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Alterations in natural flow regimes caused by dams can significantly alter the aquatic habitats of stream organisms. However, few studies have characterized flow regulation to assess its impacts on stream fauna in the context of interannually variable extreme floods. This study aims to understand the variation in stream animals along flow regulation gradients due to hydropower dams in a catchment experiencing typhoons. We observed freshwater fishes and stream insects at fully regulated sites (receiving residual flow), moderately regulated sites (receiving hydropower outflow), and nonregulated site (tributary) in the Mimi River catchment in southern Japan, in summer and winter from 2010 to 2018. We computed indicators of hydrologic alteration (IHA) in each calendar/water (July to June) year from 2007 to 2017 and selected subsets of IHA based on principal component analysis (PCA) and variance inflation factor. The largest variance was mainly explained by minimum discharge levels (e.g., 30-day annual minimum) and flow variability among IHAs, distinguishing the moderately regulated and nonregulated sites from fully regulated sites because of residual flow and suppressed high pulses in the fully regulated sites. Generalized additive models revealed that annual maxima of specific discharge were most significant predictors of fish and insect metrics while its effects were generally inconsistent between summer and winter. Non-metric multidimensional scaling revealed that insect communities were clustered into the regulation extents in both seasons. The differences in winter fauna between the regulated and nonregulated sites, characterized by Ephemeroptera-Plecoptera-Trichoptera abundance, were associated with maximum discharge and high pulse numbers. Fish community variation did not correspond to flow regime gradients. Our findings on mechanistic ecohydrological consequences of various flow regulations, supported by long-term observations, will be useful for river managers attempting to compensate for alterations in flow regime and ecological integrity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The dissemination of antimicrobial-resistant bacteria in environmental water is an emerging concern in medical and industrial settings. Here, we analysed the antimicrobial resistance of Escherichia coli isolates from river water and sewage by the use of a combined experimental phenotypic and whole-genome-based genetic approach. Among the 283 tested strains, 52 were phenotypically resistant to one or more antimicrobial agents. The E. coli isolates from the river and sewage samples were phylogenetically indistinguishable, and the antimicrobial-resistant strains were dispersedly distributed in a whole-genome-based phylogenetic tree. The prevalence of antimicrobial-resistant strains as well as the number of antimicrobials to which they were resistant were higher in sewage samples than in river samples. Antimicrobial resistance genes were more frequently detected in strains from sewage samples than in those from river samples. We also found that 16 river isolates that were classified as Escherichia cryptic clade V were susceptible to all the antimicrobials tested and were negative for antimicrobial resistance genes. Our results suggest that E. coli strains may acquire antimicrobial resistance genes more frequently and/or antimicrobial-resistant E. coli strains may have higher rates of accumulation and positive selection in sewage than in rivers, irrespective of their phylogenetic distribution.

Wastewater treatment plants could discharge Escherichia coli and antibiotic resistant bacteria to the environment adjacent to, or downstream of their discharge point. However, their discharge also contains nutrients which could promote growth of E. coli in water environments. This study was done to clarify the potential of growth and antibiotic resistance acquisition of E. coli in a river environment. Levels of E. coli were monitored in a river that receives treated sewage effluent for over four years. River water, periphyton and sediment samples were collected at sites upstream and downstream of treated sewage inflow. Concentrations of E. coli increased in river water and periphyton at the sites downstream of the treated sewage inflow, although levels of E. coli were very low or below detection limit in the treated sewage samples. Concentrations of Chlorophyll a increased at the downstream sites, likely due to nutrient input from the treated sewage. Based on pulsed field gel electrophoresis, identical genotype occurred at multiple sites both upstream and downstream of the treated sewage inflow. However, strains resistant to antibiotics such as ampicillin, cefazolin, ciprofloxacin, and chloramphenicol were more frequently obtained from the downstream sites than the upstream sites. Multidrug resistant E. coli strains were detected in periphyton and sediment samples collected at the downstream sites. Non-resistant strains with PDGE genotype identical to the multi-drug strains were also detected, indicating that E. coli might have become resistant to antibiotics by acquiring resistance genes via horizontal gene transfer. Laboratory incubation experiment showed the growth of E. coli in periphyton or sediment-fed river water samples. These results suggest that the wastewater treatment inflow did not directly provide E. coli to the river water, but could promote the growth of periphyton, which could lead to the elevated levels of E. coli and the emergence of antibiotic resistant E. coli., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this study was to rapidly and effectively analyze coliforms, which are the most fundamental indicators of water quality for fecal pollution, using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Coliform bacteria were isolated from municipal sewage, river water, and groundwater. For each sample, 100 isolates were determined by MALDI-TOF MS. In addition, these same 100 isolates were also identified via 16S rRNA gene sequence analysis. Obtained MALDI-TOF MS data were compared with the 16S rRNA sequencing analysis, and the validity of MALDI-TOF MS for classification of coliform bacteria was examined. The concordance rate of bacterial identification for the 100 isolates obtained by MALDI-TOF MS analysis and 16S rRNA gene sequence analysis for sewage, river water, and ground water were 96%, 74%, and 62% at the genus level, respectively. Among the sewage, river water, and ground water samples, the coliform bacterial flora were distinct. The dominant genus of coliforms in sewage, river water, and groundwater were Klebsiella spp., Enterobacter spp., and Serratia spp., respectively. We determined that MALDI-TOF MS is a rapid and accurate tool that can be used to identify coliforms. Therefore, without using conventional 16S rRNA sequencing, it is possible to rapidly and effectively classify coliforms in water using MALDI-TOF MS.

The concentration of MS2 phage as a model RNA virus in river water using a combined ferric colloid adsorption and foam separation-based method was examined. The MS2 phage concentrations were determined by the plaque-forming unit (PFU) method and reverse transcription quantitative PCR (RT-qPCR) analysis. When ferric colloid adsorption was performed prior to foam separation, MS2 phage was effectively removed from river water and concentrated in the generated foam within 7 min. The removal efficiency was >99% at the optimum iron and casein concentrations of 5 mg-Fe/L and 10 mg/L, respectively. Furthermore, based on the analysis of the collected ferric colloid dissolved using deferoxamine, the MS2 concentration in the colloid-dissolved solution was 190-fold higher than that found in raw water according to RT-qPCR analysis. This is a novel method for concentrating RNA viruses to facilitate their detection in river water using coagulation and foam separation combined with chelate dissolution of ferric flocs., (Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Cryptosporidium is an extremely virulent microorganism; contamination of water works systems by Cryptosporidium oocysts is a serious issue in public health. However, a great deal of effort is required to detect oocysts in source water for drinking, because the oocysts have to be concentrated from a large volume of water before detection. In this study, the concentration of oocysts in oocyst-polluted artificial river water was examined by coagulation and foam separation. The oocysts concentrated in the foam water were purified using immunomagnetic separation and assessed using epifluorescence microscopy. When ferric coagulation was performed prior to foam separation, oocysts were effectively removed from the river water and concentrated in the foam within 10 min. The removal efficiency was greater than 96% at the optimum iron and casein concentrations of 5 mg-Fe L(-1) and 20 mg L(-1), respectively. Furthermore, the number of recovered oocysts in the solution obtained by the dissolution of the collected ferric flocs from foam water was the same as that obtained using the Japanese standard procedure for detection of Cryptosporidium in water using membrane filtration. This is a novel method of concentrating Cryptosporidium oocysts for detection in river water using a coagulation and foam separation combined with acid dissolution of ferric flocs; the method is easy to use, rapid, and efficient.

As a first step for assessing the risk to human health posed by vancomycin-resistant enterococci (VRE) in the aquatic environment, we screened sewage and urban river water samples from Miyazaki, Japan for VRE. Because vancomycin-resistant organisms are not as prevalent in sewage and river water as vancomycin-susceptible organisms, the samples were screened by minimum inhibitory concentration test using the vancomycin-supplemented membrane-Enterococcus indoxyl-β-d-glucoside (mEI) agar. The isolates, presumed to be enterococci, were identified using 16S rRNA sequencing analysis. The percentages of VRE isolates screened using 4 μg mL(-1) vancomycin-supplemented mEI agar from sewage and urban river water samples were 12% and 24%, respectively. The vancomycin-resistant genes vanC1 and vanC2/3 were detected in the isolates from both samples by PCR analysis. All enterococci isolates containing vanC1, which is a specific gene for vanC-type of VRE, were identified as Enterococcus casseliflavus/gallinarum. Further, 92% enterococci isolates containing vanC2/3 were identified as E. casseliflavus/gallinarum, the remaining isolates containing vanC2/3 were E. faecium (4%) and E. faecalis (4%). Thereafter, the distribution of E. faecium and E. faecalis, which are the major types of enterococci in humans containing vanC2/3, was observed in the water samples collected.

Pseudomonas aeruginosa is responsible for a number of opportunistic and nosocomial infections. However, very little information is available on the ecology of P. aeruginosa in water environments and its association with antimicrobial resistance. In this study, the distribution of P. aeruginosa and the resistance of P. aeruginosa isolates to various antibiotics were investigated from two rivers, Kiyotake and Yae that flow through Miyazaki City, Japan. P. aeruginosa was distributed widely along the river basins with counts ranging from 2-46 cfu/100 mL. The susceptibility of P. aeruginosa isolates collected from the rivers to various antibiotics was examined by minimum inhibitory concentration. Multidrug-resistant P. aeruginosa strains were not observed or isolated from either river. However, one piperacillin-resistant P. aeruginosa was detected among a total of 516 isolates, and this isolate was also resistant to cefotaxime and showed intermediate resistance to cefitazidime. Less than 1% of all isolates (n=5) were resistant to imipenem, which is the most effective antibiotic against both Gram-negative and Gram-positive bacteria. However, all P. aeruginosa isolates were completely resistant to tetracyclines, which are the most commonly prescribed antibiotics. In advanced nations such as Japan where the majority of the population is urban and where medical services are widespread, antibiotic-resistant bacteria such as P. aeruginosa are likely to be widely distributed, even in apparently pristine rivers., (Copyright © 2013 Elsevier B.V. All rights reserved.)

This study aimed to identify specific river sources of fecal contamination by applying pulsed-field gel electrophoresis (PFGE) to environmental water samples from a recreational beach in Japan. The genotypes of all Enterococcus faecium and Enterococcus faecalis strains used as indicators of fecal pollution on the recreational beach and rivers were analyzed by PFGE, and the PFGE profiles of the strains were classified at a 0.9 similarity level using dendrogram analysis. PFGE types of E. faecium isolated from Sakai River or urban drainage were classified in the same cluster. Therefore, the probable sources of fecal pollution on the recreational beach were Sakai River and urban drainage. The approaches for microbial source tracking employed in this study used PFGE with Enterococcus species as an indicator can be a potential tool to specify the source(s) of fecal pollution and contribute to improved public health in coastal environments.

Enterococci such as Enterococcus faecalis and E. faecium are considered as the most suitable indicators of fecal pollution in an aquatic environment, and new methods for Enterococcus determination have been developed, namely, membrane filtration (MF) using membrane-Enterococcus indoxyl-β-D-glucoside agar (mEI) and defined substrate technology (DST) using Enterolert®. This study used PCR analysis to identify E. faecalis and E. faecium among Enterococcus strains in river water isolated using both mEI plates and Enterolert® trays. There was a significantly high correlation between MF and DST in terms of enterococcal counts for river water samples. The combined percentages of E. faecalis and E. faecium with respect to the total number of all strains obtained using mEI plates and Enterolert® trays were approximately 30 % and >30 %, respectively. Other than E. faecalis and E. faecium, a large number of Enterococcus species were unspecified in the actual urban river samples. A comparison of the predominance of E. faecalis and E. faecium found that the abundance of a species depended on the sampling river and date. E. faecium is a non-predominant species in intestinal and fecal Enterococci, and it was one of the main Enterococcus species detected in surface water.

Both Escherichia coli and Enterococci were collected in foam within 7 min from 500 mL of bacteria-spiked water by coagulation and foam separation using ferric chloride and milk casein. These bacterial DNA isolated in the 100 µL of extract from the foam more than 87.5% recovery using the DNeasy PowerWater® Kit. To test this method with water from three natural rivers, 0.67–2.70 µg of DNA were concentrated in 100 µL of extract from 1,000 mL of river water. When the DNA extract was subjected to 16S rRNA gene sequencing analysis, information on the bacterial flora could be obtained. [ABSTRACT FROM AUTHOR]

Rivers fulfill an essential ecological role by forming networks for material transport from upland forests to coastal areas. The way in which dams affect the organic and inorganic cycles in such systems is not well understood. Herein, we investigated the longitudinal profiles of the various components of the water chemistry across three cascade dams in Japan: the Yamasubaru Dam, Saigou Dam, and Ohuchibaru Dam, which are situated along the sediment-productive Mimi River in different flow conditions. We analyzed the following water quality components: suspended solids (SS), turbidity, total iron (TFe), dissolved iron (DFe), total organic carbon (TOC), total nitrogen (TN), total phosphorus (TP), humic substance (HS), and major ionic components (Na + , Mg 2+ , Ca 2+ , Cl − , NO 3− , and SO 4 2− ) in the downstream channels of the three dams during the low–intermediate-flow and high-flow events from 2012 to 2014. We estimated hourly loads of each component using hourly turbidity data and discharge data (i.e., L–Q model) separately, and the results are integrated to estimate the annual fluxes. The annual fluxes between the methods were compared to verify predictability of the conventional L–Q models. Annual flux of TOC, TN, DFe, and HS estimated by the turbidity displayed similar values, whereas the flux of SS, TFe, and TP tended to increase downstream of the dams. Among the dams, estimated flux proportions for TP and TFe were higher during high-flow events (74%–94%). Considering geographic conditions (e.g., absence of major tributary between the dams), the result implies that accumulated TP and TFe in the reservoirs may be flushed and transported downstream with SS over the short height dams during flood events. Assuming this process, the reservoir dams probably make only a fractional contribution to the organic and inorganic transport in the catchment studied. The percent flux errors for SS, TFe, and TP fluxes ranged from −7.2% to −97% (except for the TP flux in 2013), which highlights the risk of underestimating these components when using an L–Q model. [ABSTRACT FROM AUTHOR]