In the yeast Kluyveromyces lactis, the inactivation of structural or regulatory glycolytic and fermentative genes generates obligate respiratory mutants which can be characterized by sensitivity to the mitochondrial drug antimycin A on glucose medium (Rag(-) phenotype). Rag(-) mutations can occasionally be generated by the inactivation of genes not evidently related to glycolysis or fermentation. One such gene is the hypoxic regulatory gene KlMGA2. In this work, we report a study of the many defects, in addition to the Rag(-) phenotype, generated by KlMGA2 deletion. We analyzed the fermentative and respiratory metabolism, mitochondrial functioning and morphology in the Klmga2Δ strain. We also examined alterations in the regulation of the expression of lipid biosynthetic genes, in particular fatty acids, ergosterol and cardiolipin, under hypoxic and cold stress and the phenotypic suppression by unsaturated fatty acids of the deleted strain. Results indicate that, despite the fact that the deleted mutant strain had a typical glycolytic/fermentative phenotype and KlMGA2 is a hypoxic regulatory gene, the deletion of this gene generated defects linked to mitochondrial functions suggesting new roles of this protein in the general regulation and cellular fitness of K. lactis. Supplementation of unsaturated fatty acids suppressed or modified these defects suggesting that KlMga2 modulates membrane functioning or membrane-associated functions, both cytoplasmic and mitochondrial.

Anaerobic digestion (AD) treatment of cattle manure and slurry makes it possible to produce biogas, a renewable and storable biofuel, as well as digestate, a residual organic matter that can be used to replace chemical fertilizers. On the other hand, the intense use of antibiotics (e.g., sulfamethoxazole) in animal husbandry practices is showing increasing negative impacts resulting from the release of still metabolically active molecules into agroecosystems. In the present study, cattle manure collected from an AD plant-feeding tank was used as feedstock for AD experiments in which some batches were spiked with 5 mg L −1 of sulfamethoxazole (SMX). Adding the antibiotic affected the microbial community dynamic; in particular, the efficiency of the acidogenic and acetogenic phases of the process corresponded to higher CH 4 and H 2 production than in the control. SMX was also degraded, and at the end of the experiment (69 days), just 20% of its initial concentration was found. The relative abundance (ARG/16S) of resistance genes sul1, sul2, and the proxy intI1 initially found in the ingestate decreased during the AD in both the spiked and control batches, suggesting that this process lowers the likelihood of antibiotic resistance genes spreading.

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.

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.