River Biofilms Research Articles

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.

Dissolved organic matter does not promote glyphosate degradation in auto-heterotrophic aquatic microbial communities

Environmental dissolved organic matter (DOM) has been proved to increase microbial population sizes and stimulate the degradation of some pesticide molecules. Among these molecules, the present study investigated the biodegradation of the herbicide glyphosate depending on photoautotrophs DOM supply in a microbial consortium isolated from river biofilms. Degradation experiments in the laboratory were performed in dark and light conditions, as well as after antibiotic supply, in order to characterize the eventual interactions between photoautotrophs and heterotrophs activity during glyphosate degradation. Fifty percent of the initial concentration of glyphosate (0.6 mM) was transformed into aminomethyl phosphonic acid (AMPA) after 9 days in presence or absence of light. Accordingly, the photoautotrophic DOM supply was not stimulating glyphosate degradation by microbial heterotrophs. This lack of response was probably explained by the low net primary production values and weak dissolved organic carbon production recorded in light treatments. The supply of the antibiotic drastically stopped glyphosate transformation demonstrating the central role of bacteria in the biodegradation of the herbicide. Glyphosate also modified the structure of prokaryotes assemblages in the consortium by increasing the relative abundances of Alphaproteobacteria and slightly decreasing those of Gammaproteobacteria. The chemoorganotrophic bacteria Phenylobacterium sp. (Alphaproteobacteria) was related to the transformation of glyphosate in our microbial consortium. The present study highlights the complexity of microbial interactions between photoautotrophs and heterotrophs in microbial assemblages that can contribute to the degradation of pesticides present in aquatic environments.

Cultivation substrata differentiate the properties of river biofilm EPS and their binding of heavy metals: A spectroscopic insight

River biofilms inevitably serve as recipients of heavy metals including copper (Cu) and cadmium (Cd) following their introduction in fluvial systems. Nevertheless, the effects of cultivation substrata on the characteristics of river biofilm extracellular polymeric substances (EPS) and the binding behaviors of heavy metals on biofilms remain unclear. Integrating spectroscopic methods with chemometric analyses, we explored the binding behaviors of Cu(II) and Cd(II) onto biofilm EPS cultivated from two representative substrata at the molecular level. Chemical analysis revealed that biofilm cultivated on polyethylene (PE) pieces contained more non-fluorescent protein fractions, whereas EPS from periphyton grown on mineral, i.e., cobblestones was richer in aromatic fractions and polysaccharides. Excitation-emmision matrix combined with parallel factor analysis suggested a stronger interaction between fluorophores in periphytic EPS with Cu(II) compared to fluorophores in plastic biofilm EPS. Integrated use of infrared spectroscopy and two-dimensional correlation analyses revealed that, during the heavy metal binding processes, the amines and phenolics in plastic biofilm EPS gave the fastest responses to metal binding. While the amides and the aliphatic fractions in periphytic EPS showed a preferential binding to heavy metals. This study differentiates the effects of cultivation substrata on structuring the biofilm EPS characteristics and offers new insights into the environmental behaviors of heavy metal discharge into fluvial systems in river biofilm matrix.

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.

Microscale and molecular analyses of river biofilm communities treated with microgram levels of cerium oxide nanoparticles indicate limited but significant effects

Cerium oxide (CeO2) nanoparticles are used as in-fuel catalysts and in manufacturing processes, creating a potential for release to aquatic environments. Exposures at 1 and 10 μg/L CeO2-nanoparticles were made to assess effects during the development of river biofilm communities. Scanning transmission x-ray microscopy (STXM) indicated extensive sorption of nanoparticles to the community and co-localization with lipid moieties. Following 8 weeks of development, polycarbonate coupons were removed from the reactors and used for molecular analyses, denaturing gradient gel electrophoresis analysis (DGGE-16S rRNA) and 16S rRNA amplicon sequencing. Microscopic imaging of the biofilm communities (bacterial, photosynthetic biomass, exopolymer composition, thickness, protozoan numbers), as well as carbon substrate utilization fingerprinting was performed. There was a trend toward reduced photosynthetic biomass, but no significant effects of CeO2 exposure were found on photosynthetic and bacterial biomass or biofilm thickness. Sole carbon source utilization analyses indicated increased utilization of 10 carbon sources in the carbohydrate, carboxylic acid and amino acids categories related to CeO2 exposures; however, predominantly, no significant effects (p < 0.05) were detected. Measures of microbial diversity, lectin binding affinities of exopolymeric substances and results of DGGE analyses, indicated significant changes to community composition (p < 0.05) with CeO2 exposure. Increased binding of the lectin Canavalia ensiformis was observed, consistent with changes in bacterial-associated polymers. Whereas, no significant changes were observed in binding to residues associated with algal and cyanobacterial exopolymers. 16S rRNA amplicon sequencing of community DNA indicated changes in diversity and shifts in community composition; however, these did not trend with increasing CeO2 exposure. Counting of protozoans in the biofilm communities indicated no significant effects on this trophic level. Thus, based on biomass and functional measures, CeO2 nanoparticles did not appear to have significant effects; however, there was evidence of selection pressure resulting in significant changes in microbial community composition.

Pseudomonas strains isolated from different environmental niches exhibit different antagonistic ability

The antagonistic interactions between microorganisms in the environment seem to play a key role in determining the spatial structuring of bacterial communities. In this work, the antagonistic interactions among 82 strains belonging to different Pseudomonas species isolated from six different environmental niches have been investigated to check whether it might be related to the environmental niches and/or to the taxonomical affiliation. The bacterial panel included 20 strains from Echinacea purpurea (12 from roots, 8 from the rhizosphere), 19 from Phragmites australis, 20 from the Acquarossa river (13 and 7 from red and black epilithon, respectively), 9 strains from red epilithon of the “Infernaccio waterfalls”, 5 Antarctic strains, and 9 strains from the Amazon rainforest. Data obtained revealed that 81 out of the 82 strains exhibited an antagonistic activity (even though at a different extent) and that the environmental characteristics, which also determine different adaptative strategies, might be the main factor affecting antagonism among bacteria, even more than taxonomy. Strains isolated from heavy metal enriched river biofilms were described as the most active, while endophytic and rhizospheric strains were the most sensitive. This research, representing the first report in which Pseudomonas bacterial strains isolated from different ecological niches have been tested for their interaction abilities, opens new perspectives in discovering putative new antimicrobial compounds from different (extreme) environments.

Fate and effects of triclosan in subtropical river biofilms

Triclosan (TCS, 5-chloro-2-(2,4-dichlorophenoxy) phenol) is a broad-spectrum antimicrobial compound. Owing to its wide use, TCS has been frequently detected in river systems, especially in the (sub-)tropics. However, little information on its interaction with river biofilm in the (sub)tropics is currently available. In the present study, subtropical river biofilms were chronically exposed to TCS for 14 d at concentrations of 0.1–100 μg/L in artificial river water, which was followed by a 7 d recovery period. The results show that 100 μg/L TCS inhibited the growth of river biofilms and the no-observed-effect concentration (NOEC) of TCS on river biofilms was 10 μg/L. The affected biofilms did not completely recover within the 7 d of recovery period due to the adsorbed TCS which was not removed together with dissolved TCS. Exposure to TCS caused significant changes in prokaryotic species composition of river biofilms but no significant effects on eukaryotic species composition. In particular, the relative abundance of several TCS-tolerant bacterial species (e.g., Pseudoxanthomonas mexicana, Sphingopyxis alaskensis and Sphingomonas wittichii) in river biofilms increased following exposure to 10 and 100 μg/L TCS. River biofilm efficiently removed TCS from the liquid phase and the pH values of the aquatic system significantly affected the removal efficiency of TCS (from 36% at pH 6.5 to 60% at pH 8.5). No degradation products were detected in the liquid phase after 5 days of exposure, possibly due to strong adsorption of the hydrophobic degradation products to river biofilms and through biodegradation by bacteria utilizing TCS and its degradation products as source of carbon and energy for growth, such as Methyloversalitis universalis and Methylobacterium aquaticum.

Meta-analysis of glyphosate contamination in surface waters and dissipation by biofilms

One consequence of the intensive use of glyphosate is the contamination of rivers by the active substance and its metabolites aminomethyl phosphonic acid (AMPA) and sarcosine, inducing river eutrophication. Biofilms are the predominant lifestyle for microorganisms in rivers, providing pivotal roles in ecosystem functioning and pollutant removal. The persistence of glyphosate in these ecosystems is suspected to be mostly influenced by microbial biodegradation processes.The present study aimed to investigate the tripartite relationship among biofilms, phosphorus and glyphosate in rivers. The first part consists of a co-occurrence analysis among glyphosate, AMPA and phosphorus using an extensive dataset of measurements (n = 56,198) from French surface waters between 2013 and 2017. The second part investigated the capacity of natural river biofilms to dissipate glyphosate, depending on phosphorus availability and the exposure history of the biofilm, in a microcosm study.A strong co-occurrence among glyphosate, AMPA and phosphorus was found in surface waters. More than two-thirds of samples contained phosphorous with glyphosate, AMPA or both compounds. Seasonal fluctuations in glyphosate, AMPA and phosphorus concentrations were correlated, peaking in spring/summer shortly after pesticide spreading. Laboratory experiments revealed that natural river biofilms can degrade glyphosate. However, phosphorus availability negatively influenced the biodegradation of glyphosate and induced the accumulation of AMPA in water. An increase in alkaline phosphatase activity and phosphorus uptake was observed in glyphosate-degrading biofilms, evidencing the tight link between phosphorus limitation and glyphosate degradation by biofilms.The results of the present study show that phosphorus not only is a key driver of river eutrophication but also can reduce complete glyphosate degradation by biofilms and favour the accumulation of AMPA in river water. The predominant role of biofilms and the trophic status of rivers must therefore be considered in order to better assess the fate and persistence of glyphosate.

Visualization of the Sorption of Nickel within Exopolymer Microdomains of Bacterial Microcolonies Using Confocal and Scanning Electron Microscopy.

The sorption and distribution of nickel, a common metal contaminant in aquatic systems, were assessed in bacterial microcolonies using a combination of fluorescent staining with Newport Green and confocal laser scanning microscopy (CLSM) with confirmation by scanning electron microscopy (SEM) and X-ray microprobe analyses. CLSM with Newport Green, selected fluor-conjugated lectins, and DNA staining allowed for the discrimination of the microdomains present in the microcolony exopolymeric matrix and detection of bound nickel. This approach avoided the artefacts associated with drying and fixation required by analytical electron microscopy. The results obtained indicated that specific microcolonies within river biofilms sorbed nickel within limited microdomains present in the complex tripartite exopolymeric matrix surrounding bacterial cells. Sorption occurred such that nickel was concentrated within the exopolymeric matrix, but not directly associated with cells. These microdomains appeared to have neutral pH and be dominated by negatively charged residues favoring the sorption of nickel and other cations. These results also suggest an important role for specific community members in the sorption and concentration of metals in aquatic biofilm communities.

Sorption removal of phthalate esters and bisphenols to biofilms from urban river: From macroscopic to microcosmic investigation

River biofilms play fundamental roles in shaping the architecture of aquatic systems. Sorption to biofilms was thought to be a crucial mechanism controlling the fate and transport of trace emerging contaminants. This study focused on the role of in situ colonized river biofilms in the early fate of phthalate esters (PAEs) and bisphenols (BPs) at trace concentrations in a representative urban river. PAEs and BPs were readily sorbed to biofilms with uptakes of 38.2–162.5 μg/g for PAEs and 1787.7–4425.6 μg/g for BPs, respectively. The total mass and characteristics of the colonized biofilms varied in response to seasons and water qualities. The biofilm colonized in the downstream of a wastewater treatment plant exhibited the highest sorption capacity among the tested sites, possibly attributed to the higher organic contents of biofilms owing to the elevated availability of nutrients. Correlation analysis indicates that certain water qualities, e.g., TN and NH3N, and biofilm properties, e.g., organic and polysaccharide fractions could be selected to predict the sorption capacities of river biofilms. Hydrophobic partitioning into organic matter appears to be the dominant sorption mechanism and biofilm polysaccharides were probably responsible for the adhesion of tested compounds. The contaminant partitioning into biofilm and sediment at mass/volume ratios typical for small rivers suggests that the biofilm could serve as an important sorbing matrix for the trace organic contaminants as compared to the sediments. Our work yields new insights into the early uptake and accumulation of trace plasticizers by natural biofilms, which is of significance in understanding the subsequent transport of trace organic contaminants in fluvial systems.

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.

Use of convertible flow cells to simulate the impacts of anthropogenic activities on river biofilm bacterial communities

Bacterial attachment to surfaces and the development of biofilms are crucial processes during the self-purification of polluted rivers. Biofilm bacterial communities also are a potential indicator of the human impact on an aquatic system. Here, we used indoor reactors with 7.7cm3 transparent convertible flow cells to observe the formation of biofilms in river water from different land-use areas (i.e., an undisturbed mountainous area, a wastewater-discharge urban area, and a pesticide-fertilizer applied agricultural area). We then compared the bacterial biomass, composition, and function among the formed biofilms and explored whether the biofilm bacterial communities formed in polluted river water (urban area) could shift to those formed in unpolluted water (mountainous area) after simulating water-body remediation. After 60d of indoor biofilm cultivation, the biofilms formed with the three types of influent were markedly different. Anthropogenic activities (e.g., wastewater discharge and pesticide-fertilizer use) facilitated biofilm bacterial production and the metabolic rate and altered the composition and metabolic patterns of the biofilm bacterial communities. After switching from an urban water to mountainous water influent in the same reactor, the biofilm bacterial communities that initially formed in the polluted discharge did not shift to that formed in unpolluted water. This result indicated that even after water remediation, the composition of the river biofilm bacterial community would not recover to a community like that observed under non-polluted conditions. Our study highlights possible issues related to current pollution-remediation routines and emphasizes the importance of sustainable anthropogenic activities within river basins.

Spatial and Temporal Variability in the Development and Potential Toxicity of Phormidium Biofilms in the Tarn River, France.

Proliferation of Phormidium biofilms in rivers is becoming a worldwide sanitation problem for humans and animals, due to the ability of these bacteria to produce anatoxins. To better understand the environmental conditions that favor the development of Phormidium biofilms and the production of anatoxins, we monitored the formation of these biofilms and their toxins for two years in the Tarn River, biofilms from which are known to have caused the deaths of multiple dogs. As previously observed in New Zealand, Phormidium biofilm development occurred in riffle areas. The coverage of these biofilms at the bottom of the river exhibited strong spatial and temporal variations, but was positively correlated with water temperature and depth. Anatoxin-a was detected in less than 50% of the biofilms. The concentrations of these toxins in the biofilms exhibited high spatiotemporal variability, with the highest concentrations being recorded at the end of the summer period at the upstream sampling sites. These findings suggest that the maturity of the biofilms, combined with the local environmental conditions, have an impact on the production of anatoxin, making risk assessment for these benthic proliferations challenging.

Multiple processes acting from local to large geographical scales shape bacterial communities associated with Phormidium (cyanobacteria) biofilms in French and New Zealand rivers

River biofilms dominated by Phormidium (cyanobacteria) are receiving increased attention worldwide because of a recent expansion in their distribution and their ability to produce neurotoxins leading to animal mortalities. Limited data are available on the composition and structure of bacterial communities (BCs) associated with Phormidium biofilms despite the important role they potentially play in biofilm functioning. By using a high-throughput sequencing approach, we compared the BCs associated with Phormidium biofilms in several sampling sites of the Tarn River (France) and in eight New Zealand rivers. The structure of the BCs from both countries displayed spatial and temporal variations but were well conserved at the order level and 28% of the OTUs containing 90% of the reads were shared by these BCs. This suggests that micro-environmental conditions occurring within thick Phormidium biofilms strongly shape the associated BCs. A strong and significant distance-decay relationship (rp = 0.7; P = 0.001) was found in BCs from New Zealand rivers but the Bray-Curtis dissimilarities between French and New Zealand BCs are in the same order of magnitude of those found between New Zealand BCs. All these findings suggest that local environmental conditions seem to have more impact on BCs than dispersal capacities of bacteria.

Spatial structuring of bacterial communities in epilithic biofilms in the Acquarossa river (Italy).

Epilithic river biofilms characterize the rock surfaces along the Acquarossa river (Viterbo, Italy); they are in part red and in part black colored, maintaining a well-defined borderline. This peculiarity has raised questions about the biotic and abiotic phenomena that might avoid the mixing of the two biofilms. In this study, the structuring of bacterial communities in black and red epilithic biofilm in the Acquarossa river has been investigated with both culture dependent and independent approaches. Data obtained highlighted a (very) different taxonomic composition of black and red epilithons bacterial communities, dominated by Acinetobacter sp. and iron-oxidizing bacteria, respectively. The chemical characterization of both river water and biofilms revealed a substantial heavy metals pollution of the environment; heavy metals were also differentially accumulated in red and black epilithons. Overall, our data revealed that the structuring of red and black epilithons might be affected mainly by the antagonistic interactions exhibited by bacterial genera dominating the two biofilms. These findings suggest that biotic factors might be responsible for the structuring of natural bacterial communities, suggesting that there is a selection of populations at very small scale, and that different populations might compete for different niches.

Semi-continuous pharmaceutical and human tracer monitoring by POCIS sampling at the watershed-scale in an agricultural rural headwater river.

Pharmaceutical monitoring (37 pharmaceuticals and 3 human tracers) was conducted in a headwater streams in southwest France, an area characterized by a low population density with an elderly population (30% > 60 years old) and extensive agriculture (cow cattle breeding). Polar Organic Chemical Integrative Sampler (POCIS) were exposed for 14-day consecutive periods in 2016 at three sampling points. Three human wastewater tracers and 20 pharmaceuticals commonly used for human and/or cattle were quantified in headwaters. Succession of small Wastewater Treatment Plant (WWTP), non-collective sanitation, discharges of untreated effluents as well as the river ability to dilute discharged wastewater, mainly explain the pharmaceuticals and human tracers concentrations. Pharmaceutical loads were time-dependent and were higher during cold season due to increase of pharmaceutical consumption. In contrast, better degradation and/or sorption onto river biofilms in warm season induced the decrease of headwater pharmaceutical content. The headwaters streams were contaminated by compounds found in other type of watershed, but β-blocker were the compounds quantified in higher concentration with frequencies of 100%, which was consistent with the elderly population living in the watershed. Specific compounds (sulfamerazine and sulfamethoxazole) used to cattle medical care were detected in waters, but at a low content.

Gold nanoparticle trophic transfer from natural biofilm to grazer fish

Nanoparticle (NP) trophic transfer is reported as an important exposure route for organisms in aquatic ecosystems. This study focused on gold nanoparticle (AuNP, PEG-coated, 10 nm diameter) transfer using an experimental benthic food chain which included two trophic levels: natural river biofilm and grazer fish Hypostomus plecostomus. AuNP biofilm accumulation was assessed via water AuNP concentrations and total biofilm mass. An extended range of six AuNP concentrations in water (0, 0.0048, 0.048, 0.48, 4.8, and 48 mg Au L−1) was set. A dose-dependent relation between gold concentrations in water and natural river biofilm was observed after a 48-h exposure. This pointed out the high propensity of natural biofilms to accumulate gold. Additionally, total biofilm mass appeared to influence AuNP accumulation at the highest exposure levels. This first step enables the set-up of the transfer experiment in which grazer fish were exposed for 21 days to natural biofilms, previously contaminated by low AuNP concentrations in water (NP0.1: 0.48 and NP1: 4.8 mg Au L−1). Gold was quantified in eight fish organs, and histology was observed. Gold was transferred from biofilms to fish; bioaccumulation was organ- and exposure level-dependent. Interestingly, the brain showed significant gold accumulation at the highest exposure level (NP1). Histological observations indicated distinct inflammatory responses in fish liver, spleen, and muscle. The overall results suggest the potential hazards of subchronic nanoparticle exposure in aquatic organisms.

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.

Effects of chronic elevated nitrate concentrations on the structure and function of river biofilms

Abstract Biofilms play an important role in nutrient cycling and retention in streams and rivers, altering transport of nitrate () to downstream ecosystems. However, increased nutrient availability may alter biofilm composition and function. We assessed effects of chronic N pollution on biofilm function by incubating initially thin (i.e., early stage) and well‐developed biofilms in laboratory chambers at four concentrations (0, 0.5, 5 and 25 mg N/L). For initially thin biofilms, elevated resulted in greater photosynthesis, higher chlorophyll contents and greater biomass accrual. Elevated also increased heterotrophic activity by increasing extracellular enzyme activity and heterotrophic respiration. However, well‐developed biofilms had lower growth and were less responsive to added elevated . Uptake kinetics showed a consistent decrease in cell‐specific maximum uptake (Umax) with elevated treatments. As a result, ambient uptake at experimental concentrations was essentially constant per unit photosynthesis across the range of concentration and uptake was apparently driven by biofilm growth. Across treatments, initially thin biofilms shifted from a diatom‐dominated community to a community dominated by green algae and cyanobacteria. However, well‐developed biofilms were not affected by chronic in terms of biomass or enzyme activities. Our results show that chronic loading alters biofilm growth patterns, physiology, algal community composition and relationship between algae and bacteria. Taken together, these results provide a set of mechanisms to explain the efficiency loss of uptake as a function of stream nitrogen concentration seen in field studies.

Effects of heavy metal species, concentrations, and speciation on pentachlorophenol sorption by river biofilms.

The sorption of trace organic pollutants at solid/liquid interfaces is one of the most important processes that influence their fate and behaviours in the aquatic environment. Sorption is affected by coexisting contaminants. The process and extent to which coexisting heavy metals affect the sorption of organochlorine pesticides (OCPs), especially acid radical anion heavy metals, are still unclear. Here, the effects of the species, concentrations, and speciation of the heavy metals Cu, Pb, and Cr, and the metalloid As on the sorption of pentachlorophenol (PCP), as a model OCP, by river biofilms were investigated through batch experiments. The results show that the presence of Cu, Pb, Cr, and As decreased the maximum sorption quantity of PCP onto the biofilms by 67.7, 9.2, 58.4, and 14.4%, respectively. The inhibitory effect of heavy metals on sorption decreased as the initial concentration ratios of heavy metals to PCP increased. In addition, the impact of heavy metals on PCP sorption was attributed to differences in heavy metal speciation. Cu and Pb commonly existed as divalent cations, but Cr and As existed as anionic acid radicals under the experimental conditions. The inhibitory effects of heavy metals on PCP sorption by biofilms were enhanced as the cation valence state increased, while the effects were weakened as the anionic acid radical valence state increased. Although all four heavy metals had inhibitory effects on PCP sorption by biofilms, there were distinct differences in the mechanisms causing these effects.