Exposure to teflubenzuron reduces drought tolerance of collembolans.

Chitin synthesis inhibitors (CSIs) are commonly used insecticides compromising cuticle formation and structure in arthropods. Arthropods rely on intact cuticles to maintain water balance and cellular homeostasis to survive in different weather conditions. We hypothesized that physiological impacts of CSIs may make arthropods more vulnerable to harsh environmental conditions, such as extreme heat, cold or drought. The aim of this study was to investigate if pre-exposure to teflubenzuron (a common CSI) would influence Folsomia candida's (Collembola: Isotomidae) sensitivity to natural stressors. Here, we exposed adult collembolans to teflubenzuron through food for two weeks, then survivors were immediately divided into three groups for subsequent acute heat, cold, and drought exposure. After acute exposure to these natural stressors, the collembolans were moved to optimal conditions for a one-week recovery period during which their survival, time to regain reproduction, and egg production were examined. We analyzed the interaction between effects of teflubenzuron and natural stressors using a multiplicative model. No interaction between effects of teflubenzuron and heat was observed in any test endpoints. A synergistic interaction between effects of teflubenzuron and cold was observed in the time to regain reproduction. Both survival and egg production, on the other hand, showed synergistic interaction between effects of teflubenzuron and drought, as well as a tendency for longer reproduction recovery times. Our results suggest that pre-exposure to teflubenzuron reduces drought tolerance in F. candida, while its impact on heat or cold tolerance is minor or absent. This study is among the first to explore the combined effects of CSI and natural stressors on soil arthropods, providing more insight on potential risks posed by such chemicals in the environment.

Chronic Effects of an Insect Growth Regulator (teflubenzuron) on the Life Cycle and Population Growth Rate of Folsomia candida.

Current standard toxicity tests on nontarget soil invertebrates mainly focus on the endpoints survival and reproduction. Such results are likely insufficient to predict effects at higher organizational levels, for example, the population level. We assessed the effects of exposure to the pesticide teflubenzuron on the collembolan Folsomia candida, by performing a full life-cycle experiment exposing single individuals via contaminated food (uncontaminated control and 0.2, 0.32, 0.48, 0.72, 1.08, and 1.6 mg/kg dry yeast). Several life-history traits were considered by following the growth and development of newly hatched individuals over a period of 65 days. We assessed survival, body length, time to first oviposition, cumulative egg production, and hatchability of eggs. A two-stage model was applied to calculate the population growth rate (λ) combined with elasticity analysis to reveal the relative sensitivity of λ to the effects of teflubenzuron on each life-history parameter. Body length was the least sensitive life-history parameter (median effective concentration = 1.10 mg teflubenzuron/kg dry yeast) followed by time to first oviposition (0.96 mg/kg), survival (median lethal concentration = 0.87 mg/kg), cumulative egg production (0.32 mg/kg), and egg hatchability (0.27 mg/kg). Population growth decreased with increasing concentrations of teflubenzuron (λ = 1.162/day in control to 1.005/day in 0.72 mg/kg dry yeast, with populations going extinct at 1.08 and 1.6 mg/kg dry yeast). Elasticity analysis showed that changes in juvenile survival had a greater impact on the population growth rate compared with the other life-history traits. Our study provides a comprehensive overview of individual-level effects of long-term exposure to teflubenzuron and integrates these effects to assess the potential risk to collembolan populations. Environ Toxicol Chem 2024;43:1173-1183. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Low soil Moisture Slows Uptake and Elimination Rates of Phenanthrene in Springtails.

This study investigated the influence of soil water status on the toxicokinetics of phenanthrene in the springtail Folsomia candida allowing estimation of uptake and elimination rates at two contrasting soil water potentials. Fitting a three-phase model to the observations showed that uptake rate (ku) was almost two times higher in moist soil (-2 kPa) than in dry soil (-360 kPa). During the first days of the exposure, elimination rate (ke) was not significantly different in moist and dry soil, but after eight days ke had increased significantly more in moist soil than in dry soil. Our results confirm the general notion that the exposure route via soil pore water is important. Understanding the significance of soil moisture in exposure and effects of contaminants on soil invertebrates is crucial for assessing the ecological risks associated with soil pollution in a changing climate.

Copper-contaminated soil compromises thermal performance in the springtail Folsomia candida (Collembola).

The widespread agricultural and industrial emissions of copper-based chemicals have increased copper levels in soils worldwide. Copper contamination can cause a range of toxic effects on soil animals and influence thermal tolerance. However, toxic effects are commonly investigated using simple endpoints (e.g., mortality) and acute tests. Thus, how organisms respond to ecological realistic sub-lethal and chronic exposures across the entire thermal scope of an organism is not known. In this study, we investigated the effects of copper exposure on the thermal performance of a springtail (Folsomia candida), regarding its survival, individual growth, population growth, and the composition of membrane phospholipid fatty acids. Folsomia candida (Collembola) is a typical representative of soil arthropods and a model organism that has been widely used for ecotoxicological studies. In a full-factorial soil microcosm experiment, springtails were exposed to three levels of copper (ca. 17 (control), 436, and 1629 mg/kg dry soil) and ten temperatures from 0 to 30 °C. Results showed that three-week copper exposure at temperatures below 15 °C and above 26 °C negatively influenced the springtail survival. The body growth was significantly lower for the springtails in high-dose copper soils at temperatures above 24 °C. A high copper level reduced the number of juveniles by 50 %, thereby impairing population growth. Both temperature and copper exposure significantly impacted membrane properties. Our results indicated that high-dose copper exposure compromised the tolerance to suboptimal temperatures and decreased maximal performance, whereas medium copper exposure partially reduced the performance at suboptimal temperatures. Overall, copper contamination reduced the thermal tolerance of springtails at suboptimal temperatures, probably by interfering with membrane homeoviscous adaptation. Our results show that soil organisms living in copper-contaminated areas might be more sensitive to thermally stressful periods.

Polyproline type II helical antifreeze proteins are widespread in Collembola and likely originated over 400 million years ago in the Ordovician Period.

Antifreeze proteins (AFPs) bind to ice crystals to prevent organisms from freezing. A diversity of AFP folds has been found in fish and insects, including alpha helices, globular proteins, and several different beta solenoids. But the variety of AFPs in flightless arthropods, like Collembola, has not yet been adequately assessed. Here, antifreeze activity was shown to be present in 18 of the 22 species of Collembola from cold or temperate zones. Several methods were used to characterize these AFPs, including isolation by ice affinity purification, MALDI mass spectrometry, amino acid composition analysis, tandem mass spectrometry sequencing, transcriptome sequencing, and bioinformatic investigations of sequence databases. All of these AFPs had a high glycine content and were predicted to have the same polyproline type II helical bundle fold, a fold unique to Collembola. These Hexapods arose in the Ordovician Period with the two orders known to produce AFPs diverging around 400 million years ago during the Andean-Saharan Ice Age. Therefore, it is likely that the AFP arose then and persisted in many lineages through the following two ice ages and intervening warm periods, unlike the AFPs of fish which arose independently during the Cenozoic Ice Age beginning ~ 30 million years ago.

Tolerance of high temperature and associated effects on reproduction in euedaphic Collembola.

Studies show that tropical and mid-latitude terrestrial ectotherms are more vulnerable to global warming than species from high latitudes. However, thermal tolerance studies from these regions still lack soil invertebrates. In the present study, we investigated six euedaphic species of Collembola (of the genera Onychiurus and Protaphorura) sampled across latitudes ranging from 31° N to 64° N and determined their upper thermal limit (UTL) by static assays. In another experiment, we submitted springtails to high temperatures for exposure times, causing 5% to 30% mortality within each species. Survivors from this series of increasing heat injuries were used to determine the time-to-first-oviposition and the number of eggs produced following heat exposure. Two hypotheses are tested in this study: 1) heat tolerance of species correlates positively with the environmental temperature of their habitat; 2) the most heat-tolerant species require less time to regain reproduction and produce more eggs than the least heat-tolerant species. Results showed that the UTL positively correlates to the soil temperature of the sampling site. The sequence of UTL60 (the temperature causing 50% mortality after 60 min of exposure) from highest to least was O. yodai > P. fimata > P. armata ≈ P. tricampata > P. macfadyeni > P. pseudovanderdrifti. Heat stress inflicted on springtails can delay reproduction in all species, and two species showed a reduced egg production rate after heat exposure. For heat stress causing up to 30% mortality, the most heat-tolerant species did not have advantages over the least heat-tolerant species for what concerns the recovery of reproduction. The relation between UTL and recovery from heat stress is not linear. Our study provides evidence for a potential long-term effect of high-temperature exposure on euedaphic species of Collembola and highlights the need for further studies on the effects of global warming on soil-living organisms.

Evaluation of life-history traits in Folsomia candida exposed to combined repeated mild heat shocks with phenanthrene.

Climate change increases the frequency and intensity of extreme weather events. In nature, organisms are often exposed to climatic stressors and contaminants simultaneously, and the effects of contaminants may be modified by climate change and vice versa. Here, the effects of repeated mild heat shocks (0-5 times, 30 °C for 6 h), alone or combined with phenanthrene (PHE) (80 mg kg-1 dry soil), on life-history traits of the springtail Folsomia candida were investigated. The survival, growth, maturation, and reproduction of single juvenile springtails were assessed over a period of 37 days. Increasing number of heat bouts or PHE exposure did not have significant negative effects on overall survival at the termination of the experiment, but the interaction between the two stressors led to complex interactions for the dynamics of survival during the test. Neither body growth nor time to first oviposition was influenced by heat or PHE, but a reduction of egg production with increasing number of heat bouts was observed, and there was an interaction between the two stressors. Further, a trade-off between the number of eggs produced and egg size was observed, indicating that females invested the same amount of energy in reproduction despite exposure to stressful temperature and PHE. These results indicate that egg production (in terms of the number of eggs) was a more sensitive indicator of the combined effects of mild heat shocks and PHE than growth, and there was a trade-off between survival and egg production.

Influence of Soil Moisture on Bioaccumulation, Growth, and Recruitment of Folsomia candida Exposed to Phenanthrene-Polluted Soil.

Climate change has resulted in an increased occurrence of summer droughts in large parts of the world. Low soil moisture has marked impacts on the physiology of soil invertebrates and lowers degradation rates of organic contaminants in soil. Polycyclic aromatic hydrocarbons (PAHs) are hydrophobic contaminants that readily accumulate in the lipids of soil organisms. Here, we exposed springtails (Collembola, small soil living arthropods) to phenanthrene (a common PAH) in combination with a range of soil water contents to investigate the combined effects of these factors on the bioaccumulation, survival, recruitment, and body growth in a full factorial experiment. The results showed that phenanthrene up to 60 mg/kg dry soil had moderate effects on survival (<20%), whereas dry soil (4% soil water content) caused approximately 60% mortality. The bioaccumulation of phenanthrene increased almost 3-fold when soil water content decreased from 22 to 4%. We observed a joint effect of low soil water content and phenanthrene on recruitment, suggesting a synergistic interaction. The recruitment EC50 values of phenanthrene decreased from approximately 40 mg/kg dry soil at 22% soil water content to approximately 10 mg/kg dry soil at 12% soil water content. Our results show that the effects of phenanthrene are more pronounced in dry soil partly because bioaccumulation is enhanced when soils become dry.

Globally invariant metabolism but density-diversity mismatch in springtails.

Soil life supports the functioning and biodiversity of terrestrial ecosystems. Springtails (Collembola) are among the most abundant soil arthropods regulating soil fertility and flow of energy through above- and belowground food webs. However, the global distribution of springtail diversity and density, and how these relate to energy fluxes remains unknown. Here, using a global dataset representing 2470 sites, we estimate the total soil springtail biomass at 27.5 megatons carbon, which is threefold higher than wild terrestrial vertebrates, and record peak densities up to 2 million individuals per square meter in the tundra. Despite a 20-fold biomass difference between the tundra and the tropics, springtail energy use (community metabolism) remains similar across the latitudinal gradient, owing to the changes in temperature with latitude. Neither springtail density nor community metabolism is predicted by local species richness, which is high in the tropics, but comparably high in some temperate forests and even tundra. Changes in springtail activity may emerge from latitudinal gradients in temperature, predation and resource limitation in soil communities. Contrasting relationships of biomass, diversity and activity of springtail communities with temperature suggest that climate warming will alter fundamental soil biodiversity metrics in different directions, potentially restructuring terrestrial food webs and affecting soil functioning.

Increased daily temperature fluctuations exacerbate the toxicity of phenanthrene in Enchytraeus albidus (Enchytraeidae).

Temperature variability in soils is expected to increase due to the more frequent occurrence of heat waves, putting species under thermal stress. In addition, organic pollutants such as polycyclic aromatic hydrocarbons (PAHs) are released into the environment due to anthropogenic activities. Both stressors negatively impact terrestrial organisms and may interact with each other. Here, we subjected the soil living enchytraeid, Enchytraeus albidus, to combined exposure to phenanthrene (PHE; 0, 10, 20, 40, and 80 mg kg-1 dry soil) and a range of temperature treatments (constant temperature (CT): 10, 15 and 20 °C; different mean temperature with the same daily temperature fluctuation (DTF-5): 10 ± 5, 15 ± 5 and 20 ± 5 °C; daily temperature fluctuation with the same mean, but different amplitudes (DTF-A): 20, 20 ± 2, 20 ± 5 and 20 ± 7 °C). We measured internal PHE concentration in adults and found that an increase in mean temperature significantly increased the internal PHE concentration. The production of juveniles was measured using a standardized test. We found a synergistic interaction between the temperature amplitude (DTF-A treatments) and PHE on the reproduction of E. albidus. The EC50 of reproduction decreased with increasing amplitude. These results show that the negative effects of PHE on E. albidus can be magnified if stressful temperatures are reached (although briefly) during diurnal fluctuations of soil temperature. Our results highlight the importance and inclusion of extreme thermal events in the risk assessment of pollutants.

Climate Change Helps Polar Invasives Establish and Flourish: Evidence from Long-Term Monitoring of the Blowfly Calliphora vicina.

The isolated sub-Antarctic islands are of major ecological interest because of their unique species diversity and long history of limited human disturbance. However, since the presence of Europeans, these islands and their sensitive biota have been under increasing pressure due to human activity and associated biological invasions. In such delicate ecosystems, biological invasions are an exceptional threat that may be further amplified by climate change. We examined the invasion trajectory of the blowfly Calliphora vicina (Robineau-Desvoidy 1830). First introduced in the sub-Antarctic Kerguelen Islands in the 1970s, it is thought to have persisted only in sheltered microclimates for several decades. Here, we show that, in recent decades, C. vicina has been able to establish itself more widely. We combine experimental thermal developmental data with long-term ecological and meteorological monitoring to address whether warming conditions help explain its current success and dynamics in the eastern Kerguelen Islands. We found that warming temperatures and accumulated degree days could explain the species' phenological and long-term invasion dynamics, indicating that climate change has likely assisted its establishment. This study represents a unique long-term view of a polar invader and stresses the rapidly increasing susceptibility of cold regions to invasion under climate change.

Heat stress delays detoxification of phenanthrene in the springtail Folsomia candida.

Climate change has intensified the occurrence of heat waves, resulting in organisms being exposed to thermal and chemical stress at the same time. The effects of mild heat shock combined with sublethal concentrations of phenanthrene (PHE) on defense mechanisms in springtails Folsomia candida were investigated. The transcription of Heat Shock Protein 70 (HSP70) was significantly upregulated by heat shock but tended to reach the control levels after 42 h of recovery. The transcription of cytochrome P450 3A13 (CYP3A13) was upregulated 3-13 fold by PHE but suppressed by heat shock. The suppression by heat shock might contribute to the reduced detoxification of PHE during high-temperature exposure. In line with this, we found that the internal PHE concentration was approximately 70% higher in heat-shocked springtails than in animals kept at control temperature. In general, the transcription of genes encoding enzymes of detoxification phase Ⅱ (glutathione S-transferase 3) and phase Ⅲ (ABC transporter 1) and the activity of antioxidant defense enzymes (superoxide dismutase and catalase) were less influenced than genes encoding phase I detoxification mechanisms (CYP3A13). These results indicate that heat shock delays the detoxification of PHE in springtails.

Analysis of heat and cold tolerance of a freeze-tolerant soil invertebrate distributed from temperate to Arctic regions: evidence of selection for extreme cold tolerance.

Tolerance to thermal extremes is critical for the geographic distributions of ectotherm species, many of which are probably going to be modified by future climatic changes. To predict species distributions it is important to understand the potential of species to adapt to changing thermal conditions. Here, we tested whether the thermal tolerance traits of a common freeze-tolerant potworm were correlated with climatic conditions and if adaptation to extreme cold constrains the evolutionary potential for high temperature tolerance. Further, we tested if evolution of thermal tolerance traits is associated with costs in other fitness traits (body size and reproduction). Lastly, we tested if slopes of temperature-survival curves (i.e., the sensitivity distribution) are related to tolerance itself. Using 24 populations of the potworm, Enchytraeus albidus Henle (Enchytraeidae), collected from a wide range of climatic conditions, we established a common garden experiment in which we determined high and low temperature tolerance (using survival as endpoint), average reproductive output and adult body size. Heat tolerance was not related to environmental temperatures whereas lower lethal temperature was about 10 °C lower in Arctic populations than in populations from temperate regions. Reproduction was not related to environmental temperature, but was negatively correlated with cold tolerance. One explanation for the trade-off between cold tolerance and reproduction could be that the more cold-hardy populations need to channel energy to large glycogen reserves at the expense of less energy expenditure for reproduction. Adult body size was negatively related to environmental temperature. Finally, the slopes of temperature-survival curves were significantly correlated with critical temperature limits for heat and cold tolerance; i.e., slopes increased with thermal tolerance. Our results suggest that relatively heat-sensitive populations possess genetic variation, leaving room for improved heat tolerance through evolutionary processes, which may alleviate the effects of a warmer future climate in the Arctic. On the other hand, we observed relatively narrow sensitivity distributions (i.e., less variation) in the most heat tolerant populations. Taken together, our results suggest that both cold and heat tolerance can only be selected for (and improved) until a certain limit has been reached.

Survival and predation rate of wild-caught and commercially produced Orius majusculus (Reuter) (Hemiptera: Anthocoridae).

The quality of biological control agents used in augmentative releases may be affected by rearing conditions due to inbreeding or laboratory adaptation, or to phenotypic effects of the rearing environment. We hypothesized that individuals from a wild population would be in better body condition and kill more prey than individuals from a commercially produced population. We caught wild Orius majusculus (Reuter) in a maize field and compared their initial body mass, survival, and prey reduction capacity to commercially produced O. majusculus. Predation capacity and survival were compared in short-term Petri dish tests with Frankliniella tenuicornis (Uzel) thrips, Ephestia kuehniella (Zeller) moth eggs, or Rhopalosiphum padi (L.) aphids as prey, and in longer-term outdoor mesocosms containing live seedling wheat grass with thrips or aphids as prey. Wild-caught O. majusculus were typically heavier and overall had higher survival during tests than commercially produced O. majusculus. Females were heavier than males and typically killed more prey. However, we found no difference between wild-caught and commercially produced individuals on prey reduction, neither in Petri dishes nor in mesocosms. Our study suggests that commercially produced O. majusculus have lower body condition than wild O. majusculus due to their lower body mass and survival, but that this does not have any negative effect on the number of pest prey killed over the timelines and conditions of our tests. Commercially produced O. majusculus thus did not have a lower impact on pest prey numbers than wild-caught individuals and therefore had similar biological control value under our study conditions.

Thermal plasticity and sensitivity to insecticides in populations of an invasive beetle: Cyfluthrin increases vulnerability to extreme temperature.

Climate change increases average temperatures and the occurrence of extreme weather events, in turn accentuating the risk of organism exposure to temperature stress. When thermal conditions become stressful, the sensitivity of insects toward insecticides can be exacerbated. Likewise, exposure of insects to insecticides can subsequently influence their ability to handle stressful temperatures. Here, we investigated the effects of constant temperature and daily heat spikes, in presence/absence of insecticide treatment (cyfluthrin), on the condition (impairment of mobility) and thermal tolerance to cold (-6 °C) and heat (42.5 °C) of the terrestrial beetle Alphitobius diaperinus. The responses of insects from four populations (three farm-collected populations, one laboratory population) to different durations of extreme temperature exposure were compared. The results showed that the laboratory population was generally more sensitive to extreme cold and heat temperatures, with less than 50% of adults recovering after an exposure at -6 or +42.5 °C for 3h. Significant differences in the level of thermal tolerance were also found among insects from poultry farms. Cyfluthrin exposure incurred detrimental effects to insects' condition in all but one population. For two out of the four populations, mobility impairment was increased when adults were exposed to daily heat spikes (6 h per day at 38 °C) and cyfluthrin simultaneously, compared to cyfluthrin exposure at constant temperatures; yet, no significant interaction between the two stressors was found. Finally, using one farm collected population, effects of pre-exposure to cyfluthrin on extreme temperature tolerance provided another example of the toxicant-induced climate sensitivity in insects.

Thermal optimum for mass production of the live feed organism Enchytraeus albidus.

Live feed organisms are essential for the larval stages of many fish species grown in aquaculture, and juvenile fish reared on live feeds often exhibit higher survival and growth than those reared on formulated feed. The terrestrial enchytraeid (white worm), Enchytraeus albidus, has potential as a sustainable source of live feed because it can easily be mass produced, feeds on a wide range of organic waste materials and has high contents of protein and long-chain poly-unsaturated fatty acids. In the present study, we observed the effect of temperature on population growth over five months using soil microcosms. At the outset, each microcosm was supplied with approximately the same number of cocoons. Hatched enchytraeids were given rolled oats ad libitum as feed. We followed the population growth at seven temperatures in the range of 4-25 °C and investigated body composition in order to find optimal temperature for mass production. Results showed that E. albidus has a broad thermal optimum range and displays almost similar biomass production in the range of 15-22 °C with specific growth rates between 6.5 and 6.8%. In this temperature range, protein contents were 40-45%, glycogen contents 20-25% and total fatty acid contents 15-20% of dry weight. The temperature had a highly significant effect on fatty acid composition. In particular, the abundance of omega-3 fatty acids (18:3ω3 and 20:5ω3) was largest at low temperature. For what concerns achievable density of worms in mass cultures, our results surpassed previous results and showed that densities close to 100 g L-1 substrate are realistic. Maximum production of biomass can likely reach 80 g live worms L-1 month-1 at temperatures between 15 and 22 °C.

Temperature-Dependent Toxicokinetics of Phenanthrene in Enchytraeus albidus (Oligochaeta).

Although the toxicokinetics of organic pollutants in soil invertebrates under optimal and constant temperature has been widely reported, their uptake, elimination, and bioaccumulation under suboptimal temperatures, and especially daily fluctuating temperature (FT) regimes have received only little research attention. In this study, the uptake, elimination, and bioaccumulation of phenanthrene (PHE) in Enchytraeus albidus (Oligochaeta) under different constant temperatures, and an FT regime were investigated in a natural soil. In general, the PHE concentrations in worm tissues reached steady state within 14 days at different temperatures. The uptake (ku) and elimination (ke) rate constants and the bioaccumulation increased with increasing temperature likely because of an increased diffusivity of PHE into the worms and an increased metabolic rate. Interestingly, the bioaccumulation factor of PHE in E. albidus showed a positive relationship with temperature because the slope of the ku-temperature relationship was larger than that of the ke-temperature relationship. Further, the uptake and elimination rate constants were larger under the FT regime than at the constant average of the fluctuating temperature. These findings suggest that, climatic conditions, especially daily fluctuating temperatures, should be considered for the assessment of the toxicokinetics of organic pollutants in terrestrial organisms.

Mechanistic Effect Modeling of Earthworms in the Context of Pesticide Risk Assessment: Synthesis of the FORESEE Workshop.

Earthworms are important ecosystem engineers, and assessment of the risk of plant protection products toward them is part of the European environmental risk assessment (ERA). In the current ERA scheme, exposure and effects are represented simplistically and are not well integrated, resulting in uncertainty when the results are applied to ecosystems. Modeling offers a powerful tool to integrate the effects observed in lower tier laboratory studies with the environmental conditions under which exposure is expected in the field. This paper provides a summary of the (In)Field Organism Risk modEling by coupling Soil Exposure and Effect (FORESEE) Workshop held 28-30 January 2020 in Düsseldorf, Germany. This workshop focused on toxicokinetic-toxicodynamic (TKTD) and population modeling of earthworms in the context of ERA. The goal was to bring together scientists from different stakeholder groups to discuss the current state of soil invertebrate modeling and to explore how earthworm modeling could be applied to risk assessments, in particular how the different model outputs can be used in the tiered ERA approach. In support of these goals, the workshop aimed at addressing the requirements and concerns of the different stakeholder groups to support further model development. The modeling approach included 4 submodules to cover the most relevant processes for earthworm risk assessment: environment, behavior (feeding, vertical movement), TKTD, and population. Four workgroups examined different aspects of the model with relevance for risk assessment, earthworm ecology, uptake routes, and cross-species extrapolation and model testing. Here, we present the perspectives of each workgroup and highlight how the collaborative effort of participants from multidisciplinary backgrounds helped to establish common ground. In addition, we provide a list of recommendations for how earthworm TKTD modeling could address some of the uncertainties in current risk assessments for plant protection products. Integr Environ Assess Manag 2021;17:352-363. © 2020 SETAC.

Compartmentation and effects of lead (Pb) in the collembolan, Folsomia candida.

The impact of soil lead (Pb) pollution on survival, growth, and reproduction of the collembolan, Folsomia candida, and Pb compartmentation in its gut and remaining body parts were studied by exposing animals to laboratory-spiked soil. The survival, growth, and reproduction of F. candida were significantly reduced by increasing soil Pb concentration. The LC50 values of survival based on total and CaCl2-extractable Pb concentration in soil were 2562 mg kg-1 and 351 mg kg-1, respectively. The EC50 values of reproduction were 1244 mg kg-1 and 48 mg kg-1, respectively. The Pb concentration in whole body, gut, and remaining body parts was significantly increased with the increase of soil Pb concentration and followed an exponential increase when the soil Pb concentration was equal to or above a threshold (1000 mg kg-1 for whole body and remaining body part, 500 mg kg-1 for gut). Below this threshold, these relationships were linear. The Pb concentration in the gut was higher than whole body and remaining body part of F. candida, and the threshold of internal Pb concentration at which F. candida can compensate was in the range 7-13 mg Pb kg-1 dry animal (corresponding to soil Pb concentration 500-1000 mg Pb kg-1 dry soil). The results indicate that reproduction of F. candida was a more sensitive indicator of lead toxicity than survival and growth. Pb was mainly accumulated in the gut of F. candida. We discuss the internal Pb concentration as an indicator of adverse effects in the risk assessment of soil Pb pollution.

Mercury (Hg2+) interferes with physiological adaptations to freezing in the arctic earthworm Enchytraeus albidus.

Freezing temperatures is an important stressor in the arctic regions and has a significant influence on the population dynamics and geographic distribution of terrestrial invertebrates. Toxic metals in the environment can interfere with protective cold-acclimation responses of organisms. It is therefore important to evaluate the combined effects of cold stress and environmental contaminants. Here, we aimed to investigate the effects of Hg (HgCl2) on various physiological aspects of freeze-tolerance in the earthworm (Enchytraeus albidus). We measured the levels of the cryoprotectant glucose, the glycogen content (source of glucose molecules for cryoprotection and fuel for metabolism), and changes in the composition of membrane phospholipid fatty acids (PLFA) as an indicator of lipid peroxidation. Freezing at -6 °C had no effect on survival in uncontaminated soil, however, survival of freezing in Hg contaminated soil was clearly reduced, especially at extended exposure times. Thus, the LC50 value in frozen soil decreased from 8.3 mg Hg kg-1 (when exposed for 17 days) to only 4.2 mg Hg kg-1 after 36 days' exposure indicating that combined effects of Hg and freezing became larger at prolonged exposure times. Hg caused a depletion of glycogen reserves (almost 50% at 12 mg kg-1 dry soil), but despite this effect worms were able to maintain a constant cryoprotectant level (about 0.12 mg glucose mg-1 dry weight) at all Hg concentrations. Hg had clear negative effects on the proportion of unsaturated PLFAs, which could be an indication of lipid peroxidation. Since a high proportion of unsaturated fatty acids in the membrane is important for invertebrate freeze-tolerance, our results suggest that the negative effect of Hg on freeze-tolerance in E. albidus is related to degraded membrane functionality at low temperature.