The impact of antimicrobials on the efficiency of methane fermentation of sewage sludge, changes in microbial biodiversity and the spread of antibiotic resistance.

The study was designed to simultaneously evaluate the influence of high doses (512-1024 µg/g) the most commonly prescribed antimicrobials on the efficiency of anaerobic digestion of sewage sludge, qualitative and quantitative changes in microbial consortia responsible for the fermentation process, the presence of methanogenic microorganisms, and the fate of antibiotic resistance genes (ARGs). The efficiency of antibiotic degradation during anaerobic treatment was also determined. Metronidazole, amoxicillin and ciprofloxacin exerted the greatest effect on methane fermentation by decreasing its efficiency. Metronidazole, amoxicillin, cefuroxime and sulfamethoxazole were degraded in 100%, whereas ciprofloxacin and nalidixic acid were least susceptible to degradation. The most extensive changes in the structure of digestate microbiota were observed in sewage sludge exposed to metronidazole, where a decrease in the percentage of bacteria of the phylum Bacteroidetes led to an increase in the proportions of bacteria of the phyla Firmicutes and Proteobacteria. The results of the analysis examining changes in the concentration of the functional methanogen gene (mcrA) did not reflect the actual efficiency of methane fermentation. In sewage sludge exposed to antimicrobials, a significant increase was noted in the concentrations of ß-lactam, tetracycline and fluoroquinolone ARGs and integrase genes, but selective pressure was not specific to the corresponding ARGs.

The impact of WWTP size and sampling season on the prevalence of antibiotic resistance genes in wastewater and the river system.

There is a growing concern about the fate of antibiotic resistance genes (ARGs) during wastewater treatment and their potential impacts on the receiving water bodies. We hypothesised that the quantity of ARGs in effluents may be related to the size of wastewater treatment plants (WWTPs) and sampling season. To date, only several attempts have been made to investigate the impact of the above factors at the catchment scale. Therefore, the goal of the present study was to explore possible differences in the quantity of ARGs in treated wastewater from small, medium-sized and large WWTPs in the catchment of the Pilica River (9258 km2). The impact of treated wastewater on the concentration of ARGs was also determined along the river continuum from upland to lowland segments to the point of confluence with the Vistula (342 km). Treated effluent was sampled in 17 WWTPs, and river water was sampled in 7 sampling sites in four seasons. The concentrations of blaTEM, tet(A), ermF, sul1 and aac(6')-Ib-cr genes, the integrase gene intI1 and the 16S rRNA gene were analysed by quantitative PCR. The physical and chemical parameters and nutrient concentrations (23 various parameters) in the analysed samples were determined. The highest absolute concentrations of the studied genes were noted in effluent samples from small WWTPs (p < 0.01). The concentration of ARGs (gene copies/mL) peaked in winter and spring samples (p < 0.04). The results of statistical analyses indicate that in small WWTPs, the absolute concentration of ARGs can be predicted based on the biochemical oxygen demand, in routine water analyses. However, none of the studied parameters supported predictions of ARG abundance in medium-sized and large WWTPs or in river water.