Diurnal variation in the selected indicators of water contamination in the Białka river affected by a sewage treatment plant discharge

This study was aimed to assess the diurnal variability in the number of microbial indicators of water quality and the content of the selected main ions at 3 sites located on Białka river, Podhale. The examined sites included the stream being a receiver of sewage from the treatment plant, before the treatment plant and few kilometers downstream of the sewage discharge - at the water intake for artificial snowing of the largest ski station in the region. Twelve series of samples were collected over 36 hours period. Temperature, pH, electrical conductivity and dissolved oxygen were measured onsite, microbiological analyses included the numbers of mesophilic bacteria, fecal Escherichia coli and Enterococcus faecalis, chemical analyses determined the concentration of main ions. E. faecalis was not detected in this study, while the concentration of mesophilic bacteria and E. coli varied largely between sites and hours of sampling. The smallest content of microorganisms was observed before the treatment plant and significantly increased at the sewage discharge. It was found that the cycle of the sewage treatment plant operation affects microbial contamination of water both in the stream which receives effluent from the treatment plant and in the downstream part of the river

The autophagic response to Staphylococcus aureus provides an intracellular niche in neutrophils.

Staphylococcus aureus is a major human pathogen causing multiple pathologies, from cutaneous lesions to life-threatening sepsis. Although neutrophils contribute to immunity against S. aureus, multiple lines of evidence suggest that these phagocytes can provide an intracellular niche for staphylococcal dissemination. However, the mechanism of neutrophil subversion by intracellular S. aureus remains unknown. Targeting of intracellular pathogens by macroautophagy/autophagy is recognized as an important component of host innate immunity, but whether autophagy is beneficial or detrimental to S. aureus-infected hosts remains controversial. Here, using larval zebrafish, we showed that the autophagy marker Lc3 rapidly decorates S. aureus following engulfment by macrophages and neutrophils. Upon phagocytosis by neutrophils, Lc3-positive, non-acidified spacious phagosomes are formed. This response is dependent on phagocyte NADPH oxidase as both cyba/p22phox knockdown and diphenyleneiodonium (DPI) treatment inhibited Lc3 decoration of phagosomes. Importantly, NADPH oxidase inhibition diverted neutrophil S. aureus processing into tight acidified vesicles, which resulted in increased host resistance to the infection. Some intracellular bacteria within neutrophils were also tagged by Sqstm1/p62-GFP fusion protein and loss of Sqstm1 impaired host defense. Together, we have shown that intracellular handling of S. aureus by neutrophils is best explained by Lc3-associated phagocytosis (LAP), which appears to provide an intracellular niche for bacterial pathogenesis, while the selective autophagy receptor Sqstm1 is host-protective. The antagonistic roles of LAP and Sqstm1-mediated pathways in S. aureus-infected neutrophils may explain the conflicting reports relating to anti-staphylococcal autophagy and provide new insights for therapeutic strategies against antimicrobial-resistant Staphylococci.Abbreviations: ATG: autophagy related; CFU: colony-forming units; CMV: cytomegalovirus; Cyba/P22phox: cytochrome b-245, alpha polypeptide; DMSO: dimethyl sulfoxide; DPI: diphenyleneiodonium; EGFP: enhanced green fluorescent protein; GFP: green fluorescent protein; hpf: hours post-fertilization; hpi: hours post-infection; Irf8: interferon regulatory factor 8; LAP: LC3-associated phagocytosis; lyz: lysozyme; LWT: london wild type; Map1lc3/Lc3: microtubule-associated protein 1 light chain 3; NADPH oxidase: nicotinamide adenine dinucleotide phosphate oxidase; RFP: red fluorescent protein; ROS: reactive oxygen species; RT-PCR: reverse transcriptase polymerase chain reaction; Sqstm1/p62: sequestosome 1; Tg: transgenic; TSA: tyramide signal amplification

Antibiotics in groundwater and river water of Białka : a pristine mountain river

Antibiotics are emerging pollutants of great concern, due to detrimental effects of their sublethal concentrations on microbial communities. Monitoring of antibiotics’ presence and concentrations in the aquatic environment is of fundamental importance to the management of water resources. This study was aimed at filling the knowledge gap in terms of presence and concentration of antibiotics in surface water and groundwater in one of the mountain regions in southern Poland. The detailed aims included the assessment of whether there are spatial and/or temporal trends in antibiotic occurrence in water and the investigation of causes behind the changes in antibiotic concentrations. The study was conducted in seven sites (two groundwater and five river water) along the Białka river valley. Antibiotics were subjected to solid-phase extraction, followed by UHPLC/MS detection. Clindamycin, erythromycin, ofloxacin and trimethoprim were the most frequently detected, while the highest concentrations were observed for oxytetracycline and clindamycin. No antibiotics were detected in only one of the groundwater sites. Sewage treatment plant effluent was the hotspot of antibiotic contamination of the river downstream. The detection rates of antibiotics in the examined region seem to be driven mainly by the stability of antibiotics in the environment

Anthropogenic pollution gradient along a mountain river affects bacterial community composition and genera with potential pathogenic species

Mountain regions in Poland are among the most frequently visited tourist destinations, causing a significant anthropogenic pressure put on the local rivers. In this study, based on numbers of 9 microorganisms, content of 17 antibiotics and 17 physicochemical parameters, we determined a pollution gradient in six sites along Białka, a typical mountain river in southern Poland. The E.coli/Staphylococcus ratio varied evidently between polluted and non-polluted sites, indicating that the possible utility of this parameter in assessing the anthropogenic impact on river ecosystems is worth further investigation. Then, using next generation sequencing, we assessed the changes in bacterial community structure and diversity as a response to the pollution gradient. Proteobacteria and Bacteroidetes were the most abundant phyla in the majority of samples. Actinobacteria were the most abundant in the most pristine (groundwater) sample, while Firmicutes and Verrucomicrobia were more prevalent in polluted sites. Bacterial diversity at various levels increased with water pollution. Eleven bacterial genera potentially containing pathogenic species were detected in the examined samples, among which Acinetobacter, Rhodococcus, and Mycobacterium were the most frequent. At the species level, Acinetobacter johnsonii was most prevalent potential pathogen, detected in all surface water samples, including the pristine ones. Two bacterial taxa—genus Flectobacillus and order Clostridiales showed very distinct variation in the relative abundance between the polluted and non-polluted sites, indicating their possible potential as biomarkers of anthropogenic impact on mountain river waters

COVID-19 lockdown shows how much natural mountain regions are affected by heavy tourism

Mountain areas in Poland are among the most frequented tourist destinations and such intensive tourism negatively affects the natural environment. The COVID-19 pandemic and the resulting lockdown restricted travel for a few months in 2020, providing a unique opportunity to observe the studied mountain environment without the impact of typical tourist traffic. This study is based on the determination of antibiotic content, hydrochemical parameters, enumeration of culturable bacterial water quality indicators, antimicrobial susceptibility tests together with extended spectrum beta-lactamase (ESBL) gene detection in waterborne E. coli and NGS-based bacterial community composition at six sites along the Białka river valley (one of the most popular touristic regions in Poland) in three periods: in summer and winter tourist seasons and during the COVID-19 lockdown. The results of individual measurements showed decreased numbers of bacterial indicators of water contamination (e.g. numbers of E. coli dropped from 99 × 10(4) CFU/100 ml to 12 CFU/100 ml at the most contaminated site) and the share of antimicrobial resistant E. coli (total resistance dropped from 21% in summer to 9% during lockdown, share of multidrug resistant strains from 100 to 44%, and ESBL from 20% in summer to none during lockdown). Antibiotic concentrations were the highest during lockdown. The use of multivariate analysis (principal component analysis – PCA and heatmaps) revealed a clear pattern of tourism-related anthropogenic pressure on the water environment and positive impact of COVID-19 lockdown on water quality. PCA distinguished three major factors determining water quality: F1 shows strong effect of anthropogenic pressure; F2 describes the lockdown-related quality restoration processes; F3 is semi-natural and describes the differences between the most pristine and most anthropogenically-impacted waters