Increasing water nutrient reduces the availability of high-quality food resources for aquatic consumers and consequently simplifies river food webs.

While eutrophication has led to serious habitat degradation and biotic shifts in freshwater ecosystems, most current studies have focused on changes in community assemblages, with few considering the effect of eutrophication on food webs. We conducted a field study in subtropical headwater streams with a gradient of nutrient levels to examine the effect of increasing water nutrients on food webs by using the long-chain polyunsaturated fatty acid eicosapentaenoic acid (EPA) as a measure of the nutritional quality of food. Basal food resources (macrophytes, submerged leaf litter, and periphyton), and aquatic consumers (macroinvertebrates and fish) were collected, and their fatty acid (FA) profiles were analyzed. Our results showed that periphyton was the dominant source of EPA for macroinvertebrates and fish, and a high-quality resource for consumers. As water nutrient concentrations increased, nutritional quality of periphyton significantly decreased and, in turn, the correlation between FA profiles of periphyton and macroinvertebrates declined. However, periphyton FA profiles did not account for the variability of fish FA, which may be induced by the increasing proportions of omnivorous fish in eutrophic streams that derive EPA from other sources. Further, the reduced periphyton EPA was associated with decreased trophic links and simplified stream food webs. Our study highlights the importance of high-quality food resources for aquatic food webs as water nutrients increase in stream ecosystems and provides a nutritional perspective to understand the mechanisms how eutrophication affects aquatic ecosystems.

How do small dams alter river food webs? A food quality perspective along the aquatic food web continuum.

Damming of rivers poses a significant threat to freshwater ecosystems. Previous studies about the impact of damming on river ecosystems have mostly focused on large dams, with the impact of small dams largely unknown. Further, while the impacts of dams on aquatic communities have been widely studied, the effect on energy flow across river food webs remains unclear. In recent years, long-chain polyunsaturated fatty acid analysis (LC-PUFA) has emerged as a promising technique for assessing food quality and trophic interactions. In this study, LC-PUFA was applied to explore the nutritional effects of small dams on river food webs. A field investigation was conducted at upstream and downstream areas of three small dams in the headwaters of Dongjiang River, China, to evaluate the impact of small dams on the nutritional quality of basal food sources, and their consequent impacts on aquatic consumers and trophic links. Basal food sources (i.e., submerged leaves, macrophytes and periphyton) and aquatic consumers (i.e., macroinvertebrates and fish) were collected, and their fatty acid (FA) composition was measured. Our results showed that periphyton, rather than submerged leaves and macrophytes, was the primary high-quality food source for aquatic consumers, providing them with LC-PUFA, irrespective of whether sites were upstream or downstream. Damming the streams induced changes in aqueous nutrient concentrations (TP, PO4-P, DIN, and TN) from upstream to downstream of the dams, leading to significant variation in periphyton FA content. Compared with periphyton collected at downstream sites, periphyton at upstream sites contained higher LC-PUFA, but lower short-chain PUFA. Differences in periphyton LC-PUFA between the upstream and downstream areas of dams were reflected in the FA profiles of invertebrate grazers and filterers, and further transferred to fish. Furthermore, decreased periphyton nutritional quality at the downstream of the dams was one of the reasons for the simplification of stream food webs. Our results indicated that small dams negatively affected food webs, emphasizing the importance of high-quality food sources for stream ecosystems. We suggest that the trophic integrity of river food webs hinges on the dietary availability of periphyton supplying physiologically highly required nutrients for consumers and must thus not be compromised by damming of streams or other alterations.

The chytrid insurance hypothesis: integrating parasitic chytrids into a biodiversity-ecosystem functioning framework for phytoplankton-zooplankton population dynamics.

In temperate lakes, eutrophication and warm temperatures can promote cyanobacteria blooms that reduce water quality and impair food-chain support. Although parasitic chytrids of phytoplankton might compete with zooplankton, they also indirectly support zooplankton populations through the "mycoloop", which helps move energy and essential dietary molecules from inedible phytoplankton to zooplankton. Here, we consider how the mycoloop might fit into the biodiversity-ecosystem functioning (BEF) framework. BEF considers how more diverse communities can benefit ecosystem functions like zooplankton production. Chytrids are themselves part of pelagic food webs and they directly contribute to zooplankton diets through spore production and by increasing host edibility. The additional way that chytrids might support BEF is if they engage in "kill-the-winner" dynamics. In contrast to grazers, which result in "eat-the-edible" dynamics, kill-the-winner dynamics can occur for host-specific infectious diseases that control the abundance of dominant (in this case inedible) hosts and thus limit the competitive exclusion of poorer (in this case edible) competitors. Thus, if phytoplankton diversity provides functions, and chytrids support algal diversity, chytrids could indirectly favour edible phytoplankton. All three mechanisms are linked to diversity and therefore provide some "insurance" for zooplankton production against the impacts of eutrophication and warming. In our perspective piece, we explore evidence for the chytrid insurance hypothesis, identify exceptions and knowledge gaps, and outline future research directions.

Anomalous Temperature Effect in Weakly Coupled Superlattices: Carrier Transport in a THz Quantum Cascade Laser.

We present an investigation into the vertical transport through 13 different superlattice structures, where the well and barrier widths, doping concentration, dopant position, and contact layers were varied. Although superlattices have been extensively studied since 1970, there is a lack of publications on transport through superlattices similarly low doped as THz quantum cascade lasers (QCLs), for which the doping is in the 3-5×10^{10} cm^{-2} range. The superlattices presented are doped in the same range as THz QCLs, with contact layers and fabrication comparable to high-temperature THz QCLs. The temperature-dependent current-voltage characteristics were measured starting from 5 K and an anomalous temperature effect was observed at the first plateau. The measured current through the superlattice first decreases before increasing again with increasing temperature, resulting in the lowest current occurring at 75-110 K. This behavior is also observed in some THz QCLs. The effect disappears for thinner barriers, higher quantum well doping, or modified contact layers, indicating a strong dependency on band bending, due to the large difference in the doping of the contact layers and the superlattice, which is confirmed with multiscattering Büttiker simulations.

Evaluation of lipid extraction methods for fatty acid quantification and compound-specific δ13C and δ2Hn analyses.

Lipids, with fatty acids (FA) as a crucial subset, have become a focal point for diverse medical, physiological, and ecological studies. However, a comprehensive assessment of the various pre-analytical FA extraction methods published in the scientific literature remains lacking. In this study, we examined the efficacy of seven well-established sample preparation methods, specifically focusing on their effectiveness in total lipid and fatty acid extraction and their impact on compound-specific stable hydrogen (δ2H) and carbon (δ13C) isotope values. We also considered the repercussions of FA removal efficacy on residual bulk tissue δ2Hn analysis, because lipids typically have low δ2H values. Our findings showed that in most cases chloroform-based extraction methods outperformed those without chloroform. While discrepancies were not as evident for smaller organisms, such as plankton, marked variations were discernible in the extraction efficiencies for muscle and liver samples, which was also manifested in the residual bulk tissue δ2Hn results. Notably, most extraction methods had little effect on specific δ13C or δ2H isotope values of FA; instead, an emphasis should be on using an extraction method that achieves optimal baseline peak separation of the chromatograms for C and H isotope measurements.

Eutrophication and loss of riparian shading influence food quality and trophic relation in stream food webs.

Eutrophication induced by excessive inputs of nutrient is one of the main stressors in aquatic ecosystems. Deforestation in riparian zones alter riparian shading, which together with eutrophication is expected to exert a complex control over stream food webs. We manipulated two levels of riparian shading (open canopy vs. shading canopy) and nutrient supply (ambient vs. nutrient addition) in three headwater streams to investigate the individual and combined effects of eutrophication and loss of riparian shading on carbon sources and nutritional quality of biofilms, and the subsequent trophic effects on macroinvertebrate grazers. Nutrient enrichment increased the autochthonous carbon (i.e., algae especially diatoms) indicated by fatty acid (FA) biomarkers within biofilms and grazers. The nutritional quality indicated by eicosapentaenoic acid (EPA) content of biofilms was increased with nutrient enrichment and more so with the combined effect of an increase in riparian shading, consequently leading to an increase in the nutritional quality, density, and biomass of grazers. In particular, the trophic linkages between biofilms and grazers were mainly influenced by EPA concentration in the biofilms, and strengthened with the combined effects of riparian shading and additional nutrients. Our study emphasizes the nutritional significance of EPA for consumers at higher trophic levels and proposes its potential as an indicator for monitoring the health of aquatic ecosystems.

The importance of omega-3 polyunsaturated fatty acids as high-quality food in freshwater ecosystems with implications of global change.

Traditionally, trophic ecology research on aquatic ecosystems has focused more on the quantity of dietary energy flow within food webs rather than food quality and its effects on organisms at various trophic levels. Recent studies emphasize that food quality is central to consumer growth and reproduction, and the importance of food quality for aquatic ecosystems has become increasingly well recognized. It is timely to synthesise these findings and identify potential future research directions. We conducted a systematic review of omega-3 polyunsaturated fatty acids (ω3-PUFAs) as a crucial component of high-quality food sources in freshwater ecosystems to evaluate their impact on a variety of consumers, and explore the effects of global change on these high-quality food sources and their transfer to higher trophic consumers within and across ecosystems. In freshwater ecosystems, algae rich in ω3 long-chain PUFAs, such as diatoms, dinoflagellates and cryptophytes, represent important high-quality food sources for consumers, whereas cyanobacteria, green algae, terrestrial vascular plants and macrophytes low in ω3 long-chain PUFAs are low-quality food sources. High-quality ω3-PUFA-containing food sources usually lead to increased growth and reproduction of aquatic consumers, e.g. benthic invertebrates, zooplankton and fish, and also provide ω3 long-chain PUFAs to riparian terrestrial consumers via emergent aquatic insects. Consumers feeding on high-quality ω3-PUFA-containing foods in turn represent high-quality food for their own predators. However, the ω3-PUFA content of food sources is sensitive to global environmental changes. Warming, eutrophication, increased light intensity (e.g. from loss of riparian shading), and pollutants potentially inhibit the synthesis of algal ω3-PUFAs while at the same time promoting the growth of lower-quality foods, such as cyanobacteria and green algae. These factors combined could lead to a significant reduction in the availability of ω3-PUFAs for consumers and constrain their overall fitness. Although the effect of individual environmental factors on high-quality ω3-PUFA-containing food sources has been investigated, multiple environmental factors (e.g. climate change, human activities, pollution) will act in combination and any synergistic effects on aquatic food webs remain unclear. Identifying the sources and fate of ω3-PUFAs within and across ecosystems could represent an important approach to understand the impact of multiple environmental factors on trophic relationships and the implications for populations of freshwater and riparian consumers. Maintaining the availability of high-quality ω3-PUFA-containing food sources may also be key to mitigating freshwater biodiversity loss due to global change.

Dietary fatty acid transfer in pelagic food webs across trophic and climatic differences of Chinese lakes.

In eutrophic lake ecosystems, cyanobacteria typically lead to unbalanced phytoplankton community structure and low dietary quality for consumers at higher trophic levels. However, it still remains poorly understood how zooplankton manage to respond to seasonal and spatial differences in lake trophic gradients and temperature factors to retain highly required dietary nutrients from phytoplankton. In this field study, we investigated seston and different size classes of zooplankton of temperate and subtropical large lakes of different trophic conditions in China. We used fatty acids (FA) as dietary nutrients from seston to zooplankton to investigate how eutrophication affects the FA composition of various zooplankton size classes. This study revealed a curvilinear relationship between total phosphorus (TP) and polyunsaturated fatty acids (PUFA) contents of edible phytoplankton ("seston") across 3 seasons and 2 climatic areas. The PUFA content of seston increased until mesotrophic lake conditions (TP: 11-20 µg L-1), after which the dietary provision of PUFA for respective consumers declined. Seston FA, rather than trophic condition or water temperature, primarily predicted changes in zooplankton FA, while this predictive power decreased with zooplankton size. Despite increasing eutrophic lake conditions, LC-PUFA content of the zooplankton consistently increased per unit biomass. The results indicate that the nutritional value of phytoplankton was highest in mesotrophic lakes, and lake zooplankton selectively increased their LC-PUFA retention with body size and/or were able to convert dietary FA endogenously to meet their size-specific FA demands, independent of lake location or time (season) or the measured trophic condition of the lake (from oligo- to eutrophic).

Protocols for sample preparation and compound-specific stable-isotope analyses (δ2H, δ13C) of fatty acids in biological and environmental samples.

Compound-specific stable-isotope analysis (CSIA) of fatty acids is a powerful tool to better understand the trophic transfer of fatty acids and their biochemical fate in and across ecosystems, including tracing animal migration and understanding physiological processes. The non-exchangeable nature of C-H bonds in acyl chains, hydrogen (δ2H) and carbon (δ13C) stable-isotope values of fatty acids (FA) provide independent information about the origins of fatty acids. Several technical obstacles must be overcome to ensure accurate and reproducible measurements of FA-CSIA can be made. This protocol describes the sample preparation process for successful stable-isotope analyses of fatty acids obtained from environmental and biological samples. Numerous techniques for the preanalytical processing of fatty acid samples are available, and these often have minimal impact on δ values. Here, we provide an in-depth guide detailing our well-established laboratory protocols, ranging from the initial sample preparation, lipid extraction, and transmethylation to the instrumental arrangement, data collection, and analysis.•Protocol from obtaining a sample to standardized fatty acid specific δ2H and δ13C values.•Separate GC analysis procedures for C and H are recommended for optimal performance.

Investigating the diet source influence on freshwater fish mercury bioaccumulation and fatty acids-Experiences from Swedish lakes and Chinese reservoirs.

Dietary uptake is key for transferring potentially toxic contaminants, such as mercury (Hg) and essential dietary nutrients, such as polyunsaturated fatty acids (PUFA), to consumers at higher trophic levels of aquatic food webs. We evaluated the role of diet sources for Hg bioaccumulation and PUFA retention in fish across lake food webs in seven Swedish lakes and two Chinese reservoirs. Fish total Hg (THg) and methyl-Hg (MeHg) differed greatly between the two countries: the Chinese fish contained less than 300 ng g-1 dry weight (d.w.) THg with less than 50% as MeHg, versus the Swedish fishes which contained approximately 2000 ng g-1 d.w. THg and nearly 100% as MeHg. Fatty acids enrichment of linoleic acids (LIN) were more prevalent in the Chinese fishes regardless of size (p < 0.05). Here we examined food web length, fish growth rates, and fatty acids patterns in relation to the quality of fish as a food source for both Hg and FA. Contrary to the expectation that biodilution of Hg throughout the food chain would explain these differences, a more complex picture emerged with high levels of Hg at the base of the food web in the Chinese reservoirs, a decoupling of fatty acid and Hg bioaccumulation, and a major role for both fish stocking and fish feed. It is hoped that this work will provide a nuanced picture of fish quality as a food source in different ecosystems.

Preparing for the future offspring: European perch (Perca fluviatilis) biosynthesis of physiologically required fatty acids for the gonads happens already in the autumn.

Long-chain polyunsaturated fatty acids (PUFA) are critical for reproduction and thermal adaptation. Year-round variability in the expression of fads2 (fatty acid desaturase 2) in the liver of European perch (Perca fluviatilis) in a boreal lake was tested in relation to individual variation in size, sex, and maturity, together with stable isotopes values as well as fatty acids (FA) content in different tissues and prey items. ARA and DHA primary production was restricted to the summer months, however, perch required larger amounts of these PUFA during winter, as their ARA and DHA muscle content was higher compared to summer. The expression of fads2 in perch liver increased during winter and was higher in mature females. Mature females stored DHA in their gonads already in late summer and autumn, long before the upcoming spring spawning period in May. Lower δ13CDHA values in the gonads in September suggest that these females actively synthesized DHA as part of this reproductive investment. Lower δ13CARA values in the liver of all individuals during winter suggest that perch were synthesizing essential FA to help cope with over-wintering conditions. Perch seem able to modulate its biosynthesis of physiologically required PUFA in situations of stress (fasting or cold temperatures) or in situations of high energetic demand (gonadal development). Biosynthesis of physiologically required PUFA may be an important part of survival and reproduction in aquatic food webs with long cold periods.

Assessment of the impact of dams on aquatic food webs using stable isotopes: Current progress and future challenges.

Dams have disrupted natural river systems worldwide and although population and community level effects on aquatic biota have been well documented, food web responses remain poorly understood and difficult to characterize. The application of stable isotope analysis (SIA) provides a means to assess the effect of dams on food webs. Here we review the effect of dams on aquatic food webs using SIA, aiming to detect knowledge gaps in the field of dam impacts on aquatic food webs and propose a conceptual framework to help formulate hypotheses about dam impacts on food webs guided by food web theory. Dams can affect aquatic food webs via two pathways: a bottom-up pathway with altered basal food sources and their transfer to consumers through changes in flow, nutrients, temperature and sediment, and a top-down pathway with consumer species composition altered mainly through habitat fragmentation and related physiochemical changes. Taking these mechanisms into consideration, the impact of dams on food web attributes derived from SIA was evaluated. These studies generally apply mixing models to determine how dams alter the dominant carbon sources supporting food webs, use δ15N to examine how dams alter food-chain length, or use Layman metrics of isotope variability to assess niche changes for invertebrate and fish assemblages. Most studies compare the patterns of SIA metrics spatially (e.g. upstream vs reservoir vs downstream of dams; regulated vs unregulated rivers) and temporally (before vs after dam construction), without explicit hypotheses and/or links to theoretical concepts of food webs. We propose several steps to make SIA studies of dam impacts more rigorous and enhance their potential for producing novel insights. Future studies should quantify the shape and strength of the effect of dams on SIA-measured food web response, be conducted at larger temporal and spatial scales (particularly along the river longitudinal continuum and the lateral connected ecosystems (e.g., floodplains)), and consider effects of dams on food web resilience and tipping points.

Export of dietary lipids via emergent insects from eutrophic fishponds.

Fishponds, despite being globally abundant, have mainly been considered as food production sites and have received little scientific attention in terms of their ecological contributions to the surrounding terrestrial environment. Emergent insects from fishponds may be important contributors of lipids and essential fatty acids to terrestrial ecosystems. In this field study, we investigated nine eutrophic fishponds in Austria from June to September 2020 to examine how Chlorophyll-a concentrations affect the biomass of emergent insect taxa (i.e., quantity of dietary subsidies; n = 108) and their total lipid and long-chain polyunsaturated fatty acid content (LC-PUFA, i.e., quality of dietary subsidies; n = 94). Chironomidae and Chaoboridae were the most abundant emergent insect taxa, followed by Trichoptera, Ephemeroptera, and Odonata. A total of 1068 kg of emergent insect dry mass were exported from these ponds (65.3 hectares). Chironomidae alone exported 103 kg of total lipids and 9.4 kg of omega-3 PUFA. Increasing Chl-a concentrations were associated with decreasing biomass export and a decrease in total lipid and LC-PUFA export via emergent Chironomidae. The PUFA composition of emergent insect taxa differed significantly from dietary algae, suggesting selective PUFA retention by insects. The export of insect biomass from these eutrophic carp ponds was higher than that previously reported from oligotrophic lakes. However, lower biomass and diversity are exported from the fishponds compared to managed ponds. Nonetheless, our data suggest that fishponds provide crucial ecosystem services to terrestrial consumers by contributing essential dietary nutrients to consumer diets via emergent insects. Supplementary Information: The online version contains supplementary material available at 10.1007/s10750-022-05040-2.

Response of stable isotopes (δ2H, δ13C, δ15N, δ18O) of lake water, dissolved organic matter, seston, and zooplankton to an extreme precipitation event.

Lake ecosystems process and cycle organic substrates, thus serving as important bioreactors in the global carbon cycle. Climate change is predicted to increase extreme weather and precipitation events that can flush nutrients and organic matter from soils to streams and lakes. Here we report changes in stable isotopes (δ2H, δ13C, δ15N, or δ18O) of water, dissolved organic matter (DOM), seston, and zooplankton in a subalpine lake at short time resolution following an extreme precipitation event between early July to mid-August 2021. Water from excess precipitation and runoff remained in the lake epilimnion and coincided with increasing δ13C values of seston (-30 ‰ to -20 ‰), due to the input of carbonates and terrestrial organic matter. Particles settled into deeper lake layers after two days and contributed to the uncoupling of C and N cycling as the lake responded to this extreme precipitation event. Following the event, there was an increase in bulk δ13C values of zooplankton (from -35 ‰ to -32 ‰). Throughout this study, δ13C values of DOM remained stable throughout the water column (-29 ‰ to -28 ‰), while large isotopic fluctuations in DOM δ2H (-140 ‰ to -115 ‰) and δ18O (+9 ‰ to +15 ‰) values suggested DOM relocation and turnover. Integrating isotope hydrology, ecosystem ecology, and organic geochemistry offers an element-specific, detailed approach to investigating the impact of extreme precipitation events on freshwater ecosystems and particularly aquatic food webs.

Chytrids alleviate the harmful effect of heat and cyanobacteria diet on Daphnia via PUFA-upgrading.

Chytrid fungal parasites increase herbivory and dietary access to essential molecules, such as polyunsaturated fatty acids (PUFA), at the phytoplankton-zooplankton interface. Warming enhances cyanobacteria blooms and decreases algae-derived PUFA for zooplankton. Whether chytrids could support zooplankton with PUFA under global warming scenarios remains unknown. We tested the combined effect of water temperature (ambient: 18°C, heat: +6°C) and the presence of chytrids with Daphnia magna as the consumer, and Planktothrix rubescens as the main diet. We hypothesized that chytrids would support Daphnia fitness with PUFA, irrespective of water temperature. Heating was detrimental to the fitness of Daphnia when feeding solely on the Planktothrix diet. Chytrid-infected Planktothrix diet alleviated the negative impact of heat and could support Daphnia survival, somatic growth and reproduction. Carbon stable isotopes of fatty acids highlighted a ~3x more efficient n-3 than n-6 PUFA conversion by Daphnia feeding on the chytrid-infected diet, irrespective of temperature. The chytrid diet significantly increased eicosapentaenoic acid (EPA; 20:5n-3) and arachidonic acid (ARA; 20:4n-6) retention in Daphnia. The EPA retention remained unaffected, while ARA retention increased in response to heat. We conclude that chytrids support pelagic ecosystem functioning under cyanobacteria blooms and global warming via chytrids-conveyed PUFA toward higher trophic levels.

Chytrids enhance Daphnia fitness by selectively retained chytrid-synthesised stearidonic acid and conversion of short-chain to long-chain polyunsaturated fatty acids.

Chytrid fungal parasites convert dietary energy and essential dietary molecules, such as long-chain (LC) polyunsaturated fatty acids (PUFA), from inedible algal/cyanobacteria hosts into edible zoospores. How the improved biochemical PUFA composition of chytrid-infected diet may extend to zooplankton, linking diet quality to consumer fitness, remains unexplored.Here, we assessed the trophic role of chytrids in supporting dietary energy and PUFA requirements of the crustacean zooplankton Daphnia, when feeding on the filamentous cyanobacterium Planktothrix.Only Daphnia feeding on chytrid-infected Planktothrix reproduced successfully and had significantly higher survival and growth rates compared with Daphnia feeding on the sole Planktothrix diet. While the presence of chytrids resulted in a two-fold increase of carbon ingested by Daphnia, carbon assimilation increased by a factor of four, clearly indicating enhanced carbon transfer efficiency with chytrid presence.Bulk carbon (δ 13C) and nitrogen (δ 15N) stable isotopes did not indicate any treatment-specific dietary effects on Daphnia, nor differences in trophic position among diet sources and the consumer. Compound-specific carbon isotopes of fatty acids (δ 13CFA), however, revealed that chytrids bioconverted short-chain to LC-PUFA, making it available for Daphnia. Chytrids synthesised the ω-3 PUFA stearidonic acid de novo, which was selectively retained by Daphnia. Values of δ 13CFA demonstrated that Daphnia also bioconverted short-chain to LC-PUFA.We provide isotopic evidence that chytrids improved the dietary provision of LC-PUFA for Daphnia and enhanced their fitness. We argue for the existence of a positive feedback loop between enhanced Daphnia growth and herbivory in response to chytrid-mediated improved diet quality. Chytrids upgrade carbon from the primary producer and facilitate energy and PUFA transfer to primary consumers, potentially also benefitting upper trophic levels of pelagic food webs.

Strong temporal variation of consumer δ13C value in an oligotrophic reservoir is related to water level fluctuation.

Using stable carbon and nitrogen isotope analysis (δ13C and δ15N) to assess trophic interactions in freshwater ecosystems is a well established method, providing insight into ecosystem functioning. However, the spatial and temporal variability of isotope values, driven by environmental fluctuation is poorly understood and can complicate interpretations. We investigated how the temporal variation of stable isotopes in consumers (fish, crayfish and macrozoobenthos) of a canyon-shaped oligotrophic reservoir is associated with environmental factors such as water temperature, transparency, flooded area, and water quality measures. Consumers and their putative food sources were sampled and analyzed for carbon and nitrogen stable isotopes annually, and environmental parameters were measured monthly from 2014 to 2016. Results revealed significant differences in δ13C and δ15N values in each consumer among studied years. Over the years, fish and crayfish expressed differences in δ13C between 3 and 5‰, whereas in zoobenthos differences were 12‰. Variability in δ15N was similar across all consumers (2-4‰). Moreover, results suggest that the flooded area of the reservoir was a major driver of δ13C stable isotope values variation in consumers, while variation in δ15N was not linked to any of the studied environmental factors. Bayesian mixing models further showed significant changes in the origin of detritivorous zoobenthos carbon sources (reversal shift from terrestrial detritus to algae origin) between years with low water level to years with the standard water level. Other species showed only slight differences in food source utilization among years. Our study highlights the importance of environmental factors as sources of variation in consumer's stable isotope values which should be considered especially when studied ecosystem strongly fluctuate in some environmental factor.

Mucilage protects the planktonic desmid Staurodesmus sp. against parasite attack by a chytrid fungus.

Zoosporic fungi of the phylum Chytridiomycota are ubiquitous parasites of phytoplankton in aquatic ecosystems, but little is known about phytoplankton defense strategies against parasitic chytrid attacks. Using a model chytrid-phytoplankton pathosystem, we experimentally tested the hypothesis that the mucilage envelope of a mucilage-forming desmid species provides protection against the parasitic chytrid Staurastromyces oculus. Mucilage-forming Staurodesmus cells were not accessible to the chytrid, whereas physical removal of the mucilage envelope rendered the same Staurodesmus sp. strain equally susceptible to chytrid infections as the original non-mucilage-forming host Staurastrum sp. Epidemic spread of the parasite only occurred in Staurastrum sp., whereas non-mucilage-bearing Staurodesmus sp. allowed for co-existence of host and parasite, and mucilage-bearing Staurodesmus sp. caused parasite extinction. In addition to the mucilage defense barrier, we also demonstrate the ability of both Staurastrum sp. and Staurodesmus sp. to resist infection by preventing chytrid development while still remaining viable and being able to reproduce and thus recover from an infection. This study extends our knowledge on phytoplankton defense traits and the functional role of mucilage in phytoplankton as a physical barrier against fungal parasites.

Metal Exposure and Sex Shape the Fatty Acid Profile of Midges and Reduce the Aquatic Subsidy to Terrestrial Food Webs.

Aquatic micropollutants can be transported to terrestrial systems and their consumers by emergent aquatic insects. However, micropollutants, such as metals, may also affect the flux of physiologically important polyunsaturated fatty acids (PUFAs). As certain PUFAs have been linked to physiological fitness and breeding success of terrestrial consumers, reduced fluxes from aquatic systems could affect terrestrial populations and food webs. We chronically exposed larvae of the aquatic insect Chironomus riparius to a range of environmentally relevant sediment contents of cadmium (Cd) or copper (Cu) in a 28-day microcosm study. Since elevated water temperatures can enhance metals' toxic effects, we used two temperature regimes, control and periodically elevated temperatures (heat waves) reflecting an aspect of climate change. Cd and Cu significantly reduced adult emergence by up to 95% and 45%, respectively, while elevated temperatures had negligible effects. Both metal contents were strongly reduced (∼90%) during metamorphosis. Furthermore, the chironomid FA profile was significantly altered during metamorphosis with the factors sex and metal exposure being relevant predictors. Consequently, fluxes of physiologically important PUFAs by emergent adults were reduced by up to ∼80%. Our results suggest that considering fluxes of physiologically important compounds, such as PUFAs, by emergent aquatic insects is important to understand the implications of aquatic micropollutants on aquatic-terrestrial meta-ecosystems.

Reproducible measurements of the δ2H composition of non-exchangeable hydrogen in complex organic materials using the UniPrep2 online static vapour equilibration and sample drying system.

Non-exchangeable hydrogen-isotope (δ 2Hn) measurements of complex organic samples are used in forensics to determine sample authenticity, traceability, and provenance. However, δ 2Hn assays of organics are usually complicated by uncontrolled "exchangeable hydrogen" and residual moisture contamination; hence, δ 2Hn assays are persistently incomparable amongst laboratories. We introduce a revised technical solution (UniPrep2) to control hydrogen-isotope exchange and for robust online sample drying and vapour equilibration. The UniPrep2 device is coupled to a high-temperature thermochemical elemental analyser and continuous-flow isotope-ratio mass spectrometer. This technical solution empowers isotope analysts to:•Conduct reproducible controlled vapour equilibrations of complex organic samples and standards to determine the δ 2Hn values by controlling hydrogen-isotope exchange.•Conduct online vacuum-oven evacuation with extensive helium drying without exposure to air to reabsorb or exchange hydrogen with ambient water vapour. The protocol describes the operation of the UniPrep2 device and the step-by-step procedures needed to obtain accurate and precise δ 2Hn values for a wide range of organic sample types. Two analytical approaches are described in detail; the Dual-Vapour Equilibration (DVE) approach, intended for determining δ 2Hn for a complex organic environmental sample where matrix equivalent H isotope reference materials are not available, and the Comparative Equilibration (CE) approach, which is intended for routine high-throughput analyses of complex organic samples where at least two matrix-equivalent organic isotope reference materials with consensus δ 2Hn values are being used. These standard operating procedures are envisioned to be a sound basis for advancing hydrogen-isotope analysis for different organic environmental matrices and studies.