban area were still affected by emerging and legacy contamination. The diffuse presence of antibiotics and ARGs causes particular concern because the river water is used for drinking purposes.

Some livestock farms rely on anaerobic digestion (AD) technology for manure disposal, thus obtaining energy (biogas) and fertilizer (digestate). Mixtures of antibiotics used for animal health often occur in organic waste and their possible synergistic/antagonistic effects on microorganisms involved in AD are still poorly studied. This work focuses on the effects of adding ciprofloxacin, alone (5 mg L⁻¹) and in combination with sulfamethoxazole (2.5–5–10 mg L⁻¹), on AD efficiency and microbial community structure. The experiment consisted of 90-day cattle manure batch tests and antibiotic removal percentages were assessed. Adding antibiotics always promoted CH₄ and H₂ production compared to untreated controls; however, CH₄ production was lowered with the highest ciprofloxacin (CIP) concentrations. The overall results show antibiotic degradation caused by acidogenic Bacteria, and CH₄ was mainly produced through the hydrogenotrophic-pathway by methanogenic Archaea. Shifts in microbial community abundance (DAPI counts) and composition (Illumina-MiSeq and FISH analyses) were observed.