Total Synthesis and Structural Reassignment of the Molt-Inhibiting Marine Alkaloid Erebusinone.

The marine alkaloid erebusinone is a secondary metabolite isolated from the Antarctic sponge Isodictya erinacea. Initial biological assays have shown that erebusinone increases amphipod mortality, probably by inhibition of the biosynthesis of molting hormone (ecdysone). Herein, we report the first total synthesis of the proposed structure of erebusinone and a structural revision.

Multiple Biomarker Responses in Aegla castro Exposed to Copper: A Laboratory Approach.

Although some biomarkers have already been determined in aeglids collected in the field, data from laboratory exposures are scarce. To our knowledge, no studies have investigated oxidative stress biomarkers in aeglids exposed to metals in the laboratory, or performed hemocyte counts and the comet assay using gill and hepatopancreas of aeglids. Thus, we investigated the effects of acute Cu exposure on intermolt males of Aegla castro, collected from a reference stream, acclimated for 6 days in the laboratory, and then exposed to 11 µg L-1 of dissolved Cu (Cu 11) or only to water (CTR), for 24 h. Gill and hepatopancreas samples were used to determine Cu accumulation, DNA damage, and metallothionein content (MT), while hemolymph samples were used to determine Cu accumulation, DNA damage, and hemocyte counts. Muscle samples were used to determine Cu accumulation and acetylcholinesterase activity (AChE). Non-protein thiol content (NPSH), catalase (CAT), glutathione S-transferase activities (GST), lipoperoxidation (LPO), and protein carbonylation content (PCC) were measured only in the hepatopancreas. Aegla castro exposed to Cu accumulated this metal in gills and activated detoxification mechanisms, through increased MT content in the gill, and showed an immune response, evidenced by an increase in hyaline hemocytes. Therefore, gill and hemocytes appear to have a protective role in preventing the transport and bioavailability of Cu through the body. On the other hand, we observed a decrease in MT content in the hepatopancreas of crabs exposed to Cu, suggesting the excretion of MT in association with Cu bound to the sulfhydryl groups of this protein.

The Assessment of Methyl Methanesulfonate Absorption by Amphipods from the Environment Using Lux-Biosensors.

The ability of aquatic mesofauna representatives involved in trophic chains to sorb and accumulate toxicants is important for understanding the functioning of aquatic ecosystems and for fishing industry. This study investigated the capacity of marine amphipod Gammarus oceanicus and freshwater amphipods Eulimnogammarus vittatus and Gammarus lacustris to absorb the DNA-alkylating agent methyl methanesulfonate (MMS). The presence of alkylating agents in the environment and in the tissues of the amphipods was determined using whole-cell lux-biosensor Escherichia coli MG1655 pAlkA-lux, in which the luxCDABE genes from Photorhabdus luminescens, enabling the luminescence of the cell culture, are controlled by the PalkA promoter of DNA glycosylase. It was shown that within one day of incubation in water containing MMS at a concentration above 10 µM, the amphipods absorbed the toxicant and their tissues produce more alkylation damage to biosensor cells than the surrounding water. Concentrations of MMS above 1 mM in the environment caused the death of the amphipods before the toxicant could be significantly concentrated in their tissues. The sensitivity and the capacity to absorb MMS were found to be approximately the same for the marine amphipod G. oceanicus and the freshwater amphipods E. vittatus and G. lacustris.

Into the deep: Exploring the molecular mechanisms of hyperactive behaviour induced by three rare earth elements in early life-stages of the deep-sea scavenging amphipod Tmetonyx cicada (Lysianassidae).

With increasing socio-economic importance of the rare earth elements and yttrium (REY), Norway has laid out plans for REY mining, from land-based to deep-sea mining, thereby enhancing REY mobility in the marine ecosystem. Little is known about associated environmental consequences, especially in the deep ocean. We explored the toxicity and modes of action of a light (Nd), medium (Gd) and heavy (Yb) REY-Cl3 at four concentrations (3, 30, 300, and 3000 µg L-1) in the Arcto-boreal deep-sea amphipod Tmetonyx cicada. At the highest concentration, REY solubility was limited and increased with atomic weight (Nd < Gd < Yb). Lethal effects were practically restricted to this treatment, with the lighter elements being more acutely toxic than Yb (from ∼50 % mortality in the Gd-group at dissolved 689-504 µg L-1 to <20 % in the Yb-group at ca. 2000 µg L-1), which could be a function of bioavailability. All three REY induced hyperactivity at the low-medium concentrations. Delving into the transcriptome of T. cicada allowed us to determine a whole array of potential (neurotoxic) mechanisms underlying this behaviour. Gd induced the vastest response, affecting serotonin-synthesis; sphingolipid-synthesis; the renin-angiotensin system; mitochondrial and endoplasmic reticulum functioning (Gd, Nd); and lysosome integrity (Gd, Yb); as well as the expression of hemocyanin, potentially governing REY-uptake (Gd, Yb). While Nd and Yb shared only few pathways, suggesting a link between mode of action and atomic weight/radius, almost all discussed mechanisms imply the disruption of organismal Ca-homeostasis. Despite only fragmental genomic information available for crustaceans to date, our results provide novel insight into the toxicophysiology of REY in marine biota. The neurotoxic/behavioural effects in T. cicada at concentrations with potential environmental relevance warn about the possibility of bottom-up ecological consequences in mining exposed fjords and deep-sea ecosystems, calling for follow-up studies and regulatory measures prior to the onset of REY mining in Norway.

Contrasting alterations in brain chemistry in a crustacean intermediate host of two acanthocephalan parasites.

The dynamic properties of neural systems throughout life can be hijacked by so-called manipulative parasites. This study investigated changes in the brain chemistry of the amphipod Gammarus fossarum in response to infection with two trophically-transmitted helminth parasites known to induce distinct behavioral alterations: the bird acanthocephalan Polymorphus minutus and the fish acanthocephalan Pomphorhynchus tereticollis. We quantified brain antioxidant capacity as a common marker of homeostasis and neuroprotection, and brain total protein, on 72 pools of six brains. We analyzed the concentration of serotonin (5HT), dopamine (DA) and tyramine in 52 pools of six brains, by using ultrafast high performance liquid chromatography with electrochemical detection (UHPLC-ECD). Brain total protein concentration scaled hypo-allometrically to dry body weight, and was increased in infected gammarids compared to uninfected ones. The brain of gammarids infected with P. minutus had significantly lower total antioxidant capacity relative to total proteins. Infection with P. tereticollis impacted DA level compared to uninfected ones, and in opposite direction between spring and summer. Brain 5HT level was higher in summer compared to spring independently of infection status, and was decreased by infection after correcting for brain total protein concentration estimated from dry whole-body weight. The potential implication of 5HT/DA balance in parasite manipulation, as a major modulator of the reward-punishment axis, is discussed. Taken together, these findings highlight the need to consider both brain homeostatic and/or structural changes (antioxidant and total protein content) together with neurotransmission balance and flexibility, in studies investigating the impact of parasites on brain and behavior.

Ingestion of chitosan-starch blends: Effect on the survival of supralittoral amphipods.

Sandy beach ecosystems are particularly affected by plastic pollution. Supralittoral amphipods are important components of the food web in sandy beaches and their ability to ingest microplastics and bioplastics has been assessed. Chitosan, a polysaccharide obtained by deacetylation of chitin, the second most abundant polymer in the world, represents an interesting component to produce novel bioplastics in combination with other biopolymers like starch. Here, the possibility of ingesting chitosan-starch blends and the possible effects on the amphipod Talitrus saltator were investigated. Groups of adult individuals were fed with sheets containing mixtures of chitosan and starch in different percentages for 7 and 14 days. The results showed that chitosan ingestion is dependent on the percentage of starch present in the mixture. Moreover, FTIR analyses of both sheets and faecal pellets after consumption show that chitosan is not digested. Furthermore, the survival rate of amphipods fed with a mixture of chitosan and starch decreases after one week compared to the control groups (100 % starch and paper), and drops drastically to 0 % after two weeks the experiment began. In addition, consumption of 100 % chitosan is negligible. Therefore, the results of the experimental observations evidenced that chitosan is avoided as food resource and its consumption significantly affects the survival capacity of T. saltator. It is emphasized that the release of mixtures of chitosan and starch into the marine environment appears to be dangerous for littoral amphipods.

Designing and applying a methodology to assess sperm cell viability and DNA damage in a model amphipod.

Sperm quality is defined as the sperm cell ability to successfully fertilize eggs and allow normal embryo development⁠. Few studies explore sperm quality using aquatic invertebrates. Parhyale hawaiensis is a marine amphipod with a circumtropical distribution and considered a model for evolution, development, and ecotoxicological studies. We aimed to develop a methodology to collect sperm cells of P. hawaiensis and evaluate their viability and DNA damage (comet assay). We directly exposed the sperm cells to different mutagenic agents to optimize/develop the protocols. Then, as a proof of concept, we exposed the males to mutagenic compounds (EMS, benzo[a]pyrene (BaP), azo and anthraquinone dyes) at non-lethal concentrations verified by the proposed viability test and analyzed their sperm cells for DNA damage (comet assay). Organisms exposed to EMS presented a clear concentration response in the DNA damage response. We also showed that BaP was able to induce a statistically significant increase in DNA damage of the sperm cells. For the two dyes, although DNA damage increased, statistically differences were not observed. We believe we successfully developed a test to detect genotoxicity of chemicals in sperm cells using an invertebrate model. The protocol for sperm cell viability needs to be further explored with different chemicals to verify its utility as a toxicity endpoint. The developed genotoxicity test has the advantages to employ organisms that are easily cultivated in reduced space, use simple laboratory resources and reduced amount of material and reagents. Positive responses with this model could be used to disclose new germ cell mutagen candidates which could be further confirmed in vertebrates' systems.

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.

Acute and chronic toxicity of rare earth elements-enriched sediments from a prospective mining area: Effects on life history traits, behavioural and physiological responses of Gammarus fossarum (Crustacea Amphipoda).

Rare earth elements (REE) have an essential role and growing importance in the world's economy. They are attracting interest from society, policymakers, and scientists. The rapidly growing global demand for REE in several strategic industrial and agricultural sectors led many countries to consider the (re)-opening of mining activities for REE extraction. Hence, their increasing use led to the disruption of their biogeochemical cycles with anthropic abnormalities already observed in aquatic ecosystems. Nonetheless, REE remain less studied, and their mechanisms of toxicity actions are not fully understood. As amphipods, Gammarus fossarum represent an important part of the aquatic macroinvertebrate assemblage and are generally used in ecotoxicological studies for their high ecological relevance. However, their use for the study of REE effects has been rather limited so far. The current study aims to assess the potential effects of two naturally REE-enriched sediments (N2 and B4) on G. fossarum. Effects on life history traits, behavioural and physiological responses have been evaluated. Exposing G. fossarum males for 72h to sediments N2 and B4 led to a decrease in haemolymph osmolality and locomotion while an increase in ventilatory activity was observed. Exposing G. fossarum pre-copula pairs with females at the same reproductive stage to the naturally REE-enriched sediments, for one moult cycle duration (∼30 days) showed that sediment B4 led to i) a significant uptake of REE, ii) a significant decrease in the proportion of females with oocytes and iii) a significant reduction in the total number of juveniles. The physicochemical analyses of sediments showed that B4 contains the highest amount of REE with a higher proportion of light REE. The present study gives the first insights into the potential toxicity of REE on G. fossarum as they may have deleterious effects on G. fossarum population's dynamics, which may alter the functioning of aquatic ecosystems.

The seasonal response of metabolic rate to projected climate change scenarios in aquatic amphipods.

The responses of organisms to climate change are mediated primarily by its impact on their metabolic rates, which, in turn, drive various biological and ecological processes. Although there have been numerous seminal studies on the sensitivity of metabolic rate to temperature, little is empirically known about how this rate responds to seasonal temperature ranges and beyond under conservative IPCC climate change scenarios. Here, we measured the SMR of the aquatic amphipod, Gammarus insensibilis, which served as our subject species, with body masses ranging from 0.20 to 7.74 mg ash free weight. We assessed the response of the SMR across nine temperature levels ranging from 12 to 30.2 °C. These temperatures match seasonal temperature norms, with an incremental increase of 0.6-1.2 °C above each seasonal baseline, as projected for the years 2040 and 2100 under the modest climate change scenarios. Overall, our findings showed that the effect of temperature on SMR varies with body mass, as indicated by a negative size-temperature interaction, with larger conspecifics exhibiting less sensitivity to temperature changes than smaller ones. From the cold to warm season, the SMR increased by an average of 14% °C-1, with increases of 18.4% °C-1 in smaller individuals and 11.4% °C-1 in larger ones. The SMR of smaller individuals peaked at a 0.6 °C increase from the current summer baseline (15.08% °C-1, Q10 = 4.2), while in larger ones it peaked with a 1.2 °C increase beyond autumn temperatures (14.9% °C-1, Q10 = 3.9). However, at temperatures reflecting global warming that exceed summer temperatures, the SMR of larger individuals levelled off, while that of smaller ones continued to increase. Overall, our findings suggest that smaller-sized individuals have a broader thermal window for SMR performance, while the SMR of larger-sized ones will become increasingly constrained at summer temperatures as those summer temperatures become hotter.

Is the invasive Caprella scaura Templeton, 1836 displacing its resident congener Caprella equilibra Say, 1818 in marinas of the South Iberian Peninsula? A long-term spatio-temporal approach.

Although marine invasions are increasingly a matter of concern, the impact of invasive species in the ecosystem and their ability to replace native taxa is still little understood. Data from 2011 to 2021 in marinas of the Southern Iberian Peninsula supported that the invasive amphipod Caprella scaura is replacing the resident Caprella equilibra over time. Six marinas where C. equilibra was abundant in 2011 and C. scaura was absent, are now dominated by C. scaura. Although this displacement is more evident in Mediterranean shores than in Atlantic coasts, it is very variable between marinas. The spreading of the invasive species in marinas of the Alboran Sea mainly occurred from 2011 to 2017, preventing C. equilibra from regaining its former distribution. The ultimate factors responsible for the displacement, such as the aggressive behaviour of C. scaura, environmental influences or physiological performance in a global warming context, should be further investigated experimentally.

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.

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.

Resource utilisation and trophic niche overlap of coralline intertidal benthic amphipods: an isotopic perspective.

Benthic amphipod feeding groups are a well-established trophic classification that is mostly based on field observations and laboratory tests and are used in ecological studies to monitor the ecological state of benthic ecosystems. Globally, carbon and nitrogen stable isotope ratio investigations have provided confirmation of, and novel insights into, the trophic ecology of benthic animals, such as polychaetes. However, stable isotopic examinations of benthic amphipods have been limited. Here, we used microgram samples to compare the species-specific dietary sources, trophic positions, and isotopic niche overlap of selected benthic amphipods from the Gulf of Kachchh, Marine National Park, using elemental analyser-isotopic ratio mass spectrometry (EA-IRMS) of carbon and nitrogen. Overall, all primary carbon sources presented wide variation in the isotopic values of δ13C (6.3‰) and δ15N (greater than 13‰). Conversely, the amphipod taxa displayed relatively narrow range for δ13C (3.9‰) and wider range for δ15N (more than 10‰). The results of the Bayesian mixing model revealed that the benthic amphipods had species-specific feeding preferences. However, the predominant carbon source was organic matter in sediment which reinforced benthic pathways for energy flow for most species. According to the estimated trophic level values (1.62-3.39), these species play a significant role as primary and secondary consumers serving as crucial trophic intermediaries in the food chain, connecting the base to the top consumers. High overlapping ecological niche amongst species was detected by SIBER analysis which indicated co-existence of the benthic amphipods in their respective microhabitats. This signifies wider utilisation of resources and inter-specific feeding preferences with minimal competition amongst amphipod species.

Reproductive Capacity, but not Food Consumption, is Reduced by Continuous Exposure to Typical Genotoxic Stressor γ-Rays in the sentinel species Gammarus fossarum.

The long-term impacts of radiocontaminants (and the associated risks) for ecosystems are still subject to vast societal and scientific debate while wildlife is chronically exposed to various sources and levels of either environmental or anthropogenic ionizing radiation from the use of nuclear energy. The present study aimed to assess induced phenotypical responses in both male and female gammarids after short-term continuous γ-irradiation, acting as a typical well-characterized genotoxic stressor that can interact directly with living matter. In particular, we started characterizing the effects using standardized measurements for biological effects on few biological functions for this species, especially feeding inhibition tests, molting, and reproductive ability, which have already been proven for chemical substances and are likely to be disturbed by ionizing radiation. The results show no significant differences in terms of the survival of organisms (males and females), of their short-term food consumption which is linked to the general health status (males and females), and of the molting cycle (females). In contrast, exposure significantly affected fecundity (number of embryos produced) at the highest dose rates for irradiated females (51 mGy h-1) and males (5 and 51 mGy h-1). These results showed that, in gammarids, reproduction, which is a critical endpoint for population dynamics, is the most radiosensitive phenotypic endpoint, with significant effects recorded on male reproductive capacity, which is more sensitive than in females. Environ Toxicol Chem 2024;43:2071-2079. © 2024 SETAC.

Reduced bioavailability of Au and isotopically enriched 109Ag nanoparticles transformed through a pilot wastewater treatment plant in Hyalella azteca under environmentally relevant exposure scenarios.

Wastewater Treatment Plants (WWTP) are a major repository and entrance path of nanoparticles (NP) in the environment and hence play a major role in the final NP fate and toxicity. Studies on silver nanoparticles (AgNP) transport via the WWTP system and uptake by aquatic organisms have so far been carried out using unrealistically high AgNP concentrations, unlikely to be encountered in the aquatic environment. The use of high AgNP concentrations is necessitated by both the low sensitivity of the detection methods used and the need to distinguish background Ag from spiked AgNP. In this study, isotopically enriched 109AgNP were synthesized to overcome these shortcomings and characterized by a broad range of methods including transmission electron microscopy, dynamic and electrophoretic light scattering. 109AgNP and gold NP (AuNP) were spiked to a pilot wastewater treatment plant fed with municipal wastewater for up to 21 days. AuNP were used as chemically less reactive tracer. The uptake of the pristine and transformed NP present in the effluent was assessed using the benthic amphipod Hyalella azteca in fresh- and brackish water exposures at environmentally relevant concentrations of 30 to 500 ng Au/L and 39 to 260 ng Ag/L. The unique isotopic signature of the 109AgNP allowed to detect the material at environmentally relevant concentrations in the presence of a much higher natural Ag background. The results show that the transformations reduce the NP uptake at environmentally relevant exposure concentrations. For 109Ag, lower accumulation factors (AF) were obtained after exposure to transformed NP (250-350) compared to the AF values obtained for pristine 109AgNP (750-840). The reduced AF values observed for H. azteca exposed to effluent from the AuNP-spiked WWTP indicate that biological transformation processes (e.g. eco-corona formation) seem to be involved in addition to chemical transformation.

Non-native amphipods reach higher maximum swimming velocities than natives.

The invasion of non-native amphipods often leads to severe changes in the composition of aquatic communities and may also result in the local replacement of native species. Particularly, a lower risk of being preyed upon resulting from high swimming velocities can be an advantage in interspecific competition. Furthermore, as swimming velocities are ecologically important, they are often used in ecotoxicological studies to estimate effects of different stressors. However, knowledge on swimming velocities of native and non-native amphipods is still rather limited. We experimentally investigated the maximum swimming velocities of three native and three non-native amphipods via video analyses in the laboratory. Results showed that non-native species reach higher maximum swimming velocities compared to natives probably leading to a higher predation success and reduced risk of being preyed upon. Additionally, body length correlates positively with swimming velocities, except for the invader Dikerogammarus villosus. As D. villosus can be cannibalistic, the high swimming velocities of the small individuals may reduce the intraspecific predation and may increase the survival rates of smaller specimen. Hence, knowledge about the swimming velocities of species contribute to the understanding of interspecific competition among species and might support explanation approaches for the success of invasive species. Furthermore, it provides baselines for ecotoxicological studies of stressor impacts.

Microbial artists: the role of parasite microbiomes in explaining colour polymorphism among amphipods and potential link to host manipulation.

Parasite infections are increasingly reported to change the microbiome of the parasitized hosts, while parasites bring their own microbes to what can be a multi-dimensional interaction. For instance, a recent hypothesis suggests that the microbial communities harboured by parasites may play a role in the well-documented ability of many parasites to manipulate host phenotype, and explain why the degree to which host phenotype is altered varies among conspecific parasites. Here, we explored whether the microbiomes of both hosts and parasites are associated with variation in host manipulation by parasites. Using colour quantification methods applied to digital images, we investigated colour variation among uninfected Transorchestia serrulata amphipods, as well as amphipods infected with Plagiorhynchus allisonae acanthocephalans and with a dilepidid cestode. We then characterized the bacteriota of amphipod hosts and of their parasites, looking for correlations between host phenotype and the bacterial taxa associated with hosts and parasites. We found large variation in amphipod colours, and weak support for a direct impact of parasites on the colour of their hosts. Conversely, and most interestingly, the parasite's bacteriota was more strongly correlated with colour variation among their amphipod hosts, with potential impact of amphipod-associated bacteria as well. Some bacterial taxa found associated with amphipods and parasites may have the ability to synthesize pigments, and we propose they may interact with colour determination in the amphipods. This study provides correlational support for an association between the parasite's microbiome and the evolution of host manipulation by parasites and host-parasite interactions more generally.

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.

A dynamic energy budget (DEB) model to assess the sublethal effects of imidacloprid toward Gammarus pulex at different temperatures.

Environmental ambient temperature significantly impacts the metabolic activities of aquatic ectotherm organisms and influences the fate of various chemicals. Although numerous studies have shown that the acute lethal toxicity of most chemicals increases with increasing temperature, the impact of temperature on chronic effects - encompassing both lethal and sublethal endpoints - has received limited attention. Furthermore, the mechanisms linking temperature and toxicity, potentially unveiled by toxicokinetic-toxicodynamic models (TKTD), remains inadequately explored. This study investigated the effects of environmentally relevant concentrations of the insecticide imidacloprid (IMI) on the growth and survival of the freshwater amphipod Gammarus pulex at two different temperatures. Our experimental design was tailored to fit a TKTD model, specifically the Dynamic Energy Budget (DEB) model. We conducted experiments spanning three and six months, utilizing small G. pulex juveniles. We observed effects endpoints at least five times, employing both destructive and non-destructive methods, crucial for accurate model fittings. Our findings reveal that IMI at environmental concentrations (up to 0.3 µg/L) affects the growth and survival of G. pulex, albeit with limited effects, showing a 10% inhibition compared to the control group. These limited effects, observed in both lethal and sublethal aspects, suggest a different mode of action at low, environmentally-relevant concentrations in long-term exposure (3 months), in contrast to previous studies which applied higher concentrations and found that sublethal effects occurred at significantly lower levels than lethal effects in an acute test setting (4 days). Moreover, after parameterizing the DEB model for various temperatures, we identified a lower threshold for both lethal and sublethal effects at higher temperatures, indicating increased intrinsic sensitivity. Overall, this study contributes to future risk assessments considering temperature as a crucial factor and exemplifies the integration of the DEB model into experimental design for comprehensive toxicity evaluations.