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.

Combined effects of eutrophication and warming on polyunsaturated fatty acids in complex phytoplankton communities: A mesocosm experiment.

Climate change and eutrophication are among the main stressors of shallow freshwater ecosystems, and their effects on phytoplankton community structure and primary production have been studied extensively. However, their combined effects on the algal production of polyunsaturated fatty acids (PUFA), specifically, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are currently unresolved. Moreover, the proximate reasons for changes in phytoplankton EPA and DHA concentrations are unclear, i.e., the relative importance of ecological (changes in the community composition) vs. ecophysiological (within taxa changes in EPA and DHA levels) factors. We investigated the responses of phytoplankton EPA and DHA concentrations to warming (IPCC climate scenario) and nutrient additions in mesocosms which had been run continuously at varying temperature and nutrient levels for 15 years prior to this study. Nutrient treatment had a significant effect on phytoplankton EPA and DHA concentrations and about 59 % of the variation in EPA and DHA concentrations could be explained by changes in the phytoplankton community structure. Increased biomass of diatoms corresponded with high EPA and DHA concentrations, while cyanobacteria/chlorophyte dominated mesocosm had low EPA and DHA concentrations. Warming had only a marginal effect on the EPA and DHA concentrations in these mesocosms. However, a significant interaction was observed with warming and N:P ratio. Our findings indicate that direct nutrient/temperature effects on algal physiology and PUFA metabolism were negligible and the changes in EPA and DHA concentrations were mostly related to the phytoplankton community structure and biomass. These results also imply that in shallow temperate lakes eutrophication, leading to increased dominance of cyanobacteria, will probably be a greater threat to phytoplankton EPA and DHA production than warming. EPA and DHA are nutritionally important for upper trophic level consumers and decreased production may impair secondary production.

Longitudinal variation in the nutritional quality of basal food sources and its effect on invertebrates and fish in subalpine rivers.

There is growing recognition of the importance of food quality over quantity for aquatic consumers. In streams and rivers, most previous studies considered this primarily in terms of the quality of terrestrial leaf litter and importance of microbial conditioning. However, many recent studies suggest that algae are a more nutritional food source for riverine consumers than leaf litter. To date, few studies have quantified longitudinal shifts in the nutritional quality of basal food resources in river ecosystems and how these may affect consumers. We conducted a field investigation in a subalpine river ecosystem in Austria to investigate longitudinal variations in diet quality of basal food sources (submerged leaves and periphyton) and diet source dependence of stream consumers (invertebrate grazers, shredders, filterers and predators, and fish). Fatty acid (FA) profiles of basal food sources and their consumers were measured. Our results indicate systematic differences between the FA profiles of terrestrial leaves and aquatic biota, that is periphyton, invertebrates and fish. Submerged leaves contained very low proportions of long-chain polyunsaturated fatty acids (LC-PUFAs), which were conversely rich in aquatic biota. While the FA composition of submerged leaves remained similar among sites, the LC-PUFAs of periphyton increased longitudinally, which was associated with increasing nutrients from upstream to downstream. Longitudinal variations in periphyton LC-PUFAs were reflected in the LC-PUFAs of invertebrate grazers and shredders, and further tracked by invertebrate predators and fish. However, brown trout Salmo trutta contained a large proportion of docosahexaenoic acid (DHA, 22:6ω3), a LC-PUFA almost entirely missing in basal sources and invertebrates. The fish accumulated eicosapentaenoic acid (EPA, 20:5ω3) from invertebrate prey and may use this FA to synthesize DHA. Our results provide a nutritional perspective for river food web studies, emphasizing the importance of algal resources to consumer somatic growth and the need to account for the longitudinal shifts in the quality of these basal resources.

Polyunsaturated fatty acids in fish tissues more closely resemble algal than terrestrial diet sources.

The River Continuum Concept implies that consumers in headwater streams have greater dietary access to terrestrial basal resources, but recent studies have highlighted the dietary importance of high-quality algae. Algae provide consumers with physiologically important omega-3 (n-3) polyunsaturated fatty acids (PUFA), particularly eicosapentaenoic acid (EPA). However, terrestrial plants and most benthic stream algae lack the long-chain (LC) n-3 PUFA docosahexaenoic acid (DHA, 22:6n-3), which is essential for neural development in fish and other vertebrates. We sampled subalpine streams to investigate how the PUFA composition of neural (brain and eyes), muscle, and liver tissues of freshwater fish is related to their potential diets (macroinvertebrates, epilithon, fresh and conditioned terrestrial leaves). The PUFA composition of consumers was more similar to epilithon than to terrestrial leaves. Storage lipids of eyes most closely resembled dietary PUFA (aquatic invertebrates and algae). However, DHA and arachidonic acid (ARA, 20:4n-6) were not directly available in the diet but abundant in organs. This implies that algal PUFA were selectively retained or were produced internally via enzymatic PUFA conversion by aquatic consumers. This field study demonstrates the nutritional importance of algal PUFA for neural organs in aquatic consumers of headwater regions.

Preferential retention of algal carbon in benthic invertebrates: Stable isotope and fatty acid evidence from an outdoor flume experiment.

According to the River Continuum Concept, headwater streams are richer in allochthonous (e.g. terrestrial leaves) than autochthonous (e.g. algae) sources of organic matter for consumers. However, compared to algae, leaf litter is of lower food quality, particularly ω-3 polyunsaturated fatty acids (n-3 PUFA), and would constrain the somatic growth, maintenance, and reproduction of stream invertebrates. It may be thus assumed that shredders, such as Gammarus, receive lower quality diets than grazers, e.g. Ecdyonurus, that typically feed on algae.The objective of this study was to assess the provision of dietary PUFA from leaf litter and algae to the shredder Gammarus and the grazer Ecdyonurus. Three different diets (algae, terrestrial leaves, and an algae-leaf litter mix) were supplied to these macroinvertebrates in a flume experiment for 2 weeks. To differentiate how diet sources were retained in these consumers, algae were isotopically labelled with 13C.Both consumers became enriched with 13C in all treatments, demonstrating that both assimilated algae. For Gammarus, n-3 PUFA increased, whereas n-6 PUFA stayed constant. By contrast, the n-3 PUFA content of Ecdyonurus decreased as a consequence of declining algal supply.Results from compound-specific stable isotope analysis provided evidence that the long-chain n-3 PUFA eicosapentaenoic acid (EPA) in both consumers was more enriched in 13C than the short-chain n-3 PUFA α-linolenic acid, suggesting that EPA was taken up directly from algae and not from heterotrophic biofilms on leaf litter. Both consumers depended on algae as their carbon and EPA source and retained their EPA from high-quality algae.

Fatty acids as dietary biomarkers in mangrove ecosystems: Current status and future perspective.

The paradigm that mangrove carbon supports secondary production in mangrove and adjacent habitats has been debated in recent years. Fatty acids (FA) are one of the classic biomarkers that have been frequently applied to track mangrove carbon pathways and assess trophic relationships. However, most previous studies did not evaluate the validity, potential and limitations of FA as biomarkers. The function and metabolism of long-chain polyunsaturated FA (LC-PUFA) in mangrove fauna have been largely ignored, and overlapping single FA biomarkers were widely used to infer dietary contributions from different sources. This review aims to systematically analyze and assess the application of FA biomarkers to dietary analyses in mangrove ecosystems, with a focus on basal food sources and their consumers. Our results show that basal food sources have distinctive FA profiles, with leaves and litter rich in alpha-linolenic acids (ALA, 18:3n-3), microphytobenthos rich in eicosapentaenoic acids (EPA, 20:5n-3) and suspended particulate organic matter (SPOM), or phytoplankton rich in docosahexaenoic acids (DHA, 22:6n-3). Most consumers contain high contents of LC-PUFA, particularly DHA and EPA, but very low levels of long-chain saturated FA (e.g., 22:0, 24:0, 26:0, 28:0), a biomarker of mangrove leaf litter. Bacterial FA biomarkers are present in all consumers. Four possible carbon pathways are identified and examined, i.e., benthic feeding on mangrove leaves and litter, benthic feeding on microphytobenthos, pelagic feeding on SPOM, and benthic and pelagic feeding on bacteria. Each pathway plays a different nutritional role for consumers, together providing a diversity of carbon sources. We recommend that in future (a) a wide range of basal diet sources should be sampled rather than just "visible" sources; (b) the unique FA characteristics of each diet source and consumer should be recognized with a focus on overall FA profiles and the application of multivariate statistics; (c) controlled feeding trials should be considered for keystone or functionally important consumers before selecting certain FA biomarkers to infer animal diets, and; (d) compound-specific stable isotope analysis should be applied to provide more insights into trophic relationships as well as the FA metabolic pathways in consumers.

The influence of alum based nutrient removal process on the physical, chemical and biological characteristics of phosphorus in the paper processing facility effluent.

This study examined the physical, chemical and biological characteristics of the phosphorus (P) in the effluents from a paper processing facility before and after an alum based P removal process. The alum based treatment process reduced effluent total P concentrations from an average of 230 ± 103 (±1 SD) µg L-1 to an average of 36 ± 10 µg L-1. Ultra-filtration showed the treatment process shifted the P in these effluents from being 75 ± 9% particulate pre-removal to being 52 ± 0% dissolved and 27 ± 1% colloidal after the Trident P removal process. The alum removal process also reduced the fraction of reactive P in the dissolved and colloidal pools from 55% to 14%, respectively. Algal phosphorus bioavailability (BAP) bioassays showed the BAP of these effluents averaged 12 ± 9% of total P. Algal dissolved P uptake experiments indicated ≈ 30% of the dissolved P was converted to particulate P (which usually indicates algal uptake) during 21-day bioassays. However, treatments without algae indicated >80% of the apparent algal uptake may have been due to colloidal P forming flocs via physical-chemical processes. These analyses indicate the phosphorus contained in this paper processing facility's effluents had much lower bioavailability than the P in typical municipal wastewater treatment plant effluents.

Evaluating coral trophic strategies using fatty acid composition and indices.

The ecological success of shallow water reef-building corals has been linked to the symbiosis between the coral host and its dinoflagellate symbionts (herein 'symbionts'). As mixotrophs, symbiotic corals depend on nutrients 1) transferred from their photosynthetic symbionts (autotrophy) and 2) acquired by host feeding on particulate organic resources (heterotrophy). However, coral species differ in the extent to which they depend on heterotrophy for nutrition and these differences are typically poorly defined. Here, a multi-tracer fatty acid approach was used to evaluate the trophic strategies of three species of common reef-building coral (Galaxea fascicularis, Pachyseris speciosa, and Pocillopora verrucosa) whose trophic strategies had previously been identified using carbon stable isotopes. The composition and various indices of fatty acids were compared to examine the relative contribution of symbiont autotrophy and host heterotrophy in coral energy acquisition. A linear discriminant analysis (LDA) was used to estimate the contribution of polyunsaturated fatty acids (PUFA) derived from various potential sources to the coral hosts. The total fatty acid composition and fatty acid indices revealed differences between the more heterotrophic (P. verrucosa) and more autotrophic (P. speciosa) coral hosts, with the coral host G. fascicularis showing overlap with the other two species and greater variability overall. For the more heterotrophic P. verrucosa, the fatty acid indices and LDA results both indicated a greater proportion of copepod-derived fatty acids compared to the other coral species. Overall, the LDA estimated that PUFA derived from particulate resources (e.g., copepods and diatoms) comprised a greater proportion of coral host PUFA in contrast to the lower proportion of symbiont-derived PUFA. These estimates provide insight into the importance of heterotrophy in coral nutrition, especially in productive reef systems. The study supports carbon stable isotope results and demonstrates the utility of fatty acid analyses for exploring the trophic strategies of reef-building corals.

Feeding strategies for the acquisition of high-quality food sources in stream macroinvertebrates: Collecting, integrating, and mixed feeding.

Aquatic macroinvertebrates play an important functional role in energy transfer in food webs, linking basal food sources to upper trophic levels that include fish, birds, and humans. However, the trophic coupling of nutritional quality between macroinvertebrates and their food sources is still poorly understood. We conducted a field study in subalpine streams in Austria to investigate how the nutritional quality (measured by long-chain polyunsaturated fatty acids, LC-PUFAs) in macroinvertebrates changes relative to their basal food sources. Samples of macroinvertebrates, periphyton, and leaves were collected from 17 streams in July and October 2016 and their fatty acid (FA) composition was analyzed. Periphyton FA varied strongly with time and space, and their trophic effect on macroinvertebrate FA differed among functional feeding groups. The match between periphyton FA and macroinvertebrate FA decreased with increasing trophic levels, but LC-PUFA content increased with each trophic step from periphyton to grazers and finally predators. Macroinvertebrates fed selectively on, assimilated, and/or actively controlled their LC-PUFA, especially eicosapentaenoic acid (EPA, 20 : 5ω3) relative to their basal food sources in the face of spatial and temporal changes. Grazer FA profiles reflected periphyton FA with relatively good fidelity, and especially their EPA feeding strategy was primarily linked to periphyton FA variation across seasons. In contrast, shredders appeared to preferentially assimilate more EPA over other FA, which was determined by the availability of high-quality food over seasons. Predators may more actively control their LC-PUFA distribution with respect to different quality foods and showed less fidelity to the basal FA profiles in plants and prey. Overall, grazers and shredders showed relatively good fidelity to food FA profiles and performed as both "collectors" and "integrators" for LC-PUFA requirements across seasons, while predators at higher trophic levels were more "integrators" with added metabolic complexity leading to somewhat more divergent FA profiles. These results are potentially applicable for other aquatic consumers in freshwater and marine ecosystems.

An assessment of assumptions and uncertainty in deuterium-based estimates of terrestrial subsidies to aquatic consumers.

The deuterium ratio (2 H/1 H) in tissue is often used to estimate terrestrial subsidies to aquatic consumers because of strongly differentiated values between terrestrial and aquatic primary producers. However, quantitative deuterium-based analyses of terrestrial resource assimilation are highly dependent on several poorly defined assumptions. We explored the sensitivity of these estimates to assumptions regarding environmental water contributions to consumer deuterium content (ω) and algal photosynthetic hydrogen discrimination (εH ). We also tested whether 13 C/12 C and 2 H/1 H-based estimates of terrestrial resource assimilation give similar outcomes. The average of the 12 experiments that have directly estimated proportional contributions of environmental water to consumer tissue 2 H/1 H was 0.27 ± 0.11 (mean ± SD), with similar values for invertebrates and fish. Conversely, of the 28 field studies that have used 2 H/1 H to characterize aquatic food webs, all but one assume a value that is less than our current best estimate, usually substantially less. A reanalysis of the raw data from four recent case studies indicates the calculated terrestrial contribution to aquatic consumers is extremely sensitive to this assumption. When the authors' original assumptions were used (i.e., ω = 0.16 ± 0.05), the estimated proportional contribution of terrestrial resources to aquatic consumers (θT ) averaged 29 ± 17%, and when ω = 0.27 was used the average estimated assimilation of allochthonous resources was ≈0.00. A compilation of published photosynthetic hydrogen discrimination values for microalgae averaged εH  = -150 ± 27‰ (SD, n = 99), and a sensitivity analysis showed the outcomes of these calculations were also strongly influenced by uncertainty in εH . There was no statistical association between 13 C/12 C and 2 H/1 H-based estimates of terrestrial subsidies (r = -0.12, n = 274). This analysis indicates that the assumptions in deuterium-based estimates of terrestrial resource assimilation are highly influential but poorly constrained; therefore, the impact of these assumptions on calculated outputs must be carefully assessed and thoroughly reported. Due to the highly uncertain assumptions inherent in deuterium-based analyses, we urge much more caution when using this approach to estimate terrestrial subsidies to consumers in aquatic ecosystems.

Quantifying learning in biotracer studies.

Mixing models have become requisite tools for analyzing biotracer data, most commonly stable isotope ratios, to infer dietary contributions of multiple sources to a consumer. However, Bayesian mixing models will always return a result that defaults to their priors if the data poorly resolve the source contributions, and thus, their interpretation requires caution. We describe an application of information theory to quantify how much has been learned about a consumer's diet from new biotracer data. We apply the approach to two example data sets. We find that variation in the isotope ratios of sources limits the precision of estimates for the consumer's diet, even with a large number of consumer samples. Thus, the approach which we describe is a type of power analysis that uses a priori simulations to find an optimal sample size. Biotracer data are fundamentally limited in their ability to discriminate consumer diets. We suggest that other types of data, such as gut content analysis, must be used as prior information in model fitting, to improve model learning about the consumer's diet. Information theory may also be used to identify optimal sampling protocols in situations where sampling of consumers is limited due to expense or ethical concerns.

The bioavailability of different dissolved organic nitrogen compounds for the freshwater algae Raphidocelis subcapitata.

Understanding which factors affect the algal bioavailability of dissolved organic nitrogen (DON) compounds in natural surface waters is important for our understanding of nutrient biogeochemistry and water quality management. We used nitrogen uptake kinetics and algal cell yield to characterize the algal bioavailability of 22 dissolved DON compounds that are commonly found in natural surface waters and wastewater treatment plant effluents, including urea, amino acids, amino sugars, nucleotides, pyrimidines, oraganonitriles, polyacrylamide, EDTA, caffeine, phenolic compounds and humic acids. Twelve of these compounds were highly bioavailable, including urea, dissolved free amino acids, bovine serum albumin, DNA, RNA, ATP, AMP, acetonitrile and caffeine. Four compounds had intermediate bioavailability including two humic acids (Elliott Soil and Pahokee Peat), glycylglycine, RNA and uracil. The remaining six compounds were classified as recalcitrant, i.e., EDTA, 2,3-Dinitrophenol, aminobenzoic acid, polyacrylamide and Aldrich humic acid. For many of the compounds tested, the algal cell yield was only 60-80% of expected relative to DON uptake. These results help explain why some DON compounds are more likely to persist in natural systems, and why the DON pool is often recalcitrant in surface waters.

Dissolved organic nitrogen recalcitrance and bioavailable nitrogen quantification for effluents from advanced nitrogen removal wastewater treatment facilities.

The objective of this study was to determine the composition of nitrogen (N) in the effluents of advanced N removal (ANR) wastewater treatment plants (WWTPs). This study also tested two different experimental protocols for determining dissolved N recalcitrance. An analysis of 15 effluent samples from five WWTPs, showed effluent concentrations and especially effluent composition varied greatly from one system to the other, with total nitrogen (TN) ranging between 1.05 and 8.10 mg L-1. Nitrate (NO3-) accounted for between 38 ± 32% of TN, and ammonium accounted for a further 29 ± 28%. All of these samples were dominated by dissolved inorganic nitrogen (DIN; NO3- + NH4+), and uptake experiments indicated the DIN fraction was as expected highly bioavailable. Dissolved organic N (DON) accounted for 20 ± 11% for the total dissolved N in these effluents, and uptake experiments indicated the bioavailability of this fraction varied between 27 ± 26% depending on the WWTP assessed. These results indicate near complete DIN removal should be the primary goal of ANR treatment systems. The comparison of bioavailable nitrogen (BAN) quantification protocols showed that the dissolved nitrogen uptake bioassay approach was clearly a more reliable way to determine BAN concentrations compared to the conventional cell yield protocol. Moreover, because the nitrogen uptake experiment was much more sensitive, this protocol made it easier to detect extrinsic factors (such as biological contamination or toxicity) that could affect the accuracy of these bioassays. Based on these results, we recommend the nitrogen uptake bioassay using filtered and autoclaved samples to quantify BAN concentrations. However, for effluent samples indicating toxicity, algal bioassays will not accurately quantify BAN.

Critical Uncertainties and Gaps in the Environmental- and Social-Impact Assessment of the Proposed Interoceanic Canal through Nicaragua.

The proposed interoceanic canal will connect the Caribbean Sea with the Pacific Ocean, traversing Lake Nicaragua, the major freshwater reservoir in Central America. If completed, the canal would be the largest infrastructure-related excavation project on Earth. In November 2015, the Nicaraguan government approved an environmental and social impact assessment (ESIA) for the canal. A group of international experts participated in a workshop organized by the Academy of Sciences of Nicaragua to review this ESIA. The group concluded that the ESIA does not meet international standards; essential information is lacking regarding the potential impacts on the lake, freshwater and marine environments, and biodiversity. The ESIA presents an inadequate assessment of natural hazards and socioeconomic disruptions. The panel recommends that work on the canal project be suspended until an appropriate ESIA is completed. The project should be resumed only if it is demonstrated to be economically feasible, environmentally acceptable, and socially beneficial.

Geomorphology controls the trophic base of stream food webs in a boreal watershed .

Abstract. Physical attributes of rivers control the quantity and quality of energy sources available to consumers, but it remains untested whether geomorphic conditions of whole watersheds affect the assimilation of different resources by stream organisms. We compared the fatty acid (FA) compositions of two invertebrate taxa (caddisflies, mayflies) collected from 16 streams in southwest Alaska, USA, to assess how assimilation of terrestrial organic matter (OM) and algae varied across a landscape gradient in watershed features. We found relatively higher assimilation of algae in high-gradient streams compared with low-gradient streams, and the opposite pattern for assimilation of terrestrial OM and microbes. The strength of these patterns was more pronounced for caddisflies than mayflies. Invertebrates from low-gradient watersheds had FA markers unique to methane-oxidizing bacteria and sulfate-reducing microbes, indicating a contribution of anaerobic pathways to primary consumers. Diversity of FA composition was highest in watersheds of intermediate slopes that contain both significant terrestrial inputs as well as high algal biomass. By controlling the accumulation rate and processing of terrestrial OM, watershed features influence the energetic base of food webs in boreal streams.

Characterization of the dissolved phosphorus uptake kinetics for the effluents from advanced nutrient removal processes.

Given the importance of the watershed protection plans, direct determination of phosphorus (P) mineralization rates in advanced wastewater treatment facility effluents is crucial for developing the most protective strategies minimiz eutrophication in receiving surface waters. In this study, algal bioassays were used to determine the uptake rate of dissolved P in effluents from a broad range of advanced nutrient removal technologies (e.g., membrane biological reactor, traditional biological, tertiary membrane, Blue PRO™, etc.). Dissolved P uptake kinetics were fit to a gamma model and three first-order decay models. A traditional one-pool model correlated poorly with the experimental data (i.e., r(2) = 0.73 ± 0.09), whereas two-pool model and three-pool models performed much better (i.e., r(2) > 0.9). These models also provided strong evidence for the existence of recalcitrant P in the effluents from these tertiary facilities. The Gamma model showed the mineralization of organic P followed a reactive continuum and further suggested the partitioning of P loads with different bioavailability levels should be accounted for the future modeling practices. From a modeling perspective, the Gamma model should be considered to be the theoretically best model as it gave the most parsimonious fit to the data using the fewest terms. Our study suggested that the current Total Maximum Daily Load (TMDL) model could be easily modified with the updated mineralization kinetics, which should lead to both ecological and economic benefits.

Selective transfer of polyunsaturated fatty acids from phytoplankton to planktivorous fish in large boreal lakes.

Lake size influences various hydrological parameters, such as water retention time, circulation patterns and thermal stratification that can consequently affect the plankton community composition, benthic-pelagic coupling and the function of aquatic food webs. Although the socio-economical (particularly commercial fisheries) and ecological importance of large lakes has been widely acknowledged, little is known about the availability and trophic transfer of polyunsaturated fatty (PUFA) in large lakes. The objective of this study was to investigate trophic trajectories of PUFA in the pelagic food web (seston, zooplankton, and planktivorous fish) of six large boreal lakes in the Finnish Lake District. Docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and α-linolenic acid (ALA) were the most abundant PUFA in pelagic organisms, particularly in the zooplanktivorous fish. Our results show that PUFA from the n-3 family (PUFAn-3), often associated with marine food webs, are also abundant in large lakes. The proportion of DHA increased from ~4±3% in seston to ~32±6% in vendace (Coregonus albula) and smelt (Osmerus eperlanus), whereas ALA showed the opposite trophic transfer pattern with the highest values observed in seston (~11±2%) and the lowest in the opossum shrimp (Mysis relicta) and fish (~2±1%). The dominance of diatoms and cryptophytes at the base of the food web in the study lakes accounted for the high amount of PUFAn-3 in the planktonic consumers. Furthermore, the abundance of copepods in the large lakes explains the effective transfer of DHA to planktivorous fish. The plankton community composition in these lakes supports a fishery resource (vendace) that is very high nutritional quality (in terms of EPA and DHA contents) to humans.

Inferring Phytoplankton, Terrestrial Plant and Bacteria Bulk δ¹³C Values from Compound Specific Analyses of Lipids and Fatty Acids.

Stable isotope mixing models in aquatic ecology require δ13C values for food web end members such as phytoplankton and bacteria, however it is rarely possible to measure these directly. Hence there is a critical need for improved methods for estimating the δ13C ratios of phytoplankton, bacteria and terrestrial detritus from within mixed seston. We determined the δ13C values of lipids, phospholipids and biomarker fatty acids and used these to calculate isotopic differences compared to the whole-cell δ13C values for eight phytoplankton classes, five bacterial taxa, and three types of terrestrial organic matter (two trees and one grass). The lipid content was higher amongst the phytoplankton (9.5±4.0%) than bacteria (7.3±0.8%) or terrestrial matter (3.9±1.7%). Our measurements revealed that the δ13C values of lipids followed phylogenetic classification among phytoplankton (78.2% of variance was explained by class), bacteria and terrestrial matter, and there was a strong correlation between the δ13C values of total lipids, phospholipids and individual fatty acids. Amongst the phytoplankton, the isotopic difference between biomarker fatty acids and bulk biomass averaged -10.7±1.1‰ for Chlorophyceae and Cyanophyceae, and -6.1±1.7‰ for Cryptophyceae, Chrysophyceae and Diatomophyceae. For heterotrophic bacteria and for type I and type II methane-oxidizing bacteria our results showed a -1.3±1.3‰, -8.0±4.4‰, and -3.4±1.4‰ δ13C difference, respectively, between biomarker fatty acids and bulk biomass. For terrestrial matter the isotopic difference averaged -6.6±1.2‰. Based on these results, the δ13C values of total lipids and biomarker fatty acids can be used to determine the δ13C values of bulk phytoplankton, bacteria or terrestrial matter with ± 1.4‰ uncertainty (i.e., the pooled SD of the isotopic difference for all samples). We conclude that when compound-specific stable isotope analyses become more widely available, the determination of δ13C values for selected biomarker fatty acids coupled with established isotopic differences, offers a promising way to determine taxa-specific bulk δ13C values for the phytoplankton, bacteria, and terrestrial detritus embedded within mixed seston.

A Fatty Acid Based Bayesian Approach for Inferring Diet in Aquatic Consumers.

We modified the stable isotope mixing model MixSIR to infer primary producer contributions to consumer diets based on their fatty acid composition. To parameterize the algorithm, we generated a 'consumer-resource library' of FA signatures of Daphnia fed different algal diets, using 34 feeding trials representing diverse phytoplankton lineages. This library corresponds to the resource or producer file in classic Bayesian mixing models such as MixSIR or SIAR. Because this library is based on the FA profiles of zooplankton consuming known diets, and not the FA profiles of algae directly, trophic modification of consumer lipids is directly accounted for. To test the model, we simulated hypothetical Daphnia comprised of 80% diatoms, 10% green algae, and 10% cryptophytes and compared the FA signatures of these known pseudo-mixtures to outputs generated by the mixing model. The algorithm inferred these simulated consumers were comprised of 82% (63-92%) [median (2.5th to 97.5th percentile credible interval)] diatoms, 11% (4-22%) green algae, and 6% (0-25%) cryptophytes. We used the same model with published phytoplankton stable isotope (SI) data for δ13C and δ15N to examine how a SI based approach resolved a similar scenario. With SI, the algorithm inferred that the simulated consumer assimilated 52% (4-91%) diatoms, 23% (1-78%) green algae, and 18% (1-73%) cyanobacteria. The accuracy and precision of SI based estimates was extremely sensitive to both resource and consumer uncertainty, as well as the trophic fractionation assumption. These results indicate that when using only two tracers with substantial uncertainty for the putative resources, as is often the case in this class of analyses, the underdetermined constraint in consumer-resource SI analyses may be intractable. The FA based approach alleviated the underdetermined constraint because many more FA biomarkers were utilized (n < 20), different primary producers (e.g., diatoms, green algae, and cryptophytes) have very characteristic FA compositions, and the FA profiles of many aquatic primary consumers are strongly influenced by their diets.