Land use changes biomass and temporal patterns of insect cross-ecosystem flows.

Emergent aquatic insects constitute an important food source for higher trophic levels, linking aquatic to terrestrial ecosystems. Little is known about how land use affects the biomass or composition of insect emergence. Previous studies are limited to individual time points or seasons, hampering understanding of annual biomass export patterns and detection of phenological changes. Over 1 year's primary emergence period, we continuously determined the biomass, abundance, and identity of >45,000 aquatic insects and recorded land-use-related environmental variables in 20 stream sites using a paired design with upstream forested sites and downstream agricultural sites. Total insect biomass and abundance were 2-7 mg day-1  m-2 and 7-36 ind day-1  m-2 higher in agricultural than forested sites. However, we found turnover of families between forested and agricultural sites, with more insects with shorter generation time in agriculture, indicating lower sensitivity to land-use-related stress because of higher recovery potential. Except for stoneflies, biomass and abundance of major orders were higher in agriculture, but their phenology differed. For different orders, emergence peaked 30 days earlier to 51 days later in agriculture than forest, whereas total abundance and biomass both peaked earlier in agriculture: 3-5 and 3-19 days, respectively. The most important land-use-related drivers were pesticide toxicity and electrical conductivity, which were differentially associated with different aquatic insect order abundances and biomass. Overall, we found that land use was related to changes in composition and phenology of aquatic insect emergence, which is likely to affect food-web dynamics in a cross-ecosystem context.

Glyphosate without Co-formulants affects embryonic development of the south african clawed frog Xenopus laevis.

BACKGROUND: Glyphosate (GLY) is the most widely used herbicide in the world. Due to its mode of action as an inhibitor of the 5-enolpyruvylshikimate-3-phosphate synthase, an important step in the shikimate pathway, specifically in plants, GLY is considered to be of low toxicity to non-target organisms. However, various studies have shown the negative effects of GLY on the mortality and development of different non-target organisms, including insects, rodents, fish and amphibians. To better understand the various effects of GLY in more detail, we studied the effects of GLY without co-formulants during the embryogenesis of the aquatic model organism Xenopus laevis. RESULTS: A treatment with GLY affected various morphological endpoints in X. laevis tadpoles (body length, head width and area, eye area). Additionally, GLY interfered with the mobility as well as the neural and cardiac development of the embryos at stage 44/45. We were able to detect detailed structural changes in the cranial nerves and the heart and gained insights into the negative effects of GLY on cardiomyocyte differentiation. CONCLUSION: The application of GLY without co-formulants resulted in negative effects on several endpoints in the early embryonic development of X. laevis at concentrations that are environmentally relevant and concentrations that reflect the worst-case scenarios. This indicates that GLY could have a strong negative impact on the survival and lives of amphibians in natural waters. As a result, future GLY approvals should consider its impact on the environment.

Time between Sequential Exposures to Multiple Stress Turns Antagonism into Synergism.

Aquatic communities are exposed to repeated pulses of toxicants and environmental stressors. We hypothesize that the dose, order, and timing of stress events shape the interactions of these communities. For this, we conducted a fully-crossed, four-factorial, multiple stress exposure experiment to study the combined effects of Esfenvalerate and ultraviolet-B (UV-B) radiation related to the exposure timing and order on Daphnia magna. We revealed that initial exposure to low stress doses, independent of the stress type (UV-B or Esfenvalerate), significantly increased the resistance toward the second stressor. This beneficial effect was apparent only when the second stressor was applied immediately after the first stressor (p < 0.01). When the period between stressor applications was extended to 2 days, the antagonism between the two stressors turned into synergism. The stressor interaction could be predicted with an abstract-mechanistic model of the temporal dynamics of the early-stage stress response. With this model, the timing and order of exposures were able to successfully explain interactions observed in all treatments (model-R2 = 1.0). We conclude that especially the duration of a break between exposures and the exposure dose have a decisive influence on interactions between toxicants and environmental stressors.

Multiple Stress Reduces the Advantage of Pesticide Adaptation.

Under global change scenarios, multistress conditions may occur regularly and require adaptation. However, the adaptation to one stressor might be associated with the increased sensitivity to another stressor. Here, we investigated the ecological consequences of such trade-off under multiple stress. We compared the pesticide tolerance of the crustacean Gammarus pulex from agricultural streams with populations from reference streams. Under optimum temperature, G. pulex from agricultural streams were considerably more tolerant to pesticides as compared to the reference populations. Here, we assume that the increased tolerance in agricultural populations is the combination of acclimation, epigenetic effect, and genetic evolution. After experimental pre-exposure to very low concentration (LC50/1000), reference populations showed increased pesticide tolerance. In contrast, pre-exposure did not further increase the tolerance of agricultural populations. Moreover, these populations were more sensitive to elevated temperature alone due to the hypothesized fitness cost of genetic adaptation to pesticides. However, both reference and agricultural populations showed a similar tolerance to the combined stress of pesticides and warming due to stronger synergistic effects in adapted populations. As a result, pesticide adaptation loses its advantage. The combined effect was predicted well using the stress addition model, developed for predicting the synergistic interaction of independent stressors. We conclude that under multistress conditions, adaptation to pesticides reduces the general stress capacity of individuals and trade-off processes increase the sensitivity to additional stressors. This causes strong synergistic effects of additional stressors on pesticide-adapted individuals.

Assessing the Mixture Effects in In Vitro Bioassays of Chemicals Occurring in Small Agricultural Streams during Rain Events.

Rain events may impact the chemical pollution burden in rivers. Forty-four small streams in Germany were profiled during several rain events for the presence of 395 chemicals and five types of mixture effects in in vitro bioassays (cytotoxicity; activation of the estrogen, aryl hydrocarbon, and peroxisome proliferator-activated receptors; and oxidative stress response). While these streams were selected to cover a wide range of agricultural impacts, in addition to the expected pesticides, wastewater-derived chemicals and chemicals typical for street runoff were detected. The unexpectedly high estrogenic effects in many samples indicated the impact by wastewater or overflow of combined sewer systems. The 128 water samples exhibited a high diversity of chemical and effect patterns, even for different rain events at the same site. The detected 290 chemicals explained only a small fraction (<8%) of the measured effects. The experimental effects of the designed mixtures of detected chemicals that were expected to dominate the mixture effects of detected chemicals were consistent with predictions for concentration addition within a factor of two for 94% of the mixtures. Overall, the burden of chemicals and effects was much higher than that previously detected in surface water during dry weather, with the effects often exceeding proposed effect-based trigger values.

Environmental Stress Increases Synergistic Effects of Pesticide Mixtures on Daphnia magna.

Some widely used pesticide mixtures produce more than additive effects according to conventional combined effect models. However, synergistic effects have been so far generally observed at unrealistically high pesticide concentrations. Here, we used Daphnia magna as a test organism and investigated how food limitation-a common ecological stressor-affects the mixture toxicity of a pyrethroid insecticide and azole fungicide. We also compared three models regarding the prediction of mixture effects, including concentration addition (CA), effect addition (EA), and stress addition model (SAM). We revealed that especially under low food, the strength of synergism between esfenvalerate and prochloraz increased with an increasing concentration of prochloraz independent of the null model. Under high food conditions and at concentrations of prochloraz ≥32 µg/L, we observed a marginal synergistic effect with a model deviation ratio (MDR) = 2.1 at 32 µg/L prochloraz and 2.2 at 100 µg/L prochloraz when using CA as the null model. In contrast, the combination of both pesticides and food stress caused synergistic effects shown by an MDR = 10.9 even at 1 µg/L of prochloraz that is frequently detected in the environment. The combined effects of pesticides and food stress could be predicted best with the SAM that showed the lowest mean deviation between effect observation and prediction (mean deviation SAM = 16 [SD = 28], EA = 1072 [2105], CA = 1345 [2644]). We conclude that common environmental stressors can strongly increase the synergistic effects of toxicants. This knowledge is especially relevant considering current efforts to include the additional risk of pesticide mixtures and environmental stressors into the environmental risk assessment of pesticides.

Pesticide Body Burden of the Crustacean Gammarus pulex as a Measure of Toxic Pressure in Agricultural Streams.

Risk assessments of toxicants in aquatic environments are typically based on the evaluation of concentrations in water or sediment. However, concentrations in water are highly variable, while the body burden may provide a better time-integrated measure of pesticide exposure and potential effects in aquatic organisms. Here, we quantified pesticide body burdens in a dominant invertebrate species from agricultural streams, Gammarus pulex, compared them with pesticide concentrations in water samples, and linked the pesticide contamination with observed ecological effects on macroinvertebrate communities. In total, 19 of 61 targeted analytes were found in the organisms, ranging from 0.037 to 93.94 ng g-1 (wet weight). Neonicotinoids caused the highest toxic pressure among the pesticides detected in G. pulex. Using linear solvation energy relationships (LSERs), we derived equivalent pesticide concentrations in streamwater based on the body burden. These equivalent concentrations correlated with the concentrations in water samples collected after runoff (65% of variance explained). Pesticide pressure significantly affected the aquatic macroinvertebrate community structure, expressed as SPEARpesticides, and caused, on average, 3-fold increased insecticide tolerance in G. pulex as a result of adaptation. The toxic pressure derived from body burden and from water samples similarly explained the change in community structure (68% and 64%). However, the increased tolerance of G. pulex to pesticides was better explained by the toxicity derived from body burden (70%) than by the toxicity from water samples (53%). We conclude that the internal body burden of macroinvertebrates is suitable to assess the overall pesticide exposure and effects in agricultural streams.

Species Diversity Hinders Adaptation to Toxicants.

Environmental toxicants such as pesticides exert strong selection pressure on many species. While the resulting development of pesticide resistance in agricultural pest species is well-known, reports on the extent of adaptation in exposed nontarget species are contradictory. These contradictory reports highlight a continuing challenge in understanding the relevant ecological mechanisms that facilitate or hinder adaptation to toxicants in the field. Here we show that species diversity hinders the adaptation to toxicants. In agricultural streams with low diversity, we observed an up to 8-fold increase in insecticide tolerance in a total of 17 macroinvertebrate species that was not observed in more diverse communities under similar contamination. High species diversity occurred independently from adjacent nonpolluted refuge areas. Therefore, the low level of adaptation in diverse streams cannot be explained by an increased recolonization of sensitive individuals from refuge areas. Instead, high intraspecific competition may facilitate the selection for increased tolerance in low-diverse communities. In diverse communities, by contrast, species interactions may reduce intraspecific competition and, thus, the potential for developing toxicant resistance. We suggest that this mechanism may be the general case in adaptation to environmental stressors. Additionally, we conclude that the current framework for risk assessment of pesticides is not protective against selection for tolerant organisms and the associated risk of genetic erosion.

Sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants.

Hydrocarbons have an utmost economical importance but may also cause substantial ecological impacts due to accidents or inadequate transportation and use. Currently, freshwater biomonitoring methods lack an indicator that can unequivocally reflect the impacts caused by hydrocarbons while being independent from effects of other stressors. The aim of the present study was to develop a sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants, which can be used in hydrocarbon-specific bioindicators. We employed the Relative Sensitivity method and developed the sensitivity ranking S hydrocarbons based on literature ecotoxicological data supplemented with rapid and mesocosm test results. A first validation of the sensitivity ranking based on an earlier field study has been conducted and revealed the S hydrocarbons ranking to be promising for application in sensitivity based indicators. Thus, the first results indicate that the ranking can serve as the core component of future hydrocarbon-specific and sensitivity trait based bioindicators.

Modeling Macroinvertebrate Community Dynamics in Stream Mesocosms Contaminated with a Pesticide.

Modeling community dynamics of aquatic invertebrates is an important but challenging task, in particular in ecotoxicological risk assessment. Systematic parameter estimation and rigorous assessment of model uncertainty are often lacking in such applications. We applied the mechanistic food web model Streambugs to investigate the temporal development of the macroinvertebrate community in an ecotoxicological mesocosm experiment with pulsed contaminations with the insecticide thiacloprid. We used Bayesian inference to estimate parameters and their uncertainty. Approx. 85% of all experimental observations lie within the 90% uncertainty intervals indicating reasonably good fits of the calibrated model. However, a validation with independent data was not possible due to lacking data. Investigation of vital rates and limiting factors in the model yielded insights into recovery dynamics. Inclusion of the emergence process and sub-lethal effects turned out to be potentially relevant model extensions. Measurements of food source dynamics, individual body size (classes), and additional knowledge on sub-lethal effects would support more accurate modeling. This application of a process-based, ecotoxicological community model with uncertainty assessment by Bayesian inference increased our process understanding of toxicant effects in macroinvertebrate communities and helped identifying potential improvements in model structure and experimental design.

Pesticides reduce regional biodiversity of stream invertebrates.

The biodiversity crisis is one of the greatest challenges facing humanity, but our understanding of the drivers remains limited. Thus, after decades of studies and regulation efforts, it remains unknown whether to what degree and at what concentrations modern agricultural pesticides cause regional-scale species losses. We analyzed the effects of pesticides on the regional taxa richness of stream invertebrates in Europe (Germany and France) and Australia (southern Victoria). Pesticides caused statistically significant effects on both the species and family richness in both regions, with losses in taxa up to 42% of the recorded taxonomic pools. Furthermore, the effects in Europe were detected at concentrations that current legislation considers environmentally protective. Thus, the current ecological risk assessment of pesticides falls short of protecting biodiversity, and new approaches linking ecology and ecotoxicology are needed.

Biotic interactions govern genetic adaptation to toxicants.

The genetic recovery of resistant populations released from pesticide exposure is accelerated by the presence of environmental stressors. By contrast, the relevance of environmental stressors for the spread of resistance during pesticide exposure has not been studied. Moreover, the consequences of interactions between different stressors have not been considered. Here we show that stress through intraspecific competition accelerates microevolution, because it enhances fitness differences between adapted and non-adapted individuals. By contrast, stress through interspecific competition or predation reduces intraspecific competition and thereby delays microevolution. This was demonstrated in mosquito populations (Culex quinquefasciatus) that were exposed to the pesticide chlorpyrifos. Non-selective predation through harvesting and interspecific competition with Daphnia magna delayed the selection for individuals carrying the ace-1(R) resistance allele. Under non-toxic conditions, susceptible individuals without ace-1(R) prevailed. Likewise, predation delayed the reverse adaptation of the populations to a non-toxic environment, while the effect of interspecific competition was not significant. Applying a simulation model, we further identified how microevolution is generally determined by the type and degree of competition and predation. We infer that interactions with other species-especially strong in ecosystems with high biodiversity-can delay the development of pesticide resistance.

Analysing chemical-induced changes in macroinvertebrate communities in aquatic mesocosm experiments: a comparison of methods.

Mesocosm experiments that study the ecological impact of chemicals are often analysed using the multivariate method 'Principal Response Curves' (PRCs). Recently, the extension of generalised linear models (GLMs) to multivariate data was introduced as a tool to analyse community data in ecology. Moreover, data aggregation techniques that can be analysed with univariate statistics have been proposed. The aim of this study was to compare their performance. We compiled macroinvertebrate abundance datasets of mesocosm experiments designed for studying the effect of various organic chemicals, mainly pesticides, and re-analysed them. GLMs for multivariate data and selected aggregated endpoints were compared to PRCs regarding their performance and potential to identify affected taxa. In addition, we analysed the inter-replicate variability encountered in the studies. Mesocosm experiments characterised by a higher taxa richness of the community and/or lower taxonomic resolution showed a greater inter-replicate variability, whereas variability decreased the more zero counts were encountered in the samples. GLMs for multivariate data performed equally well as PRCs regarding the community response. However, compared to first axis PRCs, GLMs provided a better indication of individual taxa responding to treatments, as separate models are fitted to each taxon. Data aggregation methods performed considerably poorer compared to PRCs. Multivariate community data, which are generated during mesocosm experiments, should be analysed using multivariate methods to reveal treatment-related community-level responses. GLMs for multivariate data are an alternative to the widely used PRCs.

Environmental stressors can enhance the development of community tolerance to a toxicant.

Ecosystems are subject to a combination of recurring anthropogenic and natural disturbances, such as climate change and pesticide exposure. Biological communities are known to develop tolerance to recurring disturbances due to successive changes at both the community and organismal levels. However, information on how additional stressors may affect the development of such community tolerance is scarce to date. We studied the influence of hydrological disturbance on the reaction of zooplankton communities to repeated insecticide pulses in outdoor microcosms. The communities were exposed to three successive pulses of the insecticide esfenvalerate (0.03, 0.3, and 3 µg/L) and to the gradual removal of water and its subsequent replacement over three cycles or to a constant water level. Except at the highest esfenvalerate concentration, the communities developed tolerance to the toxicant, as indicated by their decreasing reaction to subsequent insecticide applications, and this development was enhanced by hydrological disturbance. The pronounced decline of the key taxa Daphnia spp. through the combined action of the two stressors was identified as the main mechanism responsible for the increase in community tolerance under a fluctuating water level. Under a constant water level, the abundance of Daphnia spp. did not decrease significantly without the insecticide treatment, indicating that other mechanisms were responsible for the observed community tolerance. The present study shows that additional stressors can facilitate the development of community tolerance and that such facilitation is propagated through community-level mechanisms.

Revealing the cascade of pesticide effects from gene to community.

Global pesticide exposure in agriculture leads to biodiversity loss, even at ultra-low concentrations below the legal limits. The mechanisms by which the effects of toxicants act at such low concentrations are still unclear, particularly in relation to their propagation across the different biological levels. In this study, we demonstrate, for the first time, a cascade of effects from the gene to the community level. At the gene level, agricultural pesticide exposure resulted in reduced genetic diversity of field-collected Gammarus pulex, a dominant freshwater crustacean in Europe. Additionally, we identified alleles associated with adaptations to pesticide contamination. At the individual level, this genetic adaptation to pesticides was linked to a lower fecundity, indicating related fitness costs. At the community level, the combined effect of pesticides and competitors caused a decline in the overall number and abundance of pesticides susceptible macroinvertebrate competing with gammarids. The resulting reduction in interspecific competition provided an advantage for pesticide-adapted G. pulex to dominate macroinvertebrate communities in contaminated areas, despite their reduced fitness due to adaptation. These processes demonstrate the complex cascade of effects, and also illustrate the resilience and adaptability of biological systems across organisational levels to meet the challenges of a changing environment.

Combined effects of herbicides and insecticides reduce biomass of sensitive aquatic invertebrates.

The structure and biomass of aquatic invertebrate communities play a crucial role in the matter dynamics of streams. However, biomass is rarely quantified in ecological assessments of streams, and little is known about the environmental and anthropogenic factors that influence it. In this study, we aimed to identify environmental factors that are associated with invertebrate structure and biomass through a monitoring of 25 streams across Germany. We identified invertebrates, assigned them to taxonomic and trait-based groups, and quantified biomass using image-based analysis. We found that insecticide pressure generally reduced the abundance of insecticide-vulnerable populations (R2 = 0.43 applying SPEARpesticides indicator), but not invertebrate biomass. In contrast, herbicide pressure reduced the biomass of several biomass aggregations. Especially, insecticide-sensitive populations, that were directly (algae feeder, R2 = 0.39) or indirectly (predators, R2 = 0.29) dependent on algae, were affected. This indicated a combined effect of possible food shortage due to herbicides and direct insecticide pressure. Specifically, all streams with increased herbicide pressure showed a reduced overall biomass share of Trichoptera from 43 % to 3 % and those of Ephemeroptera from 20 % to 3 % compared to streams grouped by low herbicide pressure. In contrast, insecticide-insensitive Gastropoda increased from 10 % to 45 %, and non-vulnerable leaf-shredding Crustacea increased from 10 % to 22 %. In summary, our results indicate that at the community level, the direct effects of insecticides and the indirect, food-mediated effects of herbicides exert a combined effect on the biomass of sensitive insect groups, thus disrupting food chains at ecosystem level.

Citizen science shows that small agricultural streams in Germany are in a poor ecological status.

Agricultural pesticides, nutrients, and habitat degradation are major causes of insect declines in lowland streams. To effectively conserve and restore stream habitats, standardized stream monitoring data and societal support for freshwater protection are needed. Here, we sampled 137 small stream monitoring sites across Germany, 83 % of which were located in agricultural catchments, with >900 citizen scientists in 96 monitoring groups. Sampling was carried out according to Water Framework Directive standards as part of the citizen science freshwater monitoring program FLOW in spring and summer 2021, 2022 and 2023. The biological indicator SPEARpesticides was used to assess pesticide exposure and effects based on macroinvertebrate community composition. Overall, 58 % of the agricultural monitoring sites failed to achieve a good ecological status in terms of macroinvertebrate community composition and indicated high pesticide exposure (SPEARpesticides status class: 29 % "moderate", 19 % "poor", 11 % "bad"). The indicated pesticide pressure in streams was related to the proportion of arable land in the catchment areas (R2 = 0.23, p < 0.001). Also with regards to hydromorphology, monitoring results revealed that 65 % of the agricultural monitoring sites failed to reach a good status. The database produced by citizen science groups was characterized by a high degree of accuracy, as results obtained by citizen scientists and professionals were highly correlated for SPEARpesticides index (R2 = 0.79, p < 0.001) and hydromorphology index values (R2 = 0.72, p < 0.001). Such citizen-driven monitoring of the status of watercourses could play a crucial role in monitoring and implementing the objectives of the European Water Framework Directive, thus contributing to restoring and protecting freshwater ecosystems.

Exploring BPA alternatives - Environmental levels and toxicity review.

Bisphenol A alternatives are manufactured as potentially less harmful substitutes of bisphenol A (BPA) that offer similar functionality. These alternatives are already in the market, entering the environment and thus raising ecological concerns. However, it can be expected that levels of BPA alternatives will dominate in the future, they are limited information on their environmental safety. The EU PARC project highlights BPA alternatives as priority chemicals and consolidates information on BPA alternatives, with a focus on environmental relevance and on the identification of the research gaps. The review highlighted aspects and future perspectives. In brief, an extension of environmental monitoring is crucial, extending it to cover BPA alternatives to track their levels and facilitate the timely implementation of mitigation measures. The biological activity has been studied for BPA alternatives, but in a non-systematic way and prioritized a limited number of chemicals. For several BPA alternatives, the data has already provided substantial evidence regarding their potential harm to the environment. We stress the importance of conducting more comprehensive assessments that go beyond the traditional reproductive studies and focus on overlooked relevant endpoints. Future research should also consider mixture effects, realistic environmental concentrations, and the long-term consequences on biota and ecosystems.

Culmination of low-dose pesticide effects.

Pesticides applied in agriculture can affect the structure and function of nontarget populations at lower doses and for longer timespans than predicted by the current risk assessment frameworks. We identified a mechanism for this observation. The populations of an aquatic invertebrate (Culex pipiens) exposed over several generations to repeated pulses of low concentrations of the neonicotinoid insecticide (thiacloprid) continuously declined and did not recover in the presence of a less sensitive competing species (Daphnia magna). By contrast, in the absence of a competitor, insecticide effects on the more sensitive species were only observed at concentrations 1 order of magnitude higher, and the species recovered more rapidly after a contamination event. The underlying processes are experimentally identified and reconstructed using a simulation model. We conclude that repeated toxicant pulse of populations that are challenged with interspecific competition may result in a multigenerational culmination of low-dose effects.