Influence of the dietary contribution of terrestrial insects to the condition factor of bleak Alburnus alburnus in a highly polluted lowland river.

Bleak Alburnus alburnus is a highly abundant but understudied fish species, and we know little about the trophic ecology of populations inhabiting rivers in central Europe. From an ecosystem perspective, this fish species is interesting as it is known to feed on surface insects, thereby linking the terrestrial with the aquatic habitat. In a previous study, we demonstrated that this flux is intensified, and dietary contribution of terrestrial insects is higher in fish inhabiting sections of the Spree River, Germany, that are polluted from iron oxides occurring from former lignite mining activities, and thus are characterized by lower abundances of aquatic insects. As terrestrial insects can be considered as food of lower quality (measured as long-chained polyunsaturated fatty acids, n-3 LC-PUFAs) compared to aquatic prey, it is reasonable to assume that the higher contribution of terrestrial insects is related to a lower body condition in fish. In this study, we explore the trophic ecology of riverine A. alburnus and their fitness consequences of feeding on terrestrial insects. We therefore modeled a terrestrial index from stable isotopes of hydrogen (δ2H) measured in the A. alburnus muscle tissue and compared individuals caught in locations upstream of a dam that were greatly influenced by iron oxides, with individuals caught in sections located downstream of a dam where passive remediation technologies are applied. The terrestrial index was significantly higher in A. alburnus caught in locations at high-iron concentrations, characterized by low abundances of aquatic prey, compared to A. alburnus caught in unpolluted habitats at low-iron concentrations. In contradiction to our hypothesis, the terrestrial index had no significant effect on the body condition of A. alburnus (measured as Fulton's condition factor K) in the sections downstream of the dam (i.e., at low-iron concentrations) and a significant positive, albeit weak, effect in sections upstream of the dam (i.e., at high-iron concentrations). However, the condition factor was generally lower in the high-iron section, potentially related to more direct effects of the iron oxide. We conclude that in A. alburnus, terrestrial insects can be considered as the less-favored food, unless the fish occur in environments where the aquatic food is of limited availability. Further research is needed to evaluate the direct and indirect effects, including the internal n-3 LC-PUFA synthesis as an adaption toward low-quality terrestrial prey on the fitness consequences of A. alburnus.

The evolutionary ecology of fatty-acid variation: Implications for consumer adaptation and diversification.

The nutritional diversity of resources can affect the adaptive evolution of consumer metabolism and consumer diversification. The omega-3 long-chain polyunsaturated fatty acids eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) have a high potential to affect consumer fitness, through their widespread effects on reproduction, growth and survival. However, few studies consider the evolution of fatty acid metabolism within an ecological context. In this review, we first document the extensive diversity in both primary producer and consumer fatty acid distributions amongst major ecosystems, between habitats and amongst species within habitats. We highlight some of the key nutritional contrasts that can shape behavioural and/or metabolic adaptation in consumers, discussing how consumers can evolve in response to the spatial, seasonal and community-level variation of resource quality. We propose a hierarchical trait-based approach for studying the evolution of consumers' metabolic networks and review the evolutionary genetic mechanisms underpinning consumer adaptation to EPA and DHA distributions. In doing so, we consider how the metabolic traits of consumers are hierarchically structured, from cell membrane function to maternal investment, and have strongly environment-dependent expression. Finally, we conclude with an outlook on how studying the metabolic adaptation of consumers within the context of nutritional landscapes can open up new opportunities for understanding evolutionary diversification.

Short-term apparent mutualism drives responses of aquatic prey to increasing productivity.

According to apparent competition theory, sharing a predator should cause indirect interactions among prey that can affect the structure and the dynamics of natural communities. Though shifts in prey dominance and predator resource use along environmental gradients are rather common, empirical evidence on the role of indirect prey-prey interactions through shared predation particularly with increasing productivity, is still scarce. In an 8-week lake mesocosm experiment, we manipulated both the addition of inorganic nutrients and the presence of generalist fish predators (crucian carp, Carassius carassius L.), to test for the effects of indirect interactions through shared predation along a productivity gradient. We found that apparent mutualism (indirect positive interaction) between benthic and pelagic prey strongly affected short-term responses of aquatic food webs to increasing productivity in the presence of a generalist fish. Increasing productivity favoured the relative abundance of benthic prey, following trends in natural productive lake systems. This led to a shift in fish selectivity from pelagic to benthic prey driven by changes in fish behaviour, which resulted in apparent mutualism due to the lower and delayed top-down control of pelagic prey at increasing productivity. Our results show empirical evidence that the coupling of multiple production pathways can lead to strong indirect interactions through shared predation, whereby prey dynamics on short time-scales are highly dependent on the foraging behaviour of generalist predators. This mechanism may play an important role in short-term responses of food webs across environmental gradients.

Fatty acid accumulation in feeding types of a natural freshwater fish population.

Fatty acids are widely used to study trophic interactions in food web assemblages. Generally, it is assumed that there is a very small modification of fatty acids from one trophic step to another, making them suitable as trophic biomarkers. However, recent literature provides evidence that many fishes possess genes encoding enzymes with a role in bioconversion, thus the capability for bioconversion might be more widespread than previously assumed. Nonetheless, empirical evidence for biosynthesis occurring in natural populations remains scarce. In this study, we investigated different feeding types of perch (Perca fluviatilis) that are specialized on specific resources with different levels of highly unsaturated fatty acids (HUFAs), and analyzed the change between HUFA proportions in perch muscle tissue compared to their resources. Perch showed matching levels to their resources for EPA, but ARA and especially DHA were accumulated. Compound-specific stable isotope analyses helped us to identify the origin of HUFA carbon. Our results suggest that perch obtain a substantial amount of DHA via bioconversion when feeding on DHA-poor benthic resources. Thus, our data indicate the capability of bioconversion of HUFAs in a natural freshwater fish population.

The CO2 -equivalent balance of freshwater ecosystems is non-linearly related to productivity.

Eutrophication of fresh waters results in increased CO2 uptake by primary production, but at the same time increased emissions of CH4 to the atmosphere. Given the contrasting effects of CO2 uptake and CH4 release, the net effect of eutrophication on the CO2 -equivalent balance of fresh waters is not clear. We measured carbon fluxes (CO2 and CH4 diffusion, CH4 ebullition) and CH4 oxidation in 20 freshwater mesocosms with 10 different nutrient concentrations (total phosphorus range: mesotrophic 39 µg/L until hypereutrophic 939 µg/L) and planktivorous fish in half of them. We found that the CO2 -equivalent balance had a U-shaped relationship with productivity, up to a threshold in hypereutrophic systems. CO2 -equivalent sinks were confined to a narrow range of net ecosystem production (NEP) between 5 and 19 mmol O2  m-3  day-1 . Our findings indicate that eutrophication can shift fresh waters from sources to sinks of CO2 -equivalents due to enhanced CO2 uptake, but continued eutrophication enhances CH4 emission and transforms freshwater ecosystems to net sources of CO2 -equivalents to the atmosphere. Nutrient enrichment but also planktivorous fish presence increased productivity, thereby regulating the resulting CO2 -equivalent balance. Increasing planktivorous fish abundance, often concomitant with eutrophication, will consequently likely affect the CO2 -equivalent balance of fresh waters.

Empirical correspondence between trophic transfer efficiency in freshwater food webs and the slope of their size spectra.

The density of organisms declines with size, because larger organisms need more energy than smaller ones and energetic losses occur when larger organisms feed on smaller ones. A potential expression of density-size distributions are Normalized Biomass Size Spectra (NBSS), which plot the logarithm of biomass independent of taxonomy within bins of logarithmic organismal size, divided by the bin width. Theoretically, the NBSS slope of multi-trophic communities is exactly -1.0 if the trophic transfer efficiency (TTE, ratio of production rates between adjacent trophic levels) is 10% and the predator-prey mass ratio (PPMR) is fixed at 104 . Here we provide evidence from four multi-trophic lake food webs that empirically estimated TTEs correspond to empirically estimated slopes of the respective community NBSS. Each of the NBSS considered pelagic and benthic organisms spanning size ranges from bacteria to fish, all sampled over three seasons in 1 yr. The four NBSS slopes were significantly steeper than -1.0 (range -1.14 to -1.19, with 95% CIs excluding -1). The corresponding average TTEs were substantially lower than 10% in each of the four food webs (range 1.0% to 3.6%, mean 1.85%). The overall slope merging all biomass-size data pairs from the four systems (-1.17) was almost identical to the slope predicted from the arithmetic mean TTE of the four food webs (-1.18) assuming a constant PPMR of 104 . Accordingly, our empirical data confirm the theoretically predicted quantitative relationship between TTE and the slope of the biomass-size distribution. Furthermore, we show that benthic and pelagic organisms can be merged into a community NBSS, but future studies have yet to explore potential differences in habitat-specific TTEs and PPMRs. We suggest that community NBSS may provide valuable information on the structure of food webs and their energetic pathways, and can result in improved accuracy of TTE-estimates.

Effects of mining activities on fish communities and food web dynamics in a lowland river.

Fish communities of streams and rivers might be substantially subsidized by terrestrial insects that fall into the water. Although such animal-mediated fluxes are increasingly recognized, little is known about how anthropogenic perturbations may influence the strength of such exchanges. Intense land use, such as lignite mining, may impact a river ecosystem due to the flocculation of iron (III) oxides, thus altering food web dynamics. We compared sections of the Spree River in North-East Germany that were greatly influenced by iron oxides with sections located downstream of a dam where passive remediation technologies are applied. Compared to locations downstream of the dam, the abundance of benthic macroinvertebrates at locations of high iron concentrations upstream of the dam was significantly reduced. Similarly, catch per unit effort of all fish was significantly higher in locations downstream of the dam compared to locations upstream of the dam, and the condition of juvenile and adult piscivorous pike Esox lucius was significantly lower in sections of high iron concentrations. Using an estimate of short-term (i.e., metabarcoding of the gut content) as well as longer-term (i.e., hydrogen stable isotopes) resource use, we could demonstrate that the three most abundant fish species, perch Perca fluviatilis, roach Rutilus rutilus, and bleak Alburnus alburnus, received higher contributions of terrestrial insects to their diet at locations of high iron concentration. In summary, lotic food webs upstream and downstream of the dam greatly differed in the overall structure with respect to the energy available for the highest tropic levels and the contribution of terrestrial insects to the diet of omnivorous fish. Therefore, human-induced environmental perturbations, such as river damming and mining activities, represent strong pressures that can alter the flow of energy between aquatic and terrestrial systems, indicating a broad impact on the landscape level.

Effects of predation pressure and resource use on morphological divergence in omnivorous prey fish.

BACKGROUND: Body shape is one of the most variable traits of organisms and responds to a broad array of local selective forces. In freshwater fish, divergent body shapes within single species have been repeatedly observed along the littoral-pelagic axes of lakes, where the structural complexity of near shore habitats provides a more diverse set of resources compared to the open-water zones. It remains poorly understood whether similar resource-driven polymorphism occurs among lakes that vary in structural complexity and predation pressure, and whether this variation is heritable. Here, we analyzed body shape in four populations of omnivorous roach (Rutilus rutilus) inhabiting shallow lakes. We tested the relationship between body shape, gradients of resources, predation pressure, and, in a subset of two lakes, diet composition. We used genome scans of 331 polymorphic AFLP markers to test whether there was a heritable component to the observed morphological diversification. RESULTS: Body shape differed among lakes and was significantly correlated to differences in predation pressure. Roach from the lake with highest predation pressure were most divergent from the average body shape of all populations, characterized by a more streamlined body and caudally inserted dorsal fins; features that facilitate predator escape. Surprisingly, diet composition was not associated with morphology. AFLP analysis revealed weak genetic differentiation among lakes and no isolation by distance (IBD). Outlier analysis detected three loci under positive selection with differing frequencies in the four populations. General linear models did not support an association of lake-specific genotypes with morphological variation. CONCLUSION: Body shape was divergent among lakes, suggesting that processes previously reported from within single lakes may also be operating at the scale of whole lakes. We found no evidence for body shape being heritable, although sample size was small in these natural populations. Rather than habitat structure and diet, we conclude that predation had a stronger effect on the prevalence of local morphotypes. A variable morphotype facilitating the efficient uptake of a variety of spatially and temporarily scattered resources seems to be favored in these small aquatic systems.

The role of vital dietary biomolecules in eco-evo-devo dynamics.

The physiological dependence of animals on dietary intake of vitamins, amino acids, and fatty acids is ubiquitous. Sharp differences in the availability of these vital dietary biomolecules among different resources mean that consumers must adopt a range of strategies to meet their physiological needs. We review the emerging work on omega-3 long-chain polyunsaturated fatty acids, focusing predominantly on predator-prey interactions, to illustrate that trade-off between capacities to consume resources rich in vital biomolecules and internal synthesis capacity drives differences in phenotype and fitness of consumers. This can then feedback to impact ecosystem functioning. We outline how focus on vital dietary biomolecules in eco-eco-devo dynamics can improve our understanding of anthropogenic changes across multiple levels of biological organization.

The interaction between metabolic rate, habitat choice, and resource use in a polymorphic freshwater species.

Resource polymorphism is common across taxa and can result in alternate ecotypes with specific morphologies, feeding modes, and behaviors that increase performance in a specific habitat. This can result in high intraspecific variation in the expression of specific traits and the extent to which these traits are correlated within a single population. Although metabolic rate influences resource acquisition and the overall pace of life of individuals it is not clear how metabolic rate interacts with the larger suite of traits to ultimately determine individual fitness.We examined the relationship between metabolic rates and the major differences (habitat use, morphology, and resource use) between littoral and pelagic ecotypes of European perch (Perca fluviatilis) from a single lake in Central Sweden.Standard metabolic rate (SMR) was significantly higher in pelagic perch but did not correlate with resource use or morphology. Maximum metabolic rate (MMR) was not correlated with any of our explanatory variables or with SMR. Aerobic scope (AS) showed the same pattern as SMR, differing across habitats, but contrary to expectations, was lower in pelagic perch.This study helps to establish a framework for future experiments further exploring the drivers of intraspecific differences in metabolism. In addition, since metabolic rates scale with temperature and determine predator energy requirements, our observed differences in SMR across habitats will help determine ecotype-specific vulnerabilities to climate change and differences in top-down predation pressure across habitats.

Intraspecific differences in metabolic rates shape carbon stable isotope trophic discrimination factors of muscle tissue in the common teleost Eurasian perch (Perca fluviatilis).

Stable isotopes represent a unique approach to provide insights into the ecology of organisms. δ13C and δ15N have specifically been used to obtain information on the trophic ecology and food-web interactions. Trophic discrimination factors (TDF, Δ13C and Δ15N) describe the isotopic fractionation occurring from diet to consumer tissue, and these factors are critical for obtaining precise estimates within any application of δ13C and δ15N values. It is widely acknowledged that metabolism influences TDF, being responsible for different TDF between tissues of variable metabolic activity (e.g., liver vs. muscle tissue) or species body size (small vs. large). However, the connection between the variation of metabolism occurring within a single species during its ontogeny and TDF has rarely been considered.Here, we conducted a 9-month feeding experiment to report Δ13C and Δ15N of muscle and liver tissues for several weight classes of Eurasian perch (Perca fluviatilis), a widespread teleost often studied using stable isotopes, but without established TDF for feeding on a natural diet. In addition, we assessed the relationship between the standard metabolic rate (SMR) and TDF by measuring the oxygen consumption of the individuals.Our results showed a significant negative relationship of SMR with Δ13C, and a significant positive relationship of SMR with Δ15N of muscle tissue, but not with TDF of liver tissue. SMR varies inversely with size, which translated into a significantly different TDF of muscle tissue between size classes.In summary, our results emphasize the role of metabolism in shaping-specific TDF (i.e., Δ13C and Δ15N of muscle tissue) and especially highlight the substantial differences between individuals of different ontogenetic stages within a species. Our findings thus have direct implications for the use of stable isotope data and the applications of stable isotopes in food-web studies.

Morphological and trophic divergence of lake and stream minnows (Phoxinus phoxinus).

Phenotypic divergence in response to divergent natural selection between environments is a common phenomenon in species of freshwater fishes. Intraspecific differentiation is often pronounced between individuals inhabiting lakes versus stream habitats. The different hydrodynamic regimes in the contrasting habitats may promote a variation of body shape, but this could be intertwined with morphological adaptations to a specific foraging mode. Herein, I studied the divergence pattern of the European minnow (Phoxinus phoxinus), a common freshwater fish that has received little attention despite its large distribution. In many Scandinavian mountain lakes, European minnows are considered as being invasive and were found to pose threats to the native fish populations due to resource competition. Minnows were recently found to show phenotypic adaptations in lake versus stream habitats, but the question remained if this divergence pattern is related to differences in resource use. I therefore studied the patterns of minnow divergence in morphology (i.e., using geometric morphometrics) and trophic niches (i.e., using stomach content analyses) in the lake Ånnsjön and its tributaries to link the changes in body morphology to the feeding on specific resources. Lake minnows showed a strong reliance on benthic Cladocera and a more streamlined body shape with a more upward facing snout, whereas stream minnows fed on macroinvertebrates (larvae and adults) to a higher degree and had a deeper body with a snout that was pointed down. Correlations showed a significant relationship of the proportion of macroinvertebrates in the gut and morphological features present in the stream minnows. The results of this study highlight the habitat-specific divergence pattern in morphology and resource use in this ubiquitous freshwater fish. Consequently, interspecific interactions of invasive minnows and the native fish population could differ in the respective food webs and resource competition could target different native fish species in the contrasting habitats.

Asymmetrical habitat coupling of an aquatic predator-The importance of individual specialization.

Predators should stabilize food webs because they can move between spatially separate habitats. However, predators adapted to forage on local resources may have a reduced ability to couple habitats. Here, we show clear asymmetry in the ability to couple habitats by Eurasian perch-a common polymorphic predator in European lakes. We sampled perch from two spatially separate habitats-pelagic and littoral zones-in Lake Erken, Sweden. Littoral perch showed stronger individual specialization, but they also used resources from the pelagic zone, indicating their ability to couple habitats. In contrast, pelagic perch showed weaker individual specialization but near complete reliance on pelagic resources, indicating their preference to one habitat. This asymmetry in the habitat coupling ability of perch challenges the expectation that, in general, predators should stabilize spatially separated food webs. Our results suggest that habitat coupling might be constrained by morphological adaptations, which in this case were not related to genetic differentiation but were more likely related to differences in individual specialization.

Decrease of Population Divergence in Eurasian Perch (Perca fluviatilis) in Browning Waters: Role of Fatty Acids and Foraging Efficiency.

Due to altered biogeochemical processes related to climate change, highly colored dissolved organic carbon (DOC) from terrestrial sources will lead to a water "brownification" in many freshwater systems of the Northern Hemisphere. This will create deteriorated visual conditions that have been found to affect habitat-specific morphological variations in Eurasian perch (Perca fluviatilis) in a previous study. So far, potential drivers and ultimate causes of these findings have not been identified. We conducted a field study to investigate the connection between morphological divergence and polyunsaturated fatty acid (PUFA) composition of perch from six lakes across a gradient of DOC concentration. We expected a decrease in the prevalence of PUFAs, which are important for perch growth and divergence with increasing DOC concentrations, due to the restructuring effects of DOC on aquatic food webs. In general, rate of morphological divergence in perch decreased with increasing DOC concentrations. Proportions of specific PUFAs (22:6n-3, 18:3n-3, 20:5n-3, and 20:4n-6) identified to primarily contribute to overall differences between perch caught in clear and brown-water lakes tended to be connected to overall decline of morphological divergence. However, no overall significant relationship was found, indicating no severe limitation of essential fatty acids for perch inhabiting brown water lakes. We further broaden our approach by conducting a laboratory experiment on foraging efficiency of perch. Therefore, we induced pelagic and littoral phenotypes by differences in habitat-structure and feeding mode and recorded attack rate in a feeding experiment. Generally, fish were less efficient in foraging on littoral prey (Ephemeroptera) when visual conditions were degraded by brown water color. We concluded that browning water may have a strong effect on the forager's ability to find particular food resources, resulting in the reduced development of evolutionary traits, such as habitat- specific morphological divergence.

Assessing the Utility of Hydrogen, Carbon and Nitrogen Stable Isotopes in Estimating Consumer Allochthony in Two Shallow Eutrophic Lakes.

Hydrogen stable isotopes (δ2H) have recently been used to complement δ13C and δ15N in food web studies due to their potentially greater power to separate sources of organic matter in aquatic food webs. However, uncertainties remain regarding the use of δ2H, since little is known about the potential variation in the amount of exchangeable hydrogen (Hex) among common sample materials or the patterns of δ2H when entire food webs are considered. We assessed differences in Hex among the typical sample materials in freshwater studies and used δ2H, δ13C and δ15N to compare their effectiveness in tracing allochthonous matter in food webs of two small temperate lakes. Our results showed higher average amounts of Hex in animal tissues (27% in fish and macroinvertebrates, 19% in zooplankton) compared to most plant material (15% in terrestrial plants and 8% in seston/periphyton), with the exception of aquatic vascular plants (23%, referred to as macrophytes). The amount of Hex correlated strongly with sample lipid content (inferred from C:N ratios) in fish and zooplankton samples. Overall, the three isotopes provided good separation of sources (seston, periphyton, macrophytes and allochthonous organic matter), particularly the δ2H followed by δ13C. Aquatic macrophytes revealed unexpectedly high δ2H values, having more elevated δ2H values than terrestrial organic matter with direct implications for estimating consumer allochthony. Organic matter from macrophytes significantly contributed to the food webs in both lakes highlighting the need to include macrophytes as a potential source when using stable isotopes to estimate trophic structures and contributions from allochthonous sources.