Bacterivory by benthic organisms in sediment: quantification using 15N enriched bacteria

International audienceThe fate of benthic bacterial biomass in benthic food webs is a topic of major importance but poorly described. This paper describes an alternative method for evaluation of bacterial grazing rate by meiofauna and macrofauna using bacteria pre-enriched with stable isotopes. Natural bacteria from the sediment of an intertidal mudflat were cultured in a liquid medium enriched with 15NH4Cl. Cultured bacteria contained 2.9 % of 15N and were enriched sufficiently to be used as tracers during grazing experiments. Cultured-bacteria presented a biovolume (0.21 µm3) and a percentage of actively respiring bacteria (10 %) similar to those found in natural communities. The number of Operational Taxon Units found in cultures fluctuated between 56 and 75 % of that found in natural sediment. Despite this change in community composition, the bacterial consortium used for grazing experiments exhibited characteristics of size, activity and diversity more representative of the natural community than usually noticed in many other grazing studies. The bacterial ingestion rates of three different grazers were in the range of literature values resulting from other methods: 1149 ngC ind-1 h-1 for the mud snail Hydrobia ulvae, 0.027 ngC ind-1 h-1 for the nematode community, and 0.067 ngC ind-1 h-1 for the foraminifera Ammonia tepida. The alternative method described in this paper overcomes some past limitations and it presents interesting advantages such as short time incubation and in situ potential utilisation

Structures of benthic prokaryotic communities and their hydrolytic enzyme activities resuspended from samples of intertidal mudflats: An experimental approach

International audienceResuspended sediment can increase plankton biomass and the growth of bacteria, thus influencing the coastal planktonic microbial food web. But little is known about resuspension itself: is it a single massive change or a whole series of events and how does it affect the quantity and quality of resuspended prokaryotic cells? We simulated the sequential erosion of mud cores to better understand the fate and role of benthic prokaryotes resus-pended in the water column. We analyzed the total, attached and free-living prokaryotic cells resuspended, their structure and the activities of their hydrolytic enzymes in terms of the biotic and abiotic factors that affect the composition of microphytobenthic biofilm. Free living prokaryotes were resuspended during the fluff layer erosion phase (for shear velocities below 5 cm · s −1) regardless of the bed sediment composition. At the higher shear velocities, resuspended prokaryotes were attached to particulate matter. Free and attached cells are thus unevenly distributed, scattered throughout the organic matter (OM) in the uppermost mm of the sediment. Only 10–27% of the total cells initially resuspended were living and most of the Bacteria were Cyanobacteria and Gamma-proteobacteria; their numbers increased to over 30% in parallel with the hydrolytic enzyme activity at highest shear velocity. These conditions released prokaryotic cells having different functions that lie deep in the sediment; the most important of them are Archaea. Finally, composition of resuspended bacterial populations varied with resuspension intensity, and intense resus-pension events boosted the microbial dynamics and enzyme activities in the bottom layers of sea water

Interactive Effects of Pesticides and Nutrients on Microbial Communities Responsible of Litter Decomposition in Streams

Global contamination of streams by a large variety of compounds, such as nutrients and pesticides, may exert a high pressure on aquatic organisms, including microbial communities and their activity of organic matter decomposition. In this study, we assessed the potential interaction between nutrients and a fungicide and herbicide [tebuconazole (TBZ) and S-metolachlor (S-Met), respectively] at realistic environmental concentrations on the structure (biomass, diversity) and decomposition activity of fungal and bacterial communities (leaf decay rates, extracellular enzymatic activities) associated with Alnus glutinosa (Alnus) leaves. A 40-day microcosm experiment was used to combine two nutrient conditions (mesotrophic and eutrophic) with four pesticide treatments at a nominal concentrations of 15 μg L-1 (control, TBZ and S-Met, alone or mixed) following a 2 × 4 full factorial design. We also investigated resulting indirect effects on Gammarus fossarum feeding rates using leaves previously exposed to each of the treatments described above. Results showed interactive effects between nutrients and pesticides, only when nutrient (i.e., nitrogen and phosphorus) concentrations were the highest (eutrophic condition). Specifically, slight decreases in Alnus leaf decomposition rates were observed in channels exposed to TBZ (0.01119 days-1) and S-Met (0.01139 days-1) than in control ones (0.01334 days-1) that can partially be explained by changes in the structure of leaf-associated microbial communities. However, exposition to both TBZ and S-Met in mixture (MIX) led to comparable decay rates to those exposed to the pesticides alone (0.01048 days-1), suggesting no interaction between these two compounds on microbial decomposition. Moreover, stimulation in ligninolytic activities (laccase and phenol oxidase) was observed in presence of the fungicide, possibly highlighting detoxification mechanisms employed by microbes. Such stimulation was not observed for laccase activity exposed to the MIX, suggesting antagonistic interaction of these two compounds on the ability of microbial communities to cope with stress by xenobiotics. Besides, no effects of the treatments were observed on leaf palatability for macroinvertebrates. Overall, the present study highlights that complex interactions between nutrients and xenobiotics in streams and resulting from global change can negatively affect microbial communities associated with leaf litter, although effects on higher trophic-level organisms remains unclear

Isolation and characterization of mesotrione-degrading Bacillus sp. from soil

International audienceDissipation kinetics of mesotrione, a new triketone herbicide, sprayed on soil from Limagne (Puy-de-Dôme, France) showed that the soil microflora were able to biotransform it. Bacteria from this soil were cultured in mineral salt solution supplemented with mesotrione as sole source of carbon for the isolation of mesotrione-degrading bacteria. The bacterial community structure of the enrichment cultures was analyzed by temporal temperature gradient gel electrophoresis (TTGE). The TTGE fingerprints revealed that mesotrione had an impact on bacterial community structure only at its highest concentrations and showed mesotrione-sensitive and mesotrione-adapted strains. Two adapted strains, identified as Bacillus sp. and Arthrobacter sp., were isolated by colony hybridization methods. Biodegradation assays showed that only the Bacillus sp. strain was able to completely and rapidly biotransform mesotrione. Among several metabolites formed, 2-amino-4-methylsulfonylbenzoic acid (AMBA) accumulated in the medium. Although sulcotrione has a chemical structure closely resembling that of mesotrione, the isolates were unable to degrade i

Impact of Leptospermone, a Natural β-Triketone Herbicide, on the Fungal Composition and Diversity of Two Arable Soils

Impact of leptospermone, a β-triketone bioherbicide, was investigated on the fungal community which supports important soil ecological functions such as decomposition of organic matter and nutrients recycling. This study was done in a microcosm experiment using two French soils, Perpignan (P) and Saint-Jean-de-Fos (SJF), differing in their physicochemical properties and history treatment with synthetic β-triketones. Soil microcosms were treated with leptospermone at recommended dose and incubated under controlled conditions for 45 days. Untreated microcosms were used as control. Illumina MiSeq sequencing of the internal transcribed spacer region of the fungal rRNA revealed significant changes in fungal community structure and diversity in both soils. Xylariales, Hypocreales, Pleosporales and Capnodiales (Ascomycota phyla) fungi and those belonging to Sebacinales, Cantharellales, Agaricales, Polyporales, Filobasidiales and Tremellales orders (Basidiomycota phyla) were well represented in treated soil microcosms compared to control. Nevertheless, while for the treated SJF a complete recovery of the fungal community was observed at the end of the experiment, this was not the case for the P treated soil, although no more bioherbicide remained. Indeed, the relative abundance of most of the saprophytic fungi were lower in treated soil compared to control microcosms whereas fungi from parasitic fungi included in Spizellomycetales and Pezizales orders increased. To the best of our knowledge, this is the only study assessing the effect of the bioherbicide leptospermone on the composition and diversity of the fungal community in soil. This study showed that leptospermone has an impact on α- and β-diversity of the fungal community. It underlines the possible interest of microbial endpoints for environmental risk assessment of biopesticide

The conserved dileucine- and tyrosine-based motifs in MLV and MPMV envelope glycoproteins are both important to regulate a common Env intracellular trafficking

BACKGROUND: Retrovirus particles emerge from the assembly of two structural protein components, Gag that is translated as a soluble protein in the cytoplasm of the host cells, and Env, a type I transmembrane protein. Because both components are translated in different intracellular compartments, elucidating the mechanisms of retrovirus assembly thus requires the study of their intracellular trafficking. RESULTS: We used a CD25 (Tac) chimera-based approach to study the trafficking of Moloney murine leukemia virus and Mason-Pfizer monkey virus Env proteins. We found that the cytoplasmic tails (CTs) of both Env conserved two major signals that control a complex intracellular trafficking. A dileucine-based motif controls the sorting of the chimeras from the trans-Golgi network (TGN) toward endosomal compartments. Env proteins then follow a retrograde transport to the TGN due to the action of a tyrosine-based motif. Mutation of either motif induces the mis-localization of the chimeric proteins and both motifs are found to mediate interactions of the viral CTs with clathrin adaptors. CONCLUSION: This data reveals the unexpected complexity of the intracellular trafficking of retrovirus Env proteins that cycle between the TGN and endosomes. Given that Gag proteins hijack endosomal host proteins, our work suggests that the endosomal pathway may be used by retroviruses to ensure proper encountering of viral structural Gag and Env proteins in cells, an essential step of virus assembly

Maize herbicides mixtures effects on soil microbial communities (microcosms studies)

Ces travaux s inscrivent dans une problématique actuelle de compréhension des effets, encore peu connus, des mélanges de pesticides dans le sol. Les formulations d herbicides S-métolachlore (Dual Gold Safeneur®), mésotrione (Callisto®) et nicosulfuron (Milagro®) ont été appliquées en mélanges et à la dose agronomique recommandée. Les molécules formulées sont plus toxiques que les molécules actives seules (Microtox®) et aucun effet synergique ou antagoniste des mélanges n a été observé. Cependant, ces mélanges impactent transitoirement et à court terme les communautés microbiennes du sol de Limagne, sans modifier les paramètres généraux de structure et d abondance. Toutefois, des effets ont été mis en évidence sur des communautés sensibles telles que les phototrophes ou celles impliquées dans les processus de nitrification et de dénitrification. De plus, une comparaison des effets des mélanges sur les communautés phototrophes de deux sols souligne l importante toxicité du Dual Gold Safeneur® et remet en cause son utilisation d un point de vue écotoxicologique.These experiments are part of the actual problematic of understanding the poorknown effects of pesticides mixtures in soil ecosystem. The herbicidal commercial formulations of S-metolachlor (Dual Gold Safeneur®), mesotrione (Callisto®) and nicosulfuron (Milagro®) were applied in mixtures at the recommended field rate. Formulated compounds are more toxic than active ingredients alone (Microtox®) and no synergistic or antagonistic effects were recorded. However, these mixtures impaired rapidly and transitory the Limagne soil microbial communities, without modifying the global parameters of structure and abundance. Effects were also recorded on sensitive communities, such as phototrophic, nitrifying and denitrifying communities. A comparison of mixtures effects on phototrophic communities from two soils underlined the high toxicity of Dual Gold Safeneur® and challenged its use from an ecotoxicological point of view..CLERMONT FD-Bib.électronique (631139902) / SudocSudocFranceF

Assessing extracellular enzymatic activities of biofilms on natural and standardized organic substrata immersed in a stream

International audienc

Structural and functional responses of leaf-associated fungal communities to chemical pollution in streams

International audienceThe decomposition of allochthonous plant litter is a key ecological process in forested streams, involving the lignolytic activities of aquatic fungi. In this study we investigated the relationship between water pollution (by a mixture of nutrients, metals and pesticides) and the biomass, structure and enzyme activities of leaf‐associated fungal communities.The experiment was performed through a microcosm approach by comparing fungal communities from a less‐polluted upstream site to a more‐polluted downstream site in the agricultural basin of the Auzon stream (Puy‐de‐Dôme region, Centre France). The resistance as well as the ability of these fungal communities to recover from pollution was also assessed through a translocation experiment.Results showed a lower laccase activity and higher band richness in fungal communities from the downstream site comparing to the upstream site, which can partially be explained by the greater pesticide toxicity and concentration of dissolved organic carbon of downstream waters.Specifically, a negative relationship between laccase activity and nitrate concentration was observed, irrespectively of the studied site, whereas the relationship between nitrate and phenol oxidase/peroxidase appeared much weaker.The translocation experiment evidenced (1) a fast decrease in laccase activity concomitant with a shift in fungal structure after stress exposure, and (2) a recovery ability in terms of laccase activity and fungal community structure 2 weeks after stress removal.This study underlines the sensitivity of leaf‐associated fungal communities in terms of laccase activity and community structure to variations in chemical stress, as well as their ability to recover once the stress is removed. This study highlights the potential use of laccase as an indicator of chemical pollution of streams