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.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]BELCA [ADD1_IRSTEA]Systèmes aquatiques soumis à des pressions multiples | International audience | Gammarus fossarum is an often-abundant crustacean detritivore that contributes importantly to leaf litter breakdown in oligotrophic, mainly heterotrophic, headwater streams. This species requires large amounts of Ca to moult, thus allowing growth and reproduction. Because resource quality is tightly coupled to the organism's growth and physiological status, we hypothesised that low Ca concentration [Ca] and low food resource quality (low phosphorus [P] and/ or reduced highly unsaturated fatty acid [HUFA] contents) would interactively impair molecular responses (gene expression) and reproduction of G. fossarum. To investigate the effects of food resources quality, we experimentally manipulated the P content of sycamore leaves and also used diatoms because they contain high amounts of HUFAs. Three resource quality treatments were tested: low quality (LQ, unmanipulated leaves: low P content), high quality 1 (HQ1; P-manipulated leaves: high P content), and high quality 2 (unmanipulated leaves supplemented with a pellet containing diatoms: high P and HUFA content). Naturally, demineralised stream water was supplemented with CaSO4 to obtain three Ca concentrations (2, 3.5, and 10.5 mg Ca L

Gammarus fossarum is an often-abundant crustacean detritivore that contributes importantly to leaf litter breakdown in oligotrophic, mainly heterotrophic, headwater streams. This species requires large amounts of Ca to moult, thus allowing growth and reproduction. Because resource quality is tightly coupled to the organism’s growth and physiological status, we hypothesised that low Ca concentration [Ca] and low food resource quality (low phosphorus [P] and/or reduced highly unsaturated fatty acid [HUFA] contents) would interactively impair molecular responses (gene expression) and reproduction of G. fossarum. To investigate the effects of food resources quality, we experimentally manipulated the P content of sycamore leaves and also used diatoms because they contain high amounts of HUFAs. Three resource quality treatments were tested: low quality (LQ, unmanipulated leaves: low P content), high quality 1 (HQ1; P-manipulated leaves: high P content), and high quality 2 (unmanipulated leaves supplemented with a pellet containing diatoms: high P and HUFA content). Naturally, demineralised stream water was supplemented with CaSO₄ to obtain three Ca concentrations (2, 3.5, and 10.5 mg Ca L⁻¹). For 21 days, pairs of G. fossarum were individually exposed to one of the nine treatments (3 [Ca] × 3 resource qualities). At the individual level, strong and significant delays in moult stage were observed in gammarids exposed to lower [Ca] and to lower resource quality, with additive effects lengthening the duration of the reproductive cycle. Effects at the molecular level were investigated by measuring expression of 12 genes involved in energy production, translation, or Ca or P homeostasis. Expression of ATP synthase beta (higher in HQ2), calcified cuticle protein (higher in HQ1 and HQ2), and tropomyosin (higher in HQ2 compared to HQ1) was significantly affected by resource quality, and significant additive effects on Ca transporting ATPase expression were induced by [Ca] and resource quality (higher for low [Ca] and higher resource quality). These results highlight the potential drastic deleterious effects of water [Ca] depletion on G. fossarum physiology, populations, and ecosystem functioning, especially in oligotrophic environments.

The risk assessment of pesticides on soil fauna is an issue to protect agroecosystem sustainability. Enchytraeids are recognized as relevant soil bioindicators of chemical stress in agroecosystems. In laboratory, the reproduction test was found to be sensitive to reveal chemical impacts on enchytraeids. However, it does not allow to assess the impacts on ecological functions in which enchytraeids are involved. The objectives of this study were (i) to explore the feasibility of the bait-lamina test with enchytraeids under laboratory conditions and (ii) to compare its sensitivity with the Enchytraeid Reproduction Test. We exposed individuals of Enchytraeus albidus to two commercial formulations of fungicides widely used in Europe. The EC50 reproduction for the Swing® Gold (50 g L−1 epoxiconazole and 133 g L−1 dimoxystrobin) and the Cuprafor micro® (50% copper oxychloride) were respectively estimated at 1.66 ± 0.93 times the recommended dose and > 496 mg kg−1 of copper. However, no impact was found on the feeding activity of enchytraeids. The bait-lamina test thus appeared less sensitive than the Enchytraeid Reproduction Test to the tested fungicides. Despite that, this test which is achievable under laboratory conditions and allows assessing indirect effects of pesticides is quick, cheap, and easy to perform. It would deserve to be used to explore longer-exposure effects through the repeated addition of bait-lamina sticks.

The risk assessment of pesticides on soil fauna is an issue to protect agroecosystem sustainability. Enchytraeids are recognized as relevant soil bioindicators of chemical stress in agroecosystems. In laboratory, the reproduction test was found to be sensitive to reveal chemical impacts on enchytraeids. However, it does not allow to assess the impacts on ecological functions in which enchytraeids are involved. The objectives of this study were (i) to explore the feasibility of the bait-lamina test with enchytraeids under laboratory conditions and (ii) to compare its sensitivity with the Enchytraeid Reproduction Test. We exposed individuals of Enchytraeus albidus to two commercial formulations of fungicides widely used in Europe. The EC50 reproduction for the Swing® Gold (50 g L⁻¹ epoxiconazole and 133 g L⁻¹ dimoxystrobin) and the Cuprafor micro® (50% copper oxychloride) were respectively estimated at 1.66 ± 0.93 times the recommended dose and > 496 mg kg⁻¹ of copper. However, no impact was found on the feeding activity of enchytraeids. The bait-lamina test thus appeared less sensitive than the Enchytraeid Reproduction Test to the tested fungicides. Despite that, this test which is achievable under laboratory conditions and allows assessing indirect effects of pesticides is quick, cheap, and easy to perform. It would deserve to be used to explore longer-exposure effects through the repeated addition of bait-lamina sticks.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [ADD1_IRSTEA]Dynamique et fonctionnement des écosystèmes | International audience | As part of the landscape, streams are influenced byland use. Here, we contributed to the understanding of the biological impacts of land use on streams, investigating how landscape effects vary with spatial scales (local vs. regional). We adopted a food web approach integrating both biological structure and functioning, to focus on the overall effect of land use on stream biocoenosis. We selected 17 sites of a small tributary of the Seine River (France) for their contrasted land use, and conducted a natural experiment by sampling three organic matter sources, three macroinvertebrate taxa, and most of the fish community. Using stable isotope analysis, we calculated three food web metrics evaluating two major dimensions of the trophic diversity displayed by the fish community: (i) the diversity of exploited resources and (ii) the trophic level richness. The idea was to examine whether (1) land-use effects varied according to spatial scales, (2) land use affected food webs through an effect on community structure and (3) land use affected food webs through an effect onavailable resources. Beside an increase in trophic diversity from upstream to downstream, our empirical data showed that food webs were influenced by land use in the riparian corridors (local scale). The effect was complex, and depended on site's position along the upstream-downstream gradient. By contrast, land use in the catchment (regional scale) did not influence stream biocoenosis. At the local scale, community structure was weakly influenced by land use, and thus played a minor role in explaining food web modifications. Our results suggested that the amount of available resources at the base of the food web was partly responsible for food web modifications. In addition, changes in biological functioning (i.e. feeding interactions) can also explain another part of the land-use effect. These results highlight the role played by the riparian corridors as a buffer zone, and advocate that riparian corridor should be at the centre of water management attention.

As part of the landscape, streams are influenced by land use. Here, we contributed to the understanding of the biological impacts of land use on streams, investigating how landscape effects vary with spatial scales (local vs. regional). We adopted a food web approach integrating both biological structure and functioning, to focus on the overall effect of land use on stream biocœnosis. We selected 17 sites of a small tributary of the Seine River (France) for their contrasted land use, and conducted a natural experiment by sampling three organic matter sources, three macroinvertebrate taxa, and most of the fish community. Using stable isotope analysis, we calculated three food web metrics evaluating two major dimensions of the trophic diversity displayed by the fish community: (i) the diversity of exploited resources and (ii) the trophic level richness. The idea was to examine whether (1) land-use effects varied according to spatial scales, (2) land use affected food webs through an effect on community structure and (3) land use affected food webs through an effect on available resources. Beside an increase in trophic diversity from upstream to downstream, our empirical data showed that food webs were influenced by land use in the riparian corridors (local scale). The effect was complex, and depended on site’s position along the upstream-downstream gradient. By contrast, land use in the catchment (regional scale) did not influence stream biocœnosis. At the local scale, community structure was weakly influenced by land use, and thus played a minor role in explaining food web modifications. Our results suggested that the amount of available resources at the base of the food web was partly responsible for food web modifications. In addition, changes in biological functioning (i.e. feeding interactions) can also explain another part of the land-use effect. These results highlight the role played by the riparian corridors as a buffer zone, and advocate that riparian corridor should be at the centre of water management attention.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]BELCA [TR2_IRSTEA]ARCEAU | International audience | extile dyes pose a significant challenge for water pollution due to the poor degradability of their complex aromatic structures (e.g., RR-120 and RBB-150). In order to minimize the harmful effects of RR-120 and RBB-150, the capacity of MgAl-layered double hydroxide for removing of these contaminants was studied herein. Batch adsorption experiments were conducted to investigate the effect of various operating parameters, such as solution pH, contact time, dye concentration, and temperature in order to provide optimal conditions for removal. Structural and morphological analyses were used to highlight the assembly and/or interaction LDH-dye. The state of equilibrium of RR-120 and RBB-150 adsorption was pH- and temperature-dependent and followed the pseudo-second-order rate model. Also, the equilibrium adsorption data of both dyes were found to adopt the Langmuir type isotherm model, which assumes a monolayer arrangement in LDH-dye. Furthermore, the effects of four major coexisting and competing mono- and divalent interlayer anions, such as NO3-, Cl-, CO32-, and SO42-, on the uptakes of RR-120 and RBB-150 were studied and the results showed that NO3- anions had insignificant effect on the uptakes of RR-120 and RBB-150 by MgAl. An equivalent study on the presence of both dyes in competitive trial adsorption/desorption from binary aqueous solution was investigated. And finally, the reuse operation of recovered material after dye adsorption was tested in up to 5 cycles of recyclability.

Textile dyes pose a significant challenge for water pollution due to the poor degradability of their complex aromatic structures (e.g., RR-120 and RBB-150). In order to minimize the harmful effects of RR-120 and RBB-150, the capacity of MgAl-layered double hydroxide for removing of these contaminants was studied herein. Batch adsorption experiments were conducted to investigate the effect of various operating parameters, such as solution pH, contact time, dye concentration, and temperature in order to provide optimal conditions for removal. Structural and morphological analyses were used to highlight the assembly and/or interaction LDH-dye. The state of equilibrium of RR-120 and RBB-150 adsorption was pH- and temperature-dependent and followed the pseudo-second-order rate model. Also, the equilibrium adsorption data of both dyes were found to adopt the Langmuir type isotherm model, which assumes a monolayer arrangement in LDH-dye. Furthermore, the effects of four major coexisting and competing mono- and divalent interlayer anions, such as NO₃⁻, Cl⁻, CO₃²⁻, and SO₄²⁻, on the uptakes of RR-120 and RBB-150 were studied and the results showed that NO₃⁻ anions had insignificant effect on the uptakes of RR-120 and RBB-150 by MgAl. An equivalent study on the presence of both dyes in competitive trial adsorption/desorption from binary aqueous solution was investigated. And finally, the reuse operation of recovered material after dye adsorption was tested in up to 5 cycles of recyclability.