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.

The evaluations of oxidative stress and neurotoxicity in threatened endemic fish Barbus meridionalis from Osor River (Spain).

Mediterranean barbel (Barbus meridionalis) an endemic species is currently facing habitat destruction and pollution in Osor River (Spain) due to mining runoff that has severely deteriorated the water quality by metals, primarily zinc (Zn). In order to assess the potential risk of metal contamination and hydrological changes in the Osor River by using oxidative stress and neurotoxicity biomarkers via IBR analyses in the barbel, five different stations were chosen: upstream (S1: reference site and S2: hydrologically changed), mine (S3), and downstream (S4 and S5). The highest tissue metal levels were measured particularly at S3 and following downstream sites. SOD activity and the GSH system parameters (GPX, GST, and GSH) were the most sensitive oxidative stress indicators among the antioxidant system parameters. The organs with the greatest changes in antioxidant biomarkers were the liver and gill. As a sign of neurotoxicity, AChE activities significantly raised in the brain and muscle but drastically lowered in the kidney, liver, and gill particularly in the area of mine and downstream compared to reference site. Integrated biomarker response index (IBR) method was applied to visualize the affect of metal and hydrological alterations with biomarker response according to sites in the Osor River. IBR analyzes together with correlations between metal levels and oxidative stress biomarkers, emphasized that S2 and S3 have the greatest impact on the biomarker levels due to mine activity and hydrological changes highlighting the vulnerability to extinction of native fish B. meridionalis. It is also critical to assess the current data based on the multi-biomarker approach for a range of detrimental effects on fish fitness at the individual level as well as population persistence from an ecological standpoint.

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.

Key factors explaining critical swimming speed in freshwater fish: a review and statistical analysis for Iberian species.

Swimming performance is a key feature that mediates fitness and survival in aquatic animals. Dispersal, habitat selection, predator-prey interactions and reproduction are processes that depend on swimming capabilities. Testing the critical swimming speed (Ucrit) of fish is the most straightforward method to assess their prolonged swimming performance. We analysed the contribution of several predictor variables (total body length, experimental water temperature, time step interval between velocity increments, species identity, taxonomic affiliation, native status, body shape and form factor) in explaining the variation of Ucrit, using linear models and random forests. We compiled in total 204 studies testing Ucrit of 35 inland fishes of the Iberian Peninsula, including 17 alien species that are non-native to that region. We found that body length is largely the most important predictor of Ucrit out of the eight tested variables, followed by family, time step interval and species identity. By contrast, form factor, temperature, body shape and native status were less important. Results showed a generally positive relationship between Ucrit and total body length, but regression slopes varied markedly among families and species. By contrast, linear models did not show significant differences between native and alien species. In conclusion, the present study provides a first comprehensive database of Ucrit in Iberian freshwater fish, which can be thus of considerable interest for habitat management and restoration plans. The resulting data represents a sound foundation to assess fish responses to hydrological alteration (e.g. water flow tolerance and dispersal capacities), or to categorize their habitat preferences.

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.