Bioconcentration and bioaccumulation of C60 fullerene and C60 epoxide in biofilms and freshwater snails (Radix sp.).

Fullerenes are carbon nanomaterials that have awaken a strong interest due to their adsorption properties and potential applications in many fields. However, there are some gaps of information about their effects and bioconcentration potential in the aquatic biota. In the present work, freshwater biofilms and snails (Radix sp.) were exposed to fullerene C60 aggregates, at concentrations in the low µg/L order, in mesocosms specifically designed to mimic the conditions of a natural stream. The bioconcentration factors of C60 fullerene and its main transformation product, [6,6]C60O epoxide, were studied to the mentioned organisms employing analyses by liquid chromatography coupled to high-resolution mass spectrometry. Our results show that C60 fullerene and its [6,6]C60O present a low bioconcentration factor (BCF) to biofilms: BCFC60 = 1.34 ±â€¯0.95 L/kgdw and BCFC60O = 1.43 ±â€¯0.72 L/kgdw. This suggests that the sorption of these aggregates to biota may be less favoured than it would be suggested by its hydrophobic character. According to our model, the surface of fullerene aggregates is saturated with [6,6]C60O molecules, which exposes the polar epoxide moieties in the surface of the aggregates and decreases their affinity to biofilms. In contrast, freshwater snails showed a moderate capacity to actively retain C60 fullerenes in their organism (BAFC60 = 2670 ±â€¯3070 L/kgdw; BAFC60O = 1330 ±â€¯1680 L/kgdw), probably through ingestion. Our results indicate that the bioaccumulation of these carbon nanomaterials can be hardly estimated using their respective octanol-water partition coefficients, and that their colloidal properties, as well as the feeding strategies of the tested organism, play fundamental roles.

Microbial Organic Matter Utilization in High-Arctic Streams: Key Enzymatic Controls.

In the Arctic, climate changes contribute to enhanced mobilization of organic matter in streams. Microbial extracellular enzymes are important mediators of stream organic matter processing, but limited information is available on enzyme processes in this remote area. Here, we studied the variability of microbial extracellular enzyme activity in high-Arctic fluvial biofilms. We evaluated 12 stream reaches in Northeast Greenland draining areas exhibiting different geomorphological features with contrasting contents of soil organic matter to cover a wide range of environmental conditions. We determined stream nitrogen, phosphorus, and dissolved organic carbon concentrations, quantified algal biomass and bacterial density, and characterized the extracellular enzyme activities involved in catalyzing the cleavage of a range of organic matter compounds (e.g., ß-glucosidase, phosphatase, ß-xylosidase, cellobiohydrolase, and phenol oxidase). We found significant differences in microbial organic matter utilization among the study streams draining contrasting geomorphological features, indicating a strong coupling between terrestrial and stream ecosystems. Phosphatase and phenol oxidase activities were higher in solifluction areas than in alluvial areas. Besides dissolved organic carbon, nitrogen availability was the main driver controlling enzyme activities in the high-Arctic, which suggests enhanced organic matter mineralization at increased nutrient availability. Overall, our study provides novel information on the controls of organic matter usage by high-Arctic stream biofilms, which is of high relevance due to the predicted increase of nutrient availability in high-Arctic streams in global climate change scenarios.

Impact of fullerenes in the bioaccumulation and biotransformation of venlafaxine, diuron and triclosan in river biofilms.

A huge variety of organic microcontaminants are presently detected in freshwater ecosystems, but there is still a lack of knowledge about their interactions, either with living organisms or with other contaminants. Actually, carbon nanomaterials like fullerenes (C60) can act as carriers of organic microcontaminants, but their relevance in processes like bioaccumulation and biotransformation of organic microcontaminants by organisms is unknown. In this study, mesocosm experiments were used to assess the bioaccumulation and biotransformation of three organic microcontaminants (venlafaxine, diuron and triclosan) in river biofilms, and to understand how much the concomitant presence of C60 at environmental relevant concentrations could impact these processes. Results indicated that venlafaxine exhibited the highest bioaccumulation (13% of the initial concentration of venlafaxine in water), while biotransformation was more evident for triclosan (5% of the initial concentration of triclosan in water). Furthermore, biotransformation products such as methyl-triclosan were also present in the biofilm, with levels up to 42% of the concentration of accumulated triclosan. The presence of C60 did not involve relevant changes in the bioaccumulation and biotransformation of microcontaminants in biofilms, which showed similar patterns. Nevertheless, the study shows that a detailed evaluation of the partition of the organic microcontaminants and their transformation products in freshwater systems are important to better understand the impact of the co-existence of others microcontaminants, like carbon nanomaterials, in their possible routes of bioaccumulation and biotransformation.

Deconvolution model to resolve cytometric microbial community patterns in flowing waters.

Flow cytometry is suitable to discriminate and quantify aquatic microbial cells within a spectrum of fluorescence and light scatter signals. Using fixed gating and operational settings, we developed a finite distribution mixture model, followed by the Voronoi tessellation, to resolve bivariate cytometric profiles into cohesive subgroups of events. This procedure was applied to outline recurrent patterns and quantitative changes of the aquatic microbial community along a river hydrologic continuum. We found five major subgroups within each of the commonly retrieved populations of cells with Low and High content of Nucleic Acids (namely, LNA, and HNA cells). Moreover, the advanced analysis allowed assessing changes of community patterns perturbed by a wastewater feed. Our approach for cytometric data deconvolution confirmed that flow cytometry could represent a prime candidate technology for assessing microbial community patterns in flowing waters. © 2017 International Society for Advancement of Cytometry.

Warmer night-time temperature promotes microbial heterotrophic activity and modifies stream sediment community.

Diel temperature patterns are changing because of global warming, with higher temperatures being predicted to be more pronounced at night. Biological reactions are temperature dependent, with some occurring only during the daylight hours (e.g., light photosynthesis) and other during the entire day (e.g., respiration). Consequently, we expect the modification of daily temperature cycles to alter microbial biological reactions in stream sediments. Here, we aimed to study the effect of warming and changes of the diel temperature patterns on stream sediment biofilm functions tied to organic carbon decomposition, as well as on biofilm meiofaunal community structure. We performed an eight-week experiment with 12 artificial streams subjected to three different diel temperature patterns: warming, warmer nights and control. Significant effects of warming on biofilm function and structure were mainly detected in the long term. Our results showed that warming altered biofilm function, especially in the warmer nights' treatment, which enhanced ß-glucosidase enzyme activity. Interestingly, clear opposite diel patterns were observed for dissolved organic carbon and ß-glucosidase activity, suggesting that, at night, sediment bacteria quickly consume the input of photosynthetic dissolved organic carbon labile compounds created during light-time. The biofilm structure was also altered by warming, as both warming and warmer night treatments enhanced copepod abundance and diminished abundances of turbellaria and nematodes, which, in turn, controlled bacterial, algal and ciliate communities. Overall, we conclude that warming has strong effect on sediment biofilm structure and enhanced microbial organic matter degradation which might, consequently, affect higher trophic levels and river carbon cycling.

Shifts in microbial community structure and function in light- and dark-grown biofilms driven by warming.

Biofilms are dynamic players in biogeochemical cycling in running waters and are subjected to environmental stressors like those provoked by climate change. We investigated whether a 2°C increase in flowing water would affect prokaryotic community composition and heterotrophic metabolic activities of biofilms grown under light or dark conditions. Neither light nor temperature treatments were relevant for selecting a specific bacterial community at initial phases (7-day-old biofilms), but both variables affected the composition and function of mature biofilms (28-day-old). In dark-grown biofilms, changes in the prokaryotic community composition due to warming were mainly related to rotifer grazing, but no significant changes were observed in functional fingerprints. In light-grown biofilms, warming also affected protozoan densities, but its effect on prokaryotic density and composition was less evident. In contrast, heterotrophic metabolic activities in light-grown biofilms under warming showed a decrease in the functional diversity towards a specialized use of several carbohydrates. Results suggest that prokaryotes are functionally redundant in dark biofilms but functionally plastic in light biofilms. The more complex and self-serving light-grown biofilm determines a more buffered response to temperature than dark-grown biofilms. Despite the moderate increase in temperature of only 2°C, warming conditions drive significant changes in freshwater biofilms, which responded by finely tuning a complex network of interactions among microbial populations within the biofilm matrix.

Resistance but not recovery is related to the role of specialist taxa in river communities submitted to hydric stress.

One of the main effects of global change is the human interference in the global water cycle, which alters river hydrological dynamics and submits their biological communities to hydric stress. Hydric stress is a pulse disturbance with potential multiple effects on biodiversity and functions in river ecosystems. The presence of habitat specialists may support the response of biological communities to pulse disturbances, maintaining ecological functions more consistently than other communities only having generalists. We tested this general hypothesis in stream communities submitted to increasing hydric stress (normal conditions vs humidity vs desiccation). We used communities with variable proportion of specialist algal and cyanobacterial taxa and tested their resistance to hydric stress by analyzing potential changes on their number of species, biovolume, proportion of intact cells, and photosynthetic variables (basal fluorescence, photosynthetic yield). We also evaluated the recovery of ecological functions (net community primary production, community respiration, phosphorus uptake) once hydric stress conditions ended. Hydric stress caused a slight decrease in the number of species and biovolume of assemblages, but the proportion of intact cells did not significantly change because of the disturbance. Basal fluorescence and photosynthetic yield under hydric stress decreased more markedly in communities without specialist taxa, while communities with habitat specialists resisted better. Metabolism did not remarkably decrease under moderate hydric stress, but dropped by half under desiccation in all communities, having or not specialist taxa. Overall, specialist taxa did provide higher resistance to stress but did not support a distinct recovery of ecological functions. We suggest that this characteristic response is related to the high plasticity of biofilm structures.

Artificial substrata to assess ecological and ecotoxicological responses in river biofilms: Use and recommendations.

River biofilms are biological consortia of autotrophs and heterotrophs colonizing most solid surfaces in rivers. Biofilm composition and biomass differ according to the environmental conditions, having different characteristics between systems and even between river habitats. Artificial substrata (AS) are an alternative for in situ or laboratory experiments to handle the natural variability of biofilms. However, specific research goals may require decisions on colonization time or type of substrata. Substrata properties (i.e., texture, roughness, hydrophobicity) and the colonization period and site are selective factors of biofilm characteristics. Here we describe the uses of artificial substrata in the assessment of ecological and ecotoxicological responses and propose a decision tree for the best use of artificial substrata in river biofilm studies. We propose departing from the purpose of the study to define the necessity of obtaining a realistic biofilm community, from which it may be defined the colonization time, the colonization site, and the type of artificial substratum. Having a simple or mature biofilm community should guide our decisions on the colonization time and type of substrata to be selected for the best use of AS in biofilm studies. Tests involving contaminants should avoid adsorbing materials while those ecologically oriented may use any AS mimicking those substrata occurring in the streambed.•We review the utilization of different artificial substrata to colonize biofilm in river ecology and ecotoxicology.•We propose a decision tree to guide on selecting the appropriate artificial substrata and colonization site and duration.•Type of artificial substrata (material, size, shape...) and colonization duration are to be decided according to the specific purpose of the study.

Extreme weather events threaten biodiversity and functions of river ecosystems: evidence from a meta-analysis.

Both gradual and extreme weather changes trigger complex ecological responses in river ecosystems. It is still unclear to what extent trend or event effects alter biodiversity and functioning in river ecosystems, adding considerable uncertainty to predictions of their future dynamics. Using a comprehensive database of 71 published studies, we show that event - but not trend - effects associated with extreme changes in water flow and temperature substantially reduce species richness. Furthermore, event effects - particularly those affecting hydrological dynamics - on biodiversity and primary productivity were twice as high as impacts due to gradual changes. The synthesis of the available evidence reveals that event effects induce regime shifts in river ecosystems, particularly affecting organisms such as invertebrates. Among extreme weather events, dryness associated with flow interruption caused the largest effects on biota and ecosystem functions in rivers. Effects on ecosystem functions (primary production, organic matter decomposition and respiration) were asymmetric, with only primary production exhibiting a negative response to extreme weather events. Our meta-analysis highlights the disproportionate impact of event effects on river biodiversity and ecosystem functions, with implications for the long-term conservation and management of river ecosystems. However, few studies were available from tropical areas, and our conclusions therefore remain largely limited to temperate river systems. Further efforts need to be directed to assemble evidence of extreme events on river biodiversity and functioning.

Green and brown stream trophic food chains show specific responses to constant or hump-shaped inputs of copper.

The brown food chain (based on decomposers) co-exists in streams with the green food chain (based on primary producers). The two trophic chains perform specific ecosystem functions which may be altered by the effect of contaminants. Copper is a common contaminant with recognized effects on several compartments of the two trophic chains. We applied it in two separate mesocosm experiments, in which we tested the effects of copper after contrasting patterns of contaminant exposure (constant vs hump-shaped). The constant input simulated a chronic contamination (average of 20 µg/L Cu), while the hump-shaped simulated the steady arrival of copper, the occurrence of a peak (reaching ca. 60 µg/L Cu), and its progressive decrease (down to 10-15 µg/L Cu). In the green trophic food chain, copper exposure decreased the total chlorophyll-a as well as the basal fluorescence and the photosynthetic yield. The treatment receiving hump-shaped inputs caused the highest mortality of the green food chain consumer, the snail Radix balthica. In the chronic copper exposure, mortality achieved a maximum of 80% by the end of the experiment but occurred later than that in the hump-shaped treatment. Effects on the brown food chain were not so pronounced; the microbial decomposition rate of leaflitter decreased nearly ca. 50% after 14 days of copper exposure. Effects on decomposition translated into the ingestion performance of detritivores, which decreased in the two copper treatments. Our results provide evidence that copper affected the two trophic food chains. The hump-shaped arrival included a peak of high concentration, which caused lethal effects on the consumers, but also a decreasing limb, which allowed a partial recovery of the algal photosynthetic variables. Our results suggest the need to consider the different compartments and functions performed within the stream trophic web when evaluating the effects of a contaminant in a river ecosystem.

River biofilms adapted to anthropogenic disturbances are more resistant to WWTP inputs.

The sensitivity and spatial recovery of river sediment biofilms along 1 km after the input of two wastewater treatment plants (WWTPs) located in two river reaches with different degrees of anthropogenic influence were investigated. First, at the upper reach, we observed an inhibition of some microbial functions (microbial respiration and extracellular enzyme activities) and strong shifts in bacterial community composition (16S rRNA gene), whereas an increase in microbial biomass and activity and less pronounced effect on microbial diversity and community composition were seen at the lower reach. Second, at the lower reach we observed a quick spatial recovery (around 200 m downstream of the effluent) as most of the functions and community composition were similar to those from reference sites. On the other hand, bacterial community composition and water quality at the upper reach was still altered 1 km from the WWTP effluent. Our results indicate that biofilms in the upstream sites were more sensitive to the effect of WWTPs due to a lower degree of tolerance after a disturbance than communities located in more anthropogenically impacted sites.

Effects of multiple stressors on river biofilms depend on the time scale.

Global change exposes ecosystems to a myriad of stressors differing in their spatial (i.e. surface of stressed area) and temporal (i.e. exposure time) components. Among freshwater ecosystems, rivers and streams are subject to physical, chemical and biological stressors, which interact with each other and might produce diverging effects depending on exposure time. We conducted a manipulative experiment using 24 artificial streams to examine the individual and combined effects of warming (1.6 °C increase in water temperature), hydrological stress (simulated low-flow situation) and chemical stress caused by pesticide exposure (15.1-156.7 ng L-1) on river biofilms. We examined whether co-occurring stressors could lead to non-additive effects, and if these differed at two different exposure times. Specifically, structural and functional biofilm responses were assessed after 48 hours (short-term effects) and after 30 days (long-term effects) of exposure. Hydrological stress caused strong negative impacts on river biofilms, whereas effects of warming and pesticide exposure were less intense, although increasing on the long term. Most stressor combinations (71%) resulted in non-significant interactions, suggesting overall additive effects, but some non-additive interactions also occurred. Among non-additive interactions, 59% were classified as antagonisms after short-term exposure to the different stressor combinations, rising to 86% at long term. Our results indicate that a 30-day exposure period to multiple stressors increases the frequency of antagonistic interactions compared to a 48-hour exposure to the same conditions. Overall, the impacts of multiple-stressor occurrences appear to be hardly predictable from individual effects, highlighting the need to consider temporal components such as duration when predicting the effects of multiple stressors.

Ecotoxicological effects of carbon based nanomaterials in aquatic organisms.

An increasing amount of carbon-based nanomaterials (CNM) (mostly fullerenes, carbon nanotubes and graphene) has been observed in aquatic systems over the last years. However, the potential toxicity of these CNM on aquatic ecosystems remains unclear. This paper reviews the existing literature on the toxic effects of CNM in aquatic organisms as well as the toxic effects of CNM through influencing the toxicity of other micro-pollutants, and outlines a series of research needs to reduce the uncertainty associated with CNMs toxic effects. The results show that environmental concentrations of CNM do not pose a threat on aquatic organisms on their own. The observed concentrations of CNM in aquatic environments are in the order of ngL-1 or even lower, much below than the lowest observed effect concentrations (LOEC) on different aquatic organisms (in the order of mgL-1). Toxic effects have been mainly observed in short-term experiments at high concentrations, and toxicity principally depends on the type of organisms, exposition time and CNM preparation methods. Moreover, we observed that CNM interact (establishing synergistic and/or antagonistic effects) with other micro-pollutants. Apparently, the resulting interaction is highly dependent on the chemical properties of each micro-pollutant, CNM acting either as carriers or as sorbents, thereby modifying the original toxicity of the contaminants. Results stress the need of studying the interactive effects of CNM with other micro-pollutants at environmental relevant concentrations, as well as their effects on biological communities in the long-term.

Value of glycolic acid analysis in ethylene glycol poisoning: A clinical case report and systematic review of the literature.

OBJECTIVE: To evaluate the clinical utility of glycolic acid (GA) determination in the diagnosis and prognosis of ethylene glycol (EG) intoxications. METHOD: Systematic review of serum and/or urine GA concentrations available in the literature in cases of EG poisoning. Present a clinical case in which the determination of the GA was decisive. RESULTS: In total, 137 patients were included. Serum GA concentrations (but not EG) of patients who survive are different from those who die. The optimal cut-off of serum GA to predict mortality was 990.5mg/L (sensitivity 85.2%, specificity 54.3%) with an Odds Ratio of 6.838 (2.868-16.302). In our clinical case, serum EG was negative; however, urine GA was positive (1230.7mg/L). CONCLUSIONS: In all suspected cases of EG poisoning, it is advisable to carry out the simultaneous analysis of EG and GA.

Fullerenes Influence the Toxicity of Organic Micro-Contaminants to River Biofilms.

Organic micro-contaminants (OMCs) enter in freshwaters and interact with other contaminants such as carbon nanoparticles, becoming a problem of unknown consequences for river ecosystems. Carbon nanoparticles (as fullerenes C60) are good adsorbents of organic contaminants and their interaction can potentially affect their toxicity to river biofilms. We tested the C60 interactions with selected OMCs and their effects on river biofilms in different short-term experiments. In these, river biofilms were exposed to C60 and three OMCs (triclosan, diuron, or venlafaxine) and their respective mixtures with fullerenes (C60 + each OMC). The effects were evaluated on structural, molecular, and functional descriptors of river biofilms. Our results showed that C60 did not cause toxic effects in river biofilms, whereas diuron and triclosan significantly affected the heterotrophic and phototrophic components of biofilms and venlafaxine affected only the phototrophic component. The joint exposure of C60 with venlafaxine was not producing differences with respect to the former response of the toxicant, but the overall response was antagonistic (i.e., decreased toxicity) with diuron, and synergistic (i.e., increased toxicity) with triclosan. We suggest that differences in the toxic responses could be related to the respective molecular structure of each OMC, to the concentration proportion between OMC and C60, and to the possible competition between C60 pollutants on blocking the receptors of the biological cell membranes. We conclude that the presence of C60 at low concentrations modified the toxicity of OMC to river biofilms. These interactions should therefore be considered when predicting toxicity of OMC in river ecosystems.

Quality and reactivity of dissolved organic matter in a Mediterranean river across hydrological and spatial gradients.

Understanding DOM transport and reactivity in rivers is essential to having a complete picture of the global carbon cycle. In this study, we explore the effects of hydrological variability and downstream transport on dissolved organic matter (DOM) dynamics in a Mediterranean river. We sampled the main stem of the river Tordera from the source to the sea, over a range of fifteen hydrological conditions including extreme events (flood and drought). By exploring spatial and temporal gradients of DOM fluorescence properties, river hydrology was found to be a significant predictor of DOM spatial heterogeneity. An additional space-resolved mass balance analysis performed on four contrasting hydrological conditions revealed that this was due to a shift in the biogeochemical function of the river. Flood conditions caused a conservative transport of DOM, generating a homogeneous, humic-like spatial profile of DOM quality. Lower flows induced a non-conservative, reactive transport of DOM, which enhanced the spatial heterogeneity of DOM properties. Moreover, the downstream evolution of DOM chemostatic behaviour revealed that the role of hydrology in regulating DOM properties increased gradually downstream, indicating an organised inter-dependency between the spatial and the temporal dimensions. Overall, our findings reveal that riverine DOM dynamics is in constant change owing to varying hydrological conditions, and emphasize that in order to fully understand the role of rivers in the global carbon cycle, it is necessary to take into account the full range of hydrological variability, from floods to droughts.