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Global change drives multiple facets of biodiversity including interaction diversity, which is fundamental for ecosystem functioning. However, studying trophic interactions is challenging in meta‐ecosystems, that is ecosystems connected by spatial flows of energy, materials and organisms across ecosystem boundaries. While analytical methods based on abundances of polyunsaturated fatty acids (PUFAs) and stable isotopes of amino acids (AAs) are being increasingly used, it has never been explored if both approaches could be: (i) combined in mixing models to enhance precision in dietary inference (ii) compared to disentangle transfers of various PUFAs and proteins in food webs in the wild.We explore the utility of analytical approaches based on PUFA abundances and AA isotopes to resolve resource transfers in a natural riparian food web. We focus on spiders and their potential prey from blue, green and brown sources to address three important and persisting methodological issues in food‐web ecology, namely whether (i) essential AA carbon isotopes can resolve protein origin from blue, green and brown resources, (ii) PUFA relative abundance and AA isotopes can be combined in a mixing model to provide higher precision estimates (i.e. narrower intervals) and (iii) combining the two approaches can unveil the coupling of protein and PUFA transfers in food webs.Our research demonstrates the power of AA isotopes and PUFAs to distinguish blue, green, and brown sources and their transfer up to consumers. We show that combining PUFA relative abundance and AA isotopes in a mixing model provides overall estimates similar to the individual estimates but significantly increases precision. In addition, we showcase how combining approaches unveil the coupling of protein and PUFA transfers. For instance, we show that most PUFAs are less concentrated from prey to predators, relative to proteins, highlighting uncoupling of PUFAs and protein transfer along food chains.We show for the first time the effectiveness of combining AA isotopes and PUFA abundances, particularly relevant for complex trophic interactions in a meta‐ecosystem context. Our study illustrates the trophic uncoupling of proteins and PUFAs, highlighting the necessity in combining both approaches. [ABSTRACT FROM AUTHOR]

Functional traits are measurable characteristics of an organism that have an impact on its fitness. Variation in functional traits between and among species has been suggested to represent the basis for competition and selection, thus allowing for evolution in natural populations.In freshwater ecosystems, the availability of essential polyunsaturated fatty acids (PUFAs), in particular ω3‐ and ω6‐PUFAs, determines the food quality of phytoplankton for the herbivorous zooplankton Daphnia, an unselective filter feeder. The content of such essential PUFAs in the phytoplankton is thus a functional phytoplankton trait affecting the trophic transfer efficiency and dynamics at the pelagic plant–herbivore interface.In turn, the susceptibility of consumers to become limited by the availability of essential PUFAs is a fitness‐determining trait of Daphnia genotypes, and variability of this herbivore trait may thus affect the daphnids' intrapopulation competition. To estimate the intrapopulation variation in susceptibility, we isolated clonal lines of Daphnia longispina from a natural population and compared the strength of their limitation by dietary PUFA availability via standardised laboratory growth assays. We used a liposome supplementation technique to enrich a PUFA‐poor green alga with essential ω3‐ and ω6‐PUFAs and determined juvenile somatic growth rate of different D. longispina genotypes as a fitness proxy.As expected, D. longispina genotypes that coexisted in a natural population differed markedly in their specific patterns of susceptibility to dietary PUFA availability. On average, the D. longispina population was more strongly susceptible to limitations in the availability of the ω6‐PUFA arachidonic acid (20:4ω6) than to limitations in the availability of ω3‐PUFAs α‐linolenic acid (18:3ω3) and eicosapentaenoic acid (20:5ω3).The ability to cope with PUFA limitation is thus a crucial trait that can probably affect intraspecific competition and Daphnia population structure. Therefore, we suggest that such intrapopulation variation in susceptibility to absence of dietary PUFAs might be one of the driving forces of natural selection and local adaptation among freshwater zooplankton. [ABSTRACT FROM AUTHOR]

Essential polyunsaturated fatty acids (PUFAs) have been recognized as a crucial factor that determines the trophic transfer efficiency in plankton communities. As many animals cannot synthesize the classes of ω3‐ and ω6‐PUFAs, the dietary availability of these PUFAs can constrain the fitness of freshwater zooplankton such as Daphnia spp. In particular, eicosapentaenoic acid (EPA, 20:5ω3) is considered to be a crucial determinant of the transfer of biomass at the freshwater plant‐herbivore interface. In contrast to ω3‐PUFAs, the group of ω6‐PUFAs has previously been considered to be of less ecological relevance, although the potential role of the ω6‐PUFA arachidonic acid (ARA, 20:4ω6) remains controversial. To investigate its potential role, we conducted dose‐response growth experiments with two Daphnia species, D. pulex and D. magna, supplemented with EPA or ARA, which allowed us to calculate EPA and ARA saturation thresholds for growth and reproduction of both Daphnia species. Our results provide evidence that not only the availability of ω3‐PUFAs, but also the availability of a ω6‐PUFA, namely ARA, can limit both the growth and reproduction of Daphnia spp. to an equal extent. The saturation thresholds for growth and reproduction were consistently, but not significantly, higher for EPA than for ARA in both Daphnia species. As shifts in phytoplankton community composition might result in environmental fluctuations in the dietary availability of ω3‐ and ω6‐PUFAs, our findings present a significant step in understanding the consequences of the ongoing global biodiversity loss for trophic transfer efficiency at the phytoplankton‐zooplankton interface. [ABSTRACT FROM AUTHOR]

For instance, algae often produce less polyunsaturated fatty acids (PUFAs) in warmer conditions (Fuschino et al., 2011; Von Elert & Fink, 2018), which can have implications for consumers, such as the herbivorous grazer I Daphnia i . Temperature reaction norms of Daphnia magna: The effect of food concentration. A test of the role of polyunsaturated fatty acids in phytoplankton food quality for Daphnia using liposome supplementation. Climate change threatens all ecosystems and the organisms within them. [Extracted from the article]