Exposure and recovery: The effect of different dilution factors of treated and untreated metal mining effluent on freshwater biofilm function and structure.

Abandoned mines generate effluents rich in heavy metals, and these contaminants are released uncontrolled into the nearby aquatic ecosystems, causing severe pollution. However, no real solution exists, leaving a legacy of global pollution. In this study, the efficiency of the treatment technologies in reducing the ecological impacts of mining effluents to freshwater ecosystems with different dilution capacities was tested using biofilm communities as biological indicators. The functional and structural recovery capacity of biofilm communities after 21 days of exposure was assessed. With this aim, we sampled aquatic biofilms from a pristine stream and exposed them to treated (T) and untreated (U) metal mining effluent from Frongoch abandoned mine (Mid Wales, UK). Additionally, we simulated two different flow conditions for the receiving stream: high dilution (HD) and low dilution (LD). After exposure, the artificial streams were filled with artificial water for 14 days to assess the biofilm recovery. Unexposed biofilm served as control for biofilm responses (functional and structural) measured throughout time. During the exposure, short term effects on biofilm functioning (photosynthetic efficiency, nutrient uptake) were observed in T-LD, U-HD, and U-LD, whereas long term effects (community composition, chl-a, and diatom metrics) were observed on the structure of all biofilms exposed to the treated and untreated mining effluent. On the other hand, metal accumulation occurred in biofilms exposed to the mining effluents. However, a functional recovery was observed for all treatments, except in the U-LD in which biofilm structure did not present a significant recovery after the exposure period. The results presented here highlight the need to consider the dilution capacity of the receiving stream to assess the real efficiency of treatment technologies applied to mining effluents to mitigate the ecological impact on freshwater ecosystems.

What happens when salinization meets eutrophication? A test using stream microcosms.

Nutrient and salt pollution often co-occur in rivers and streams due to human activities (e.g., agriculture, urbanization). Thus, understanding the interactive effects of nutrients and salinity on freshwater ecosystems is critical for environmental management. We experimentally assessed the interactive effects of nutrient and salt pollution on stream microcosms using biofilm and macroinvertebrates as model systems. Six treatments were performed in triplicate: control (C: N-NH4+ = 0.05; P- PO43- = 0.037; Cl- = 33.5 mg L-1), intermediate nutrient (IN: N-NH4+ = 0.4; P- PO43- = 0.271; Cl- = 33. 5 mg L-1), high nutrient (HN: N-NH4+ = 0.84; P- PO43- = 0.80; Cl- = 33.5 mg L-1), salt (S: N-NH4+ = 0.05; P- PO43- = 0.037; Cl- = 3000 mg L-1), salt with intermediate nutrient (SIN: N-NH4+ = 0.4; P- PO43- = 0.27; Cl- = 3000 mg L-1) and salt with high nutrient (SHN: N-NH4+ = 0.84; P- PO43- = 0.80; Cl- = 3000 mg L-1). After 14 days of exposure, biofilm chlorophyll-a increased across all treatments, with cyanobacteria replacing diatoms and green algae. Treatments with no added nutrients (C and S) had more P uptake capacity than the rest. The indicator species analysis showed 8 significant taxa, with Orthocladius (Orthocladius) gr. Wetterensis and Virganytarsus significantly associated with the salinity treatment. Overall, salt pollution led to a very strong decline in macroinvertebrate richness and diversity. However, salt toxicity seemed to be ameliorated by nutrient addition. Finally, both structural equation models and biotic-abiotic interaction networks showed that complex biological interactions could be modulating the response of the biological communities to our treatments. Thus, our study calls for species-level assessments of salt and nutrient effects on river ecosystems and advocates for better management of co-occurring pollutants.

Dissipation of pesticides by stream biofilms is influenced by hydrological histories.

To evaluate the effects of hydrological variability on pesticide dissipation capacity by stream biofilms, we conducted a microcosm study. We exposed biofilms to short and frequent droughts (daily frequency), long and less frequent droughts (weekly frequency) and permanently immersed controls, prior to test their capacities to dissipate a cocktail of pesticides composed of tebuconazole, terbuthylazine, imidacloprid, glyphosate and its metabolite aminomethylphosphonic acid. A range of structural and functional descriptors of biofilms (algal and bacterial biomass, extracellular polymeric matrix (EPS) concentration, microbial respiration, phosphorus uptake and community-level physiological profiles) were measured to assess drought effects. In addition, various parameters were measured to characterise the dynamics of pesticide dissipation by biofilms in the different hydrological treatments (% dissipation, peak asymmetry, bioconcentration factor, among others). Results showed higher pesticide dissipation rates in biofilms exposed to short and frequent droughts, despite of their lower biomass and EPS concentration, compared to biofilms in immersed controls or exposed to long and less frequent droughts. High accumulation of hydrophobic pesticides (tebuconazole and terbuthylazine) was measured in biofilms despite the short exposure time (few minutes) in our open-flow microcosm approach. This research demonstrated the stream biofilms capacity to adsorb hydrophobic pesticides even in stressed drought environments.

Experimental assessment of salinization effects on freshwater zooplankton communities and their trophic interactions under eutrophic conditions.

Freshwater ecosystems are becoming saltier due to human activities. The effects of increased salinity can lead to cascading trophic interactions, affecting ecosystem functioning and energy transfer, through changes in community and size structure. These effects can be modulated by other environmental factors, such as nutrients. For example, communities developed under eutrophic conditions could be less sensitive to salinization due to cross-tolerance mechanisms. In this study, we used a mesocosm approach to assess the effects of a salinization gradient on the zooplankton community composition and size structure under eutrophic conditions and the cascading effects on algal communities. Our results showed that zooplankton biomass, size diversity and mean body size decreased with increased chloride concentration induced by salt addition. This change in the zooplankton community did not have cascading effects on phytoplankton. The phytoplankton biomass decreased after the chloride concentration threshold of 500 mg L-1 was reached, most likely due to direct toxic effects on the osmotic regulation and nutrient uptake processes of certain algae rather than as a response to community turnover or top-down control. Our study can help to put in place mitigation strategies for salinization and eutrophication, which often co-occur in freshwater ecosystems.

Hypersaline mining effluents affect the structure and function of stream biofilm.

The salinisation of freshwater ecosystems is a global environmental problem that threatens biodiversity, ecosystem functioning and human welfare. The aim of this study was to investigate the potential impact of a realistic salinity gradient on the structure and functioning of freshwater biofilms. The salinity gradient was based on the real ion concentration of a mining effluent from an abandoned mine in Germany. We exposed biofilm from a pristine stream to 5 increasing salinities (3 to 100 g L-1) under controlled conditions in artificial streams for 21 days. We evaluated its functional (photosynthetic efficiency, nutrient uptake, and microbial respiration) and structural responses (community composition, algal biomass and diatom, cyanobacteria and green algae metrics) over time. Then we compared their responses with an unexposed biofilm used as control. The functionality and structure of the biofilm exposed to the different salinities significantly decreased after short-term and long-term exposure, respectively. The community composition shifted to a new stable state where the most tolerant species increased their abundances. At the same time, we observed an increase in the community tolerance (measured as Pollution-Induced Community Tolerance) along the salinity gradient. This study provides relevant information on the salt threshold concentrations that can substantially damage algal cells (i.e., between 15 and 30 g L-1). The results provide new insights regarding the response and adaptation of stream biofilm to salinity and its potential implications at the ecosystem level.

Effects of an hypersaline effluent from an abandoned potash mine on freshwater biofilm and diatom communities.

Potash abandoned mines cause severe environmental damage to their bordering environment, with significant impacts on freshwater ecosystems mostly through uncontrolled discharge of hypersaline effluents. This study aimed to evaluate the ecological impact caused by a hypersaline effluent from an abandoned potash mine (Menteroda, Germany) on freshwater biofilms and, specifically, on diatom communities. Biofilm from a pristine stream was exposed under controlled conditions in microcosms to a mining effluent (ME), and its structural (algal biomass, community composition, diatom metrics) and functional (photosynthetic activity, nutrient uptake) responses were evaluated over time and compared with unexposed biofilms used as control. Biofilm exposed to ME showed drastic functional responses after one day of exposure, with a significant decrease in photosynthetic efficiency and nutrient uptake, that were recovered over time. Biofilm exposed to ME showed a progressive increase in diatom metrics (abundance, density and growth rate) over time, compared to the control. However, a significant decrease in diatom species diversity, richness and cell size was also observed in biofilm exposed to ME. This study revealed that the ME affected the biofilm causing short-term functional responses, which were recovered simultaneously with a drastic diatom community structure shift.

Effects of the interaction between nutrient concentration and DIN:SRP ratio on geosmin production by freshwater biofilms.

The global increase of cyanobacterial blooms occurrence has been associated with the presence of compounds that generate earthy and musty odor in freshwater systems, among which geosmin stands out. The lack of information on the factors associated to geosmin production by benthic organisms has driven the development of this study, whose main goal is to determine the effects of nutrient concentration and DIN:SRP ratio on geosmin formation and release. The experiment was performed in 18 microcosms under controlled conditions for 21 days, using a natural biofilm suspension from Ter river (NE, Spain) to promote biofilm settlement. Six treatments were set crossing three DIN:SRP ratios (A = 4:1, B = 16:1 and C = 64:1) with two nutrient concentrations (Low and High). After 7 days of experiment, geosmin was detected in biofilm, being higher under high nutrient concentration and low DIN:SRP ratio conditions. In this treatment, geosmin in biofilm reached its maximum concentration at day 16 (3.8 ± 0.9 ng/mg), decreasing at the end of the experiment (21d) due to cyanobacteria detachment and geosmin release into the water (136 ± 6 ng/L). Overall, this experimental study showed that high nutrient concentration and low DIN:SRP ratio favored the Oscillatoria genus development within biofilm communities, generating the optimal conditions for geosmin production. The interaction between these two factors was demonstrated to be a potential driver of benthic geosmin production and release, and should be monitored and controlled in rivers exploited for drinking water purposes.

Driving Factors of Geosmin Appearance in a Mediterranean River Basin: The Ter River Case.

In recent decades, human activity coupled with climate change has led to a deterioration in the quality of surface freshwater. This has been related to an increase in the appearance of algal blooms, which can produce organic compounds that can be toxic or can affect the organoleptic characteristics of the water, such as its taste and odor. Among these latter compounds is geosmin, a metabolite produced by certain cyanobacteria that confers an earthy taste to water and which can be detected by humans at very low concentrations (nanogram per liter). The difficulty and cost of both monitoring the presence of this compound and its treatment is a problem for drinking water treatment companies, as the appearance of geosmin affects consumer confidence in the quality of the drinking water they supply. In this field study, the evaluation of four sampling sites with different physicochemical conditions located in the upper part of the Ter River basin, a Mediterranean river located in Catalonia (NE Spain), has been carried out, with the aim of identifying the main triggers of geosmin episodes. The results, obtained from 1 year of sampling, have made it possible to find out that: (i) land uses with a higher percentage of agricultural and industrial activity are related to high nutrient conditions in river water, (ii) these higher nutrient concentrations favor the development of benthic cyanobacteria, (iii) in late winter-early spring, when these cyanobacteria are subjected to both an imbalance of the dissolved inorganic nitrogen and soluble reactive phosphorus ratio, guided by a phosphorus concentration increase, and to cold-mild temperatures close to 10°C, they produce and release geosmin, and (iv) 1-2 weeks after cyanobacteria reach a high relative presence in the whole biofilm, an increase in geosmin concentration in water is observed, probably associated with the cyanobacteria detachment from cobbles and consequent cell lysis. These results could serve as a guide for drinking water treatment companies, indicating under what conditions they can expect the appearance of geosmin episodes and implement the appropriate treatment before it reaches consumers' tap.

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.

Quality and quantity of leaf litter: Both are important for feeding preferences and growth of an aquatic shredder.

The study of leaf litter as a resource for shredders has emerged as a key topic in trophic links in ecology. However, thus far, most studies have emphasized the leaf quality as one of the main determinants of shredder behaviour and growth without simultaneously considering the leaf quantity availability. Nevertheless, the combined effects of leaf quantity and quality on shredder behaviour and growth is particularly crucial to further understand how ecosystem functioning may respond to the increasing flow intermittency due to climate change. In this study, we explore how changes in the leaf litter quality and quantity influence the feeding preferences and growth of an invertebrate shredder (Potamophylax latipennis). To do so, we used black poplar leaves conditioned in two streams with different flow regimens as a food resource. Afterwards, using a microcosm approach, we offered leaf discs that varied in terms of leaf quantity and quality to P. latipennis. Our results showed that flow intermittency had a negative effect on the quality of the food resource, and a lower quality had a negative effect on the consumption and growth rates of P. latipennis. Furthermore, we found that P. latipennis fed selectively on higher quality leaves even though the availability (quantity) of this resource was lower. In the context of climate change, with higher aridity/drier conditions/scenarios, our findings suggest that a decrease in the availability (quantity) of high-quality resources could potentially threaten links in global fluvial food webs and thus threaten ecosystem functioning.

Heterogeneity in leaf litter decomposition in a temporary Mediterranean stream during flow fragmentation.

In temporary Mediterranean streams, flow fragmentation during summer droughts originates an ephemeral mosaic of terrestrial and aquatic habitat types. The heterogeneity of habitat types implies a particular ecosystem functioning in temporary streams that is still poorly understood. We assessed the initial phases of leaf litter decomposition in selected habitat types: running waters, isolated pools and moist and dry streambed sediments. We used coarse-mesh litter bags containing Populus nigra leaves to examine decomposition rates, microbial biomass, macroinvertebrate abundance and dissolved organic carbon (DOC) release rates in each habitat type over an 11-day period in late summer. We detected faster decomposition rates in aquatic (running waters and isolated pools) than in terrestrial habitats (moist and dry streambed sediments). Under aquatic conditions, decomposition was characterized by intense leaching and early microbial colonization, which swiftly started to decompose litter. Microbial colonization in isolated pools was primarily dominated by bacteria, whereas in running waters fungal biomass predominated. Under terrestrial conditions, leaves were most often affected by abiotic processes that resulted in small mass losses. We found a substantial decrease in DOC release rates in both aquatic habitats within the first days of the study, whereas DOC release rates remained relatively stable in the moist and dry sediments. This suggests that leaves play different roles as a DOC source during and after flow fragmentation. Overall, our results revealed that leaf decomposition is heterogeneous during flow fragmentation, which has implications related to DOC utilization that should be considered in future regional carbon budgets.

Effects of water flow regulation on ecosystem functioning in a Mediterranean river network assessed by wood decomposition.

Mediterranean rivers are extensively modified by flow regulation practises along their courses. An important part of the river impoundment in this area is related to the presence of small dams constructed mainly for water abstraction purposes. These projects drastically modified the ecosystem morphology, transforming lotic into lentic reaches and increasing their alternation along the river. Hydro-morphologial differences between these reaches indicate that flow regulation can trigger important changes in the ecosystem functioning. Decomposition of organic matter is an integrative process and this complexity makes it a good indicator of changes in the ecosystem. The aim of this study was to assess the effect caused by flow regulation on ecosystem functioning at the river network scale, using wood decomposition as a functional indicator. We studied the mass loss from wood sticks during three months in different lotic and lentic reaches located along a Mediterranean river basin, in both winter and summer. Additionally, we identified the environmental factors affecting decomposition rates along the river orders. The results revealed differences in decomposition rates between sites in both seasons that were principally related to the differences between stream orders. The rates were mainly related to temperature, nutrient concentrations (NO2(-), NO3(2-)) and water residence time. High-order streams with higher temperature and nutrient concentrations exhibited higher decomposition rates compared with low-order streams. The effect of the flow regulation on the decomposition rates only appeared to be significant in high orders, especially in winter, when the hydrological characteristics of lotic and lentic habitats widely varied. Lotic reaches with lower water residence time exhibited greater decomposition rates compared with lentic reaches probably due to more physical abrasion and differences in the microbial assemblages. Overall, our study revealed that in high orders the reduction of flow caused by flow regulation affects the wood decomposition indicating changes in ecosystem functioning.