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Changes in the macroinvertebrate community were investigated over 10 months at four sites along a 19 km salinity gradient (0.21-2.60 g l )1 ) in a sixth-order stream, the Meurthe River, northeastern France. Abiotic characteristics other than salinity were similar between the sites. Macroinvertebrate taxonomic richness decreased by 30% downstream of the 1.4 g l )1 sites while diversity, evenness or total abundance of taxa did not change along the gradient. In terms of functioning, a slight change in relative abundances of invertebrate feeding groups followed the salinity gradient. Eight invertebrate assemblages occurred within specific salinity distributions were identified. The exotics Gammarus tigrinus, Dreissena polymorpha, Corbicula fluminalis and Corophium curvispinum, were more abundant at the highest salinity site. These results suggest that rising salinity concentrations drastically affect the species composition, including favouring exotic species.

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.

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. In comparison to COI, the 16S marker amplified more insect species and amplified more equally, probably due to decreased primer bias. Rough estimation of biomass might thus be less biased with 16S than with COI. 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 species will be missed using COI Folmer primers. If the goal is to obtain a more comprehensive survey in a context where it is possible to build a local reference database, the 16S marker could be more appropriate.

Knowledge of characteristics helpful in screening potential invaders and in elaborating strategies to limit their success is highly desirable. We focused on gammarid amphipods from Western Europe and North America to discover biological and/or ecological traits that may explain successful invasion by these species. Two typologies were considered: an analytical one, with groups built on the basis of biological or ecological similarities, and an empirical one, with groups constituted a priori according to a species' invasive status and its fresh or brackish water origin. The results obtained are discussed in the light of three hypotheses that may influence invasiveness: biotic potential, species size and euryoeciousness. The analysis revealed a particular ecological profile for invaders, with a strong influence of salinity tolerance, but no typology was found based on biological characteristics. Invasiveness cannot be predicted from a limited number of criteria, and is the result of a combination of several characteristics. Invasive species therefore exhibit a particular ecological profile rather than a biological one, contrary to most classical explanations.

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Evenness indices are numerous but the lack of knowledge of their properties is a limitation to their biological usefulness. 15 evenness indices, two of them being recently proposed, were studied in this work. We investigated the sensitivity of each index using (1) 189 macroinvertebrate communities sampled in the field, and (2) a set of communities modified in a controlled way.There is no single way to measure evenness. We demonstrated that a measure should be chosen considering (1) the kind of data analysed and, (2) the index properties wanted by users. Depending on the ecological data set the index should be more sensitive to variations in rare, median or abundant species. For most of macroinvertebrate community analyses, a convenient evenness index requirs to be symmetric, unsensitive to variation on rare taxa, with a large range of variation and can be compared with a diversity index. Depending on diversity measure used, five indices correspond to these criteria: E Pielou , E Hurlbert, E -ln(D) , E 1-D, E MI. Our results were summarized in a table which may help users to select a convenient evenness measure according to their specific data. Concerning index properties, three main features that an evenness index should meet are briefly discussed: dependence with richness, symmetry criteria and variation range. A revue of this controversial subject allowed a best understanding of values obtained with evenness measures depending on their own features.

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