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.

Emerging Midges Transport Pesticides from Aquatic to Terrestrial Ecosystems: Importance of Compound- and Organism-Specific Parameters.

Emerging aquatic insects have the potential to retain aquatic contaminants after metamorphosis, potentially transporting them into adjacent terrestrial food webs. It is unknown whether this transfer is also relevant for current-use pesticides. We exposed larvae of the nonbiting midge, Chironomus riparius, to a sublethal pulse of a mixture of nine moderately polar fungicides and herbicides (logKow 2.5-4.7) at three field relevant treatment levels (1.2-2.5, 17.5-35.0, or 50.0-100.0 µg/L). We then assessed the pesticide bioaccumulation and bioamplification over the full aquatic-terrestrial life cycle of both sexes including the egg laying of adult females. By applying sensitive LC-MS/MS analysis to small sample volumes (∼5 mg, dry weight), we detected all pesticides in larvae from all treatment levels (2.8-1019 ng/g), five of the pesticides in the adults from the lowest treatment level and eight in the higher treatment levels (1.5-3615 ng/g). Retention of the pesticides through metamorphosis was not predictable based solely on pesticide lipophilicity. Sex-specific differences in adult insect pesticide concentrations were significant for five of the pesticides, with greater concentrations in females for four of them. Over the duration of the adults' lifespan, pesticide concentrations generally decreased in females while persisting in males. Our results suggest that a low to moderate daily dietary exposure to these pesticides may be possible for tree swallow nestlings and insectivorous bats.

Microbial community history and leaf species shape bottom-up effects in a freshwater shredding amphipod.

Arable land use and the associated application of agrochemicals can affect local freshwater communities with consequences for the entire ecosystem. For instance, the structure and function of leaf-associated microbial communities can be affected by pesticides, such as fungicides. Additionally, the leaf species on which these microbial communities grow reflects another environmental filter for community structure. These factors and their interaction may jointly modify leaves' nutritional quality for higher trophic levels. To test this assumption, we studied the structure of leaf-associated microbial communities with distinct exposure histories (pristine [P] vs vineyard run off [V]) colonising two leaf species (black alder, European beech, and a mixture thereof). By offering these differently colonised leaves as food to males and females of the leaf-shredding amphipod Gammarus fossarum (Crustacea; Amphipoda) we assessed for potential bottom-up effects. The growth rate, feeding rate, faeces production and neutral lipid fatty acid profile of the amphipod served as response variable in a 2 × 3 × 2-factorial test design over 21d. A clear separation of community history (P vs V), leaf species and an interaction between the two factors was observed for the leaf-associated aquatic hyphomycete (i.e., fungal) community. Sensitive fungal species were reduced by up to 70 % in the V- compared to P-community. Gammarus' growth rate, feeding rate and faeces production were affected by the factor leaf species. Growth was negatively affected when Gammarus were fed with beech leaves only, whereas the impact of alder and the mixture of both leaf species was sex-specific. Overall, this study highlights that leaf species identity had a more substantial impact on gammarids relative to the microbial community itself. Furthermore, the sex-specificity of the observed effects (excluding fatty acid profile, which was only measured for male) questions the procedure of earlier studies, that is using either only one sex or not being able to differentiate between males and females. However, these results need additional verification to support a reliable extrapolation.

Phenotypic adaptation of Chironomus riparius to chronic Bti exposure: effects on emergence time and nutrient content.

Insects with aquatic larval and terrestrial adult life stages are a key component of coupled aquatic-terrestrial ecosystems. Thus, stressors applied to water bodies adversely affecting those larvae have the potential to influence the riparian zone through altered emergence, with differences in prey availability, timing, or nutrition. In this study, the common model organism Chironomus riparius, a species of Chironomidae (Diptera), was used. This selection was further motivated by its wide distribution in European freshwaters and its importance as prey for terrestrial predators. A stressor of high importance in this context is the globally used mosquito control agent Bacillus thuringiensis var. israelensis (Bti) which has been shown to affect Chironomidae. Here, we investigated the ability of chironomid populations to adapt to a regularly applied stressor, leading to a reduced impact of Bti. Therefore, the initial sensitivity of laboratory populations of C. riparius was investigated under the influence of field-relevant Bti treatments (three doses × two application days) and different food sources (high-quality TetraMin vs. low-quality Spirulina). Following a chronic exposure to Bti over six months, the sensitivity of pre-exposed and naïve populations was re-evaluated. Food quality had a strong impact on emergence timing and nutrient content. In addition, alterations in emergence time as well as protein and lipid contents of chronically exposed populations indicated a selection for individuals of advantageous energetics, potentially leading to a more efficient development while combating Bti. Signs of adaptation could be confirmed in five out of 36 tested scenarios suggesting adaptation to Bti at the population level. Adaptive responses of one or several species could theoretically (via eco-evolutionary dynamics) result in a community shift, favouring the prevalence of Bti-tolerant species. (In)direct effects of Bti and the adaptive responses at both population and community levels could affect higher trophic levels and may determine the fate of meta-ecosystems.

Trophic effects of Bti-based mosquito control on two top predators in floodplain pond mesocosms.

Chironomid (Diptera: Chironomidae) larvae play a key role in aquatic food webs as prey for predators like amphibian and dragonfly larvae. This trophic link may be disrupted by anthropogenic stressors such as Bacillus thuringiensis var. israelensis (Bti), a biocide widely used in mosquito control. In a companion study, we recorded a 41% reduction of non-target larval chironomids abundance in outdoor floodplain pond mesocosms (FPMs) treated with Bti. Therefore, we examined the diet of two top predators in the FPMs, larvae of the palmate newt (Salamandridae: Lissotriton helveticus) and dragonfly (Aeshnidae: predominantly Anax imperator), using bulk stable isotope analyses of carbon and nitrogen. Additionally, we determined neutral lipid fatty acids in newt larvae to assess diet-related effects on their physiological condition. We did not find any effects of Bti on the diet proportions of newt larvae and no significant effects on the fatty acid content. We observed a trend in Aeshnidae larvae from Bti-FPMs consuming a higher proportion of large prey (Aeshnidae, newt, damselfly larvae; ~42%), and similar parts of smaller prey (chironomid, mayfly, Libellulidae, and zooplankton), compared to controls. Our findings may suggest bottom-up effects of Bti on aquatic predators but should be further evaluated, for instance, by using compound-specific stable isotope analyses of fatty acids or metabarcoding approaches.

Subsidy Quality Affects Common Riparian Web-Building Spiders: Consequences of Aquatic Contamination and Food Resource.

Anthropogenic stressors can affect the emergence of aquatic insects. These insects link aquatic and adjacent terrestrial food webs, serving as high-quality subsidy to terrestrial consumers, such as spiders. While previous studies have demonstrated that changes in the emergence biomass and timing may propagate across ecosystem boundaries, the physiological consequences of altered subsidy quality for spiders are largely unknown. We used a model food chain to study the potential effects of subsidy quality: Tetragnatha spp. were exclusively fed with emergent Chironomus riparius cultured in the absence or presence of either copper (Cu), Bacillus thuringiensis var. israelensis (Bti), or a mixture of synthetic pesticides paired with two basal resources (Spirulina vs. TetraMin®) of differing quality in terms of fatty acid (FA) composition. Basal resources shaped the FA profile of chironomids, whereas their effect on the FA profile of spiders decreased, presumably due to the capacity of both chironomids and spiders to modify (dietary) FA. In contrast, aquatic contaminants had negligible effects on prey FA profiles but reduced the content of physiologically important polyunsaturated FAs, such as 20:4n-6 (arachidonic acid) and 20:5n-3 (eicosapentaenoic acid), in spiders by approximately 30% in Cu and Bti treatments. This may have contributed to the statistically significant decline (40%-50%) in spider growth. The observed effects in spiders are likely related to prey nutritional quality because biomass consumption by spiders was, because of our experimental design, constant. Analyses of additional parameters that describe the nutritional quality for consumers such as proteins, carbohydrates, and the retention of contaminants may shed further light on the underlying mechanisms. Our results highlight that aquatic contaminants can affect the physiology of riparian spiders, likely by altering subsidy quality, with potential implications for terrestrial food webs. Environ Toxicol Chem 2023;42:1346-1358. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Contaminant fluxes across ecosystems mediated by aquatic insects.

Metals and organic contaminants in aquatic systems affect the coupling of aquatic and terrestrial ecosystems through two pathways: contaminant-induced effects on insect emergence and emergence-induced contaminant transfer. Consequently, the impact of aquatic contaminants on terrestrial ecosystems can be driven by modifications in the quantity and quality of adult aquatic insects serving as prey or contaminants entering terrestrial food webs as part of the diet of terrestrial predators. Here, we provide an overview of recent advances in the field, separating metals from organic contaminants due to their differential propensity to bioaccumulate and thus their potential contribution to either of the two pathways. Finally, this review highlights the knowledge gap in the relative impact of these pathways on terrestrial insectivores.

Mixture effects of a fungicide and an antibiotic: Assessment and prediction using a decomposer-detritivore system.

Antimicrobials, such as fungicides and antibiotics, pose a risk for microbial decomposers (i.e., bacteria and aquatic fungi) and invertebrate detritivores (i.e., shredders) that play a pivotal role in the ecosystem function of leaf litter breakdown. Although waterborne toxicity and diet-related effects (i.e., dietary exposure and microorganism-mediated alterations in food quality for shredders) of fungicides and antibiotics on decomposer-detritivore systems have been increasingly documented, their joint effect is unknown. We therefore assessed waterborne and dietary effects of an antimicrobial mixture consisting of the fungicide azoxystrobin (AZO) and the antibiotic ciprofloxacin (CIP) on microbial decomposers and the shredder Gammarus fossarum using a tiered approach. We compared effect sizes measured in the present study with model predictions (i.e., independent action) based on published data. During a 7-day feeding activity assay quantifying waterborne toxicity in G. fossarum, the leaf consumption of gammarids was reduced by ∼60 % compared to the control when subjected to the mixture at concentrations of each component causing a 20 % reduction in the same response variable when applied individually. Moreover, the selective feeding of gammarids during the food choice assay indicated alterations in food quality induced by the antimicrobial mixture. The food selection and, in addition, the decrease in microbial leaf decomposition is likely linked to changes in leaf-associated bacteria and fungi. During a long-term assay, energy processing, growth and energy reserves of gammarids were increased in presence of 15 and 500 µg/L of AZO and CIP, respectively, through the dietary pathway. These physiological responses were probably driven by CIP-induced alterations in the gut microbiome or immune system of gammarids. In general, model predictions matched observed effects caused by waterborne exposure on the leaf consumption, energy processing and growth of gammarids during short- and long-term assays, respectively. However, when complex horizontal (bacteria and aquatic fungi) and vertical (leaf-associated microorganisms and shredders) interactions were involved, model predictions partly over- or underestimated mixture effects. Therefore, the present study identifies uncertainties of mixture effect predictions for complex biological systems calling for studies targeting the underlying processes and mechanisms.

Relative importance of dietary uptake and waterborne exposure for a leaf-shredding amphipod exposed to thiacloprid-contaminated leaves.

Systemic neonicotinoids are commonly used in forest pest management programs. Senescent leaves containing neonicotinoids may, however, fall from treated trees into nearby streams. There, leaf-shredding invertebrates are particularly exposed due to their diet (feeding on neonicotinoid-contaminated leaves) or collaterally via the water phase (leaching of a neonicotinoid from leaves) - a fact not considered during aquatic environmental risk assessment. To unravel the relevance of these pathways we used leaves from trees treated with the neonicotinoid thiacloprid to subject the amphipod shredder Gammarus fossarum for 21 days (n = 40) either to dietary, waterborne or a combined (dietary + waterborne) exposure. Dietary exposure caused - relative to the control - similar reductions in gammarids' leaf consumption (~35%) and lipid content (~20%) as observed for the waterborne exposure pathway (30 and 22%). The effect sizes observed under combined exposure suggested additivity of effects being largely predictable using the reference model "independent action". Since gammarids accumulated - independent of the exposure pathway - up to 280 ng thiacloprid/g, dietary exposure may also be relevant for predators which prey on Gammarus. Consequently, neglecting dietary exposure might underestimate the environmental risk systemic insecticides pose for ecosystem integrity calling for its consideration during the evaluation and registration of chemical stressors.