Summary: The bacterioplankton diversity in large rivers has thus far been under‐sampled despite the importance of streams and rivers as components of continental landscapes. Here, we present a comprehensive dataset detailing the bacterioplankton diversity along the midstream of the D anube River and its tributaries. Using 16 S rRNA ‐gene amplicon sequencing, our analysis revealed that bacterial richness and evenness gradually declined downriver in both the free‐living and particle‐associated bacterial communities. These shifts were also supported by beta diversity analysis, where the effects of tributaries were negligible in regards to the overall variation. In addition, the river was largely dominated by bacteria that are commonly observed in freshwaters. Dominated by the acI lineage, the freshwater SAR 11 ( LD 12) and the P olynucleobacter group, typical freshwater taxa increased in proportion downriver and were accompanied by a decrease in soil and groundwater‐affiliated bacteria. Based on views of the meta‐community and River Continuum Concept, we interpret the observed taxonomic patterns and accompanying changes in alpha and beta diversity with the intention of laying the foundation for a unified concept for river bacterioplankton diversity.

Hydrological observatories bear a lot of resemblance to the more traditional research catchment concept, but tend to differ in providing more long-term facilities that transcend the lifetime of individual projects, are more strongly geared towards performing interdisciplinary research, and are often designed as networks to assist in performing collaborative science. This paper illustrates how the experimental and monitoring set-up of an observatory, the 66 ha Hydrological Open Air Laboratory (HOAL) in Petzenkirchen, Lower Austria, has been established in a way that allows meaningful hypothesis testing. The overarching science questions guided site selection, identification of dissertation topics and the base monitoring. The specific hypotheses guided the dedicated monitoring and sampling, individual experiments, and repeated experiments with controlled boundary conditions. The purpose of the HOAL is to advance the understanding of water-related flow and transport processes involving sediments, nutrients and microbes in small catchments. The HOAL catchment is ideally suited for this purpose, because it features a range of different runoff generation processes (surface runoff, springs, tile drains, wetlands), the nutrient inputs are known, and it is convenient from a logistic point of view as all instruments can be connected to the power grid and a high-speed glassfibre local area network (LAN). The multitude of runoff generation mechanisms in the catchment provides a genuine laboratory where hypotheses of flow and transport can be tested, either by controlled experiments or by contrasting sub-regions of different characteristics. This diversity also ensures that the HOAL is representative of a range of catchments around the world, and the specific process findings from the HOAL are applicable to a variety of agricultural catchment settings. The HOAL is operated jointly by the Vienna University of Technology and the Federal Agency for Water Management and takes advantage of the Vienna Doctoral Programme on Water Resource Systems funded by the Austrian Science Funds. The paper presents the science strategy of the set-up of the observatory, discusses the implementation of the HOAL, gives examples of the hypothesis testing and summarises the lessons learned. The paper concludes with an outlook on future developments.

Numerous bacterial genetic markers are available for the molecular detection of human sources of fecal pollution in environmental waters. However, widespread application is hindered by a lack of knowledge regarding geographical stability, limiting implementation to a small number of well-characterized regions. This study investigates the geographic distribution of five human-associated genetic markers (HF183/BFDrev, HF183/BacR287, BacHum-UCD, BacH, and Lachno2) in municipal wastewaters (raw and treated) from 29 urban and rural wastewater treatment plants (750 – 4,400,000 population equivalents) from 13 countries spanning six continents. In addition, genetic markers were tested against 280 human and non-human fecal samples from domesticated, agricultural and wild animal sources. Findings revealed that all genetic markers are present in consistently high concentrations in raw (median log10 7.2 - 8.0 marker equivalents (ME) 100 ml-1) and biologically treated wastewater samples (median log10 4.6 - 6.0 ME 100 ml-1) regardless of location and population. The false positive rates of the various markers in non-human fecal samples ranged from 5% to 47%. Results suggest that several genetic markers have considerable potential for measuring human-associated contamination in polluted environmental waters. This will be helpful in water quality management and pollution modeling and health risk assessment across the globe (as demonstrated by QMRAcatch).

This was a detailed investigation of the seasonal occurrence, dynamics, removal and resistance of human-associated genetic Bacteroidetes faecal markers (GeBaM) compared with ISO-based standard faecal indicator bacteria (SFIB), human-specific viral faecal markers and one human-associated Bacteroidetes phage in raw and treated wastewater of municipal and domestic origin. Characteristics of the selected activated sludge wastewater treatment plants (WWTPs) from Austria and Germany were studied in detail (WWTPs, n = 13, connected populations from 3 to 49000 individuals), supported by volume-proportional automated 24-h sampling and chemical water quality analysis. GeBaM were consistently detected in high concentrations in raw (median log10 8.6 marker equivalents (ME) 100 ml(-1)) and biologically treated wastewater samples (median log10 6.2-6.5 ME 100 ml(-1)), irrespective of plant size, type and time of the season (n = 53-65). GeBaM, Escherichia coli, and enterococci concentrations revealed the same range of statistical variability for raw (multiplicative standard deviations s* = 2.3-3.0) and treated wastewater (s* = 3.7-4.5), with increased variability after treatment. Clostridium perfringens spores revealed the lowest variability for raw wastewater (s* = 1.5). In raw wastewater correlations among microbiological parameters were only detectable between GeBaM, C. perfringens and JC polyomaviruses. Statistical associations amongst microbial parameters increased during wastewater treatment. Two plants with advanced treatment were also investigated, revealing a minimum log10 5.0 (10th percentile) reduction of GeBaM in the activated sludge membrane bioreactor, but no reduction of the genetic markers during UV irradiation (254 nm). This study highlights the potential of human-associated GeBaM to complement wastewater impact monitoring based on the determination of SFIB. In addition, human-specific JC polyomaviruses and adenoviruses seem to be a valuable support if highly specific markers are needed.

Changes in climatic conditions will directly affect the quality and quantity of water resources. Further on, they will affect them indirectly through adaptation in land use which ultimately influences diffuse nutrient emissions to rivers and therefore potentially the compliance with good ecological status according to the EU Water Framework Directive (WFD). We present an integrated impact modelling framework (IIMF) to track and quantify direct and indirect pollution impacts along policy-economy-climate-agriculture-water interfaces. The IIMF is applied to assess impacts of climatic and socio-economic drivers on agricultural land use (crop choices, farming practices and fertilization levels), river flows and the risk for exceedance of environmental quality standards for determination of the ecological water quality status in Austria. This article also presents model interfaces as well as validation procedures and results of single models and the IIMF with respect to observed state variables such as land use, river flow and nutrient river loads. The performance of the IIMF for calculations of river nutrient loads (120 monitoring stations) shows a Nash-Sutcliffe Efficiency of 0.73 for nitrogen and 0.51 for phosphorus. Most problematic is the modelling of phosphorus loads in the alpine catchments dominated by forests and mountainous landscape. About 63% of these catchments show a deviation between modelled and observed loads of 30% and more. In catchments dominated by agricultural production, the performance of the IIMF is much better as only 30% of cropland and 23% of permanent grassland dominated areas have a deviation of >30% between modelled and observed loads. As risk of exceedance of environmental quality standards is mainly recognized in catchments dominated by cropland, the IIMF is well suited for assessing the nutrient component of the WFD ecological status.