Identification of ecological thresholds from variations in phytoplankton communities among lakes: contribution to the definition of environmental standards.

In aquatic ecosystems, the identification of ecological thresholds may be useful for managers as it can help to diagnose ecosystem health and to identify key levers to enable the success of preservation and restoration measures. A recent statistical method, gradient forest, based on random forests, was used to detect thresholds of phytoplankton community change in lakes along different environmental gradients. It performs exploratory analyses of multivariate biological and environmental data to estimate the location and importance of community thresholds along gradients. The method was applied to a data set of 224 French lakes which were characterized by 29 environmental variables and the mean abundances of 196 phytoplankton species. Results showed the high importance of geographic variables for the prediction of species abundances at the scale of the study. A second analysis was performed on a subset of lakes defined by geographic thresholds and presenting a higher biological homogeneity. Community thresholds were identified for the most important physico-chemical variables including water transparency, total phosphorus, ammonia, nitrates, and dissolved organic carbon. Gradient forest appeared as a powerful method at a first exploratory step, to detect ecological thresholds at large spatial scale. The thresholds that were identified here must be reinforced by the separate analysis of other aquatic communities and may be used then to set protective environmental standards after consideration of natural variability among lakes.

Assessing the impact of chloroacetanilide herbicides and their metabolites on periphyton in the Leyre River (SW France) via short term growth inhibition tests on autochthonous diatoms.

The Leyre River is the main tributary to the Bassin d'Arcachon lagoon. Herbicides belonging to the chloroacetanilide class have been found in the river (S-metolachlor and acetochlor) as well as some of their metabolites at higher concentrations. As the environmental toxicity of these molecules is not well known, ecotoxicological tests have been carried out on river periphyton at different levels of biological diversity: from the clone of one diatom species (Nitzschia nana) to the population of the same species (several clones) up to the multi-specific species community dominated by diatoms. Moreover, tests were performed on diatoms coming from an unpolluted upstream site and from a contaminated downstream site, in order to investigate possible tolerance acquisition to pollutants. The method consisted in measuring diatom growth inhibition at different doses of each substance from the increase of chlorophyll-a concentration after 4 days. It resulted that acetochlor was clearly more toxic than S-metolachlor at all levels of biological diversity. EC(50) values estimated from the tests suggest no effect of contaminants on diatom growth or biomass in the river. The toxicity of the metabolites appeared very low compared to that of their parent compounds. No difference in tolerance to the herbicides was demonstrated between summer diatom communities from the two sites in spite of different specific compositions. However, concerning the populations of N. nana isolated in winter following the highest herbicide concentrations in the river (about 0.5 µg L(-1)), the downstream population showed a higher tolerance to acetochlor but there was no co-tolerance to S-metolachlor. Thus, it appeared that acetochlor represents the highest toxic pressure on periphyton among the other contaminants in the Leyre River.

Variations of periphytic diatom sensitivity to the herbicide diuron and relation to species distribution in a contamination gradient: implications for biomonitoring.

Diatoms are commonly used as bioindicators of trophic and saprobic pollution in rivers. However, more knowledge is needed concerning their sensitivity to toxicants such as agricultural herbicides. In this study, seven species of periphytic diatoms were isolated from the Morcille River (Beaujolais area, France) which presents a streamward contamination gradient by pesticides and particularly diuron. The sensitivity of these species to diuron was assessed through ecotoxicological tests based on short-term growth inhibition of monospecific cultures. After application of an appropriate toxicological model, EC50 were determined and the species were ranked according to their tolerance. EC50 values ranged from 4.5 to 19 µg L(-1). Finally, the results were related to field periphyton samples from an upstream and a downstream site in order to check if variations in specific relative abundance between sites are consistent with differences in tolerance to diuron. Species distribution between sites was only partially in accordance with toxicological results suggesting that other factors (toxic or trophic) have an important influence on diatom communities in the river. Nevertheless, diatoms showed their potential to indicate water contamination by pesticides and toxic indices could be developed in complement to existing trophic indices.

Characterization of Freshwater Diatom Communities: Comparing Taxonomic and Genetic-Fingerprinting Approaches.

Benthic diatom assemblages from five sampling sites located on two rivers were characterized simultaneously by means of traditional microscopic observations and PCR-DGGE fingerprinting with primers specifically designed for Bacillariophyceae. Community structure, richness, and diversity assessed by both methods were compared. Diatom lists obtained from morphological identification were separated into subsets, depending on (i) the taxonomic level considered (genus, species, variety) and, for each of them, (ii) the relative abundance (RA) of each component (the whole data set, RA > 1%, RA > 2%). These data were then compared to genetic fingerprinting data. Clusters based on taxonomic composition and DGGE banding patterns were very similar, showing good correspondence of community structure between the two methods. Data were compared by linear regressions between indices (richness, diversity) and by Mantel tests on dissimilarity matrices generated for each community composition data set. Statistical analysis indicated that the most reliable correlations with fingerprinting were obtained for genera representing more than 1% RA or species representing more than 2% RA. The results reveal that the PCR-DGGE protocol described here offers a satisfactory alternative for performing preliminary screening of coarse differences in diatom global community structure between samples. It can be regarded as a good complement to taxonomic analyses, which still remain necessary to detect precise changes in richness and diversity, especially when considering species with low abundance in natural assemblages.

Evaluation of single and joint toxic effects of diuron and its main metabolites on natural phototrophic biofilms using a pollution-induced community tolerance (PICT) approach.

This study assessed the single and joint acute toxicity of diuron and two of its metabolites (DCPMU and 3,4-DCA) on natural phototrophic biofilms using a PICT approach with photosynthesis bioassays. River biofilm communities were collected at three sampling stations exhibiting increasing concentrations of diuron, DCPMU and 3,4-DCA from upstream to downstream. Applied individually, the parent compound was more toxic than its metabolites, with DCPMU being more toxic than 3,4-DCA which only inhibited photosynthesis at very high concentrations (EC25 at about 5 mg/l). Sensitivity of biofilm communities to diuron and DCPMU decreased from upstream to downstream, revealing tolerance induction in contaminated sections of the river, as expected from the PICT concept. Nevertheless, PICT was not applicable for 3,4-DCA, which similarly affected upstream, intermediate and downstream biofilm communities. Chemical mixtures of diuron and DCPMU demonstrated additive effects whereas combinations with 3,4-DCA enhanced the observed effects. Our results reveal that the individual and combined presence of diuron and DCPMU in lotic ecosystems can have both short-term effects (as shown with bioassays) and long-term effects (as shown through the PICT approach) on phototrophic biofilms, whereas environmental concentrations of 3,4-DCA may not affect biofilm photosynthetic activity.