Speed it up: How temperature drives toxicokinetics of organic contaminants in freshwater amphipods.

The acceleration of global climate change draws increasing attention towards interactive effects of temperature and organic contaminants. Many studies reported a higher sensitivity of aquatic invertebrates towards contaminant exposure with increasing or fluctuating temperatures. The hypothesis of this study was that the higher sensitivity of invertebrates is associated with the changes of toxicokinetic processes that determine internal concentrations of contaminants and consequently toxic effects. Therefore, the influence of temperature on toxicokinetic processes and the underlying mechanisms were studied in two key amphipod species (Gammarus pulex and Hyalella azteca). Bioconcentration experiments were carried out at four different temperatures with a mixture of 12 exposure relevant polar organic contaminants. Tissue and medium samples were taken in regular intervals and analysed by online solid-phase extraction liquid chromatography high-resolution tandem mass spectrometry. Subsequently, toxicokinetic rates were modelled and analysed in dependence of the exposure temperature using the Arrhenius equation. An exponential relationship between toxicokinetic rates versus temperature was observed and could be well depicted by applying the Arrhenius equation. Due to a similar Arrhenius temperature of uptake and elimination rates, the bioconcentration factors of the contaminants were generally constant across the temperature range. Furthermore, the Arrhenius temperature of the toxicokinetic rates and respiration was mostly similar. However, in some cases (citalopram, cyprodinil), the bioconcentration factor appeared to be temperature dependent, which could potentially be explained by the influence of temperature on active uptake mechanisms or biotransformation. The observed temperature effects on toxicokinetics may be particularly relevant in non-equilibrated systems, such as exposure peaks in summer as exemplified by the exposure modelling of a field measured pesticide peak where the internal concentrations increased by up to fourfold along the temperature gradient. The results provide novel insights into the mechanisms of chemical uptake, biotransformation and elimination in different climate scenarios and can improve environmental risk assessment.

Elucidating the spatial distribution of organic contaminants and their biotransformation products in amphipod tissue by MALDI- and DESI-MS-imaging.

The application of mass spectrometry imaging (MSI) is a promising tool to analyze the spatial distribution of organic contaminants in organisms and thereby improve the understanding of toxicokinetic and toxicodynamic processes. MSI is a common method in medical research but has been rarely applied in environmental science. In the present study, the suitability of MSI to assess the spatial distribution of organic contaminants and their biotransformation products (BTPs) in the aquatic invertebrate key species Gammarus pulex was studied. Gammarids were exposed to a mixture of common organic contaminants (carbamazepine, citalopram, cyprodinil, efavirenz, fluopyram and terbutryn). The distribution of the parent compounds and their BTPs in the organisms was analyzed by two MSI methods (MALDI- and DESI-HRMSI) after cryo-sectioning, and by LC-HRMS/MS after dissection into different organ compartments. The spatial distribution of contaminats in gammarid tissue could be successfully analyzed by the different analytical methods. The intestinal system was identified as the main site of biotransformation, possibly due to the presence of biotransforming enzymes. LC-HRMS/MS was more sensitive and provided higher confidence in BTP identification due to chromatographic separation and MS/MS. DESI was found to be the more sensitive MSI method for the analyzed contaminants, whereas additional biomarkers were found using MALDI. The results demonstrate the suitability of MSI for investigations on the spatial distribution of accumulated organic contaminants. However, both MSI methods required high exposure concentrations. Further improvements of ionization methods would be needed to address environmentally relevant concentrations.

The use of Gammarus pulex as a model organism for ecotoxicological assessment of ibuprofen and propranolol at environmental relevant concentrations.

The aim of this study is to assess the toxicity of ibuprofen (IBU) and propranolol (PRO) drugs usingGammarus pulex as a model organism. Firstly, the 96 h LC50 values of IBU and PRO were determined and then three sublethal concentrations of the drugs were exposed to G. pulex. The activities of superoxide dismutase (SOD), catalase (CAT) and acetylcholinesterase (AChE) were evaluated. SOD activity decreased in G. pulex exposed to IBU and PRO compared to control. In all groups exposed to IBU, CAT activity increased at different concentrations at 24 and 96 h. In the groups exposed to different PRO concentrations, CAT activities increased after 24 h compared to the control group (p < 0.05). AChE activities increased in all application groups exposed to IBU for 96 hours (p < 0.05). In conclusion, exposure to IBU and PRO resulted in increased oxidative damage. PRO has been found to cause neurotoxicity.

Multiple Stress Reduces the Advantage of Pesticide Adaptation.

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

Feeding Behavioural Studies with Freshwater Gammarus spp.: The Importance of a Standardised Methodology.

Freshwater Gammarids are common leaf-shredding detritivores, and they usually feed on naturally conditioned organic material, in other words leaf litter that is characterised by an increased palatability, due to the action and presence of microorganisms (Chaumot et al. 2015; Cummins 1974: Maltby et al. 2002). Gammarus spp. are biologically omnivorous organisms, so they are involved in shredding leaf litter and are also prone to cannibalism, predation behaviour (Kelly et al. 2002) and coprophagy when juveniles (McCahon and Pascoe 1988). Gammarus spp. is a keystone species (Woodward et al. 2008), and it plays an important role in the decomposition of organic matter (Alonso et al. 2009; Bundschuh et al. 2013) and is also a noteworthy prey for fish and birds (Andrén and Eriksson Wiklund 2013; Blarer and Burkhardt-Holm 2016). Gammarids are considered to be fairly sensitive to different contaminants (Ashauer et al. 2010; Bloor et al. 2005; Felten et al. 2008a; Lahive et al. 2015; Kunz et al. 2010); in fact Amphipods have been reported to be one of the most sensitive orders to metals and organic compounds (Wogram and Liess 2001), which makes them representative test organisms for ecotoxicological studies and valid sentinel species for assessing water quality status (Garcia-Galan et al. 2017).

Parasites influence cannibalistic and predatory interactions within and between native and invasive amphipods.

In Northern Ireland, the amphipods Gammarus duebeni celticus (native) and G. pulex (invasive) coexist in some places, whilst in others the native species has been replaced by the invader. We explored the role of parasites in mediating interactions between these amphipods, which demonstrate mutual intraguild predation (IGP: predation between animals that also compete for prey). IGP and cannibalism can be important factors in structuring populations and communities. We investigated the effects of parasitism on rates of IGP between G. d. celticus and G. pulex and on cannibalism within each species by comparing functional responses (FRs: relationships between the use of a prey resource and its availability). Infection with the microsporidian Pleistophora mulleri caused an increase in IGP and cannibalism by G. d. celticus, which showed increased attack rates and reduced prey handling times. In contrast, infection with the acanthocephalan parasite Echinorhynchus truttae did not alter IGP or cannibalism by G. pulex. A prey preference experiment revealed that both amphipods were more likely to feed on heterospecific rather than conspecific prey, and this was also corroborated by the fact that overall IGP FRs were higher than cannibalism FRs. This may be selectively advantageous, as feeding on heterospecific prey removes possible competitors without the risk of consuming juvenile kin or acquiring parasites from infected conspecifics. Infection of the native G. d. celticus with P. mulleri enhanced IGP on the invasive G. pulex, which is likely to facilitate the coexistence of the 2 species.

Acanthocephalan infection patterns in amphipods: a reappraisal in the light of recently discovered host cryptic diversity.

Amphipods are model species in studies of pervasive biological patterns such as sexual selection, size assortative pairing and parasite infection patterns. Cryptic diversity (i.e. morphologically identical but genetically divergent lineages) has recently been detected in several species. Potential effects of such hidden diversity on biological patterns remain unclear, but potentially significant, and beg the question of whether we have missed part of the picture by involuntarily overlooking the occurrence and effects of cryptic diversity on biological patterns documented by previous studies. Here we tested for potential effects of cryptic diversity on parasite infection patterns in amphipod populations and discuss the implications of our results in the context of previously documented host-parasite infection patterns, especially amphipod-acanthocephalan associations. We assessed infection levels (prevalence and abundance) of 3 acanthocephalan species (Pomphorhynchus laevis, P. tereticollis and Polymorphus minutus) among cryptic lineages of the Gammarus pulex/G. fossarum species complex and G. roeseli from sampling sites where they occur in sympatry. We also evaluated potential differences in parasite-induced mortality among host molecular operational taxonomic units (MOTUs)-parasite species combinations. Acanthocephalan prevalence, abundance and parasite-induced mortality varied widely among cryptic MOTUs and parasite species; infection patterns were more variable among MOTUs than sampling sites. Overall, cryptic diversity in amphipods strongly influenced apparent infection levels and parasite-induced mortality. Future research on species with cryptic diversity should account for potential effects on documented biological patterns. Results from previous studies may also need to be reassessed in light of cryptic diversity and its pervasive effects.

Gammarus pulex as a Model Organism to Assess the Residual Toxicity of Slaughterhouse Wastewater Treated by Electrocoagulation Process.

It was assessed the efficiency of the electrocoagulation (EC) in slaughterhouse wastewater (SW) treatment by using antioxidant parameters of Gammarus pulex. The SW was treated by EC. Superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) activities and malondialdehyde (MDA) levels in G. pulex exposed to pre- and post-treated of the SW during 24 h and 96 h were analysed. Standard methods were applied during the analysing process of the physicochemical quality parameters for both untreated and treated SW. All measured physicochemical parameters were decreased following the treatment process via EC. After the treatment process, it was observed that while SOD activities and MDA levels were decreased, CAT activities were increased and GPx activities did not exhibit any change. In conclusion, the present study demonstrated the abilities of untreated SW to promote oxidative stress in model organism. The SOD, CAT activities and MDA levels in G. pulex revealed that EC process were efficient in the SW treatment.

Synergistic impacts by an invasive amphipod and an invasive fish explain native gammarid extinction.

BACKGROUND: Worldwide freshwater ecosystems are increasingly affected by invasive alien species. In particular, Ponto-Caspian gobiid fishes and amphipods are suspected to have pronounced effects on aquatic food webs. However, there is a lack of systematic studies mechanistically testing the potential synergistic effects of invasive species on native fauna. In this study we investigated the interrelations between the invasive amphipod Dikerogammarus villosus and the invasive fish species Neogobius melanostomus in their effects on the native amphipod Gammarus pulex. We hypothesized selective predation by the fish as a driver for displacement of native species resulting in potential extinction of G. pulex. The survival of G. pulex in the presence of N. melanostomus in relation to the presence of D. villosus and availability of shelter was analyzed in the context of behavioural differences between the amphipod species. RESULTS: Gammarus pulex had a significantly higher susceptibility to predation by N. melanostomus compared to D. villosus in all experiments, suggesting preferential predation by this fish on native gammarids. Furthermore, the presence of D. villosus significantly increased the vulnerability of G. pulex to fish predation. Habitat structure was an important factor for swimming activity of amphipods and their mortality, resulting in a threefold decrease in amphipods consumed with shelter habitat structures provided. Behavioral differences in swimming activity were additionally responsible for higher predation rates on G. pulex. Intraguild predation could be neglected within short experimental durations. CONCLUSIONS: The results of this study provide evidence for synergistic effects of the two invasive Ponto-Caspian species on the native amphipod as an underlying process of species displacements during invasion processes. Prey behaviour and monotonous habitat structures additionally contribute to the decline of the native gammarid fauna in the upper Danube River and elsewhere.

Suspended particles only marginally reduce pyrethroid toxicity to the freshwater invertebrate Gammarus pulex (L.) during pulse exposure.

Current ecotoxicological research on particle-associated pyrethroids in freshwater systems focuses almost exclusively on sediment-exposure scenarios and sediment-dwelling macroinvertebrates. We studied how suspended particles influence acute effects of lambda-cyhalothrin and bifenthrin on the epibenthic freshwater amphipod Gammarus pulex (L.) using brief pulse exposures followed by a 144 h post exposure recovery phase. Humic acid (HA) and the clay mineral montmorillonite (MM) were used as model sorbents in environmentally realistic concentrations (5, 25 and 125 mg L(-1)). Mortality of G. pulex was recorded during the post exposure recovery phase and locomotor behavior was measured during exposure to lambda-cyhalothrin. We found that HA in concentrations ≥25 mg L(-1) adsorbed the majority of pyrethroids but only reduced mortality of G. pulex up to a factor of four compared to pyrethroid-only treatments. MM suspensions adsorbed a variable fraction of pyrethroids (10% for bifenthrin and 70% for lambda-cyhalothrin) but did not significantly change the concentration-response relationship compared to pure pyrethroid treatments. Behavioral responses and immobilisation rate of G. pulex were reduced in the presence of HA, whereas behavioral responses and immobilisation rate were increased in the presence of MM. This indicates that G. pulex was capable of sensing the bioavailable fraction of lambda-cyhalothrin. Our results imply that suspended particles reduce to only a limited extent the toxicity of pyrethroids to G. pulex and that passive uptake of pyrethroids can be significant even when pyrethroids are adsorbed to suspended particles.

Gender differences in responses in Gammarus pulex exposed to BDE-47: A gel-free proteomic approach.

Very few ecotoxicological studies have considered differences in toxic effects on male and female organisms. Here, we investigated protein expression differences in caeca of Gammarus pulex males and females under control conditions (unexposed) and after 96h exposure to BDE-47. Using gel-free proteomic analysis, we have identified 45 proteins, of which 25 were significantly differently expressed according to sex and/or BDE-47 exposure. These proteins were involved in several biological processes such as energy metabolism, chaperone proteins, or transcription/translation. In unexposed amphipods, 11 proteins were significantly over-expressed in females, and 6 proteins were over-expressed in males. Under BDE-47 stress, 7 proteins were differently impacted according to sex. For example, catalase was over-expressed in exposed females and under-expressed in exposed males, as compared to respective controls. Conversely, proteins involved in energy metabolism were up-regulated in males and down-regulated in females. Our proteomic study showed differences in responses of males and females to BDE-47 exposure, emphasizing that sex is a confounding factor in ecotoxicological assessment. However, due to the limited information existing in databases on Gammarids, it was difficult to define a BDE-47 mechanism of action. The gel-free proteomic seems to be a promising method to develop in future ecotoxicological studies and thus, to improve our understanding of the mechanism of action of xenobiotics.

Quantifying diet-borne metal uptake in Gammarus pulex using stable isotope tracers.

Gammarids are aquatic amphipods widely used for water quality monitoring. To investigate the copper and cadmium diet-borne metal uptake in Gammarus pulex, we adapted the pulse-chase stable isotopes-based approach to determine the food ingestion rate (IR), the gut retention time (GRT) and the metal assimilation efficiencies (AE). G. pulex were fed with (65)Cu-, (106)Cd-, and (53)Cr-labeled alder leaves for 7.5h and then with unlabeled leaves for 5d. The metal stable isotope contents in the gammarids, leaves, filtered water and periodically collected feces were determined. Chromium was poorly assimilated by the gammarids; thus, Cr was used as an unassimilated tracer. The first tracer defecation occurred before the first feces harvest, indicating a gut passage time of less than 9h. A 24-h GRT and a 0.69gg(-1)d(-1) IR were estimated. The Cd AE value was estimated as 5-47%, depending on the assimilation determination method applied. The Cu AE value could not be evaluated regardless of the determination method used, most likely because of the rapid Cu regulation in gammarids in addition to analytical uncertainties when determining the Cu content in leaves. Application of the Cd AE value in the framework of the biodynamic bioaccumulation model shows that the diet-borne uptake of Cd significantly contributes (66-95%) to the metal bioaccumulation in G. pulex fed with alder leaves.

Description and prevalence of gregarines infecting the amphipod Gammarus pulex, in the Water of Leith, Scotland, UK.

Gregarines are symbiotic protists that are found in a broad spectrum of invertebrates, including insects, crustaceans, and annelids. Among these the globally distributed amphipod Gammarus pulex is one of the earliest recognized hosts for aquatic gregarines and is prevalent among macroinvertebrates in freshwater environments. In this study, samples of G. pulex were collected in the Water of Leith river, Scotland, UK. Gregarines were identified using light and scanning electron microscopy as well as standard molecular techniques. We identified three septate eugregarine symbionts-Heliospora longissima, Cephaloidophora gammari, and the here newly characterized Cephaloidophora conus n. sp. (formerly Cephaloidophora sp.) associated with Gammarus pulex in the Water of Leith. Prevalences for identified gregarine species were calculated and seasonal dynamics of gregarine infections/colonization were analyzed. Prevalences were highest in autumn and spring reaching almost 50 %. While the two Cephaloidophora species showed similar colonization patterns, the prevalence of Heliospora showed an opposite trend. Identifying gregarine infection/colonization patterns is one step towards better understanding the gregarine-host relationship, as well as possible impacts of the gregarines on their hosts.

Determination of oxidative stress responses caused by aluminum oxide (γ-Al2O3 and α-Al2O3) nanoparticles in Gammarus pulex.

The rapid growth in the use of aluminum oxide nanoparticles (Al2O3 NPs) in various fields such as medicine, pharmacy, cosmetics industries and engineering, and the fact that these NPs and their wastes mix with the aquatic environment and damage the aquatic ecosystem, affect the organisms in the water, enter the food chain and reach humans is a major problem is cause for concern. The aim of this study is to investigate the oxidative stress caused by two separate forms of aluminum oxide, γ-Al2O3 and α-Al2O3, in Gammarus pulex, which is a good indicator species, with biochemical parameters. For this purpose, G. pulex was exposed to different concentrations (0, 10, 20, 40 ppm) of γ-Al2O3 and α-Al2O3 separately. The experiments were carried out for 24 and 96 h by creating 3 repeated experimental groups consisting of 4 groups. For biomarker analysis, superoxide dismutase (SOD), catalase (CAT) activities and glutathione (GSH) and thiobarbituric acid (TBARS) levels were performed using an ELISA kit. As a result of the in experimental study, it was observed that both nanoparticles affected oxidative stress and antioxidant parameters after 96 h compared to the control group. Increases in SOD activity were observed, γ-Al2O3 caused a decrease in CAT activity at 24 h, and α- Al2O3 caused increases in CAT activity at 96 h. Decreases in GSH levels and increases in TBARS levels have been observed.

Effects of the antidepressant fluoxetine on the swimming behaviour of the amphipod Gammarus pulex: Comparison of short-term and long-term toxicity in the laboratory and the semi-field.

Fluoxetine is one of the worlds most prescribed antidepressant, and frequently detected in surface waters. Once present in the aquatic environment, fluoxetine has been shown to disrupt the swimming behaviour of fish and invertebrates. However, swimming behaviour is also known to be highly variable according to experimental conditions, potentially concealing relevant effects. Therefore, the aims of this study were two-fold: i) investigate the swimming and feeding behaviour of Gammarus pulex after exposure to the antidepressant fluoxetine (0.2, 2, 20, and 200 µg/L), and ii) assess to what degree the experimental test duration (short-term and long-term) and test location (laboratory and semi-field conditions) affect gammarid's swimming behaviour. We used automated video tracking and analysis to asses a range of swimming behaviours of G. pulex, including swimming speed, startle responses after light transition, acceleration, curvature and thigmotaxis. We found larger effects on the swimming behaviour of G. pulex due to experimental conditions than due to tested antidepressant concentrations. Gammarids swam faster, more straight and showed a stronger startle response during light transition when kept under semi-field conditions compared to the laboratory. Effects found for different test durations were opposite in the laboratory and semi-field. In the laboratory gammarids swam slower and spent more time at the inner zone of the arena after 2 days compared to 21 days while for the semi-field the reverse was observed. Fluoxetine had only minor impacts on the swimming behaviour of G. pulex, but experimental conditions influenced behavioural outcomes in response to fluoxetine exposure. Overall, our results highlight the importance of standardizing and optimizing experimental protocols that assess behaviour to achieve reproducible results in ecotoxicology.

A novel approach for the assessment of invertebrate behavior and its use in behavioral ecotoxicology.

Sublethal effects are becoming more relevant in ecotoxicological test methods due to their higher sensitivity compared to lethal endpoints and their preventive nature. Such a promising sublethal endpoint is the movement behavior of invertebrates which is associated with the direct maintenance of various ecosystem processes, hence being of special interest for ecotoxicology. Disturbed movement behavior is often related to neurotoxicity and can affect drift, mate-finding, predator avoidance, and therefore population dynamics. We show the practical implementation of the ToxmateLab, a new device that allows monitoring the movement behavior of up to 48 organisms simultaneously, for behavioral ecotoxicology. We quantified behavioral reactions of Gammarus pulex (Amphipoda, Crustacea) after exposure to two pesticides (dichlorvos and methiocarb) and two pharmaceuticals (diazepam and ibuprofen) at sublethal, environmentally relevant concentrations. We simulated a short-term pulse contamination event that lasted 90 min. Within this short test period, we successfully identified behavioral patterns that were most pronounced upon exposure to the two pesticides: Methiocarb initially triggered hyperactivity, after which baseline behavior was restored. On the other hand, dichlorvos induced hypoactivity starting at a moderate concentration of 5 µg/L - a pattern we also found at the highest concentration of ibuprofen (10 µg/L). An additional acetylcholine esterase inhibition assay revealed no significant impact of the enzyme activity that would explain the altered movement behavior. This suggests that in environmentally realistic scenarios chemicals can induce stress - apart from mode-of-action - that affects non-target organisms' behavior. Overall, our study proves the practical applicability of empirical behavioral ecotoxicological approaches and thus represents a next step towards routine practical use.

Database on eukaryotic symbionts of native and invasive gammarids (Crustacea, Amphipoda) in the Baltic region of Poland with information on water parameters for sampling sites.

This dataset documents the diversity of eukaryotic endo- and epibiotic organisms from 612 host individuals of seven gammarid (Amphipoda) species (Gammarus pulex, Gammarus zaddachi, Gammarus roeselii, Gammarus tigrinus, Dikerogammarus villosus, Pontogammarus robustoides, Echinogammarus ischnus) of native and invasive origin in the Baltic region of Poland. We identify 60 symbiotic species of nine phyla from 16 localities of freshwater and brackish habitats. Twenty-nine symbiotic species belonged to the Ciliophora, 12 to Apicomplexa, 8 to Microsporidia, 3 to Platyhelminthes, 2 to Acanthocephala, 2 to Nematoda, 2 to Rotifera, 1 to Choanozoa and 1 to Nematomorha. The material in this Data in Brief paper is composed of three Microsoft® Excel files. The first file represents the raw data on the number of individuals (infrapopulation size) of each eukaryotic symbiont taxa recorded in each host individual and location. The data set contains information on the assemblage of symbionts per host individual in one table-matrix; macro- (host) and symbiont taxa name, host length, the date of collection, the geographic coordinates and locality name in columns; and amphipod host specimens in lines. The second file reports the symbiont species list (the species breakdown by phyla in spreadsheets) with information on host species, sampling date, locality and geographic coordinates, infection site, obtained sequences (if the case), brief morphological characteristics and microphotographs. The third file reports measured water parameters, habitat features and host density per sample. We generate the present dataset to evaluate the richness, diversity, population and community features of symbiotic organisms in native and invasive gammarid hosts in Poland. Biological sciences: ParasitologyEnvironmental Science: Ecology; Hydrology and Water Quality.

Disentangling the determinants of symbiotic species richness in native and invasive gammarids (Crustacea, Amphipoda) of the Baltic region.

Dispersal of alien species is a global problem threatening native biodiversity. Co-introduction of non-native parasites and pathogens adds to the severity of this threat, but this indirect impact has received less attention. To shed light on the key factors determining the richness of microorganisms in native and invasive host species, we compared symbiotic (parasitic and epibiotic) communities of gammarids across different habitats and localities along the Baltic coast of Poland. Seven gammarid species, two native and five invasive, were sampled from 16 freshwater and brackish localities. Sixty symbiotic species of microorganisms of nine phyla were identified. This taxonomically diverse species assemblage of symbionts allowed us to assess the effect of host translocation and regional ecological determinants driving assembly richness in the gammarid hosts. Our results revealed that (i) the current assemblages of symbionts of gammarid hosts in the Baltic region are formed by native and co-introduced species; (ii) species richness of the symbiotic community was higher in the native Gammarus pulex than in the invasive hosts, probably reflecting a process of species loss by invasive gammarids in the new area and the distinct habitat conditions occupied by G. pulex and invasive hosts; (iii) both host species and locality were key drivers shaping assembly composition of symbionts, whereas habitat condition (freshwater versus brackish) was a stronger determinant of communities than geographic distance; (iv) the dispersion patterns of the individual species richness of symbiotic communities were best described by Poisson distributions; in the case of an invasive host, the dispersion of the rich species diversity may switch to a right-skewed negative binomial distribution, suggesting a host-mediated regulation process. We believe this is the first analysis of the symbiotic species richness in native and invasive gammarid hosts in European waters based on original field data and a broad range of taxonomic groups including Microsporidia, Choanozoa, Ciliophora, Apicomplexa, Platyhelminthes, Nematoda, Nematomorha, Acanthocephala and Rotifera, to document the patterns of species composition and distribution.

Reduced genetic diversity of freshwater amphipods in rivers with increased levels of anthropogenic organic micropollutants.

Anthropogenic chemicals in freshwater environments contribute majorly to ecosystem degradation and biodiversity decline. In particular anthropogenic organic micropollutants (AOM), a diverse group of compounds, including pesticides, pharmaceuticals, and industrial chemicals, can significantly impact freshwater organisms. AOM were found to impact genetic diversity of freshwater species; however, to which degree AOM cause changes in population genetic structure and allelic richness of freshwater macroinvertebrates remains poorly understood. Here, the impact of AOM on genetic diversity of the common amphipod Gammarus pulex (Linnaeus, 1758) (clade E) was investigated on a regional scale. The site-specific AOM levels and their toxic potentials were determined in water and G. pulex tissue sample extracts for 34 sites along six rivers in central Germany impacted by wastewater effluents and agricultural run-off. Population genetic parameters were determined for G. pulex from the sampling sites by genotyping 16 microsatellite loci. Genetic differentiation among G. pulex from the studied rivers was found to be associated with geographic distance between sites and to differences in site-specific concentrations of AOM. The genetic diversity parameters of G. pulex were found to be related to the site-specific AOM levels. Allelic richness was significantly negatively correlated with levels of AOM in G. pulex tissue (p < 0.003) and was reduced by up to 22% at sites with increased levels of AOM, despite a positive relationship of allelic richness and the presence of waste-water effluent. In addition, the inbreeding coefficient of G. pulex from sites with toxic AOM levels was up to 2.5 times higher than that of G. pulex from more pristine sites. These results indicate that AOM levels commonly found in European rivers significantly contribute to changes in the genetic diversity of an ecologically relevant indicator species.

Acanthocephalan parasites reflect ecological status of freshwater ecosystem.

Acanthocephalans' position in food webs, in close interaction with free-living species, could provide valuable information about freshwater ecosystem health through the viability of the parasites' host populations. We explored Pomphorhynchus laevis cystacanths' and adults' intensities of infection, and the prevalence of infected hosts respectively in their Gammarus pulex intermediate hosts and Squalius cephalus definitive hosts in a Mediterranean river. First, we analysed the relationship between P. laevis intensity of infection, its two hosts populations and the other acanthocephalan species found (Pomphorhynchus tereticollis and Polymorphus minutus). Second, we characterised the influence of bacteriological, physicochemical and biological water parameters on these acanthocephalans, and their intermediate and definitive hosts. This research highlights that P. laevis infection was closely related to their two preferential hosts population in the river. Moreover, P. laevis intensity of infection was positively correlated with organic pollution in the river but negatively correlated with biodiversity and with ecological indexes of quality. Pomphorhynchus laevis could thus benefit from moderate freshwater pollution, which promotes their tolerant intermediate and definitive hosts.