Distinct dynamics in mountain watersheds: Exploring mercury and microplastic pollution-Unraveling the influence of atmospheric deposition, human activities, and hydrology.

The intensification of human activities all around the globe has led to the spread of micropollutants in high-mountain freshwater environments. We therefore aimed to assess the geospatial distribution and determine the potential sources of (total-) mercury (THg) and microplastics (MPs) in mountain freshwater ecosystems. To do so, we analyzed THg and MP concentrations in brown trout, biofilm, and sediments from lotic and lentic ecosystems in the Pyrenees - all subjected to different types of human pressure. Additionally, we assessed the potential impacts of these pollutants on fish, and explored the bioindication capacity of brown trout (Salmo trutta fario) and biofilm regarding THg and MP pollution. For the first time, we measured concentrations of MPs trapped in the matrix of freshwater biofilm. Our results suggest that THg in the Pyrenees might be explained by both legacy (regional) and distant sources, in combination with environmental characteristics such as the presence of peatlands or streamwater physicochemistry, while MPs in fish are linked to recent local pollution sources such as single-use plastics. In contrast, MPs in biofilm matrix and sediments indicate a combination of distant (i.e., atmospheric deposition) and recent local pollution sources. Moreover, hydrodynamics and plastic density likely control MP distribution in rivers. Based on Fulton's condition factor, we also found that higher THg concentrations caused a negative impact on fish health (K < 1), while no impact of MPs could be seen. Therefore, we suggest that brown trout and biofilm can serve as bioindicators of atmospheric deposition of THg in high-altitude lakes and that biofilm is a reliable bioindicator to assess MP pollution in remote environments. Brown trout may also act as a bioindicator of MP pollution, but only efficiently in more polluted areas.

A trait-based approach to determine the risks of Zn to the overall health status of native fish species Barbus meridionalis.

Fish adapt to changing environments by maintaining homeostasis or making energy trade-offs that impact fitness. We investigated the effect of Zn on the fitness and physiology of Barbus meridionalis, a native cyprinid fish species, under two exposure scenarios. The Osor stream's mine-effluent reach represented long-term (chronic) exposure, while the upstream reach served as a control/acute exposure. Acute exposure involved exposing B. meridionalis to 1mg/L Zn for 96 h in the laboratory. We examined physiological traits (Standard metabolic rate SMR, Maximum metabolic rate MMR, Absolute Aerobic scope AAS, Critical swimming capacity Ucrit) and antioxidant system, AS (Superoxide dismutase, SOD; Catalase, CAT; Glutathione peroxidase, GPX; Glutathione-S-transferase, GST; Glutathione, GSH; Thiobarbaturic acid reactive substances, TBARS) biomarkers. The results indicated that Zn had no significant effect on osmoregulatory cost (SMR) in either exposure scenario but impaired energetically costly exercise (low MMR). AAS reduction in both exposure groups suggested compromised energy allocation for life-history traits, evidenced by decreased locomotor performance (Ucrit) after acute exposure. Tissue-specific and time-dependent responses were observed for AS biomarkers. The fish exhibited ineffective control of oxidative damage, as evidenced by high TBARS levels in the liver and gills, despite increased CAT and GSH in the liver under acute conditions. Our findings demonstrate differential responses at the subcellular level between the two exposure scenarios, while trait-based endpoints followed a similar pattern. This highlights the utility of a trait-based approach as a supplementary endpoint in biomonitoring studies, which provides insights into impacts on individual fitness and population demography.

Size-related effects and the influence of metabolic traits and morphology on swimming performance in fish.

Energy metabolism fuels swimming and other biological processes. We compared the swimming performance and energy metabolism within and across eight freshwater fish species. Using swim tunnel respirometers, we measured the standard metabolic rate (SMR) and maximum metabolic rate (MMR) and calculated the critical swimming speed (U crit). We accounted for body size, metabolic traits, and some morphometric ratios in an effort to understand the extent and underlying causes of variation. Body mass was largely the best predictor of swimming capacity and metabolic traits within species. Moreover, we found that predictive models using total length or SMR, in addition to body mass, significantly increased the explained variation of U crit and MMR in certain fish species. These predictive models also underlined that, once body mass has been accounted for, U crit can be independently affected by total length or MMR. This study exemplifies the utility of multiple regression models to assess within-species variability. At interspecific level, our results showed that variation in U crit can partly be explained by the variation in the interrelated traits of MMR, fineness, and muscle ratios. Among the species studied, bleak Alburnus alburnus performed best in terms of swimming performance and efficiency. By contrast, pumpkinseed Lepomis gibbosus showed very poor swimming performance, but attained lower mass-specific cost of transport (MCOT) than some rheophilic species, possibly reflecting a cost reduction strategy to compensate for hydrodynamic disadvantages. In conclusion, this study provides insight into the key factors influencing the swimming performance of fish at both intra- and interspecific levels.

Antioxidant system status in threatened native fish Barbus meridionalis from the Osor River (Iberian Peninsula): I. Characterization and II. In vitro Zn assays.

The evaluation of antioxidant system capacity is important in aquatic toxicology. It was aimed to characterize the liver antioxidant enzymes (SOD, CAT, GPX, GR, and GST) and to test the in vitro Zn effect (200 and 400 ZnSO4 µg/L) in native fish Barbus meridionalis obtained from the Osor River (NE, Spain) influenced by Zn contamination. The maximal enzyme activities were at pH 7.0-7.5 and 100 mM phosphate buffer. Barbel showed high catalytic activity (high Vmax and low Km) indicating the efficient antioxidant detoxification ability. Direct Zn effect caused an antioxidant system imbalance. Mostly upon lower Zn concentration, GPX activity decreased (95-100 %) though CAT, GR, and GST increased (36-1543 %). GSH values either stimulated (290 %) or inhibited (85-93 %) due to tissue differences. The first record of barbel antioxidant enzyme characterization and in vitro data presenting an unbalanced antioxidant pattern could be significant to evaluate the metal pollution in the Osor River for further in vivo studies.

Biotic and Abiotic Factors Influencing Arsenic Biogeochemistry and Toxicity in Fluvial Ecosystems: A Review.

This review is focused on the biogeochemistry of arsenic in freshwaters and, especially, on the key role that benthic microalgae and prokaryotic communities from biofilms play together in through speciation, distribution, and cycling. These microorganisms incorporate the dominant iAs (inorganic arsenic) form and may transform it to other arsenic forms through metabolic or detoxifying processes. These transformations have a big impact on the environmental behavior of arsenic because different chemical forms exhibit differences in mobility and toxicity. Moreover, exposure to toxicants may alter the physiology and structure of biofilms, leading to changes in ecosystem function and trophic relations. In this review we also explain how microorganisms (i.e., biofilms) can influence the effects of arsenic exposure on other key constituents of aquatic ecosystems such as fish. At the end, we present two real cases of fluvial systems with different origins of arsenic exposure (natural vs. anthropogenic) that have improved our comprehension of arsenic biogeochemistry and toxicity in freshwaters, the Pampean streams (Argentina) and the Anllóns River (Galicia, Spain). We finish with a briefly discussion of what we consider as future research needs on this topic. This work especially contributes to the general understanding of biofilms influencing arsenic biogeochemistry and highlights the strong impact of nutrient availability on arsenic toxicity for freshwater (micro) organisms.

Responses of resident (DNA) and active (RNA) microbial communities in fluvial biofilms under different polluted scenarios.

Pollution from human activities is a major threat to the ecological integrity of fluvial ecosystems. Microbial communities are the most abundant organisms in biofilms, and are key indicators of various pollutants. We investigated the effects some human stressors (nutrients and heavy metals) have on the structure and activity of microbial communities in seven sampling sites located in the Ter River basin (NE Spain). Water and biofilm samples were collected in order to characterize physicochemical and biofilm parameters. The 16S rRNA gene was analysed out from DNA and RNA extracts to obtain α and ß diversity. Principal coordinates analyses (PCoA) of the operational taxonomic units (OTUs) in the resident microbial community revealed that nutrients and conductivity were the main driving forces behind the diversity and composition. The effects of mining have had mainly seen on the taxonomic composition of the active microbial community, but also at the OTUs level. Remarkably, metal-impacted communities were very active, which would indicate a close link with the stress faced, that is probably related to the stimulation of detoxification.

Water Flow and Light Availability Influence on Intracellular Geosmin Production in River Biofilms.

Hydro-morphological alterations in water bodies caused by climate change and human activities affects the ecosystem functioning and generate important water quality problems. Some of these alterations can generate an increase in cyanobacterial blooms, which are associated with the appearance of bad taste and odorous compounds such as geosmin. The factors that trigger their production are still unclear, and this inability to predict geosmin episodes provokes economic problems for water supply companies. This study aims to evaluate the effects of water flow and light availability on biofilm development and intracellular geosmin formation. A mesocosm experiment was performed between February-April, 2019. The mesocosms were a set of 10 outdoor 3 m long flumes, with a continuous water supply from the Ter river (Catalonia, NE Spain). Two light intensities were established: natural light and light reduced to 80%, combined with five gradual water flows from 0.09 to 1.10 L/s. Water samples were taken to analyze nutrients, and biofilm samples, to analyze geosmin concentration, chlorophyll a and the community. Geosmin in biofilm was detected in those treatments in which Oscillatoria sp. appeared. The concentration of intracellular geosmin was higher at lower water flows (0.09 and 0.18 L/s), and the highest (2.12 mg/g) was found in the flume with the lowest water flow (0.09 L/s) and irradiation (20%). This flume was the one that presented a greater concentration of Oscillatoria sp. (21% of the community). It stands out that, when geosmin in biofilm was found, the dissolved inorganic nitrogen and soluble reactive phosphorus ratio decreased, from an average of 417:1 to 14:1. This was mainly due to an increase in inorganic phosphorus concentration generated by a change in the nutrient uptake capacity of the community's biofilm. The results obtained in this study indicated the potential implications for stream ecosystem management to control geosmin appearance. Likewise, they could be used as an early warning system, establishing that in times of drought, which lead to a general decrease in river water flow, the situation could be optimal for the appearance and development of geosmin producing cyanobacteria in low-flow areas near the river banks.

Mutual interaction between arsenic and biofilm in a mining impacted river.

Gold mining activities in fluvial systems may cause arsenic (As) pollution, as is the case at the Anllóns River (Galicia, NW Spain), where high concentrations of arsenate (AsV) in surface sediments (up to 270 mg kg-1) were found. A 51 day-long biofilm-translocation experiment was performed in this river, moving some biofilm-colonized substrata from upstream (less As-polluted) to downstream the mine area (more As-polluted site), to explore the effect of As on benthic biofilms, as well as their role on As retention and speciation in the water-sediment interface. Eutrophic conditions (range: 0.07-0.38 mg L-1 total phosphorus, TP) were detected in water in both sites, while sediments were not considered P-polluted (below 600 mg kg-1). Dimethylarsenate (DMAV) was found intracellularly and in the river water, suggesting a detoxification process by biofilms. Since most As in sediments and water was AsV, the high amount of arsenite (AsIII) detected extracellularly may also confirm AsV reduction by biofilms. Furthermore, translocated biofilms accumulated more As and showed higher potential toxicity (higher As/P ratio). In concordance, their growth was reduced to half that observed in those non-translocated, became less nutritive (less nitrogen content), and with higher bacterial and dead diatom densities. Besides the high As exposure, other environmental conditions such as the higher riparian cover at the more As-polluted site could contribute to those effects. Our study provides new arguments to understand the contribution of microorganisms to the As biogeochemistry in freshwater environments.

Structural and functional responses of periphyton and macroinvertebrate communities to ferric Fe, Cu, and Zn in stream mesocosms.

Two mesocosm experiments were conducted to examine effects of ferric iron (Fe) and mixtures of ferric Fe with aqueous metals (Cu, Zn) on stream benthic communities. Naturally colonized benthic communities were exposed to a gradient of ferric Fe (0, 0.4, 1.0, 2.5, 6.2, and 15.6 mg/L) that bracketed the current US Environmental Protection Agency water quality criterion value (1.0 mg/L). After 10 d of exposure to ferric Fe, total macroinvertebrate abundance, number of taxa, and abundance of all major macroinvertebrate groups (Ephemeroptera, Plecoptera, Trichoptera, and Diptera) were significantly reduced. Heptageniid mayflies and chironomids were especially sensitive to Fe oxide deposition and were significantly reduced at 0.4 and 1.0 mg/L total Fe, respectively. In a second mesocosm experiment, periphyton and macroinvertebrate communities were exposed to ferric Fe (0.60 mg/L) with or without aqueous Cu and Zn at 2 treatment levels: low (0.01 mg/L Cu + 0.1 mg/L Zn) and high (0.05 mg/L Cu + 0.5 mg/L Zn). In contrast to previous research, we observed no evidence of a protective effect of Fe on toxicity of metals. Growth rates and protein content of periphyton were significantly reduced by both ferric Fe and aqueous metals, whereas abundance of heptageniid mayflies (Cinygmula) and whole community metabolism were significantly reduced by ferric Fe alone. We hypothesize that Fe oxides inhibited algal growth and enhanced metal accumulation, leading to a reduction in the quantity and quality of food resources for grazers. Mesocosm experiments conducted using natural benthic communities provide a unique opportunity to quantify the relative importance of indirect physical effects and to develop a better understanding of the relationship between basal food resources and consumers in natural stream ecosystems. Environ Toxicol Chem 2018;37:1320-1329. © 2017 SETAC.

Combined effects of hydrologic alteration and cyprinid fish in mediating biogeochemical processes in a Mediterranean stream.

Flow regimes are important drivers of both stream community and biogeochemical processes. However, the interplay between community and biogeochemical responses under different flow regimes in streams is less understood. In this study, we investigated the structural and functional responses of periphyton and macroinvertebrates to different densities of the Mediterranean barbel (Barbus meridionalis, Cyprinidae) in two stream reaches differing in flow regime. The study was conducted in Llémena Stream, a small calcareous Mediterranean stream with high nutrient levels. We selected a reach with permanent flow (permanent reach) and another subjected to flow regulation (regulated reach) with periods of flow intermittency. At each reach, we used in situ cages to generate 3 levels of fish density. Cages with 10 barbels were used to simulate high fish density (>7indm-2); cages with open sides were used as controls (i.e. exposed to actual fish densities of each stream reach) thus having low fish density; and those with no fish were used to simulate the disappearance of fish that occurs with stream drying. Differences in fish density did not cause significant changes in periphyton biomass and macroinvertebrate density. However, phosphate uptake by periphyton was enhanced in treatments lacking fish in the regulated reach with intermittent flow but not in the permanent reach, suggesting that hydrologic alteration hampers the ability of biotic communities to compensate for the absence of fish. This study indicates that fish density can mediate the effects of anthropogenic alterations such as flow intermittence derived from hydrologic regulation on stream benthic communities and associated biogeochemical processes, at least in eutrophic streams.

Examining predictors of chemical toxicity in freshwater fish using the random forest technique.

Chemical pollution is one of the main issues globally threatening the enormous biodiversity of freshwater ecosystems. The toxicity of substances depends on many factors such as the chemical itself, the species affected, environmental conditions, exposure duration, and concentration. We used the random forest technique to examine the factors that mediate toxicity in a set of widespread fishes and analyses of covariance to further assess the importance of differential sensitivity among fish species. Among 13 variables, the 5 most important predictors of toxicity with random forests were, by order of importance, the chemical substance itself (i.e., Chemical Abstracts Service number considered as a categorical factor), octanol-water partition coefficient (log P), pollutant prioritization, ecological structure-activity relationship (ECOSAR) classification, and fish species for 50% lethal concentrations (LC50) and the chemical substance, fish species, log P, ECOSAR classification, and water temperature for no observed effect concentrations (NOECs). Fish species was a very important predictor for both endpoints and with the two contrasting statistical techniques used. Different fish species displayed very different relationships with log P, often with different slopes and with as much importance as the partition coefficient. Therefore, caution should be exercised when extrapolating toxicological results or relationships among species. In addition, further research is needed to determine species-specific sensitivities and unravel the mechanisms behind them.

Short-term arsenic exposure reduces diatom cell size in biofilm communities.

Arsenic (As) pollution in water has important impacts for human and ecosystem health. In freshwaters, arsenate (As(V)) can be taken up by microalgae due to its similarity with phosphate molecules, its toxicity being aggravated under phosphate depletion. An experiment combining ecological and ecotoxicological descriptors was conducted to investigate the effects of As(V) (130 µg L(-1) over 13 days) on the structure and function of fluvial biofilm under phosphate-limiting conditions. We further incorporated fish (Gambusia holbrooki) into our experimental system, expecting fish to provide more available phosphate for algae and, consequently, protecting algae against As toxicity. However, this protection role was not fully achieved. Arsenic inhibited algal growth and productivity but not bacteria. The diatom community was clearly affected showing a strong reduction in cell biovolume; selection for tolerant species, in particular Achnanthidium minutissimum; and a reduction in species richness. Our results have important implications for risk assessment, as the experimental As concentration used was lower than acute toxicity criteria established by the USEPA.

Ecotoxicological risk assessment of chemical pollution in four Iberian river basins and its relationship with the aquatic macroinvertebrate community status.

Ecotoxicological risk assessment of chemical pollution in four Iberian river basins (Llobregat, Ebro, Júcar and Guadalquivir) was performed. The data set included more than 200 emerging and priority compounds measured at 77 sampling sites along four river basins studied. The toxic units (TU) approach was used to assess the risk of individual compounds and the concentration addition model (CA) to assess the site specific risk. Link between chemical pollution and aquatic macroinvertebrate communities in situ was examined by using four biological indexes; SPEAR ("Species at Risk Index") as the indicator of decline of sensitive species in relation to general organic (SPEARorganic) and pesticides (SPEARpesticides) pollution; and Shannon and Margalef biodiversity indexes. The results of the study suggested that organic chemicals posed the risk of acute effects at 42% of the sampling sites and the risk of chronic effects at all the sites. Metals posed the acute risk at 44% of the sites. The main drivers of the risk were mainly pesticides and metals. However, several emerging contaminants (e.g. the antidepressant drug sertraline and the disinfectant triclosan) were contributing to the chronic effects risk. When risk associated with metals and organic chemicals was compared, the latter dominated in 2010, mainly due to the presence of highly toxic pesticides, while metals did in 2011. Compounds that are not regulated on the European level were posing the risk of chronic effects at 23% of the sites. The decline of sensitive macroinvertebrate taxa expressed in terms of SPEAR index was correlated with the increase of toxic stress related to organic compounds Biodiversity indexes were negatively correlated with the metals and the urban land use type in the catchment.

Effects of low arsenic concentration exposure on freshwater fish in the presence of fluvial biofilms.

Arsenic (As) is a highly toxic element and its carcinogenic effect on living organisms is well known. However, predicting real effects in the environment requires an ecological approach since toxicity is influenced by many environmental and biological factors. The purpose of this paper was to evaluate if environmentally-realistic arsenic exposure causes toxicity to fish. An experiment with four different treatments (control (C), biofilm (B), arsenic (+As) and biofilm with arsenic (B+As)) was conducted and each one included sediment to enhance environmental realism, allowing the testing of the interactive effects of biofilm and arsenic on the toxicity to fish. Average arsenic exposure to Eastern mosquitofish (Gambusia holbrooki) was 40.5 ± 7.5 µg/L for +As treatment and 34.4 ± 1.4 µg/L for B+As treatment for 56 days. Fish were affected directly and indirectly by this low arsenic concentration since exposure did not only affect fish but also the function of periphytic biofilms. Arsenic effects on the superoxide dismutase (SOD) and glutathione reductase (GR) activities in the liver of mosquitofish were ameliorated in the presence of biofilms at the beginning of exposure (day 9). Moreover, fish weight gaining was only affected in the treatment without biofilm. After longer exposure (56 days), effects of exposure were clearly seen. Fish showed a marked increase in the catalase (CAT) activity in the liver but the interactive influence of biofilms was not further observed since the arsenic-affected biofilm had lost its role in water purification. Our results highlight the interest and application of incorporating some of the complexity of natural systems in ecotoxicology and support the use of criterion continuous concentration (CCC) for arsenic lower than 150 µg/L and closer to the water quality criteria to protect aquatic life recommended by the Canadian government which is 5 µg As/L.

Arsenic toxicity effects on microbial communities and nutrient cycling in indoor experimental channels mimicking a fluvial system.

The toxicity of chemicals in the environment is influenced by many factors, such as the adsorption to mineral particles, active biological surfaces, biotransformation and/or nutrient concentration. In the present study, a simplified fluvial system including fish, periphyton and sediment was used to investigate the fate and effects of environmentally realistic concentration of arsenic (As) on biofilm growth and nutrient cycling. Total dissolved arsenic concentration decreased exponentially from 120µg/L to 28.0±1.5µg/L during the experiment (60 days), mostly sinking to the sediment and a smaller percentage accumulated in the periphytic biofilm. Most P and N, which was provided by fish, was also retained in the epipsammic biofilm (growing on sediment grains). We conclude that exposure to this concentration of arsenic under oligotrophic conditions is changing the quality and quantity of the base of the aquatic food chain and its respective contribution to nutrient cycling, and normal functioning of the ecosystem. The effects include lowering the total biomass of biofilm and its potential ability to use organic P (i.e., phosphatase activity), inhibiting algal growth, especially that of diatoms, decreasing nitrogen content, and making the epipsammic biofilm more heterotrophic, thus reducing its ability to oxygenate the aquatic environment.

Cumulative Stressors Trigger Increased Vulnerability of Diatom Communities to Additional Disturbances.

Chronic, non-lethal stressors occurring gradually (in space or time) can result in cumulative impacts that are more dramatic than higher intensities or occasional critical levels of any single one of these stressors. The negative effects of the chronic stressors trigger lasting impacts that may grow in intensity and become problematic over time and/or to higher trophic levels. In rivers, aquatic organisms experience this type of cumulative stress along the up- to downstream gradient in natural and anthropogenic contaminants generally observed in inhabited watersheds. Diatoms are a major component of the periphyton in rivers; their richness and diversity in natural communities are directly related to their varied ecological preferences and sensitivity to disturbance. In this study, we monitored from 2003 to 2008 the changes in the diversity of taxonomic and non-taxonomic features along a small river (Riou-Mort, South West France), at three sites: one site upstream considered as a reference for this watershed, one intermediate site with high nutrient load, and one downstream site exposed to both nutrient and metal pollution. The cumulative impacts of nutrients plus metals led to a gradual decrease in species richness and diversity, and in a potential capacity to cope with additional stresses, e.g., climate change-related ones. This is reflected by a decrease in species richness downstream, more dramatic in the hot summer of 2003 than in cooler summers. With the increasingly protective environmental regulations (e.g., Water Framework Directive in Europe), accumulation of stresses on aquatic resources are recommended to receive increasing attention, in particular considering the expected changes in climate.

Is the toxicity of pesticide mixtures on river biofilm accounted for solely by the major compounds identified?

Comparative effects of long-term exposure to Polar Organic Chemical Integrative Sampler (POCIS) extracts (PE) and to a reconstituted mixture based on the major compounds quantified in the PE were evaluated on river biofilm communities. The study aimed to characterize the effects of long-term and low-dose exposure to pesticides on natural biofilm communities and to evaluate if the effects due to PE exposure could be explained solely by the major compounds identified in the extracts. Biofilms from an uncontaminated site were exposed in artificial channels to realistic environmental concentrations using diluted PE, with the 12 major compounds quantified in the extracts (Mix) or with water not containing pesticides (Ctr). Significant differences between biofilms exposed to pesticides or not were observed with regard to diatom density, biomass (dry weight and ash-free dry mass), photosynthetic efficiency (ΦpsII) and antioxidant enzyme activities. After 14 days of exposure to the different treatments, the observed trend towards a decrease of mean diatom cell biovolumes in samples exposed to pesticides was related to the control biofilms' higher relative abundance of large species like Cocconeis placentula or Amphora copulata and lower relative abundance of small species like Eolimna minima compared to the contaminated ones. Principal component analyses clearly separated contaminated (PE and Mix) from non-contaminated (Ctr) biofilms; on the contrary, the analyses did not reveal separation between biofilms exposed to PE or to the 12 major compounds identified in the extract.

Pharmaceuticals and pesticides in reclaimed water: Efficiency assessment of a microfiltration-reverse osmosis (MF-RO) pilot plant.

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.

Effects of flow intermittency and pharmaceutical exposure on the structure and metabolism of stream biofilms.

Increasing concentrations of pharmaceutical compounds occur in many rivers, but their environmental risk remains poorly studied in stream biofilms. Flow intermittency shapes the structure and functions of ecosystems, and may enhance their sensitivity to toxicants. This study evaluates the effects of a long-term exposure of biofilm communities to a mixture of pharmaceutical compounds at environmental concentrations on biofilm bioaccumulation capacity, the structure and metabolic processes of algae and bacteria communities, and how their potential effects were enhanced or not by the occurrence of flow intermittency. To assess the interaction between those two stressors, an experiment with artificial streams was performed. Stream biofilms were exposed to a mixture of pharmaceuticals, as well as to a short period of flow intermittency. Results indicate that biofilms were negatively affected by pharmaceuticals. The algal biomass and taxa richness decreased and unicellular green algae relatively increased. The structure of the bacterial (based on denaturing gradient gel electrophoresis of amplified 16S rRNA genes) changed and showed a reduction of the operational taxonomic units (OTUs) richness. Exposed biofilms showed higher rates of metabolic processes, such as primary production and community respiration, attributed to pharmaceuticals stimulated an increase of green algae and heterotrophs, respectively. Flow intermittency modulated the effects of chemicals on natural communities. The algal community became more sensitive to short-term exposure of pharmaceuticals (lower EC50 value) when exposed to water intermittency, indicating cumulative effects between the two assessed stressors. In contrast to algae, the bacterial community became less sensitive to short-term exposure of pharmaceuticals (higher EC50) when exposed to water intermittency, indicating co-tolerance phenomena. According to the observed effects, the environmental risk of pharmaceuticals in nature is high, but different depending on the flow regime, as well as the target organisms (autotrophs vs heterotrophs).

Influence of the interaction between phosphate and arsenate on periphyton's growth and its nutrient uptake capacity.

Periphyton communities grown in microcosms were studied under the exposure to different arsenate (As) and phosphate (P) regimes with the aim of revealing the effect of chronic exposure to As on periphyton physiological and structural characteristics. Also, we aimed to study periphyton changes on sensitivity to As, exposed to different P and As regimes. As affected structural and functional parameters of periphyton communities starved of P, inhibiting algal growth, photosynthetic capacity, changing community composition and reducing the ability of the community to retain P. The effects of As on these parameters were only detected in P starved communities, showing that chronic exposure to As led to changes in the photosynthetic apparatus under the conditions of P-limitation, but not when P-availability was higher. This fact reveals a lower toxicity and/or a higher adaptation of the P-amended community. Intracellular As contents were higher in communities starved of P. However, As tolerance was only induced by the combination of As and P but not by As or P alone indicating that tolerance induction may be an ATP-dependent mechanism. This study reveals that chronic exposure of natural communities to environmentally realistic As concentrations will damage periphyton communities affecting key ecosystem processes, as P uptake, leading to changes in stream ecosystems, as these organisms play a key role in nutrient cycling through nutrient uptake and transfer to higher trophic levels.