Most of known troglobiotic species occur in caves and subterranean environments from great depths. However, recently more attention has been given to other subterranean environments, such as the hypothelminorheic habitats. It comprises the most superficial among all subterranean habitats. This kind of environment is characterized by the constant presence of wet spots, absence of light and very particular abiotic characteristics, comprising unique species. The first hypothelminorheic Amphipoda from South America is here described, a new species of the genus Hyalella which occurs in a wetland on Southern Brazil. The new species differs from other troglobiotics of the genus by the presence of a curved seta on the inner ramus of uropod 1 and elongation of appendices, as the first pair of antennae and peraeopods 6 and 7. However, human impacts in the area where the new species occurs have changed heavily their habitat, which may have led the species to a critical level of threat or even extinction, demonstrating the fragility of this environment.

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.

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.

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.

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.

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.

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.

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.

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.

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.