Predicting the Combined Effects of Multiple Stressors and Stress Adaptation in Gammarus pulex.

Global change confronts organisms with multiple stressors causing nonadditive effects. Persistent stress, however, leads to adaptation and related trade-offs. The question arises: How can the resulting effects of these contradictory processes be predicted? Here we show that Gammarus pulex from agricultural streams were more tolerant to clothianidin (mean EC50 148 µg/L) than populations from reference streams (mean EC50 67 µg/L). We assume that this increased tolerance results from a combination of physiological acclimation, epigenetic effects, and genetic evolution, termed as adaptation. Further, joint exposure to pesticide mixture and temperature stress led to synergistic interactions of all three stressors. However, these combined effects were significantly stronger in adapted populations as shown by the model deviation ratio (MDR) of 4, compared to reference populations (MDR = 2.7). The pesticide adaptation reduced the General-Stress capacity of adapted individuals, and the related trade-off process increased vulnerability to combined stress. Overall, synergistic interactions were stronger with increasing total stress and could be well predicted by the stress addition model (SAM). In contrast, traditional models such as concentration addition (CA) and effect addition (EA) substantially underestimated the combined effects. We conclude that several, even very disparate stress factors, including population adaptations to stress, can act synergistically. The strong synergistic potential underscores the critical importance of correctly predicting multiple stresses for risk assessment.

Chronic metal contamination shapes the size structure of Gammarus fossarum populations in French headwater rivers.

Assessing the effects of multigenerational exposure of aquatic animal populations to chemical contamination is essential for ecological risk assessment. However, beyond rare examples reporting the sporadic emergence of a toxicological tolerance within populations that persist in contaminated environments, conclusive results are even more limited from field studies when it comes to the alteration of life-history traits. Here, we investigated whether long-term exposure to cadmium (Cd) influences size-related life-history traits (i.e., size at puberty, median adult size, maximum size) in Gammarus fossarum, a keystone species of European stream ecosystems. We studied 13 field populations of G. fossarum (cryptic lineage B) living in headwater rivers located in natural areas scattered at a large geographical scale and exposed to contrasted bioavailable Cd contamination levels due to different local geochemical backgrounds. We achieved a detailed description of the physical and physicochemical conditions of the river reaches investigated. Land-use parameters, hydrological characteristics (flow, slope, river width, flow structure, mosaic of substrates), and physicochemical conditions (temperature, conductivity, dissolved oxygen) were measured. Metallic bioavailable contamination was assessed using a standardized active biomonitoring procedure (Gammarus caging). Based on the field demographic census of the 13 populations, our results demonstrated that chronic Cd contamination significantly influences life-history in the G. fossarum species, with a significant reduction in all size traits of populations (size at puberty, median adult size, maximum size). In addition, we confirmed Cd-tolerance in contaminated populations during exposure tests in the laboratory. Various hypotheses can be then put forward to explain the modification of size-related life-history traits: a direct toxic effect of Cd, a cost of Cd-tolerance, or an adaptive evolution of life-history exposed to toxic pressure.

Attenuation Effect of Azolla spp. on Lambda-Cyhalothrin Toxicity to Aquatic Organisms.

Wetlands play a crucial role in providing valuable ecosystem services, including the removal of various pollutants. In agricultural basins, wetlands are exposed to agrochemical loads. This study aims to assess the attenuation effect of the ubiquitous macrophyte Azolla spp. on the toxicity of lambda-cyhalothrin to sensitive aquatic organisms. An indoor mesocosm experiment was conducted to compare the concentration of lambda-cyhalothrin at different time points after pesticide application in vegetated and unvegetated treatments, including a control without pesticide addition. Toxicity tests were performed throughout the experiment on three organisms: a fish (Cnesterodon decemmaculatus), a macroinvertebrate (Hyalella curvispina), and an amphibian (Boana pulchella). The results demonstrated that lambda-cyhalothrin concentration and toxicity in water were significantly lower in the Azolla spp. treatment. Furthermore, the half-life of lambda-cyhalothrin decreased from 1.2 days in the unvegetated treatment to 0.4 days in the vegetated treatment. The vegetated treatment also resulted in a significantly lower mortality rate for both H. curvispina and C. decemmaculatus. However, no mortality was observed in B. pulchella for any of the treatments. Sublethal effects were observed in this organism, such as lateral bending of the tail and impairment of the ability to swim, which were attenuated in the vegetated treatment. We conclude that Azolla spp. can effectively reduce the concentration and toxicity of lambda-cyhalothrin, suggesting its potential use in farm-scale best management practices to mitigate the effects of pesticide loads from adjacent crops.

Insights into In Situ Benthic Caging Tests for Ecotoxicity Assessments Targeting Discharging Groundwater Contaminant Plumes.

While in situ toxicity testing with caged organisms has been used to assess surface water and sediment contamination, no successful application to benthic organisms exposed to highly contaminated groundwater plumes discharging to surface waters has been reported. The objective of this study was to demonstrate and evaluate this application using four sets of tests performed at three previously reported contaminated groundwater sites, which include one river site affected by volatile organic contaminant plumes, and two sites, one pond and one small urban stream, impacted by landfill plumes. The study examined multiple cage designs and orientations and two test organisms: an amphipod (Hyalella azteca) and midge larvae (Chironomus riparius; only one study). Cages were deployed for between 5 and 28 days and assessed for organism survival and growth. At all sites and for some deployment conditions, cages exposed to high contaminant concentrations in the plume footprint had greater mortality compared to those exposed to lower or background concentrations. Organism growth was less clear as a metric of toxicity. Vertically oriented cages typically showed high mortality to plume contaminants, but some were also affected by other non-target groundwater conditions (e.g., low dissolved oxygen, other contaminant sources), while horizontally oriented cages were rarely responsive to either groundwater influence. A hybrid cage design showed much promise in its single study. Useful observations on the test organisms and on potentially problematic site conditions were also made. The informed use of in situ toxicity cages could be an additional beneficial tool for groundwater contaminated site assessments.

Effects of acute arsenic exposure in two different populations of Hyalella curvispina amphipods from North Patagonia Argentina.

Arsenic (As) is a toxic metalloid present in high levels in diverse regions of Argentina. The aim of this study was to determine acute As-mediated toxicity in two different populations of autochthonous Hyalella curvispina amphipods from a reference site (LB) and an agricultural one (FO) within North Patagonia Argentina. Previously, both populations exhibited significant differences in pesticide susceptibility. Lab assays were performed to determine acute lethal concentrations, as well as some biochemical parameters. Lethal concentration (LC50) values obtained after 48 and 96 hr As exposure were not significantly different between these populations, although FO amphipods appeared slightly less susceptible. LC50-48 hr values were 3.33 and 3.92 mg/L As, while LC50-96 hr values were 1.76 and 2.14 mg/L As for LB and FO amphipods. The no observed effect concentration (NOEC) values were 0.5 mg/L As. Cholinesterase (ChE) activity was significantly diminished by As acute exposure (0.5-1.5 mg/L As), indicative of a significant neurotoxic action for this metalloid in both amphipod populations. Activities of catalase (CAT) and glutathione S-transferase (GST) and levels of reduced glutathione (GSH) were differentially altered following As exposure. CAT activity was increased after 96 hr As exposure. GST activity and GSH levels were significantly elevated followed by either a decrease or a return to control values after 96 hr treatment. However, additional studies are necessary to understand the mechanisms underlying the As-mediated oxidative effects in H. curvispina. Our findings suggest that measurement of ChE activity in H. curvispina amphipods might serve as a useful biomarker of As exposure and effect.

Chronic effects of the strobilurin fungicide azoxystrobin in the leaf shredder Gammarus fossarum (Crustacea; Amphipoda) via two effect pathways.

Fungicides pose a risk for crustacean leaf shredders serving as key-stone species for leaf litter breakdown in detritus-based stream ecosystems. However, little is known about the impact of strobilurin fungicides on shredders, even though they are presumed to be the most hazardous fungicide class for aquafauna. Therefore, we assessed the impact of the strobilurin azoxystrobin (AZO) on the survival, energy processing (leaf consumption and feces production), somatic growth (growth rate and molting activity), and energy reserves (neutral lipid fatty and amino acids) of the amphipod crustacean Gammarus fossarum via waterborne exposure and food quality-mediated (through the impact of leaf colonizing aquatic microorganisms) and thus indirect effects using 2 × 2-factorial experiments over 24 days. In a first bioassay with 30 µg AZO/L, waterborne exposure substantially reduced survival, energy processing and affected molting activity of gammarids, while no effects were observed via the dietary pathway. Furthermore, a negative growth rate (indicating a body mass loss in gammarids) was induced by waterborne exposure, which cannot be explained by a loss in neutral lipid fatty and amino acids. These energy reserves were increased indicating a disruption of the energy metabolism in G. fossarum caused by AZO. Contrary to the first bioassay, no waterborne AZO effects were observed during a second experiment with 15 µg AZO/L. However, an altered energy processing was determined in gammarids fed with leaves microbially colonized in the presence of AZO, which was probably caused by fungicide-induced effects on the microbial decomposition efficiency ultimately resulting in a lower food quality. The results of the present study show that diet-related strobilurin effects can occur at concentrations below those inducing waterborne toxicity. However, the latter seems to be more relevant at higher fungicide concentrations.

Sub-chronic effects of AgNPs and AuNPs on Gammarus fossarum (Crustacea Amphipoda): From molecular to behavioural responses.

The aim of the present study was the assessment of the sub-chronic effects of silver (AgNPs) and gold nanoparticles (AuNPs) of 40 nm primary size either stabilised with citrate (CIT) or coated with polyethylene glycol (PEG) on the freshwater invertebrate Gammarus fossarum. Silver nitrate (AgNO3) was used as a positive control in order to study the contribution of silver ions potentially released from AgNPs on the observed effects. A multibiomarker approach was used to assess the long-term effects of AgNPs and AuNPs 40 nm on molecular, cellular, physiological and behavioural responses of G. fossarum. Specimen of G. fossarum were exposed for 15 days to 0.5 and 5 µgL-1 of CIT and PEG AgNPs and AuNPs 40 nm in the presence of food. A significant uptake of both Ag and Au was observed in exposed animals but was under the toxic threshold leading to mortality of G. fossarum. Silver nanoparticles (CIT-AgNPs and PEG-AgNPs 40 nm) led to an up-regulation of Na+K+ATPase gene expression. An up-regulation of Catalse and Chitinase gene expressions due to exposure to PEG-AgNPs 40 nm was also observed. Gold nanoparticles (CIT and PEG-AuNPs 40 nm) led to an increase of CuZnSOD gene expression. Furthermore, both AgNPs and AuNPs led to a more developed digestive lysosomal system indicating a general stress response in G. fossarum. Both AgNPs and AuNPs 40 nm significantly affected locomotor activity of G. fossarum while no effects were observed on haemolymphatic ions and ventilation.

The anti-lipidemic drug simvastatin modifies epigenetic biomarkers in the amphipod Gammarus locusta.

The adverse effects of certain environmental chemicals have been recently associated with the modulation of the epigenome. Although changes in the epigenetic signature have yet to be integrated into hazard and risk assessment, they are interesting candidates to link environmental exposures and altered phenotypes, since these changes may be passed across multiple non-exposed generations. Here, we addressed the effects of simvastatin (SIM), one of the most prescribed pharmaceuticals in the world, on epigenetic regulation using the amphipod Gammarus locusta as a proxy, to support its integration into hazard and environmental risk assessment. SIM is a known modulator of the epigenome in mammalian cell lines and has been reported to impact G. locusta ecological endpoints at environmentally relevant levels. G. locusta juveniles were exposed to three SIM environmentally relevant concentrations (0.32, 1.6 and 8 µg L-1) for 15 days. Gene transcription levels of selected epigenetic regulators, i.e., dnmt1, dmap1, usp7, kat5 and uhrf1 were assessed, along with the quantification of DNA methylation levels and evaluation of key ecological endpoints: survival and growth. Exposure to 0.32 and 8 µg L-1 SIM induced significant downregulation of DNA methyltransferase 1 (dnmt1), concomitant with global DNA hypomethylation and growth impacts. Overall, this work is the first to validate the basal expression of key epigenetic regulators in a keystone marine crustacean, supporting the integration of epigenetic biomarkers into hazard assessment frameworks.

Long term bioassessment multiple stressors study in a residential California stream.

The relationship of various benthic metrics to physical habitat metrics, pyrethroids, metals and sediment parameters was evaluated for a 10 year data set in Pleasant Grove Creek (Roseville, California) using univariate linear models, stepwise multiple regressions, and canonical correlation analysis. In general, total physical habitat scores in this residential stream were considered to be marginal to suboptimal. The most dominant benthic taxa were generally considered to be tolerant of environmental stressors and the benthic communities were rated as impaired based on a benthic index. Potentially toxic sediment concentrations of cadmium, chromium, copper, nickel and zinc were reported at various sites based on a comparison with existing threshold effect levels. The sum of pyrethroid Toxics Units (TUs) indicated that 10 of 21 sites based on a sensitive Hyalella laboratory toxicity test had TUs greater than one thus suggesting toxicity at various sites. In summary, the effects of the physical habitat, as reflected by certain habitat metrics that were indicative of stream-flow, hydrology, habitat diversity, and substrate quality overshadowed any apparent effects of pyrethroids and metals on shaping resident benthic communities when all environmental variables were considered in multivariate analyses.

Can Toxicities Induced by Insecticide Methomyl be Remediated Via Soil Bacteria Ochrobactrum thiophenivorans and Sphingomonas melonis?

The research study was about revealing the biochemical response of Gammarus pulex related to insecticide methomyl before and after bioremediation by two soil bacteria species, Ochrobactrum thiophenivorans and Sphingomonas melonis. Catalase (CAT), glutathione S-transferase.(GST), cytochrome. P4501A1 (CYP1A1) activities in G. Pulex related to methomyl solution were investigated in 24 h and 96 h. ELISA method was used for test studies. CAT enzyme was decreased in Gammarus pulex that was exposed to methomyl after all exposure period (P < 0.05). CAT activities were returned to control results after bioremediation assays. GST enzyme activity was decreased depending on methomyl exposure during 24 h but increased during 4 days (P < 0.05). After 8 days of bioremediation period, GST activity increased again during 24 h while decreased during 4 days (P < 0.05). CYP1A1 activity increased in Gammarus pulex that was exposed to methomyl after all exposure period (P > 0.05). After bioremediation, statistically significant changes were not revealed in CYP1A1 activities (P > 0.05). According to the results of our study, CYP1A1, CAT, and GST activities in G. pulex sanctioned the capability of Ochrobactrum thiophenivorans and Sphingomonas melonis in methomyl bioremediation. Isolated and enriched Ochrobactrum thiophenivorans and Sphingomonas melonis that were added to 2.5 ppb concentrations of methomyl for 8 days. Each day, chemical oxygen demand (COD) and biochemical oxygen demand (BOD5), pH and dissolved oxygen parameters were monitored. At the final phase of the bioremediation step, it was determined that these bacteria have efficient methomyl bioremediation properties in a mixed corsortia at a rate of 86%. These results show that these bacteria can be used for bioremediate the receiving environments that are polluted by these kinds of insecticides.

The chronic toxicity of emamectin benzoate to three marine benthic species using microcosms.

The commercial farming of Atlantic salmon, Salmo salar, may require the periodic application of emamectin benzoate (EB) treatments to reduce the effects of biological pests, such as sea lice. As a result, EB is detected in sediments beneath these fish farms at considerable levels. Literature sediment toxicity data for EB for marine benthic species is only available for 10-day sediment toxicity tests, which might be too short to assess field effects. Here, we present a sediment toxicity test to determine 28-day mortality and growth effect concentrations for the non-target polychaete worm Arenicola marina, the crustacean Corophium volutator and the mollusk Cerastoderma edule using a marine microcosm setup. Results indicate that no concentration-dependent increase of mortality and growth rate was apparent to A. marina and C. edule. But for C. volutator, a concentration-dependent increase in mortality was observed, resulting in a calculated 28-d LC50 of 316 µg/kg dry sediment (95% confidence interval: 267-373 µg/kg dry sediment). There were significant effects on C. volutator growth rate at concentrations of 100 µg/kg dry sediment and above (NOEC = 30 µg/kg dry sediment). These observations show that C. volutator is more sensitive to EB than A. marina, which differs from results reported in previous studies. Comparison to the most sensitive NOEC (30 µg/kg dry sediment) found for C. volutator (organisms of 8-11 mm length), shows that the Environmental Quality Standard, derived by the Scottish Environment Protection Agency in 2017 which based on freshwater species data (NOEC = 1.175 µg/kg dry sediment), are relatively strict and is sufficiently protective for the marine species tested in this paper.

Population- and sex-specific sensitivity of the marine amphipod Allorchestes compressa to metal exposure.

The sensitivity to contaminants of natural populations varies greatly depending on their historical exposure and on the sex of the individual. These factors result in great uncertainty in ecotoxicological risk assessments and challenge the protection of marine biodiversity. This study investigated the role of background pollution in the environment in shaping the sensitivity of males and females of the common marine amphipod Allorchestes compressa to the common trace marine pollutant, copper (Cu). Female and male amphipods were collected from two sites: Geelong (the polluted site) and Clifton Springs (the clean site). Amphipods were exposed to Cu treatments of 0, 50, 100, and 250 µg/L for 10 days, followed by a 10-day recovery period. Cu-exposed males from Geelong showed a reduction in feeding rate at a higher Cu concentration than males from Clifton Springs, suggesting that they have a higher tolerance to Cu than males from Clifton Springs. This can be explained by their higher base level of metallothioneins (MTs) and glutathione-S-transferase (GST), the key physiological responses for detoxification and defence against damages from Cu toxicity. Males showed a higher tolerance to Cu than females. This pattern was similar in both populations, which may be associated with a higher level of GST. During the recovery period, only males from Geelong fully recovered to the control level. Our results emphasize the importance of considering population- and sex-specific sensitivity of invertebrates to contaminants in ecotoxicological risk assessments.

Influence of pH on the toxicity of ionisable pharmaceuticals and personal care products to freshwater invertebrates.

The majority of pharmaceuticals and personal health-care products are ionisable molecules at environmentally relevant pHs. The ionization state of these molecules in freshwater ecosystems may influence their toxicity potential to aquatic organisms. In this study we evaluated to what extent varying pH conditions may influence the toxicity of the antibiotic enrofloxacin (ENR) and the personal care product ingredient triclosan (TCS) to three freshwater invertebrates: the ephemeropteran Cloeon dipterum, the amphipod Gammarus pulex and the snail Physella acuta. Acute toxicity tests were performed by adjusting the water pH to four nominal levels: 6.5, 7.0, 7.5 and 8.0. Furthermore, we tested the efficiency of three toxicity models with different assumptions regarding the uptake and toxicity potential of ionisable chemicals with the experimental data produced in this study. The results of the toxicity tests indicate that pH fluctuations of only 1.5 units can influence EC50-48 h and EC50-96 h values by a factor of 1.4-2.7. Overall, the model that only focuses on the fraction of neutral chemical and the model that takes into account ion-trapping of the test molecules showed the best performance, although present limitations to perform risk assessments across a wide pH range (i.e., well above or below the substance pKa). Under such conditions, the model that takes into account the toxicity of the neutral and the ionized chemical form is preferred. The results of this study show that pH fluctuations can have a considerable influence on toxicity thresholds, and should therefore be taken into account for the risk assessment of ionisable pharmaceuticals and personal health-care products. Based on our results, an assessment factor of at least three should be used to account for toxicity differences between standard laboratory and field pH conditions. The models evaluated here can be used to perform refined risk assessments by taking into account the influence of temporal and spatial pH fluctuations on aquatic toxicity.

Biomarkers in bivalve mollusks and amphipods for assessment of effects linked to cyanobacteria and elodea: Mesocosm study.

The effects of cyanobacteria (Aphanizomenon flos-aquae (90%), Microcystis aeruginosa) and dense Elodea canadensis beds on the health endpoints of the amphipod Gmelinoides fasciatus and bivalve mollusc Unio pictorum were examined in mesocosms with simulated summer conditions (July-August 2018) in the environment of the Rybinsk Reservoir (Volga River Basin, Russia). Four treatments were conducted, including one control and three treatments with influencing factors, cyanobacteria and dense elodea beds (separately and combined). After 20 days of exposure, we evaluated the frequency of malformed and dead embryos in amphipods, heart rate (HR) and its recovery (HRR) after stress tests in molluscs as well as heat tolerance (critical thermal maximum or CTMax) in both amphipods and molluscs. The significant effect, such as elevated number of malformed embryos, was recorded after exposure with cyanobacteria (separately and combined with elodea) and presence of microcystins (MC) in water (0.17 µg/l, 40% of the most toxic MC-LR contribution). This study provided evidence that an elevated number (>5% of the total number per female) of malformed embryos in amphipods showed noticeable toxicity effects in the presence of cyanobacteria. The decreased oxygen under the influence of dense elodea beds led to a decrease in HR (and an increase in HRR) in molluscs. The notable effects on all studied biomarkers, embryo malformation frequency and heat tolerance in the amphipod G. fasciatus, as well as the heat tolerance and heart rate in the mollusc U. pictorum, were found when both factors (elodea and cyanobacteria) were combined. The applied endpoints could be further developed for environmental monitoring, but the obtained results support the importance of the combined use of several biomarkers and species, especially in the case of multi-factor environmental stress.

Sediment toxicity of the fungicide fludioxonil to benthic macroinvertebrates -evaluation of the tiered effect assessment procedure.

28-Day sediment-spiked laboratory toxicity tests with eight benthic macroinvertebrates and the lipophilic fungicide fludioxonil were conducted to verify the proposed tiered sediment effect assessment procedure as recommended by the European Food Safety Authority (EFSA). The test species were the oligochaetes Lumbriculus variegatus and Tubifex tubifex, the insects Chironomus riparius and Caenis horaria, the crustaceans Hyalella azteca and Asellus aquaticus and the bivalves Corbicula fluminalis and Pisidium amnicum. Toxicity estimates were expressed in terms of total concentration of dry sediment as well as in pore water concentration. Field-collected sediment, also used in a previously performed sediment-spiked microcosm experiment, was used in tests with all species. L. variegatus and C. riparius had similar lowest 28d-L(E)C10 values when expressed in terms of total sediment concentration, but in terms of pore water concentration L. variegatus was more sensitive. Three of the six additional benthic test species (A. aquaticus, C. horaria, C. fluminalis) had 28d-EC10 values a factor of 2-6 lower than that of L. variegatus. Comparing different effect assessment tiers for sediment organisms, i.e. Tier-0 (Modified Equilibrium Partitioning approach), Tier-1 (Standard Test Species approach), Tier-2 (Species Sensitivity Distribution (SSD) approach) and Tier-3 (Model Ecosystem approach), it is concluded that the tiers based on sediment-spiked laboratory toxicity tests provide sufficient protection when compared with the Tier-3 Regulatory Acceptable Concentration (RAC). Differences between Tier-1 and Tier-2 RACs, however, appear to be relatively small and not always consistent, irrespective of expressing the RAC in terms of total sediment or pore water concentration. Derivation of RACs by means of the SSD approach may be a challenge, because it is difficult obtaining a sufficient number of valid chronic EC10 values with appropriate 95% confidence bands for sediment-dwelling macroinvertebrates. Therefore, this paper proposes a Tier-2 Weight-of-Evidence approach to be used in case an insufficient number of valid additional toxicity data is made available. Similar studies with pesticides that differ in fate properties and toxic mode-of-action are necessary for further validation of the tiered effect assessment approach for sediment organisms.

Comparisons Between Laboratory Sediment Toxicity Test Results and Assessment of Benthic Community Changes for a Lake with Mild Metal Contamination.

Effects of moderate-to-severe sediment contamination may be readily discernable in both sediment toxicity bioassays and benthic community assessments. However, the impact may be less obvious under conditions of relatively mild contamination and significant variation of natural environmental variables. This study evaluated sediment toxicity and benthic community impacts for a shallow eutrophic lake system with relatively low levels of sediment metal contamination. We evaluated selected sediment physical and chemical properties, as well as benthic community structure, for 50 sites along a sediment Pb contamination gradient in the lake. We tested the toxicity of sediment from 20 of the sites, using a standardized 42-days sediment bioassay with Hyalella azteca survival, reproduction, and growth endpoints. Using principal component and correlation analyses we found negative relationships for both Pb and Cu, between sediment metal concentrations and the diversity and abundance of benthic macroinvertebrates. Taxa known to be metal-sensitive (e.g., Hyalella sp. and Physa sp.) were less abundant at sites with relatively higher Pb and Cu concentrations. However, amphipod performance in the chronic toxicity test was not related to sediment Pb or Cu concentrations (but was influenced by sediment organic content). Our results demonstrate that an assessment of community-level effects may be warranted even when sediment metal concentrations are too low to elicit detectable toxic effects in standardized laboratory tests.

A "Population Dynamics" Perspective on the Delayed Life-History Effects of Environmental Contaminations: An Illustration with a Preliminary Study of Cadmium Transgenerational Effects over Three Generations in the Crustacean Gammarus.

We explore the delayed consequences of parental exposure to environmentally relevant cadmium concentrations on the life-history traits throughout generations of the freshwater crustacean Gammarus fossarum. We report the preliminary results obtained during a challenging one-year laboratory experiment in this environmental species and propose the use of population modeling to interpret the changes in offspring life-history traits regarding their potential demographic impacts. The main outcome of this first long-term transgenerational assay is that the exposure of spawners during a single gametogenesis cycle (3 weeks) could result in severe cascading effects on the life-history traits along three unexposed offspring generations (one year). Indeed, we observed a decrease in F1 reproductive success, an early onset of F2 offspring puberty with reduced investment in egg yolk reserves, and finally a decrease in the growth rate of F3 juveniles. However, the analysis of these major transgenerational effects by means of a Lefkovitch matrix population model revealed only weak demographic impacts. Population compensatory processes mitigating the demographic consequences of parental exposure seem to drive the modification of life-history traits in offspring generations. This exploratory study sheds light on the role of population mechanisms involved in the demographic regulation of the delayed effects of environmental toxicity in wild populations.

Multiple Stressors in Aquatic Ecosystems: Sublethal Effects of Temperature, Dissolved Organic Matter, Light and a Neonicotinoid Insecticide on Gammarids.

Whether and to which extent the effects of chemicals in the environment interact with other factors remains a scientific challenge. Here we assess the combined effects of temperature (16 vs. 20°C), light conditions (darkness vs. 400 lx), dissolved organic matter (DOM; 0 vs. 6 mg/L) and the model insecticide thiacloprid (0 vs. 3 µg/L) in a full-factorial experiment on molting and leaf consumption of Gammarus fossarum. Thiacloprid was the only factor significantly affecting gammarids' molting. While DOM had low effects on leaf consumption, temperature, light and thiacloprid significantly affected this response variable. The various interactions among these factors were not significant suggesting additivity. Only the interaction of the factors temperature and thiacloprid suggested a tendency for antagonism. As most stressors interacted additively, their joint effects may be predictable with available models. However, synergistic interactions are difficult to capture while being central for securing ecosystem integrity.

Comparison between transcriptomic responses to short-term stress exposures of a common Holarctic and endemic Lake Baikal amphipods.

BACKGROUND: Lake Baikal is one of the oldest freshwater lakes and has constituted a stable environment for millions of years, in stark contrast to small, transient bodies of water in its immediate vicinity. A highly diverse endemic endemic amphipod fauna is found in one, but not the other habitat. We ask here whether differences in stress response can explain the immiscibility barrier between Lake Baikal and non-Baikal faunas. To this end, we conducted exposure experiments to increased temperature and the toxic heavy metal cadmium as stressors. RESULTS: Here we obtained high-quality de novo transcriptome assemblies, covering mutiple conditions, of three amphipod species, and compared their transcriptomic stress responses. Two of these species, Eulimnogammarus verrucosus and E. cyaneus, are endemic to Lake Baikal, while the Holarctic Gammarus lacustris is a potential invader. CONCLUSIONS: Both Baikal species possess intact stress response systems and respond to elevated temperature with relatively similar changes in their expression profiles. G. lacustris reacts less strongly to the same stressors, possibly because its transcriptome is already perturbed by acclimation conditions.

Acute and chronic toxicity of neonicotinoid and butenolide insecticides to the freshwater amphipod, Hyalella azteca.

Neonicotinoids are the most widely used insecticides in the world. They are preferentially toxic to insects while displaying a low toxicity toward vertebrates, and this selective toxicity has resulted in the rapid and ubiquitous use of these compounds. However, neonicotinoids have been detected in agricultural surface waters and are known to cause adverse effects in non-target aquatic organisms. A wide range of toxicity has been reported for aquatic crustaceans, but most of the studies focus on the acute effects of imidacloprid, and few data are available regarding chronic effects of other neonicotinoids or neonicotinoid replacements (e.g., butenolides). The objective of this study was to assess the acute and chronic toxicity of six neonicotinoids (imidacloprid, thiamethoxam, acetamiprid, clothianidin, thiacloprid, and dinotefuran) and one butenolide (flupyradifurone) to the freshwater amphipod Hyalella azteca. Chronic (28-d), water-only, static-renewal tests were conducted. Survival was assessed weekly, and growth was measured at the end of the exposure. Effects of neonicotinoids varied depending on the compound. Acute (7-d) LC50s were 4.0, 4.7, 60, 68, 230, and 290 µg/L for clothianidin, acetamiprid, dinotefuran, thiacloprid, imidacloprid, and thiamethoxam, respectively. Chronic (28-d) survival and growth were reduced at similar concentrations to acute (7-d) survival for thiamethoxam, acetamiprid, clothianidin, and dinotefuran. However, chronic survival and growth of amphipods exposed to imidacloprid and thiacloprid were reduced at lower concentrations than acute survival, with respective 28-d LC50s of 90 and 44 µg/L, and EC50s of 4 and 3 µg/L. Flupyradifurone was intermediate in toxicity compared to the neonicotinoids: 7-d LC50, 28-d LC50, and 28-d EC50 were 26, 20, and 16 µg/L, respectively. The concentrations of imidacloprid and clothianidin reported for North American surface waters fall within the effect ranges observed in this study, indicating the potential for these compounds to cause adverse effects to indigenous populations of H. azteca.