[Departement_IRSTEA]Eaux [TR1_IRSTEA]BELCA [ADD1_IRSTEA]Systèmes aquatiques soumis à des pressions multiples | International audience | Wastewater treatment plant effluents from urban area are a well-known source of chronic multiple micropollution to the downstream living organisms. In this study, ecologically relevant laboratory-bred freshwater gastropods, Lymnaea stagnalis, were exposed for 29 days to raw effluents of a wastewater treatment plant in Lyon area (France). A time-course analysis of individual markers of immunocompetence (hemocyte density and viability, hemocyte NADPH activity, phenol oxidase activity, and capacity of phagocytosis) has shown slight trends of inflammatory-like responses induced by the 100% effluents. So far, no short-term hazard for L. stagnalis can be revealed. However, over the long term, such environmental stress-stimulating immune responses could provoke deleterious life history trade-offs because the immune system is known to be highly energy-consuming.

Wastewater treatment plant effluents from urban area are a well-known source of chronic multiple micropollution to the downstream living organisms. In this study, ecologically relevant laboratory-bred freshwater gastropods, Lymnaea stagnalis, were exposed for 29 days to raw effluents of a wastewater treatment plant in Lyon area (France). A time-course analysis of individual markers of immunocompetence (hemocyte density and viability, hemocyte NADPH activity, phenol oxidase activity, and capacity of phagocytosis) has shown slight trends of inflammatory-like responses induced by the 100% effluents. So far, no short-term hazard for L. stagnalis can be revealed. However, over the long term, such environmental stress-stimulating immune responses could provoke deleterious life history trade-offs because the immune system is known to be highly energy-consuming.

International audience | The antifungal activity of seven essential oils (eucalyptus, clove, mint, oregano, savory, tea tree, and thyme) was studied on Venturia inaequalis, the fungus responsible for apple scab. The composition of the essential oils was checked by gas chromatography-mass spectrometry. Each essential oil had its main compound. Liquid tests were performed to calculate the IC50 of essential oils as well as their majority compounds. The tests were made on two strains with different sensitivities to tebuconazole: S755, the sensitive strain, and rs552, the strain with reduced sensitivity. Copper sulfate was selected as the reference mineral fungicidal substance. IC50 with confidence intervals were calculated after three independent experiments. The results showed that all essential oils and all major compounds had in vitro antifungal activities. Moreover, it was highlighted that the effectiveness of four essential oils (clove, eucalyptus, mint, and savory) was higher than copper sulfate on both strains. For each strain, the best activity was obtained using clove and eucalyptus essential oils. For clove, the IC50 obtained on the sensitive strain (5.2 mg/L [4.0–6.7 mg/L]) was statistically lower than the IC50 of reduced sensitivity strain (14 mg/L [11.1–17.5 mg/L]). In contrast, for eucalyptus essential oil, the IC50 were not different with respectively 9.4–13.0 and 12.2–17.9 mg/L for S755 and rs552 strains. For mint, origano, savory, tea tree, and thyme, IC50 were always the best on rs552 strain. The majority compounds were not necessarily more efficient than their corresponding oils; only eugenol (for clove) and carvacrol (for oregano and savory) seemed to be more effective on S755 strain. On the other hand, rs552 strain seemed to be more sensitive to essential oils than S755 strain. In overall, it was shown that essential oils have different antifungal activities but do not have the same antifungal activities depending on the fungus strain used.

The antifungal activity of seven essential oils (eucalyptus, clove, mint, oregano, savory, tea tree, and thyme) was studied on Venturia inaequalis, the fungus responsible for apple scab. The composition of the essential oils was checked by gas chromatography-mass spectrometry. Each essential oil had its main compound. Liquid tests were performed to calculate the IC₅₀ of essential oils as well as their majority compounds. The tests were made on two strains with different sensitivities to tebuconazole: S755, the sensitive strain, and rs552, the strain with reduced sensitivity. Copper sulfate was selected as the reference mineral fungicidal substance. IC₅₀ with confidence intervals were calculated after three independent experiments. The results showed that all essential oils and all major compounds had in vitro antifungal activities. Moreover, it was highlighted that the effectiveness of four essential oils (clove, eucalyptus, mint, and savory) was higher than copper sulfate on both strains. For each strain, the best activity was obtained using clove and eucalyptus essential oils. For clove, the IC₅₀ obtained on the sensitive strain (5.2 mg/L [4.0–6.7 mg/L]) was statistically lower than the IC₅₀ of reduced sensitivity strain (14 mg/L [11.1–17.5 mg/L]). In contrast, for eucalyptus essential oil, the IC₅₀ were not different with respectively 9.4–13.0 and 12.2–17.9 mg/L for S755 and rs552 strains. For mint, origano, savory, tea tree, and thyme, IC₅₀ were always the best on rs552 strain. The majority compounds were not necessarily more efficient than their corresponding oils; only eugenol (for clove) and carvacrol (for oregano and savory) seemed to be more effective on S755 strain. On the other hand, rs552 strain seemed to be more sensitive to essential oils than S755 strain. In overall, it was shown that essential oils have different antifungal activities but do not have the same antifungal activities depending on the fungus strain used.

A utilisé MicroScope Platform | International audience | Several studies have suggested the existence of a close relationship between antibiotic-resistant phenotypes and resistance to other toxic compounds such as heavy metals, which involve co-resistance or cross-resistance mechanisms. A metagenomic library was previously constructed in Escherichia coli with DNA extracted from the bacterial community inhabiting an acid mine drainage (AMD) site highly contaminated with heavy metals. Here, we conducted a search for genes involved in antibiotic resistance using this previously constructed library. In particular, resistance to antibiotics was observed among five clones carrying four different loci originating from CARN5 and CARN2, two genomes reconstructed from the metagenomic data. Among the three CARN2 loci, two carry genes homologous to those previously proposed to be involved in antibiotic resistance. The third CARN2 locus carries a gene encoding a membrane transporter with an unknown function and was found to confer bacterial resistance to rifampicin, gentamycin, and kanamycin. The genome of Thiomonas delicata DSM 16361 and Thiomonas sp. X19 were sequenced in this study. Homologs of genes carried on these three CARN2 loci were found in these genomes, two of these loci were found in genomic islands. Together, these findings confirm that AMD environments contaminated with several toxic metals also constitute habitats for bacteria that function as reservoirs for antibiotic resistance genes.

Several studies have suggested the existence of a close relationship between antibiotic-resistant phenotypes and resistance to other toxic compounds such as heavy metals, which involve co-resistance or cross-resistance mechanisms. A metagenomic library was previously constructed in Escherichia coli with DNA extracted from the bacterial community inhabiting an acid mine drainage (AMD) site highly contaminated with heavy metals. Here, we conducted a search for genes involved in antibiotic resistance using this previously constructed library. In particular, resistance to antibiotics was observed among five clones carrying four different loci originating from CARN5 and CARN2, two genomes reconstructed from the metagenomic data. Among the three CARN2 loci, two carry genes homologous to those previously proposed to be involved in antibiotic resistance. The third CARN2 locus carries a gene encoding a membrane transporter with an unknown function and was found to confer bacterial resistance to rifampicin, gentamycin, and kanamycin. The genome of Thiomonas delicata DSM 16361 and Thiomonas sp. X19 were sequenced in this study. Homologs of genes carried on these three CARN2 loci were found in these genomes, two of these loci were found in genomic islands. Together, these findings confirm that AMD environments contaminated with several toxic metals also constitute habitats for bacteria that function as reservoirs for antibiotic resistance genes.

The applicability of radon (222Rn) measurements to delineate non-aqueous phase liquids (NAPL) contamination in subsoil is discussed at a site with lithological discontinuities through a blind test. Three alpha spectroscopy monitors were used to measure radon in soil air in a 25,000-m2 area, following a regular sampling design with a 20-m2 grid. Repeatability and reproducibility of the results were assessed by means of duplicate measurements in six sampling positions. Furthermore, three points not affected by oil spills were sampled to estimate radon background concentration in soil air. Data histograms, Q-Q plots, variograms, and cluster analysis allowed to recognize two data populations, associated with the possible path of a fault and a lithological discontinuity. Even though the concentration of radon in soil air was dominated by this discontinuity, the characterization of the background emanation in each lithological unit allowed to distinguish areas potentially affected by NAPL, thus justifying the application of radon emanometry as a screening technique for the delineation of NAPL plumes in sites with lithological discontinuities.

The applicability of radon (²²²Rn) measurements to delineate non-aqueous phase liquids (NAPL) contamination in subsoil is discussed at a site with lithological discontinuities through a blind test. Three alpha spectroscopy monitors were used to measure radon in soil air in a 25,000-m² area, following a regular sampling design with a 20-m² grid. Repeatability and reproducibility of the results were assessed by means of duplicate measurements in six sampling positions. Furthermore, three points not affected by oil spills were sampled to estimate radon background concentration in soil air. Data histograms, Q-Q plots, variograms, and cluster analysis allowed to recognize two data populations, associated with the possible path of a fault and a lithological discontinuity. Even though the concentration of radon in soil air was dominated by this discontinuity, the characterization of the background emanation in each lithological unit allowed to distinguish areas potentially affected by NAPL, thus justifying the application of radon emanometry as a screening technique for the delineation of NAPL plumes in sites with lithological discontinuities.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [ADD1_IRSTEA]Systèmes aquatiques soumis à des pressions multiples | International audience | We used historical sources from the end of the nineteenth to the mid-twentieth century and current data to retrace fish assemblage trajectories for the past 150 years on 29 river stretches distributed throughout the Seine River basin. We based our analyses on species presence/absence, occurrence of amphidromous and non-native species, and species ecological traits related to habitat preference, oxygen and temperature requirements, and diet. In accordance with general trends observed for Western Europe, we detected a general decline of amphidromous species and an increase in nonnative species, even if, at some sites, several non-native species were extirpated. These changes affecting amphidromous and non-native species led to a weak increase in beta-diversity in fish assemblages. Independently of amphidromous and non-native species, for a reduced set of sites, we noted that changes in the balance of ecological traits over time, trace, in a consistent way, the major steps that affected watercourses like waterway development, increasing pollution, dam construction, or, locally, the recent wastewater treatment improvement. Despite local variations, we found contrasted trends between, on one hand, large rivers and/or catchments which had experienced strong expansion in human population, where fish assemblages had deteriorated, and, on the other hand, upstream catchments, with a declining human population, where fish assemblages showed signs of improvement. Because our results suggested that long-term changes affecting fish assemblages cannot be summarized as an unequivocal gradual degradation, we questioned the use of historical data to define ecological reference conditions for river assessment and management purposes.

We used historical sources from the end of the nineteenth to the mid-twentieth century and current data to retrace fish assemblage trajectories for the past 150 years on 29 river stretches distributed throughout the Seine River basin. We based our analyses on species presence/absence, occurrence of amphidromous and non-native species, and species ecological traits related to habitat preference, oxygen and temperature requirements, and diet. In accordance with general trends observed for Western Europe, we detected a general decline of amphidromous species and an increase in non-native species, even if, at some sites, several non-native species were extirpated. These changes affecting amphidromous and non-native species led to a weak increase in beta-diversity in fish assemblages. Independently of amphidromous and non-native species, for a reduced set of sites, we noted that changes in the balance of ecological traits over time, trace, in a consistent way, the major steps that affected watercourses like waterway development, increasing pollution, dam construction, or, locally, the recent wastewater treatment improvement. Despite local variations, we found contrasted trends between, on one hand, large rivers and/or catchments which had experienced strong expansion in human population, where fish assemblages had deteriorated, and, on the other hand, upstream catchments, with a declining human population, where fish assemblages showed signs of improvement. Because our results suggested that long-term changes affecting fish assemblages cannot be summarized as an unequivocal gradual degradation, we questioned the use of historical data to define ecological reference conditions for river assessment and management purposes.

International audience

The rhizosphere hosts a considerable microbial community. Among that community, bacteria called plant growth-promoting rhizobacteria (PGPR) can promote plant growth and defense against diseases using diverse distinct plant-beneficial functions. Crop inoculation with PGPR could allow to reduce the use of pesticides and fertilizers in agrosystems. However, microbial crop protection and growth stimulation would be more efficient if cooperation between rhizosphere bacterial populations was taken into account when developing biocontrol agents and biostimulants. Rhizospheric bacteria live in multi-species biofilms formed all along the root surface or sometimes inside the plants (i.e., endophyte). PGPR cooperate with their host plants and also with other microbial populations inside biofilms. These interactions are mediated by a large diversity of microbial metabolites and physical signals that trigger cell–cell communication and appropriate responses. A better understanding of bacterial behavior and microbial cooperation in the rhizosphere could allow for a more successful use of bacteria in sustainable agriculture. This review presents an ecological view of microbial cooperation in agrosystems and lays the emphasis on the main microbial metabolites involved in microbial cooperation, plant health protection, and plant growth stimulation. Eco-friendly inoculant consortia that will foster microbe–microbe and microbe–plant cooperation can be developed to promote crop growth and restore biodiversity and functions lost in agrosystems.

Biochemical indicators are potent tools to assess ecosystem functioning under anthropic and global pressures. Nevertheless, additional work is needed to improve the methods used for the measurement of these indicators, and for a more relevant interpretation of the obtained results. To face these challenges, the platform Biochem-Env aims at providing innovative and standardized measurement protocols, as well as database and information system favoring result interpretation and opening. Its skills and tools are also offered for expertise, consulting, training, and standardization. In addition, the platform is a service of a French Research Infrastructure for Analysis and Experimentation on Ecosystems, for research in environmental and agricultural sciences.

Biochemical indicators are potent tools to assess ecosystem functioning under anthropic and global pressures. Nevertheless, additional work is needed to improve the methods used for the measurement of these indicators, and for a more relevant interpretation of the obtained results. To face these challenges, the platform Biochem-Env aims at providing innovative and standardized measurement protocols, as well as database and information system favoring result interpretation and opening. Its skills and tools are also offered for expertise, consulting, training, and standardization. In addition, the platform is a service of a French Research Infrastructure for Analysis and Experimentation on Ecosystems, for research in environmental and agricultural sciences.