A global systematic map of knowledge of inland commercial navigation effects on freshwater ecosystems.

Inland navigation is one of the most sustainable transport alternatives to help decarbonise the world economy. However, the likely impacts of intensifying inland navigation on freshwater ecosystems are difficult to predict. A global map of knowledge that considers both abiotic and biotic responses to increasing shipping traffic and developing infrastructures is lacking. Deriving general evidence-based assessments is challenging, because most studies on inland navigation impacts are merely descriptive and either consist of local case studies, or address single navigation stressors or specific taxa only. We conducted a systematic mapping of the published literature (1908-2021) to provide a global synthesis of the effects of inland navigation on the biotic and abiotic components of freshwater ecosystems. We show that only half of the reported navigation-related impacts were statistically tested. Navigation itself (vessel operation) had mainly negative effects on native taxa (57%), followed by waterway management (40%), and navigation infrastructures (35%). Navigation has direct negative impacts caused by physical disturbances such as vessel-induced waves, and indirect impacts that facilitate the spread of aquatic invasive species, and altering the abiotic habitat conditions. Thirty percent of the tested relationships showed non-significant impacts on the biotic environment, while in 10% of cases impacts were context-dependent. We identified the main gaps of knowledge, namely (i) impacts of waterway management on communities, (ii) underlying processes of navigation impacts on river ecosystems; and (iii) interactions between multiple navigation factors and cascading effects on multi-taxa responses. These future research directions should improve the diagnosis, mitigate the negative impacts of navigation on rivers and provide guidelines for improving navigated river management.

Morphological vs. DNA metabarcoding approaches for the evaluation of stream ecological status with benthic invertebrates: Testing different combinations of markers and strategies of data filtering.

Macroinvertebrate assemblages are the most common bioindicators used for stream biomonitoring, yet the standard approach exhibits several time-consuming steps, including the sorting and identification of organisms based on morphological criteria. In this study, we examined if DNA metabarcoding could be used as an efficient molecular-based alternative to the morphology-based monitoring of streams using macroinvertebrates. We compared results achieved with the standard morphological identification of organisms sampled in 18 sites located on 15 French wadeable streams to results obtained with the DNA metabarcoding identification of sorted bulk material of the same macroinvertebrate samples, using read numbers (expressed as relative frequencies) as a proxy for abundances. In particular, we evaluated how combining and filtering metabarcoding data obtained from three different markers (COI: BF1-BR2, 18S: Euka02 and 16S: Inse01) could improve the efficiency of bioassessment. In total, 140 taxa were identified based on morphological criteria, and 127 were identified based on DNA metabarcoding using the three markers, with an overlap of 99 taxa. The threshold values used for sequence filtering based on the "best identity" criterion and the number of reads had an effect on the assessment efficiency of data obtained with each marker. Compared to single marker results, combining data from different markers allowed us to improve the match between biotic index values obtained with the bulk DNA versus morphology-based approaches. Both approaches assigned the same ecological quality class to a majority (86%) of the site sampling events, highlighting both the efficiency of metabarcoding as a biomonitoring tool but also the need for further research to improve this efficiency.

A minimalist macroparasite diversity in the round goby of the Upper Rhine reduced to an exotic acanthocephalan lineage.

The round goby, Neogobius melanostomus, is a Ponto-Caspian fish considered as an invasive species in a wide range of aquatic ecosystems. To understand the role that parasites may play in its successful invasion across Western Europe, we investigated the parasitic diversity of the round goby along its invasion corridor, from the Danube to the Upper Rhine rivers, using data from literature and a molecular barcoding approach, respectively. Among 1666 parasites extracted from 179 gobies of the Upper Rhine, all of the 248 parasites barcoded on the c oxidase subunit I gene were identified as Pomphorhynchus laevis. This lack of macroparasite diversity was interpreted as a loss of parasites along its invasion corridor without spillback compensation. The genetic diversity of P. laevis was represented by 33 haplotypes corresponding to a haplotype diversity of 0·65 ± 0·032, but a weak nucleotide diversity of 0·0018 ± 0·00015. Eight of these haplotypes were found in 88·4% of the 248 parasites. These haplotypes belong to a single lineage so far restricted to the Danube, Vistula and Volga rivers (Eastern Europe). This result underlines the exotic status of this Ponto-Caspian lineage in the Upper Rhine, putatively disseminated by the round goby along its invasion corridor.

Increase in developmental instability in a field-collected Chironomus population maintained under laboratory conditions.

In order to be a relevant indicator of exposure towards teratogenic stressors, morphological defects should not be passed on to the next generation. In this study, we compare morphological variations in Chironomids collected from a contaminated river stretch with those of their progeny, reared in uncontaminated sediment under laboratory conditions. We focused on mentum defects (deformities, fluctuating asymmetry and mean shape change), measured by geometric morphometrics. We observed no significant variation in deformity rate between the parental generation and its progeny. On the contrary, we observed a significant increase in fluctuating asymmetry and a significant decrease in mentum centroid size in the offspring. Our results suggest that shape defects are not caused by direct exposure to teratogenic stressors alone. We propose four hypotheses to explain this: (a) teratogenic contaminants are present in egg-clutches, (b) contaminants at the sampling site have mutagenic effects, (c) costs of tolerance, and (d) contamination-induced genetic impoverishment.

Finding the perfect pairs: A matchmaking of plant markers and primers for multi-marker eDNA metabarcoding.

As the scope of plant eDNA metabarcoding diversifies, so do the primers, markers and methods. A wealth of primers exists today, but their comparative evaluation is lacking behind. Similarly, multi-marker approaches are recommended but debates persist regarding barcode complementarity and optimal combinations. After a literature compilation of used primers, we compared in silico 102 primer pairs based on amplicon size, coverage and specificity, followed by an experimental evaluation of 15 primer pairs on a mock community sample covering 268 plant species and genera, and about 100 families. The analysis was done for the four most common plant metabarcoding markers, rbcL, trnL, ITS1 and ITS2 and their complementarity was assessed based on retrieved species. By focusing on existing primers, we identify common designs, promote alternatives and enhance prior-supported primers for immediate applications. The ITS2 was the best-performing marker for flowering vascular plants and was congruent to ITS1. However, the combined taxonomic breadth of ITS2 and rbcL surpassed any other combination, highlighting their high complementarity across Streptophyta. Overall, our study underscores the significance of comprehensive primer and barcode evaluations tailored to metabarcoding applications.

A comparative analysis of eDNA metabarcoding and field surveys: Exploring freshwater plant communities in rivers.

While environmental DNA (eDNA) metabarcoding holds promise as a holistic approach to assess vegetation changes and community composition across diverse spatial and temporal scales, systematic investigations of its efficacy compared to conventional field surveys remain scarce in the literature. The present study explores the differences in plant diversity recovered from field surveys and captured with a multi-marker eDNA metabarcoding approach (two nrDNA ITS1 and ITS2, and two cpDNA rbcL and trnL) from river water samples. The eDNA metabarcoding approach retrieved 46 aquatic plants (hydrophytes and helophytes) and 245 terrestrial plants, compared to 24 and 127 species identified from field surveys. On average, eDNA samples collected immediately downstream of the survey sites recovered 43 % and 39 % of the aquatic and terrestrial species observed, respectively. Discrepancies were explained by differences in taxonomic resolution, the stochasticity of the retrieval of rare and elusive species, and the presence of reference sequences. We found a significant positive correlation between spatial and community distances at scales ranging from 2 to 9 km and identified turnover as the driving force of these differences. Metabarcoding demonstrated sensitivity to community changes and both approaches converge on a similar community structure. Interestingly, eDNA samples collected immediately upstream of the survey sites exhibited significant species overlap with the downstream samples (c. 100 m apart). Overall, our results demonstrate that within-site species mismatches between the methods are nonnegligible, and they question the use of eDNA for generating complete species lists at scales comparable to our field surveys (< 100-m transects). However, with adequate sampling and a multi-marker metabarcoding approach, eDNA has the potential to approximate catchment gamma diversity with less sampling effort than conventional surveys.

Inland navigation and land use interact to impact European freshwater biodiversity.

Inland navigation in Europe is proposed to increase in the coming years, being promoted as a low-carbon form of transport. However, we currently lack knowledge on how this would impact biodiversity at large scales and interact with existing stressors. Here we addressed this knowledge gap by analysing fish and macroinvertebrate community time series across large European rivers comprising 19,592 observations from 4,049 sampling sites spanning the past 32 years. We found ship traffic to be associated with biodiversity declines, that is, loss of fish and macroinvertebrate taxonomic richness, diversity and trait richness. Ship traffic was also associated with increases in taxonomic evenness, which, in concert with richness decreases, was attributed to losses in rare taxa. Ship traffic was especially harmful for benthic taxa and those preferring slow flows. These effects often depended on local land use and riparian degradation. In fish, negative impacts of shipping were highest in urban and agricultural landscapes. Regarding navigation infrastructure, the negative impact of channelization on macroinvertebrates was evident only when riparian degradation was also high. Our results demonstrate the risk of increasing inland navigation on freshwater biodiversity. Integrative waterway management accounting for riparian habitats and landscape characteristics could help to mitigate these impacts.

Water temperature dynamics in a headwater forest stream: Contrasting climatic, anthropic and geological conditions create thermal mosaic of aquatic habitats.

The thermal regime of streams is a relevant driver of their ecological functioning. As this regime is presently submitted to numerous alterations (among others, impoundments, and climate change), it seems important to study both their effects and potential recovery from the latter. Thus, we investigated the surface and hyporheic water temperature along a small headwater stream with contrasting environmental contexts: forest landscape, open grassland landscape without riparian vegetation, several artificial run-of-the-river impoundments and one discharge point of a by-pass impoundment. The main objectives were to study the influence of these contrasting contexts on surface and subsurface water temperature at a local scale. Contrasting contexts were supposed to create effects on both surface and hyporheic thermal regimes at a local scale. Differences of thermal regimes between surface and hyporheos were expected, as well as between geological contexts. Sensors located at multiple stations allowed monitoring of stream and hyporheos temperature along the stream, while comparison with adjacent reference stream allowed for surface water thermal regime benchmark. Impoundments and landscapes significantly influenced stream thermal regime at a local scale (impoundments created up to +3.7°C temperature increase in average). Their effect on hyporheos thermal regime was less marked than the ones generated by solar radiation or geological features. Hyporheos thermal regime varies from stream one by temperature dynamics delay (up to 18h) and decrease (up to -7°C between surface and hyporheos temperature in average). These coupled effects create a mosaic of thermal habitats, which could be used for river biodiversity preservation and restoration.

Influence of gender and season on reduced glutathione concentration and energy reserves of Gammarus roeseli.

As biomarkers are known to be influenced by biotic and abiotic factors (e.g. gender, temperature), we investigated over a one-year long sampling period, the influence of season and gender on reduced glutathione concentrations and its synthesis in the crustacean amphipod Gammarus roeseli. At the same time, we assessed energy reserves and malondialdehyde levels as toxic biomarker. Results have shown that, in both genders, reduced glutathione concentrations were inversely correlated to water temperature, and higher in females than in males whatever the season. Total lipid and glycogen contents were higher in females than in males, allowing females to have enough energy to assume the reproductive period and maintain high GSH concentrations for detoxification processes. Conversely, females have lower cell damages than males. These differences between genders could induce differential sensitivity in a contamination context, and thus affect the population. Females could resist better than males in contaminated environments, especially in spring when reduced glutathione concentration is the highest.

Is there a link between shell morphology and parasites of zebra mussels?

The shell morphology of zebra mussels, Dreissena polymorpha, was analyzed to determine if alterations in shell shape and asymmetry between valves were related to its infection status, i.e. infected or not by microparasites like ciliates Ophryoglena spp. or intracellular bacteria Rickettsiales-like organisms (RLOs), and by macroparasites like trematodes Phyllodistomum folium and Bucephalus polymorphus. For microparasites, two groups of mussels were observed depending on shell measurements. Mussels with the more concave shells were the most parasitized by ciliates. This could be more a consequence than a cause and we hypothesized that a modification of the water flow through the mantle cavity could promote the infection with a ciliate. There were more RLOs present in the most symmetrical individuals. A potential explanation involved a canalization of the left-right asymmetry as a by-product of the parasite infection. Trematode infections were associated with different responses in valve width. Females infected by P. folium displayed significantly higher symmetry in valve width compared with non-infected congeners, whereas the infection involved an opposite pattern in males. B. polymorphus was also linked to a decrease in valve width asymmetry. This study suggested that a relationship exists between parasitism and shell morphology through the physiological condition of host zebra mussels.

Evolution of relict floodplain forest in river stretches of Western and Central Europe as affected by river infrastructure networks.

We studied the impact of infrastructure networks on relict floodplain forest along three stretches of the Upper Rhine (Kembs-Efringen-Kirchen, Strasbourg-Kehl and Beinheim-Iffezheim) and the Inn-Danube (Mulheim-Obernberg, Passau-Ingling and Engelhartszell-Jochenstein), each on the border between two countries. We analysed land use patterns within a 500 m wide buffer area along the main channel using photo-interpretation and compared the situations between the 1950s, 1980's and 2010's. Temporal changes were assessed with transition matrices and selected spatial metrics, including fragmentation indices. Over this period, forest area remained similar at three sites, increased slightly at two sites and decreased at one site. However, on average, 12.5% of floodplain forest had changed location (range: 7.3% (Engelhartszell-Jochenstein)- 26.5% (Kembs-Efringen-Kirchen)). The natural development of unmanaged areas and agricultural abandonment after World War II has led to the emergence of young riparian forests along rivers. In the Upper Rhine region, the results showed asymmetry in these two factors, with unmanaged natural areas most important on the French side and agricultural abandonment on the German side. Along the Inn-Danube, agricultural abandonment has led to an increase or stagnation of floodplain forest areas. In most cases, development of transport infrastructure between the 1950s and 2010s has caused fragmentation of the forest area, reducing the relict forest to a patchy green corridor with reduced functionality and interfacing. To go further and improve the management of these relict forests, we have to investigate the interdependency between practices related to infrastructure operation and the role that biodiversity plays for stakeholders.

Restoring fluvial forms and processes by gravel augmentation or bank erosion below dams: A systematic review of ecological responses.

Aquatic biological communities have directly undergone human-induced changes. Altered hydrological and morphological processes in running waters have caused the degradation of main habitats for biotas and have disturbed ecosystem functionality. The latest advances in river restoration concerned the rise in far-reaching hydromorphological restoration actions that have been implemented below dams to reverse well-known negative impacts of anthropogenic pressures. Some authors emphasized the enhancement of sediment supply and habitat diversity using gravel augmentation or bank erosion to restore morphodynamics, and thus improve biodiversity. We explored the Web of Science database for empirical research papers that specifically addressed such hydromorphological river restoration actions. Articles were examined using a text-content analysis tool to determine the major concepts or ideas they deal with. It has also been proved as useful in defining interrelationships and degree of interdisciplinary. Results showed that a low number of published scientific articles exist about such projects, mainly condensed in the North hemisphere. Divergent ecological issues were highlighted by the word co-occurrence networks: (i) gravel augmentation was used to improve spawning habitats for fish of economic interest whereas (ii) erodible corridor was designed to safeguard natural riparian systems, approaching morphological goals of channel widening. Overall, ecological responses were consistent with those expected, leading however rather to functional shifts than richness increase. Gravel augmentation or bank erosion were not usually combined with in-channel structure management. However, this might be an option to consider since the biological communities seem to be sensitive during first restorations with such combination. This review demonstrates the value of word co-occurrence networks in exploring a high number of previous publications, keys for formulating guidance to manage gravel augmentation or bank erosion along ecological purposes.

Field evidence for non-host predator avoidance in a manipulated amphipod.

Manipulative parasites are known to alter the spatial distribution of their intermediate hosts in a way that enables trophic transmission to definitive hosts. However, field data on the ecological implications of such changes are lacking. In particular, little is known about the spatial coexistence between infected prey and dead-end predators after a parasite-induced habitat shift. Here, we used an Amphipoda (Gammarus roeseli)-Acanthocephala (Polymorphus minutus) association to investigate how infection with a manipulative parasite affects the predation risk by non-hosts within the invertebrate community. First, we collected invertebrates by sampling various natural habitats and calculated the distribution amplitude of amphipods according to their infection status. Infection with P. minutus significantly reduced the habitat breadth in G. roeseli, parasitised individuals being mainly found in floating materials whereas uninfected ones were widespread throughout the sampled habitats. Second, to test if these changes also affect the risk for P. minutus to be ingested by non-hosts, we estimated the predation risk experienced by G. roeseli within the macro-invertebrate community. The habitat overlap between potential invertebrate predators and G. roeseli showed that the spatial probability of encounter was lower for P. minutus-infected amphipods than for uninfected conspecifics. For the first time, to our knowledge, a study used ecological tools to bring field evidence for the spatial avoidance of dead-end predators in a manipulated amphipod.

High food quality increases infection of Gammarus pulex (Crustacea: Amphipoda) by the acanthocephalan parasite Pomphorhynchus laevis.

Parasitism is an important process in ecosystems, but has been largely neglected in ecosystem research. However, parasites are involved in most trophic links in food webs with, in turn, a major role in community structure and ecosystem processes. Several studies have shown that higher nutrient availability in ecosystems tends to increase the prevalence of parasites. Yet, most of these studies focused on resource availability, whereas studies investigating resource quality remain scarce. In this study, we tested the impact of the quality of host food resources on infection by parasites, as well as on the consequences for the host. Three resources were used to individually feed Gammarus pulex (Crustacea: Amphipoda) experimentally infected or not infected with the acanthocephalan species Pomphorhynchus laevis: microbially conditioned leaf litter without phosphorus input (standard resource); microbially conditioned leaf litter enriched in phosphorus; and microbially conditioned leaf litter without phosphorus input but complemented with additional inputs of benthic diatoms rich in both phosphorus and eicosapentaenoic acid. During the 110 day experiment, infection rate, parasite load, host survival, and parasite-mediated behavioral traits implicated in trophic transmission were measured (refuge use, geotaxis and locomotor activity). The resources of higher quality, regardless of the infection status, reduced gammarid mortality and increased gammarid growth. In addition, higher quality resources increased the proportion of infected gammarids, and led to more cases of multi-infections. While slightly modifying the geotaxis behavior of uninfected gammarids, resource quality did not modulate the impact of parasites on host behavior. Finally, for most parameters, consumption of algal resources had a greater impact than did phosphorus-enriched leaf litter. Therefore, manipulation of resource quality significantly affected host-parasite relationships, which stressed the need for future research to investigate in natura the relationships between resource availability, resource quality and parasite prevalence.

Host manipulation of a freshwater crustacean (Gammarus roeseli) by an acanthocephalan parasite (Polymorphus minutus) in a biological invasion context.

Several gammarid species serve as intermediate hosts for the acanthocephalan parasite Polymorphus minutus. This parasite influences gammarid behaviour in order to favour transmission to its ultimate host, generally a bird. We investigated this host manipulation in Gammarus roeseli, a gammarid species introduced in France 150 years ago which now coexists with several exotic species from different origins. In the field, vertical distribution of G. roeseli revealed a higher proportion of infected individuals close to the water's surface and the size distribution of infected gammarids revealed predation pressure on infected individuals. However, under laboratory conditions both infected and non-infected individuals remained benthic. The addition of a second gammarid, Dikerogammarus villosus, to the experimental device involved a vertical displacement of infected G. roeseli towards the water's surface. Dikerogammarus villosus, originating from the Ponto-Caspian basin, can be considered as an aggressive predator. The substitution of D. villosus with Atyaephyra desmarestii, a planktivore decapod, did not alter the gammarids' distribution, with both infected and uninfected G. roeseli staying benthic. Thus, biotic interactions between D. villosus and G. roeseli represent selective pressure encouraging the expression of manipulated behaviour in infected amphipods. Through manipulation, P. minutus was found to increase the survival of infected G. roeseli when faced with non-host predators and to make it more vulnerable to predation by the parasite's definitive host.

Testing the potential of a ribosomal 16S marker for DNA metabarcoding of insects.

Cytochrome c oxidase I (COI) is a powerful marker for DNA barcoding of animals, with good taxonomic resolution and a large reference database. However, when used for DNA metabarcoding, estimation of taxa abundances and species detection are limited due to primer bias caused by highly variable primer binding sites across the COI gene. Therefore, we explored the ability of the 16S ribosomal DNA gene as an alternative metabarcoding marker for species level assessments. Ten bulk samples, each containing equal amounts of tissue from 52 freshwater invertebrate taxa, were sequenced with the Illumina NextSeq 500 system. The 16S primers amplified three more insect species than the Folmer COI primers and amplified more equally, probably due to decreased primer bias. Estimation of biomass might be less biased with 16S than with COI, although variation in read abundances of two orders of magnitudes is still observed. According to these results, the marker choice depends on the scientific question. If the goal is to obtain a taxonomic identification at the species level, then COI is more appropriate due to established reference databases and known taxonomic resolution of this marker, knowing that a greater proportion of insects will be missed using COI Folmer primers. If the goal is to obtain a more comprehensive survey the 16S marker, which requires building a local reference database, or optimised degenerated COI primers could be more appropriate.

Decrease of fluctuating asymmetry among larval instars in an aquatic, holometabolous insect.

The use of fluctuating asymmetry (FA) to reveal environmental stress in natural populations has been investigated extensively over the past decade. In aquatic ecosystems, many investigations involve amphibiotic insects with larval development stages in freshwater. To explain the important variability in study results, we investigated the FA level of a natural Hydropsyche exocellata (Insecta: Trichoptera) population at four different stages, from first larval instar to adult. The level of FA decreased between larval stages, but remained constant between the last larval instar and the adult. The presence of a compensational growth mechanism and the elimination of asymmetric individuals by natural selection are two hypotheses that could explain this observation.

Exposure to sediments from polluted rivers has limited phenotypic effects on larvae and adults of Chironomus riparius.

Laboratory studies have sometimes failed to detect a relationship between toxic stress and morphological defects in invertebrates. Several hypotheses have been proposed to account for this lack of effect. (1) It was suggested that only a combination of stressful conditions - rather than a single one - would affect the phenotype. (2) Phenotypic defects should be detected on adult individuals, rather than on juveniles. (3) Phenotypic abnormalities might mostly affect the progeny of the exposed individuals, some contaminants exhibiting trans-generational effects. In the present study, we test those three hypotheses. We first examined the effects of a multiple exposure by using laboratory Chironomus riparius larvae cultured on two sediments sampled in contaminated rivers and those containing a mixture of mineral and organic compounds. On the larvae, we investigated mentum phenotypes: the frequency of phenodeviants, the shape fluctuating asymmetry and the mean shape. To test whether adult's morphology was more sensitive than the larval's, we also measured asymmetry and mean shape of the adult wings. Finally, to test for a trans-generational phenotypic effect, we measured mentum shape variations in the offspring derived from the measured adults. Overall, our results point out a very limited phenotypic response to contaminated sediments, suggesting that a multiple exposure is not necessarily sufficient to generate phenotypic defects. Adult traits were no more affected than larval traits, discarding the hypothesis that adult phenotypes would be more sensitive biomarkers. Finally, no effect was detected on the offspring generation, suggesting that no trans-generational effect occurs. This general lack of effect suggests that the use of phenotypic defects in C. riparius as an indicator of sediment contamination should be considered cautiously.

Biochemical and morphological responses in Chironomus riparius (Diptera, Chironomidae) larvae exposed to lead-spiked sediment.

The aim of the present study was to assess the potential use of biochemical markers and mentum deformities as indicators of long-term exposure to lead (Pb) in Chironomus riparius larvae. To do this, the authors measured 3 biochemical markers (i.e., malondialdehyde level, metallothionein concentration, and energy reserve content) as well as larval growth and mentum deformities after 16-d exposure to sediment containing Pb. The concentrations studied ranged from 3.5 mg/kg to 505.5 mg/kg dry weight. Despite the bioaccumulation of Pb in C. riparius bodies, frequencies of both mentum deformities and the dry weight were not significantly different between the control and stressed groups. On the contrary, Pb exposure caused a significant increase of both malondialdehyde level and metallothionein concentration. The increase of body Pb concentrations did not significantly modify body copper and zinc concentrations. Moreover, we observed a decrease of total lipid content and an increase of glycogen content as a function of a dose-response relationship, while no variation in protein concentrations was observed. Despite the adverse effects observed at the biochemical level, larval development was not affected. These results suggested that measurements of malondialdehyde level, metallothionein concentration, and energy reserve content can be used as relevant biomarkers of long-term sublethal exposure to Pb in C. riparius larvae.

Parasitism can be a confounding factor in assessing the response of zebra mussels to water contamination.

Biological responses measured in aquatic organisms to monitor environmental pollution could be also affected by different biotic and abiotic factors. Among these environmental factors, parasitism has often been neglected even if infection by parasites is very frequent. In the present field investigation, the parasite infra-communities and zebra mussel biological responses were studied up- and downstream a waste water treatment plant in northeast France. In both sites, mussels were infected by ciliates and/or intracellular bacteria, but prevalence rates and infection intensities were different according to the habitat. Concerning the biological responses differences were observed related to the site quality and the infection status. Parasitism affects both systems but seemed to depend mainly on environmental conditions. The influence of parasites is not constant, but remains important to consider it as a potential confounding factor in ecotoxicological studies. This study also emphasizes the interesting use of integrative indexes to synthesize data set.