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Freshwater ecosystems are becoming saltier due to human activities. The effects of increased salinity can lead to cascading trophic interactions, affecting ecosystem functioning and energy transfer, through changes in community and size structure. These effects can be modulated by other environmental factors, such as nutrients. For example, communities developed under eutrophic conditions could be less sensitive to salinization due to cross-tolerance mechanisms. In this study, we used a mesocosm approach to assess the effects of a salinization gradient on the zooplankton community composition and size structure under eutrophic conditions and the cascading effects on algal communities. Our results showed that zooplankton biomass, size diversity and mean body size decreased with increased chloride concentration induced by salt addition. This change in the zooplankton community did not have cascading effects on phytoplankton. The phytoplankton biomass decreased after the chloride concentration threshold of 500 mg L-1 was reached, most likely due to direct toxic effects on the osmotic regulation and nutrient uptake processes of certain algae rather than as a response to community turnover or top-down control. Our study can help to put in place mitigation strategies for salinization and eutrophication, which often co-occur in freshwater ecosystems., This study is part of the Global Lakes Ecological Observatory Network (GLEON), Global Salt Initiative. We thank FEHM and CT-BETA for allowing us to use their research facilities. LP has received funding from the Beatriu de Pinós Postdoctoral Fellowships Programme, funded by the Secretary of Universities and Research (Government of Catalonia) and by the Horizon 2020 Programme of Research and Innovation of the European Union under the Marie Skłodowska-Curie Grant Agreement No. 801370 and by a Ramón y Cajal contract funded by the Spanish Ministry of Science and Innovation (RYC 2020-029829-I). MCA was supported by the Serra-Hunter programme funded by the Generalitat of Catalunya and by a Ramón y Cajal contract funded by the Spanish Ministry of Science and Innovation (RYC 2020-029829-I). We also thank three anonymous reviewers for their constructive comments and suggestions

Human-induced salinization caused by the use of road deicing salts, agricultural practices, mining operations, and climate change is a major threat to the biodiversity and functioning of freshwater ecosystems. Yet, it is unclear if freshwater ecosystems are protected from salinization by current water quality guidelines. Leveraging an experimental network of land-based and in-lake mesocosms across North America and Europe, we tested how salinization—indicated as elevated chloride (Cl−) concentration—will affect lake food webs and if two of the lowest Cl− thresholds found globally are sufficient to protect these food webs. Our results indicated that salinization will cause substantial zooplankton mortality at the lowest Cl− thresholds established in Canada (120 mg Cl−/L) and the United States (230 mg Cl−/L) and throughout Europe where Cl− thresholds are generally higher. For instance, at 73% of our study sites, Cl− concentrations that caused a ≥50% reduction in cladoceran abundance were at or below Cl− thresholds in Canada, in the United States, and throughout Europe. Similar trends occurred for copepod and rotifer zooplankton. The loss of zooplankton triggered a cascading effect causing an increase in phytoplankton biomass at 47% of study sites. Such changes in lake food webs could alter nutrient cycling and water clarity and trigger declines in fish production. Current Cl− thresholds across North America and Europe clearly do not adequately protect lake food webs. Water quality guidelines should be developed where they do not exist, and there is an urgent need to reassess existing guidelines to protect lake ecosystems from human-induced salinization. This article contains supporting information online at https://www.pnas.org/lookup/suppl/doi:10.1073/pnas.2115033119/-/DCSupplemental

The salinisation of freshwater ecosystems is a global environmental problem that threatens biodiversity, ecosystem functioning and human welfare. The aim of this study was to investigate the potential impact of a realistic salinity gradient on the structure and functioning of freshwater biofilms. The salinity gradient was based on the real ion concentration of a mining effluent from an abandoned mine in Germany. We exposed biofilm from a pristine stream to 5 increasing salinities (3 to 100 g L-1) under controlled conditions in artificial streams for 21 days. We evaluated its functional (photosynthetic efficiency, nutrient uptake, and microbial respiration) and structural responses (community composition, algal biomass and diatom, cyanobacteria and green algae metrics) over time. Then we compared their responses with an unexposed biofilm used as control. The functionality and structure of the biofilm exposed to the different salinities significantly decreased after short-term and long-term exposure, respectively. The community composition shifted to a new stable state where the most tolerant species increased their abundances. At the same time, we observed an increase in the community tolerance (measured as Pollution-Induced Community Tolerance) along the salinity gradient. This study provides relevant information on the salt threshold concentrations that can substantially damage algal cells (i.e., between 15 and 30 g L-1). The results provide new insights regarding the response and adaptation of stream biofilm to salinity and its potential implications at the ecosystem level., We thank T.S and A.S from the Dresden Groundwater Research Center for the information about Menteroda abandoned mine provided. Lorenzo Proia that has received funding from the Postdoctoral Fellowships Programme Beatriu de Pinos, funded by the Secretary of Universities and Research (Government of Catalonia) and by the Horizon 2020 Programme of Research and Innovation of the European Union under the Marie Skłodowska-Curie Grant Agreement No. 801370 and by a Ramón y Cajal contract funded by the Spanish Ministry of Science and Innovation (RYC2020-029829-I). Miguel Cañedo-Argüelles was supported by the Serra-Hunter programme funded by the Generalitat of Catalunya and by a Ramón y Cajal contract funded by the Spanish Ministry of Science and Innovation (RYC2020-029829-I).

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Determination of the levels of heavy metal ions would support assessment of sources and pathways of water pollution. However, traditional spatial assessment by manual sampling and off-site detection in the laboratory is expensive and time-consuming and requires trained personnel. Aiming to fill the gap between on-site automatic approaches and laboratory techniques, we developed an autonomous sensing boat for on-site heavy metal detection using square-wave anodic stripping voltammetry. A fluidic sensing system was developed to integrate into the boat as the critical sensing component and could detect ≤1 μg/L Pb, ≤6 μg/L Cu, and ≤71 μg/L Cd simultaneously in the laboratory. Once its integration was completed, the autonomous sensing boat was tested in the field, demonstrating its ability to distinguish the highest concentration of Pb in an effluent of a galena-enriched mine compared to those at other sites in the stream (Osor Stream, Girona, Spain)., An autonomous sensing boat for heavy metal spatial assessment can distinguish the highest concentration of Pb in an effluent of a galena-enriched mine compared with those of other sites in the stream.

The global increase of cyanobacterial blooms occurrence has been associated with the presence of compounds that generate earthy and musty odour in freshwater systems, among which geosmin stands out. The lack of information on the factors associated to geosmin production by benthic organisms has driven the development of this study, whose main goal is to determine the effects of nutrient concentration and DIN:SRP ratio on geosmin formation and release. The experiment was performed in 18 microcosms under controlled conditions for 21 days, using a natural biofilm suspension from Ter river (NE, Spain) to promote biofilm settlement. Six treatments were set crossing three DIN:SRP ratios (A = 4:1, B = 16:1 and C = 64:1) with two nutrient concentrations (Low and High). After 7 days of experiment, geosmin was detected in biofilm, being higher under high nutrient concentration and low DIN:SRP ratio conditions. In this treatment, geosmin in biofilm reached its maximum concentration at day 16 (3.8 ± 0.9 ng/mg), decreasing at the end of the experiment (21d) due to cyanobacteria detachment and geosmin release into the water (136 ± 6 ng/L). Overall, this experimental study showed that high nutrient concentration and low DIN:SRP ratio favoured the Oscillatoria genus development within biofilm communities, generating the optimal conditions for geosmin production. The interaction between these two factors was demonstrated to be a potential driver of benthic geosmin production and release, and should be monitored and controlled in rivers exploited for drinking water purposes. We would like to acknowledge the water utilities Aigües de Vic S.A. and Aigües d'Osona S.A. (Catalonia, Spain), for their funding support in the study presented in this publication. We also want to thank the help provided by Dr. Helena Guasch and Ms. Núria Pou in identifying the algal community present in the biofilm samples, and the help provided by Dr. Victoria Osorio and Dr. Joan Colon in the development of the analytical method for geosmin. The research leading to these results has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program – H2020 INTCATCH (grant agreement n° 689341 ). LP has received funding from the Postdoctoral Fellowships Programme Beatriu de Pinós, funded by the Secretary of Universities and Research (Government of Catalonia) and by the Horizon 2020 Programme of Research and Innovation of the European Union under the Marie Skłodowska-Curie Grant Agreement No. 801370

Potash abandoned mines cause severe environmental damage to their bordering environment, with significant impacts on freshwater ecosystems mostly through uncontrolled discharge of hypersaline effluents. This study aimed to evaluate the ecological impact caused by a hypersaline effluent from an abandoned potash mine (Menteroda, Germany) on freshwater biofilms and, specifically, on diatom communities. Biofilm from a pristine stream was exposed under controlled conditions in microcosms to a mining effluent (ME), and its structural (algal biomass, community composition, diatom metrics) and functional (photosynthetic activity, nutrient uptake) responses were evaluated over time and compared with unexposed biofilms used as control. Biofilm exposed to ME showed drastic functional responses after one day of exposure, with a significant decrease in photosynthetic efficiency and nutrient uptake, that were recovered over time. Biofilm exposed to ME showed a progressive increase in diatom metrics (abundance, density and growth rate) over time, compared to the control. However, a significant decrease in diatom species diversity, richness and cell size was also observed in biofilm exposed to ME. This study revealed that the ME affected the biofilm causing short-term functional responses, which were recovered simultaneously with a drastic diatom community structure shift. This work was supported by the European Comissio LIFE program throughout the LIFE DEMINE project (LIFE16 ENV/ES/000218).We thank T.S and A.S from the Dresden Groundwater Research Center for the information about Menteroda abandoned mine provided. Lorenzo Proia that has received funding from the Postdoctoral Fellowships Programme Beatriu de Pin?s, funded by the Secretary of Universities and Research (Government of Catalonia) and by the Horizon 2020 Programme of Research and Innovation of the European Union under the Marie Sk?odowska-Curie Grant Agreement No. 801370. This study has been conducted with financial support from the French National Research Agency (ANR) in the framework of the Investments for the Future Programme, within the COTE Cluster of Excellence (ANR-10-LABX-45).

Spatial heterogeneity along river networks is interrupted by dams, affecting the transport, processing, and storage of organic matter, as well as the distribution of biota. We here investigated the structure of planktonic (free-living, FL), particle-attached (PA) and sediment-associated (SD) bacterial and archaeal communities within a small reservoir. We combined targeted-amplicon sequencing of bacterial and archaeal 16S rRNA genes in the DNA and RNA community fractions from FL, PA and SD, followed by imputed functional metagenomics, in order to unveil differences in their potential metabolic capabilities within the reservoir (tail, mid, and dam sections) and lifestyles (FL, PA, SD). Both bacterial and archaeal communities were structured according to their life-style preferences rather than to their location in the reservoir. Bacterial communities were richer and more diverse when attached to particles or inhabiting the sediment, while Archaea showed an opposing trend. Differences between PA and FL bacterial communities were consistent at functional level, the PA community showing higher potential capacity to degrade complex carbohydrates, aromatic compounds, and proteinaceous materials. Our results stressed that particle-attached prokaryotes were phylogenetically and metabolically distinct from their free-living counterparts, and that performed as hotspots for organic matter processing within the small reservoir Tis study was funded by the project of the Spanish Ministry of Economy and Competitiveness SPACESTREAM (CGL2017-88640-C2-1-R). The authors acknowledge the support from the Economy and Knowledge Department of the Catalan Government through Consolidated Research Group (ICRA-ENV 2017 SGR 1124), and to R. Marcé, D.von Schiller, C. Gutierrez, J. P. Casas-Ruiz, L. Gómez-Gener, V. Acuña and B. Obrador for their help during the sampling. ICRA is part of the CERCA program

Abandoned mines cause serious environmental damage to their surroundings with considerable impacts on freshwater ecosystems. These impacts occur mainly due to the uncontrolled discharge of polluted effluents, which may contain high concentrations of heavy metals. Currently, no real solution exists for this important environmental problem, leaving a legacy of global pollution. This study aimed to assess the impact of a metal mining effluent from an abandoned mine on freshwater ecosystems, using aquatic biofilms as an ecological indicator. At the same time, the efficiency of different innovative treatment technologies in reducing the ecological impacts caused by mining effluents was evaluated, consisting of nanofiltration and nanofiltration combined with electrocoagulation. To do that, aquatic biofilms obtained from a pristine stream, were exposed, under microcosms conditions, to a metal mining effluent, untreated or treated by the innovative treatment technologies and responses were compared with unexposed biofilm which served as control. The structural and functional responses of the biofilm were measured with throughout time. Biofilms that were exposed to the untreated mining effluent showed significant differences respect to the rest of treatments and the control, particularly exhibiting inhibitory effects on photosynthetic efficiency just after 24 h of exposure and a progressive shift of the photosynthetic community composition throughout the exposure period. The treatment technologies significantly reduced the ecological impact caused by the metal mining effluent. However, metal bioaccumulation in biofilm revealed a potential long-term impact. These observations evidenced the biofilm as a useful ecological indicator to assess the ecological impact caused by metal mining effluents on freshwaters and the efficiency of different treatment technologies to reduce it. This work was supported by the European Commission LIFE program throughout the LIFE DEMINE project (LIFE16 ENV/ES/000218).

Controls on the degradation of dissolved organic matter (DOM) are complex but key to understand the role of freshwaters in the carbon cycle. Both the origin and previous degradation history have been suggested to determine DOM reactivity, but it is still a major challenge to understand the links between DOM composition and biodegradation kinetics. An appropriate context to study these links are intermittent rivers, as summer drought naturally diversifies DOM sources and sinks. Here we investigated the biodegradation kinetics of DOM in the main aquatic environments present in a temporary river. During dark incubations we traced the dynamics of bulk DOM and its main chromatographic fractions defined using LC-OCD: high molecular weight substances (HMWS), low molecular weight substances (LMWS), and humic substances and building blocks. Bulk DOM decay patterns were successfully fitted to the reactivity continuum (RC) biodegradation model. The RC parameters depicted running waters as the sites presenting a more reactive DOM, and temporary pools, enriched in leaf litter, as the ones with slowest DOM decay. The decay patterns of each DOM fraction were consistent throughout sites. LMWS and HMWS decayed in all cases and could be modeled using the RC model. Notably, the dynamics of LMWS controlled the bulk DOM kinetics. We discuss the mechanistic basis for the chromatographic fractions' kinetics during biodegradation and the implications that preconditioning and summer drought can have for DOM biodegradation in intermittent rivers.

Water reuse is becoming a common practice in several areas in the world, particularly in those impacted by water scarcity driven by climate change and/or by rising human demand. Since conventional wastewater treatment plants (WWTPs) are not able to efficiently remove many organic contaminants and pathogens, more advanced water treatment processes should be applied to WWTP effluents for water reclamation purposes. In this work, a pilot plant based on microfiltration (MF) followed by reverse osmosis (RO) filtration was applied to the effluents of an urban WWTP. Both the WWTP and the pilot plant were investigated with regards to the removal of a group of relevant contaminants widely spread in the environment: 28 pharmaceuticals and 20 pesticides. The combined treatment by the MF–RO system was able to quantitatively remove the target micropollutants present in the WWTP effluents to values either in the low ng/L range or below limits of quantification. Monitoring of water quality of reclaimed water and water reclamation sources is equally necessary to design the most adequate treatment procedures aimed to water reuse for different needs AcknowledgmentsThis research work has been co-financed by the SpanishMinistry of Economy and Competitiveness through the projectSCARCE (Consolider-Ingenio 2010 CSD2009-00065) and the Euro-pean Union through the European Regional Development Fund(ERDF) and partly supported by the Generalitat de Catalunya (Con-solidated Research Group: Catalan Institute for Water Research2014 SGR 291)

This study aims to investigate the prevalence of clinically relevant carbapenemases genes (blaKPC, blaNDM and blaOXA-48) in water samples collected over one-year period from hospital (H), raw and treated wastewater of two wastewater treatment plants (WWTPs) as well as along the Zenne River (Belgium). The genes were quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed that absolute abundances were the highest in H waters. Although absolute abundances were significantly reduced in WWTP effluents, the relative abundance (normalized per 16S rRNA) was never lowered through wastewater treatment. Particularly, for the PAB the relative abundances were significantly higher in the effluents respect to the influents of both WWTPs for all the genes. The absolute abundances along the Zenne River increased from upstream to downstream, peaking after the release of WWTPs effluents, in both fractions. Our results demonstrated that blaKPC, blaNDM and blaOXA-48 are widely distributed in the Zenne as a consequence of chronic discharge from WWTPs. To conclude, the levels of carbapenemases genes are significantly lower than other genes conferring resistance to more widely used antibiotics (analyzed in previous studies carried out at the same sites), but could raise up to the levels of high prevalent resistance genes.

During summer drought, Mediterranean fluvial networks are transformed into highly heterogeneous landscapes characterized by different environments (i.e., running and impounded waters, isolated river pools and dry beds). This hydrological setting defines novel biogeochemically active areas that could potentially increase the rates of carbon emissions from the fluvial network to the atmosphere. Using chamber methods, we aimed to identify hot spots for carbon dioxide (CO2) and methane (CH4) emissions from two typical Mediterranean fluvial networks during summer drought. The CO2 efflux from dry beds (mean ± SE = 209 ± 10 mmolCO2m-2 d-1) was comparable to that from running waters (120 ± 33 mmol m-2 d-1) and significantly higher than from impounded waters (36.6 ± 8.5 mmol m-2 d-1) and isolated pools (17.2 ± 0.9 mmol m-2 d-1). In contrast, the CH4 efflux did not significantly differ among environments, although the CH4 efflux was notable in some impounded waters (13.9 ± 10.1 mmol CH4 m-2 d-1) and almost negligible in the remaining environments (mean\0.3 mmol m-2 d-1). Diffusion was the only mechanism driving CO2 efflux in all environments and was most likely responsible for CH4 efflux in running waters, isolated pools and dry beds. In contrast, the CH4 efflux in impounded waters was primarily ebullition-based. Using a simple heuristic approach to simulate potential changes in carbon emissions from Mediterranean fluvial networks under future hydrological scenarios, we show that an extreme drying out (i.e., a four-fold increase of the surface area of dry beds) would double the CO2 efflux from the fluvial network. Correspondingly, an extreme transformation of running waters into impounded waters (i.e., a twofold increase of the surface area of impounded waters) would triple the CH4 efflux. Thus, carbon emissions from dry beds and impounded waters should be explicitly considered in carbon assessments of fluvial networks, particularly under predicted global change scenarios, which are expected to increase the spatial and temporal extent of these environments This research was funded by the Spanish Ministry of Economy and Competitiveness through the projects CGL2011-30474-C02-01 and CGL2014-58760-C3-1-R

This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in Environmental science and technology, copyright © American Chemical Society after peer review. To access the final edited and published work see [insert ACS Articles on Request author-directed link to Published Work, see http://pubs.acs.org/doi/abs/10.1021/acs.est.6b06506 Physical heterogeneity determines interstitial fluxes in porous media. Nutrients and organic matter distribution in depth influence physicochemical and microbial processes occurring in subsurface. Columns 50 cm long were filled with sterile silica sand following five different setups combining fine and coarse sands or a mixture of both mimicking potential water treatment barriers. Water was supplied continuously to all columns during 33 days. Hydraulic conductivity, nutrients and organic matter, biofilm biomass, and activity were analyzed in order to study the effect of spatial grain size heterogeneity on physicochemical and microbial processes and their mutual interaction. Coarse sediments showed higher biomass and activity in deeper areas compared to the others; however, they resulted in incomplete denitrification, large proportion of dead bacteria in depth, and low functional diversity. Treatments with fine sediment in the upper 20 cm of the columns showed high phosphorus retention. However, low hydraulic conductivity values reported in these sediments seemed to constraint biofilm activity and biomass. On the other hand, sudden transition from coarse-to-fine grain sizes promoted a hot-spot of organic matter degradation and biomass growth at the interface. Our results reinforce the idea that grain-size disposition in subsurface sandy sediments drives the interstitial fluxes, influencing microbial processes.

Antibiotics are emerging contaminants, which wing to their bioactivity, may lead to short-term and long-term alterations of natural microbial communities in aquatic environment. We investigated the effects of antibiotics on biofilm bacterial communities in the Llobregat River (Northeast Spain). Three sampling sites were selected: two less polluted sites and one hotspot. River water was collected from each site and used both as inoculum and medium for growing biofilms in independent mesocosms. After 25 d of biofilm colonization, we exposed the colonized biofilms to river waters from the downstream sites (progressively contaminated by antibiotics). A control from each site was maintained where the growing biofilm was always exposed to water from the same site. The bacterial community composition, bacterial live/dead ratio and extracellular enzyme activities of the biofilms were measured before and 9 d after exposing the biofilms to increasing contaminated waters. Sixteen antibiotic compounds were detected in the water from the three sampling sites. At each site, the antibiotics present in the highest concentrations were sulfonamides, followed by quinolones and macrolides. Bacterial communities of biofilms grown with the three river waters differed markedly in their structure, but less so in terms of functional descriptors. After switching the medium water to increasing pollution, biofilms exhibited increased levels of actinobacteria (HGC), a trend that was associated to the higher antibiotic concentrations in the water. These biofilms also showed increased bacterial mortality, and decreased extracellular leucine-aminopeptidase and alkaline phosphatase. There was a significant correlation between antibiotic concentrations and biofilm responses. Our results indicate that the continuous entrance of antibiotics in running waters cause significant structural and functional changes in microbial attached communities.

Biofilms are a key component in the nutrient removal from the water column. However, nutrient uptake by biofilms may be hampered by the occurrence of pollutants or other stressors. This study aimed: (i) to investigate the biofilm phosphorus (P) uptake capacity as a relevant process for the maintenance of fluvial water quality and (ii) to explore the sensitivity of this process to different chemical and environmental stressors. We conducted chamber experiments to test for the relevance of biofilm P uptake capacity (PUC) as a tool to detect effects of pollutants on river self-depuration. PUC was calculated by measuring P temporal decay after performing controlled P-spikes in chambers with biofilm-colonized tiles. Four different experiments were conducted to evaluate the response of PUC to: (a) several river waters from increasing polluted sites; (b) the effect of the bactericide triclosan (TCS); (c) the combined effect of TCS and grazers; and (d) the effect of TCS after a drought episode that affected the biofilms. These experiments showed that biofilms decreased their PUC along the pollution gradient. The biofilm PUC was significantly reduced after receiving high TCS concentrations, though lower TCS concentrations also affected the biofilm when this was submitted to grazing pressure. PUC decrease was induced by flow interruption which further enhanced the TCS negative effects. Overall, PUC was sensitive to the effects of pollutants like TCS as well as to the action of biological (grazing) and environmental (drought) factors. The study also showed that multiple stressors enhance the negative effects of pollutants on the PUC of biofilms. Our study values the use of biofilms’ PUC as a sensitive ecological-based tool to assess the effects of chemicals on freshwater communities and their derived functioning in river ecosystems.

Nutrients and light are the most determinant factors for microbial benthic assemblages in oligotrophic forested streams. We investigated the importance of nutrients and light availability on the structure and the function of epilithic biofilms in a Mediterranean forested stream (Fuirosos, Spain). Biofilms grew on artificial substrata in both enriched and unenriched reaches where shade conditions were simulated. Four different treatments were generated: higher light unenriched, lower light unenriched, higher light enriched (HL-E) and lower light enriched. Chlorophyll a, bacterial density, extracellular polymeric substances (EPS), extracellular leucine aminopeptidase (LAmP) and alkaline phosphatase (APase) activities were analysed during the colonisation at days 4, 9, 16, 22 and 52. At day 52, confocal laser scanning microscopy (CLSM) was used to determine differences in biofilm architecture. CLSM evidenced differences in thickness and structural complexity of biofilms grown in different conditions. Biofilms in HL-E were the thickest and had the most complex structure. The CLSM highlighted that the EPS was agglomerated in the upper layer of enriched-grown biofilms, but evenly distributed through the biofilm in unenriched biofilms. CLSM 3D images suggested that cyanobacteria increased under higher nutrient conditions. Nutrient enrichment caused the decrease of APase activity. Interaction between the two factors affected LAmP activity. HL-E had the highest LAmP and the lowest APase activities, an indication that biofilm responses to nutrients mostly occurred with high-light availability. Our results revealed that the conjoint availability of light and nutrients caused the highest changes in biofilm spatial organisation, microbial structure and functioning in oligotrophic forested streams.

Summary Nutrient retention efficiency, an indicator of stream function, has become a significant focus of research in fluvial biogeochemistry. The most popular method used to measure nutrient retention efficiency is constant rate addition (CRA), although studies using pulse addition (PA) as an alternative have been published. The similarities of the results produced by the two methods have not yet been explored. With the aim of clarifying this uncertainty, the present study assessed the similarities between the results of the CRA and PA experiments. Thirteen phosphate CRA–PA paired additions were performed in two different Mediterranean streams. We used ANOVA to compare the results of the uptake rate coefficients from the CRA ( K tCRA ) and the PA ( K tPA ) experiments. Results from CRA and PA were significantly different, with K tCRA being higher than K tPA in most cases, however, the difference between K tCRA and K tPA decreased as specific water residence time increased. When specific water residence time exceeded a specific threshold value, they became equal ( K tCRA ≈ K tPA ). Results suggested that the differences in nutrient retention estimates between the two methods may be a consequence of the difference in solute spreading along the stream reaches that shape the duration and magnitude of solute interaction across the surface water and streambed interface.

The Water Framework Directive of the European Union requires achievement of good chemical and ecological status in river systems. Risk assessment for toxicants is mostly based on individual single-species tests, though extrapolation of such results to ecosystem evaluation is often afflicted with uncertainties. Advances include higher-tier testing, which comprises use of multi-faceted ecotoxicity tests, and community (multi-species) tests. Including several trophic levels in pollutant testing shows that both direct and indirect effects occur in biological communities. Direct effects are expressed as changes in community composition, growth rates, metabolism and death rate. Indirect effects at other trophic levels (primary producers, consumers, predators) are exerted through trophic links. Biofilm-grazers comprise a simple food chain, where potential and negative effects affecting one of them could be magnified through the food web (bottom-up versus top-down effects), thus reinforcing or decreasing the ability of biofilms to be indicators of the effects of toxicants in aquatic ecosystems.

We present an overview of possible links between chemical contamination and the diversity of fluvial communities, in order to assess how conventional and more recently developed methodologies used to estimate the diversity of rivers and streams could be used to explore this relationship. We analyze different trophic levels (bacteria, autotrophic biofilm, benthic macroinvertebrates and fish) and discuss their usefulness in diversity-related studies. We present some practical examples of their application. We strongly recommend that future research should involve a multi-index approach and multidisciplinary studies that integrate chemical and biological analyses to assess effectively the relationship between biological diversity and chemical contamination.

6 figuras, 5 tablas, Large-scale factors associated with the environmental context of streams can explain a notable amount of variability in patterns of stream N cycling at the reach scale. However, when environmental factors fail to accurately predict stream responses at the reach level, focusing on emergent properties from small-scale heterogeneity in N cycling rates may help understand observed patterns in stream N cycling. To address how small-scale heterogeneity may contribute to shape patterns in whole-reach N uptake, we examined the drivers and variation in microbial N uptake at small spatial scales in two stream reaches with different environmental constraints (i.e., riparian canopy). Our experimental design was based on two 15N additions combined with a hierarchical sampling design from reach to microhabitat scales. Regardless of the degree of canopy cover, small-scale heterogeneity of microbial N uptake ranged by three orders of magnitude, and was characterized by a low abundance of highly active microhabitats (i.e., hot spots). The presence of those hot spots of N uptake resulted in a nonlinear spatial distribution of microbial N uptake rates within the streambed, especially in the case of epilithon assemblages. Small-scale heterogeneity in N uptake and turnover rates at the microhabitat scale was primarily driven by power relationships between N cycling rates and stream water velocity. Overall, FBOM assemblages responded clearly to changes in the degree of canopy cover, overwhelming small-scale heterogeneity in its N uptake rates, and suggesting that FBOM contribution to wholereach N uptake was principally imposed by environmental constraints from larger scales. In contrast, N uptake rates by epilithon showed no significant response to different environmental influences, but identical local drivers and spatial variation in each study reach. Therefore, contribution of epilithon assemblages to whole-reach N uptake was mainly associated with emerging properties from small-scale heterogeneity at lower spatial scales., Financial support was provided by the Spanish Ministry of Science and Innovation through the ISONEF (ref: CGL2008-05504-C02-02/BOS) and MED_FORESTREAM (ref: CGL2011-30590-C02-02) projects. M. Peipoch and E. Bastias were granted by two FPI PhD fellowships from the Spanish Ministry of Science and Innovation through the ISONEF and MED-FORESTREAM projects, respectively. M. Ribot was supported by a technical training contract also associated to ISONEF project.

Interactions between emerging contaminants and other drivers of community structure and function are poorly known. We used laboratory microcosms to investigate the single and combined effects of grazing and triclosan toxicity on the structure and function of stream periphyton. Grazing alone strongly reduced algal biomass, but also reduced oxidative stress and increased periphyton productivity and phosphorus (P) uptake capacity. This suggests that grazed algal communities can compensate for reduced biomass by enhanced growth after being released from grazing. Triclosan exposure (11.6 ± 1.1 μg L−1) reduced the detoxification capacity and P-uptake capacity of periphyton and altered diatom taxonomic composition. This indicates that triclosan at environmentally relevant concentrations affects the capacity of periphyton to remove dissolved nutrients and to cope with toxicant mixtures commonly occurring in streams. Triclosan exposure and grazing pressure had negative synergistic effects on algal size-class distribution and diatom mortality, since the effects of triclosan were higher than expected when periphyton was subject to grazing. Periphyton exposed to toxic substances such as triclosan had a lower capacity to cope with grazing than unexposed communities, because toxicity can limit algal regrowth after release from grazing and promote the loss of less abundant species. This synergism may have important implications because grazing pressure will magnify the negative effects of toxicants on community structure and ecosystem functions such as primary production and nutrient cycling This study was supported by the European Commission through the project Modelkey (Project 511237-2 GOCE). Additional funds were provided by the European Keybioeffects Project (MRTN-CT-2006-035695), VIECO (009/RN08/011), SCARCE (Consolider-Ingenio 2010, CSD2009- 00065) and Fluvialmultistress (CTM2009-14111-CO2-01) and MPCUdG2016/120

Algae in rivers are affected by light, water turbulence, and nutrient availability. These environmental factors ultimately affect algae according to their habitat, growth form, and specific physiological abilities. Water flow imposes limitations in the diffusion and availability of gases and resources, also in relation to algal size and growth form. Algae adapt physiologically to light scarcity or excess via photosynthetic mechanisms, as well as by modifying their pigment composition. The algal ability to obtain and keep resources is mediated by enzymes, and its ability to use and store materials is specific of the different algal groups. Toxicants impose a limit to algal performance and may affect photosynthesis as well as nutrient uptake, amongst other effects on algal cells.

7 páginas, 3 figuras, Stream microbial communities and associated processes are influenced by environmental fluctuations that may ultimately dictate nutrient export. Discharge fluctuations caused by intermittent stream flow are increasing worldwide in response to global change. We examined the impact of flow cessation and drying on in-stream nitrogen cycling. We determined archaeal (AOA) and bacterial ammonia oxidizer (AOB) abundance and ammonia oxidation activity in surface and deep sediments from different sites along the Fuirosos stream (Spain) subjected to contrasting hydrological conditions (i.e., running water, isolated pools, and dry streambeds). AOA were more abundant than AOB, with no major changes across hydrological conditions or sediment layers. However, ammonia oxidation activity and sediment nitrate content increased with the degree of stream drying, especially in surface sediments. Upscaling of our results shows that ammonia oxidation in dry streambeds can contribute considerably (~50%) to the high nitrate export typically observed in intermittent streams during first-flush events following flow reconnection. Our study illustrates how the dry channels of intermittent streams can be potential hotspots of ammonia oxidation. Consequently, shifts in the duration, spatial extent and severity of intermittent flow can play a decisive role in shaping nitrogen cycling and export along fluvial networks in response to global change., This research was granted by the projects DARKNESS (CGL2012-32747, MINECO) to E. O. Casamayor and MED_FORESTREAM (CGL2011-30590- CO2- 02, MINECO) and REFRESH (244121 FP7 EU Commission) to E. Martí.

13 pages, 6 figures, 2 tables, supplementary material http://dx.doi.org/10.3389/fmicb.2015.01293, Deductions about the ecology of high taxonomic bacterial ranks (i.e., phylum, class, order) are often based on their abundance patterns, yet few studies have quantified how accurately variations in abundance of these bacterial groups represent the dynamics of the taxa within them. Using 454-pyrosequencing of the 16S rRNA gene, we investigated whether the changes in abundance of six dominant bacterial classes (Actinobacteria, Beta-/Alpha-/Gamma-proteobacteria, Flavobacteria, and Sphingobacteria) along a large dam-regulated river are reflected by those of their constituent Operational Taxonomic Units (OTUs; 97% similarity level). The environmental impact generated by the reservoirs promoted clear compositional shifts in all bacterial classes that resulted from changes in the abundance of individual OTUs rather than from the appearance of new taxa along the river. Abundance patterns at the class level represented the dynamics of only a small but variable proportion of their constituting OTUs, which were not necessarily the most abundant ones. Within most classes, we detected sub-groups of OTUs showing contrasting responses to reservoir-induced environmental changes. Overall, we show that the patterns observed at the class level fail to capture the dynamics of a significant fraction of their constituent members, calling for caution when the ecological attributes of high-ranks are to be interpreted, This study has been financially supported by the EU through the FP7 project GLOBAQUA (Grant agreement No 603629). It reflects only the author's views. [...] Additional research funds were provided by the Spanish Ministry of Economy and Competitiveness (MINECO) through projects STORM (CTM2009-09352/MAR) and SCARCE (Consolider-Ingenio 2010 CSD2009-00065), as well as by the Confederación Hidrográfica del Ebro (CHE)

The extensive use of antibiotics in human and veterinary medicine and their subsequent release into the environment may have direct consequences for autochthonous bacterial communities, especially in freshwater ecosystems. In small streams and rivers, local inputs of wastewater treatment plants (WWTPs) may become important sources of organic matter, nutrients and emerging pollutants, such as antibiotic resistance genes (ARGs). In this study, we evaluated the effect of WWTP effluents as a source of ARGs in river biofilms. The prevalence of genes conferring resistance to main antibiotic families, such as beta-lactams (blaCTX-M), fluoroquinolones (qnrS), sulfonamides (sul I), and macrolides (ermB), was determined using quantitative PCR (qPCR) in biofilm samples collected upstream and downstream WWTPs discharge points in four low-order streams. Our results showed that the WWTP effluents strongly modified the hydrology, physico-chemistry and biological characteristics of the receiving streams and favoured the persistence and spread of antibiotic resistance in microbial benthic communities. It was also shown that the magnitude of effects depended on the relative contribution of each WWTP to the receiving system. Specifically, low concentrations of ARGs were detected at sites located upstream of the WWTPs, while a significant increase of their concentrations was observed in biofilms collected downstream of the WWTP discharge points (particularly ermB and sul I genes). These findings suggest that WWTP discharges may favour the increase and spread of antibiotic resistance among streambed biofilms. The present study also showed that the presence of ARGs in biofilms was noticeable far downstream of the WWTP discharge (up to 1 km). It is therefore reasonable to assume that biofilms may represent an ideal setting for the acquisition and spread of antibiotic resistance determinants and thus be considered suitable biological indicators of anthropogenic pollution by active pharmaceutical compounds.

The consequences of global change on rivers include altered flow regime, and entrance of compounds that may be toxic to biota. When water is scarce, a reduced dilution capacity may amplify the effects of chemical pollution. Therefore, studying the response of natural communities to compromised water flow and to toxicants is critical for assessing how global change may affect river ecosystems. This work aims to investigate how an episode of drought might influence the response of river biofilms to pulses of triclosan (TCS). The objectives were to assess the separate and combined effects of simulated drought (achieved through drastic flow alteration) and of TCS exposure on biofilms growing in artificial channels. Thus, three-week-old biofilms were studied under four conditions: Control (normal water flow); Simulated Drought (1 week reduced flow + 2 days interrupted flow); TCS only (normal water flow plus a 48-h pulse of TCS); and Simulated Drought + TCS. All channels were then left for 2 weeks under steady flow conditions, and their responses and recovery were studied. Several descriptors of biofilms were analyzed before and after each step. Flow reduction and subsequent interruption were found to provoke an increase in extracellular phosphatase activity, bacterial mortality and green algae biomass. The TCS pulses severely affected biofilms: they drastically reduced photosynthetic efficiency, the viability of bacteria and diatoms, and phosphate uptake. Latent consequences evidenced significant combined effects caused by the two stressors. The biofilms exposed only to TCS recovered far better than those subjected to both altered flow and subsequent TCS exposure: the latter suffered more persistent consequences, indicating that simulated drought amplified the toxicity of this compound. This finding has implications for river ecosystems, as it suggests that the toxicity of pollutants to biofilms may be exacerbated following a drought. Fil: Proia, L.. Universidad de Girona; España Fil: Vilches, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján. Departamento de Ciencias Básicas; Argentina. Universidad de Girona; España Fil: Boninneau, C.. Universidad de Girona; España Fil: Kantiani, L.. Instituto Catalán de Investigaciones en Agua; España Fil: Farré, Maria Marcela. Instituto Catalán de Investigaciones en Agua; España Fil: Romaní, Anna M.. Fil: Sabater Cortés, Sergi. Universidad de Girona; España. Instituto Catalán de Investigaciones en Agua; España Fil: Guasch, H.. Universidad de Girona; España

We investigated the effects of pharmaceuticals and pesticides detected in a Mediterranean river, on fluvial biofilms by translocation experiments performed under controlled conditions. Water was sampled from three sites along a pollution gradient. Biofilms grown in mesocosms containing relatively clean water were translocated to heavily polluted water. Several biofilm descriptors were measured before and after translocations. Fifty-seven pharmaceuticals and sixteen pesticides compounds were detected in river waters. The translocation from less to more polluted site was the most effective. Autotrophic biomass and peptidase increased while phosphatase and photosynthetic efficiency decreased. Multivariate analysis revealed that analgesics and anti-inflammatories significantly affected biofilm responses. Ibuprofen and paracetamol were associated with negative effects on photosynthesis, and with the decrease of the green algae/cyanobacteria ratio, while diclofenac was associated with phosphatase activity. The effects of these emerging compounds on biofilms structure and function may cause important alterations in river ecosystem functioning.© 2013 Elsevier Ltd. All rights reserved., Financial support was provided by the EU projects, MODELKEY (SSPI-CT-2003-511237-2) and KEYBIOEFFECTS (MRTN-CT-2006–035695), and the Spanish projects, SCARCE (Consolider-Ingenio CSD2009-00065), FLUMED-HOT-SPOTS(CGL2011-30151-C02-01) and VIECO (009/RN08/011).

This chapter aims to review the current literature on field studies assessing the effects of pharmaceutically active compounds (PhACs) on fluvial biological communities. The importance of the use of these communities in the ecological risk assessment of PhACs in river ecosystems is tackled. The chapter reviews the literature about the potential effects of PhACs on the communities commonly belonging to the bottom and the top of river food chain (i.e., microbial and fish communities). In this framework, the Llobregat river is presented as a greatly PhAC-polluted case, where numerous field studies have been performed. Particularly, evidences of PhACs effects on biofilm, macroinvertebrate, and fish communities of the Llobregat, reported by diverse investigations, are reviewed in this chapter. The conclusions suggest that the continuous population growth and the subsequent increase of sewage waste discharge on the aquatic systems may be reflected in even more pronounced effects of PhACs on biological communities.

16 pages, 5 figures, 3 tables, Large rivers are commonly regulated by damming, yet the effects of such disruption on prokaryotic communities have seldom been studied. We describe the effects of the three large reservoirs of the Ebro River (NE Iberian Peninsula) on bacterioplankton assemblages by comparing several sites located before and after the impoundments on three occasions. We monitored the abundances of several bacterial phylotypes identified by rRNA gene probing, and those of two functional groups (picocyanobacteria and aerobic anoxygenic phototrophic bacteria-AAPs). Much greater numbers of particles colonized by bacteria were found in upstream waters than downstream sites. Picocyanobacteria were found in negligible numbers at most sites, whereas AAPs constituted up to 14% of total prokaryotes, but there was no clear effect of reservoirs on the spatial dynamics of these two groups. Instead, damming caused a pronounced decline in Betaproteobacteria, Gammaproteobacteria and Bacteroidetes from upstream to downstream sites, whereas Alphaproteobacteria and Actinobacteria significantly increased after the reservoirs. Redundancy analysis revealed that conductivity, temperature and dissolved inorganic nitrogen were the environmental predictors that best explained the observed variability in bacterial community composition. Our data show that impoundments exerted significant impacts on bacterial riverine assemblages and call attention to the unforeseen ecological consequences of river regulation, This study was funded by the Confederaci on Hidrogr afica del Ebro. Additional funds were provided by the project SCARCE (Consolider-Ingenio 2010, CSD2009-00065). We acknowledge the support of Concha Duran throughout the study, and the help from Elisabet Tornes and Carmen Guti errez in the field and laboratory

The aim of this chapter is to highlight the importance of microbial attached communities in the assessment of the effects of pollutants on freshwater ecosystems. We particularly focus on the role of heterotrophs in biofilms developing on different substrata. Firstly, an overview of the importance of microbial communities for the whole ecosystem processes is given, focusing on bacteria and fungi either living in consortia with autotrophs or as the microbial decomposing community on plant litter in river ecosystems. A series of detailed examples of direct effects of priority and emerging pollutants on bacteria in epilithic biofilms and on attached decomposers are included. Microbial ecological interactions between organisms in heterogeneous complex communities are highlighted describing the indirect effects observed in a series of study cases. A collection of laboratory and field study data is used to demonstrate the relevance of natural heterogeneous communities to obtain a more realistic approach to ecosystem processes. Finally, an upscaling from the effects observed at the microbial scale to the potential implication for ecosystems health and risk is included.

Nowadays, the term biomarker has become widespread in environmental sciences and ecology as shown by the increasing amount of research articles published on this topic in these research areas (2,042 articles published between 2000 and 2010 over a total of 2,352 published since 1985, WOK, ISI Web of Knowledge, http://www.isiknowledge.com). Despite this increasing enthusiasm for biomarkers, they are still poorly used in regulatory monitoring or ecological risk assessment. Indeed, the interest and pertinence of using biomarkers in such approaches have been questionned. While biomarkers of various toxicants for many species can be found in the literature, the limited extrapolation of biomarker results is particularly criticized and may prevent routine application. In this chapter, we discuss how to link biomarker responses to chemical exposure but also how to correlate them to effects at higher levels of biological organization. In the first part, the word biomarker is defined in the context of risk assessment. Then, the historical and current use of biomarkers in river biofilms, macroinvertebrates, and fish is shortly presented focusing on the specificity of biomarkers’ utilization in each of these biological entities. Next, laboratory and field studies are used to exemplify relationships between stressors and biomarkers but also between biomarkers at different levels of biological organization. Finally, current limitations of biomarkers are discussed and some propositions are made to overcome these limitations and to apply a multibiomarker approach to environmental risk assessment.

Bacterial community composition was assessed during riverine biofilm development by the Catalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) in combination with Confocal Laser Scanning Microscopy. Using artificial substrates, it was possible to follow the dynamics of specific bacterial clusters, while maintaining the unaltered structure and architecture of the biofilm.

The effects of the herbicide Diuron (DIU) and the bactericide Triclosan (TCS) were assessed on laboratory-grown stream biofilms. Four week-old biofilms were exposed in mesocosms to 48-hours of short pulses of either DIU or TCS. The direct and indirect effects of each toxicant on the biofilms, and the subsequent recovery of the biofilms, were evaluated according to structural and functional biomarkers. These parameters were analyzed immediately before exposure, immediately after exposure, and 9 and 16days post-exposure. DIU caused an increase in diatom mortality (+79%), which persisted until the end of the experiment. TCS also affected diatom mortality (+41%), although the effect did not appear until 1week post-exposure. TCS caused an increase in bacterial mortality (+45%); however, this parameter returned to normal values 1week post-exposure. TCS compromised the cellular integrity of the green alga Spirogyra sp., whereas DIU did not. TCS also strongly inhibited phosphate uptake (-71%), which did not return to normal values until 2weeks post-exposure. DIU directly affected algae, but barely affected the heterotrophs, whereas TCS seriously impaired bacteria (direct effect) as well as autotrophs (indirect effect). However, the biofilms recovered their normal structure and function within only a few days to a few weeks. These findings demonstrate the capacity of biofilms to cope with periodic inputs of toxicants, but also the risks associated to repeated exposure or multi-contamination in aquatic ecosystems.

Triclosan is a commonly used bactericide that survives several degradation steps in WWTP (wastewater treatment plants) and potentially reaches fluvial ecosystems. In Mediterranean areas, where water scarcity results in low dilution capacity, the potential environmental risk of triclosan is high. A set of experimental channels was used to examine the short-term effects of triclosan (from 0.05 to 500 μg L−1) on biofilm algae and bacteria. Environmentally relevant concentrations of triclosan caused an increase of bacterial mortality with a no effect concentration (NEC) of 0.21 μg L−1. Dead bacteria accounted for up to 85% of the total bacterial population at the highest concentration tested. The toxicity of triclosan was higher for bacteria than algae. Photosynthetic efficiency was inhibited with increasing triclosan concentrations (NEC = 0.42 μg L−1), and non-photochemical quenching mechanisms decreased. Diatom cell viability was also affected with increasing concentrations of triclosan. Algal toxicity may be a result of indirect effects on the biofilm toxicity, but the clear and progressive reduction observed in all the algal-related endpoints suggest the existence of direct effects of the bactericide. The toxicity detected on the co-occurring non-target components of the biofilm community, the capacity of triclosan to survive through WWTP processes and the low dilution capacity that characterizes Mediterranean systems extend the relevance of triclosan toxicity beyond bacteria in aquatic habitats., This study was supported by the European Commission project Modelkey (Project 511237-2 GOCE). Additional funds were provided by the European Project Keybioeffects (MRTN-CT-2006-035695), the Spanish projects Fluvialfitomarc (GCL 2006-12785/HID), VIECO (009/RN08/011), SCARCE (Consolider-Ingenio 2010, CSD2009-00065) and Fluvialmultiestres (CTM2009-14111-CO2-01), as well as by Consorci de la Costa Brava and Empresa Mixta d’Aigües de la Costa Brava, S.A.

Among increasingly used pharmaceutical products, beta-blockers have been commonly reported at low concentrations in rivers and littoral waters of Europe and North America. Little is known about the toxicity of these chemicals in freshwater ecosystems while their presence may lead to chronic pollution. Hence, in this study the acute toxicity of 3 beta-blockers: metoprolol, propranolol and atenolol on fluvial biofilms was assessed by using several biomarkers. Some were indicative of potential alterations in biofilm algae (photosynthetic efficiency), and others in biofilm bacteria (peptidase activity, bacterial mortality). Propranolol was the most toxic beta-blocker, mostly affecting the algal photosynthetic process. The exposure to 531microg/L of propranolol caused 85% of inhibition of photosynthesis after 24h. Metoprolol was particularly toxic for bacteria. Though estimated No-Effect Concentrations (NEC) were similar to environmental concentrations, higher concentrations of the toxic (503microg/L metoprolol) caused an increase of 50% in bacterial mortality. Atenolol was the least toxic of the three tested beta-blockers. Effects superior to 50% were only observed at very high concentration (707mg/L). Higher toxicity of metoprolol and propranolol might be due to better absorption within biofilms of these two chemicals. Since beta-blockers are mainly found in mixtures in rivers, their differential toxicity could have potential relevant consequences on the interactions between algae and bacteria within river biofilms.

The Llobregat is a Mediterranean river that is severely impacted by anthropogenic pressures. It is characterized by high flow variability which modulates its chemical and biological status. The present work evaluates the effects of flow changes on the concentration of pharmaceutically active compounds (PhACs) and their relationship to cellular parameters of river biofilms. To this end, at two selected sampling sites at the lower course of the Llobregat river, surface water samples were collected twice a week over two hydrologically different periods exhibiting low and high river flows. Higher levels of PhACs were detected at the downstream sampling site. Irrespective of the flow regime, analgesics, anti-inflammatories and lipid regulators were the most abundant substances at both sampling sites with total concentrations of up to1000 ng/L and 550 ng/L at the upstream and downstream sites, respectively. Antibiotics (fluoroquinolones) and psychiatric treatment drugs were also detected at high levels in the second campaign achieving concentrations of up to 500 ng/L. The principal component analysis (PCA) performed with the PhACs concentrations of the two campaigns revealed differences in the various therapeutic groups depending on sampling site and period. After a flash flood event during the second sampling period, dilution of PhACs occurred, but their average concentrations measured before the flood were restored within two weeks. For the majority of compounds, PhAC concentrations displayed an inverse relationship with river discharge The effects of water containing different concentrations of PhACs on biofilm communities were evaluated and related to flow regime variations. Translocation of biofilm communities from a less to a more polluted site of the river demonstrated an increase in bacteria mortality in the translocated biofilms. After the flood, extracellular peptidase activity and chlorophyll-a concentration were significantly reduced, and biofilm growth rate was significantly lower.