In freshwaters, microbial communities are of outstanding importance both from ecological and public health perspectives, however, they are threatened by the impact of global warming. To reveal how different prokaryotic communities in a large temperate river respond to environment conditions related to climate change, the present study provides the first detailed insight into the composition and spatial and year-round temporal variations of planktonic and epilithic prokaryotic community. Microbial diversity was studied using high-throughput next generation amplicon sequencing. Sampling was carried out monthly in the midstream and the littoral zone of the Danube, upstream and downstream from a large urban area. Result demonstrated that river habitats predominantly determine the taxonomic composition of the microbiota; diverse and well-differentiated microbial communities developed in water and epilithon, with higher variance in the latter. The composition of bacterioplankton clearly followed the prolongation of the summer resulting from climate change, while the epilithon community was less responsive. Rising water temperatures was associated with increased abundances of many taxa (such as phylum Actinobacteria, class Gammaproteobacteria and orders Synechococcales, Alteromonadales, Chitinophagales, Pseudomonadales, Rhizobiales and Xanthomonadales), and the composition of the microbiota also reflected changes of several further environmental factors (such as turbidity, TOC, electric conductivity, pH and the concentration of phosphate, sulphate, nitrate, total nitrogen and the dissolved oxygen). The results indicate that shift in microbial community responding to changing environment may be of crucial importance in the decomposition of organic compounds (including pollutants and xenobiotics), the transformation and accumulation of heavy metals and the occurrence of pathogens or antimicrobial resistant organisms.

Freshwaters play an essential role in providing ecosystem services worldwide, however, the water quality of different water bodies is strongly influenced by human activities such as urbanization, industry and agriculture. In this study, water and biofilm samples were collected from the main channel of the Danube River upstream and downstream of a metropolitan, from a regulated side arm within an urbanized area, and from two differently separated oxbow lakes located in nature conservation areas. The taxonomic diversity of bacterial communities was revealed by 16S rRNA gene-based amplicon sequencing using Illumina MiSeq platform. The results showed that all samples were dominated by phyla Pseudomonadota, Actinobacteriota and Bacteroidota. The bacterial community structures, however, clearly differentiated according to planktonic and epilithic or epiphytic habitats, as well as by riverine body types (main channel, side arm, oxbow lakes). The taxonomic diversity of biofilm communities was higher than that of planktonic ones in all studied habitats. Human impacts were mainly reflected in the slowly changing biofilm composition compared to the planktonic ones. Genera with pollution tolerance and/or degradation potential, such as Acinetobacter, Pseudomonas and Shewanella were mainly detected in biofilm communities of the highly urbanized section of the river side arm.

In drinking water supply, riverbank filtration (RBF) is an efficient and cost-effective way of eliminating pathogens and micropollutants using a combination of biotic and abiotic processes. Microbial communities in the hyporheic zone both contribute to and are shaped by these processes. Microbial water quality at the point of consumption is in turn influenced by the source water microbiome, water treatment and distribution system. Understanding microbial community shifts from source to tap and the factors behind them is instrumental in maintaining safe drinking water delivery. To this end, microbial communities of an RBF-based drinking water supply system were investigated by metabarcoding in a one-year sampling campaign. Samples were collected from the river, RBF wells, treated water, and a consumer’s tap. Metabarcoding data were analysed in the context of physicochemical and hydrological parameters. Microbial diversity as well as cell count decreased consistently from the surface water to the tap. While Proteobacteria were dominant throughout the water supply system, typical river water microbiome phyla Bacteroidota, Actinobacteria, and Verrucomicrobiota were replaced by Nitrospira, Patescibacteria, Chloroflexi, Acidobacteriota, Methylomicrobilota, and the archaeal phylum Nanoarcheota in well water. Well water communities were differentiated by water chemistry, in wells with high concentration groundwater derived iron, manganese, and sulphate, taxa related to iron and sulphur biogeochemical cycle were predominant, while methane oxidisers characterised the more oxic wells. Chlorine-resistant and filtration-associated taxa (Acidobacteria, Firmicutes, and Bdellovibrionota) emerged after water treatment, and no potentially pathogenic taxa were identified at the point of consumption. River discharge had a distinct impact on well water microbiome indicative of vulnerability to climate change. Low flow conditions were characterised by anaerobic heterotrophic taxa (Woesarchaeales, Aenigmarchaeales, and uncultured bacterial phyla MBNT15 and WOR-1), implying reduced efficiency in the degradation of organic substances. High flow was associated the emergence of typical surface water taxa. Better understanding of microbial diversity in RBF water supply systems contributes to preserving drinking water safety in the future changing environment.

The adaptability of plant populations to a changing environment depends on their genetic diversity, which in turn is influenced by the degree of sexual reproduction and gene flow from distant areas. Aquatic macrophytes can reproduce both sexually and asexually, and their reproductive fragments are spread in various ways (e.g. by water). Although these plants are obviously exposed to hydrological changes, the degree of vulnerability may depend on the types of their reproduction and distribution, as well as the hydrological differences of habitats. The aim of this study was to investigate the genetic diversity of the cosmopolitan macrophyte Ceratophyllum demersum in hydrologically different aquatic habitats, i.e. rivers and backwaters separated from the main river bed to a different extent. For this purpose, the first microsatellite primer set was developed for this species. Using 10 developed primer pairs, a high level of genetic variation was explored in C. demersum populations. Overall, more than 80% of the loci were found to be polymorphic, a total of 46 different multilocus genotypes and 18 private alleles were detected in the 63 individuals examined. The results demonstrated that microsatellite polymorphism in this species depends on habitat hydrology. The greatest genetic variability was revealed in populations of rivers, where flowing water provides constant longitudinal connections with distant habitats. The populations of the hydrologically isolated backwaters showed the lowest microsatellite polymorphism, while plants from an oxbow occasionally flooded by the main river had medium genetic diversity. The results highlight that in contrast to species that spread independently of water flow or among hydrologically isolated water bodies, macrophytes with exclusive or dominant hydrochory may be most severely affected by habitat fragmentation, for example due to climate change.

Riverbank filtration is a cost-effective and efficient method for drinking water production, using the natural filtration capacity of the river gravelbed. Removal efficiency for organic micropollutants (OMP) in field studies is generally calculated by comparing the concentrations measured in surface water and in the wells either on the same day or with a shift of fixed time interval, neither of which can account for the variability of surface water quality and travel time in the aquifer. The present study proposes a novel method based on travel time distribution determined by a numerical transport model with a hypothesis that it will provide more reliable estimate for OMP removal. The model was developed for two production sites of Budapest Waterworks, Hungary on Danube River. River water and riverbank filtered well water were sampled regularly for one year (158 samples each) and analysed for 41 OMPs (pesticides, pharmaceutical residues and industrial pollutants). Nineteen pollutants were detected in >50 % of the well water samples. Median removal rates were 4-97 %, while the concentration of five compounds increased in some wells. Removal rates of telmisartan, tramadol, sulfamethoxazole, 4-methyl-benzotriazole, 5-methyl-benzotriazole and desethyl-terbuthylazine correlated negatively to redox potential (|r|=0.456-0.805). Median travel time increased after high flow events resulting in reduced removal of telmisartan, tramadol, 4-methyl-benzotriazole and desethyl-terbuthylazine (|r|= 0.435-0.661). Removal of diatrizoate, iopamidol, tramadol and benzotriazole increased with distance from the shore (148 vs 395 m) by 25 %, 28 %, 8 %, 16 %, respectively. Background groundwater contamination increased pesticide concentration in the wells located in agricultural areas 1.5-5-fold compared to river water. The model-based method gave more consistent results compared to traditional calculations for OMP removal efficiency during the sampling campaign and allowed for estimating the impact of various environmental factors.

The detection of non-point pollution in large rivers requires high-frequency sampling over a longer period of time, which, however presumably provides data with large spatial and temporal variance. Variability may mean that data sets recorded upstream and downstream from a densely populated area overlap, suggesting at first glance that the urban area did not affect water quality. This study presents a simple way to explore trend-like effects of non-point pollution in the Danube based on data that varied strongly in space and time. For one year, biweekly sampling was carried out upstream and downstream from a large city with negligible emission of untreated wastewater and the surrounding settlements, industrial and agricultural areas.