Search

International audience; Nitrogen is an important element that controls plant growth and yield. Recently we showed that protein farnesylation is involved in nitrate-driven root architecture organization in Arabidopsis. In this study, we aimed to deeper investigate the role of this post-translational maturation in plant responses to nitrogen, by studying the metabolic and phenotypic adaptations of the era1.8 mutant (enhanced response to abscisic acid 1; i.e. lacking protein farnesylation activity) to various NH4NO3 supplies. WT and era1.8 plants were grown on low-nitrogen substrate supplemented with 0, 2 or 10 mM NH4NO3, and were analyzed for biomass production and carbonand nitrogen-related metabolites. Our results showed that compared to WT, era1.8 developed exacerbated responses to N starvation, i.e. a higher root/shoot ratio, a higher anthocyanins content and a higher carbon/nitrogen ratio. Moreover, transcriptomic analyzes revealed an over-expression of several genes related to nitrogen starvation adaptation in era1.8 and pointed out two genes encoding uncharacterized farnesylated proteins. At 2 and 10 mM NH4NO3, although the era1.8 mutant displayed a higher biomass, its metabolism and gene expression are comparable to that of WT. Overall, our results show that era1.8 develops exacerbated responses to nitrogen starvation with specific metabolic balances, highlighting that protein farnesylation participates to plant adaptation to nitrogen nutritional stress.

International audience; Biodiversity citizen science projects are both valued for their contribution to scientific research and for their impact on participants' science learning and engagement towards the environment. In this paper, we assess the impact of participation in a biodiversity citizen science project (the Spipoll, dedicated to pollinators' monitoring) through the analysis of online interactions within the program's data sharing platform. By drawing on a previous qualitative analysis of the comments exchanged by the participants within this platform, we focus on those comments which share items on aspects of biology and ecology related to the Spipoll program. This sample gathers 2279 comments from 2010 to 2018. We first classified the different constitutive elements from these comments into seven categories following the topics they deal with. We then studied the temporal change in occurrence of each of these topics from 2010 to 2018. We show that long-term participation is associated with the growing expression of scientific procedural skills: formulation of hypothesis and explanation, proposition of new research questions. To our knowledge, our study is the first one that detects the acquisition of such procedural skills in biodiversity citizen science. We also show that long-term participation is associated with the growing attention to natural seasonal cycles. This study finally illustrates the value of the online traces of citizen scientists' activities to analyze participants' outcomes of citizen science. Consequently, it should encourage the development of such online communication spaces within contributory projects, without restricting them to online citizen science.

International audience; Tryptophan at concentrations higher than 0.1 mM, triggered characteristic early physiological effects such as rapid (within 5 min) dose-dependent membrane hyperpolarization in Mimosa pudica motor cells and modification of the time course of the spontaneous proton efflux monitored in the incubation medium of pulvinar tissues. The rapid modifications of the leaf turgor-mediated movements seen on the primary pulvini of M. pudica following a shock and on Cassia fasciculata leaflets during a transition from light to darkness indicate that tryptophan disturbed the ionic migrations involved in the electrophysiological events and in the osmocontractile reaction of the motor cells. These reactions were specific to tryptophan compared to those induced by serine and 5-hydroxytryptophan. The tryptophan mode of action cannot be linked to a direct modification of the plasma membrane H+-ATPase activity as monitored on purified pulvinar plasma membrane vesicles. The tryptophan metabolism-linked products tryptamine and indole also inhibited the motile reactions, activated in a continuous manner the H+ secretion of pulvinar tissues and showed properties of a protonophore and an ATPase activity inhibitor on plasma membrane vesicles, respectively. The specific behavior of tryptophan in the reaction studies here is discussed in light of the previously reported action of phytohormones.

International audience; We report the discovery of a new enveloped, spherical virus belonging to the Phenuiviridae family of negative ssRNA viruses associated with a massive outbreak in a French population of the endangered white-clawed crayfish Austropotamobius pallipes. We call this virus Bunya-like Brown Spot Virus (BBSV) and characterize it using transmission electronic microscopy, genome sequencing and clinical signs. Infected specimens show discolored brown spots on the cuticle. Using RNA-seq data we assembled a partial sequence for the L, M and S genome segments of BBSV. Phylogenetic analyses using all three segments show this virus is closely related to the Wenling crustacean virus 7 (China) and to a bunya-like virus found in feces of the European otter. Our survey of the mass mortality event indicates the virus is less virulent than the crayfish plague caused by Aphanomyces astaci. Overall, the discovery of BBSV provides a new important asset to monitor A. pallipes populations.

Bacteria of the order Legionellales, such as Legionella pneumophila, the agent of Legionnaires' disease, and Coxiella burnetii, the agent of Q fever, are widely recognized as human pathogens. While our view of the Legionellales is often limited to clinical isolates, ecological surveys are continually uncovering new members of the Legionellales that do not fall into the recognized pathogenic species. Here we emphasize that most of these Legionellales are nonpathogenic forms that have evolved symbiotic lifestyles with nonvertebrate hosts. The diversity of nonpathogenic forms remains, however, largely underexplored. We conjecture that its characterization, once contrasted with the data on pathogenic species, will reveal novel highlights on the mechanisms underlying lifestyle transitions of intracellular bacteria, including the emergence of pathogenesis and mutualism, transmission routes, and host specificity.

International audience; Maintaining the equilibrium between saturated and unsaturated fatty acids within membrane phospholipids (PLs) is crucial to sustain the optimal membrane biophysical properties, compatible with selective organellebased processes. Lipointoxication is a pathological condition under which saturated PLs tend to accumulate within the cell at the expense of unsaturated species, with major impacts on organelle function. Here, we show that human bronchial epithelial cells extracted from lungs of patients with Obstructive Pulmonary Diseases (OPDs), i. e. Cystic Fibrosis (CF) individuals and Smokers, display a characteristic lipointoxication signature, with excessive amounts of saturated PLs. Reconstitution of this signature in cellulo and in silico revealed that such an imbalance results in altered membrane properties and in a dramatic disorganization of the intracellular network of bronchial epithelial cells, in a process which can account for several OPD traits. Such features include Endoplasmic Reticulum-stress, constitutive IL8 secretion, bronchoconstriction and, ultimately, epithelial cell death by apoptosis. We also demonstrate that a recently-identified lipid-like molecule, which has been shown to behave as a "membrane-reshaper", counters all the lipointoxication hallmarks tested. Altogether, these insights highlight the modulation of membrane properties as a potential new strategy to heal and prevent highly detrimental symptoms associated with OPDs.

International audience; Fungal communities associated to three epiphytic lichens active against Candida, were investigated using culture-based methods We hypothetized that associated fungi would contribute to lichens activities. The ability of specific fungi to grow inside or outside lichens was investigated. To detect biogenesis pathways involved in the production of secondary metabolites, genes coding for nonribosomal peptide synthetase (NRPS) and polyketide synthase I (PKS I) were screened by PCR from fungal DNA extracts. Both endo and epilichenic communities were isolated from two fructicose (Evernia prunastri and Ramalina fastigiata) and one foliose (Pleurosticta acetabulum) lichens. A total of 86 endolichenic and 114 epilichenic isolates were obtained, corresponding to 18 and 24 phylogenetic groups respectively suggesting a wide diversity of fungi. The communities and the species richness were distinct between the three lichens which hosted potentially new fungal species. Additionally, the endo- and epilichenic communities differed in their composition: Sordariomycetes were particularly abundant among endolichenic fungi and Dothideomycetes among epilichenic fungi. Only a few fungi colonized both habitats, such as S. fimicola, Cladosporium sp1 and Botrytis cinerea. Interestingly, Nemania serpens (with several genotypes) was the most abundant endolichenic fungus (53% of isolates) and was shared by the three lichens. Finally, 12 out of 36 phylogenetic groups revealed the presence of genes coding for nonribosomal peptide synthetase (NRPs) and polyketide synthase I (PKS I). This study shows that common lichens are reservoirs of diverse fungal communities, which could potentially contribute to global activity of the lichen and, therefore, deserve to be isolated for further chemical studies.

International audience; Acanthamoeba castellanii is a free-living amoeba commonly found in aquatic environment. It feeds on bacteria even if some bacteria resist amoebal digestion. Thus, A. castellanii is described as a Trojan horse able to harbor pathogenic bacteria. L. pneumophila is one of the amoeba-resisting bacteria able to avoid host degradation by phagocytosis and to multiply inside the amoeba. When infecting its host, L. pneumophila injects hundreds of effectors via a type IV secretion system that change physiology of the amoeba to its profit. In this study, we assess mobility of A. castellanii upon infection with L. pneumophila. Electron-microscopy analysis of amoebae revealed a reduction of acanthopodia on cells infected with L. pneumophila. Analysis of velocity showed that migration of A. castellanii infected with L. pneumophila was significantly impaired compare to uninfected cells. Taken together, infection with L. pneumophila could prevent formation of cytoplasmic extensions such as acanthopodia with consequences on the shape, adherence and mobility of A. castellanii.

International audience; Klebsiella pneumoniae is a bacterium that can be in relation with free living amoebae like Acanthamoeba castellanii in natural environments such as soil and water. This pathogen, which is responsible for community-acquired pneumonia and for nosocomial infections, also has interactions with host defense mechanisms like macrophages. As it has been shown that A. castellanii shares some traits with macrophages, in particular the ability to phagocyte bacteria, we have studied the uptake and the fate of the bacteria after contact with the two phagocytic cells. In our conditions, K. pneumoniae growth was increased in coculture in presence of A. castellanii or Thp-1 macrophagic cells and bacterial development was also increased by A. castellanii supernatant. In addition, we showed that the presence of the bacteria had a negative effect on the macrophages whereas it does not affect amoeba viability. Using gentamicin, which kills bacteria outside cells, we showed that only macrophages were able to internalize K. pneumoniae. This result was confirmed by electron microscopy. We have consequently reported some differences in bacterial uptake and internalization between a free living amoeba and macrophagic cells, highlighting the fact that results obtained with this amoebal model should not be extrapolated to the relationships between K. pneumoniae and macrophages.

Stressors are omnipresent in the environment and trigger specific physiological and/or behavioural responses that remain relatively less known in invertebrates. Among the parameters used to quantify the impact of stress on physiology, the circulating level of glucose is commonly used in aquatic crustacean species. Terrestrial isopods are small crustaceans commonly used as bioindicators of soil quality; however, their response to environmental stressors remains poorly documented. In laboratory conditions, we investigated how different stressors influence the physiological response of two terrestrial isopod species. Thus, we exposed Armadillidium vulgare and Porcellio dilatatus to a series of both acute (physical) and chronic (temperature, social isolation, and exposure to herbicide) stressors that mimic potentially stressful situations encountered in their environment. Our results revealed that, for both species, glycaemia is not impacted by thermic shock (1 h at 4 °C), mechanical disturbances (1 min of shaking or upside-down motion) or social isolation (4 weeks) at the population level. Nevertheless, we observed a significantly higher sensitivity in male P. dilatatus exposed to upside-down movements and in males of both species exposed to social isolation. Acute (96 h) and chronic (21 days) exposure to glyphosate (1.8–2.5 kg.ha−1) did not impact glycaemia levels in A. vulgare. However, at both tested doses, individuals exposed to glyphosate for 25 days and subjected to an additional stressor (haemolymph sampling at 96 h) had significantly higher glycaemia in comparison to their control. Survival was also impacted by the joint application of chronic glyphosate exposure and intermediate haemolymph sampling. We conclude that glucose is a potentially good indicator to study the stress response of terrestrial isopods. Additional studies for this parameter, combined with other physiological indicators, are needed to better characterize this response.

International audience; Autophagy is a eukaryotic process responsible for the degradation of intracellular content such as damaged organelles. Several putative autophagy-related genes have been identified within the annotated genome of the free-living amoeba Acanthamoeba castellanii. However, the involvement of the corresponding proteins in the autophagy pathway had not been formerly established. Here, we report that AcAtg8 cDNA can complement ATG8-deficient Saccharomyces cerevisiae.

In the last decades, dispersal studies have benefited from the use of molecular markers for detecting patterns differing between categories of individuals and have highlighted sex-biased dispersal in several species. To explain this phenomenon, several hypotheses implying mating systems, intrasexual competition or sex-related handicaps have been proposed. In this context, we investigated sex-biased dispersal in Armadillidium vulgare, a terrestrial isopod with a promiscuous mating system. As a proxy for effective dispersal, we performed a fine-scale investigation of the spatial genetic structure in males and females, using individuals originating from five sampling points located within 70 meters of each other. Based on microsatellite markers and spatial autocorrelation analyses, our results revealed that while males did not present a significant genetic structure at this geographic scale, females were significantly and genetically more similar to each other when they were collected in the same sampling point. As females invest more parental care than males in A. vulgare, but also because this species is promiscuous and males experience a high intrasexual competition, our results meet the predictions of most classical hypotheses for sex-biased dispersal. We suggest that widening dispersal studies to other isopods or crustaceans, differing in their ecology or mating system and displaying varying levels of parental care, might shed light on the processes underlying the evolution of sex-biased dispersal., 23 pages (including 2 figures and one table) and two supplementary files containing 5 pages with 3 tables S1 to S3 and one figure S1, Last two authors have contributed equally to this study

International audience; Transposable elements (TEs) are DNA pieces that are present in almost all the living world at variable genomic density. Due to their mobility and density, TEs are involved in a large array of genomic modifications. In eukaryotes, TE expression has been studied in detail in several species. In prokaryotes, studies of IS expression are generally linked to particular copies that induce a modification of neighboring gene expression. Here we investigated global patterns of IS transcription in the Alphaproteobacterial endosymbiont Wolbachia wVulC, using both RT-PCR and bioinformatic analyses. We detected several transcriptional promoters in all IS groups. Nevertheless, only one of the potentially functional IS groups possesses a promoter located upstream of the transposase gene, that could lead up to the production of a functional protein. We found that the majority of IS groups are expressed whatever their functional status. RT-PCR analyses indicate that the transcription of two IS groups lacking internal promoters upstream of the transposase start codon may be driven by the genomic environment. We confirmed this observation with the transcription analysis of individual copies of one IS group. These results suggest that the genomic environment is important for IS expression and it could explain, at least partly, copy number variability of the various IS groups present in the wVulC genome and, more generally, in bacterial genomes.

International audience; Biofilms cause chronic infections in tissues or by developing on the surfaces of medical devices. Biofilm infections persist despite both antibiotic therapy and the innate and adaptive defence mechanisms of the patient. Biofilm infections are characterized by persisting and progressive pathology due primarily to the inflammatory response surrounding the biofilm. For this reason, many biofilm infections may be difficult to diagnose and treat efficiently. It is the purpose of the guideline to bring the current knowledge of biofilm diagnosis and therapy to the attention of clinical microbiologists and infectious disease specialists. Selected hallmark biofilm infections in tissues (e.g. cystic fibrosis with chronic lung infection, patients with chronic wound infections) or associated with devices (e.g. orthopaedic alloplastic devices, endotracheal tubes, intravenous catheters, indwelling urinary catheters, tissue fillers) are the main focus of the guideline, but experience gained from the biofilm infections included in the guideline may inspire similar work in other biofilm infections. The clinical and laboratory parameters for diagnosing biofilm infections are outlined based on the patient's history, signs and symptoms, microscopic findings, culture-based or culture-independent diagnostic techniques and specific immune responses to identify microorganisms known to cause biofilm infections. First, recommendations are given for the collection of appropriate clinical samples, for reliable methods to specifically detect biofilms, for the evaluation of antibody responses to biofilms, for antibiotic susceptibility testing and for improvement of laboratory reports of biofilm findings in the clinical microbiology laboratory. Second, recommendations are given for the prevention and treatment of biofilm infections and for monitoring treatment effectiveness. Finally, suggestions for future research are given to improve diagnosis and treatment of biofilm infections.

A study of the structure-activity relationship carried out on several benzoic acid-related phenolics indicates that this type of compounds hinders the osmocontractile reaction of pulvinar cells in the range of 0-100%. Tentatively, we tried to find a way that could explain this differential action. With this aim, the relationship between the inhibitory effect and important molecular physico-chemical parameters (namely lipophilicity and degree of dissociation) was drawn. In addition, the effect of a variety of these compounds was investigated on their capacity to modify the electrical transmembrane potential and induce modifications in proton fluxes. Finally, using plasma membrane vesicles purified from pulvinar tissues, we examined the effects of some selected compounds on the proton pump activity and catalytic activity of the plasma membrane H(+)-ATPase. Taken together, the results indicate that a modification of the molecular structure of phenolics may induce important variation in the activity of the compound on these early membrane events. Among the tested phenolics, salicylic acid (SA) and acetylsalicylic acid (ASA, aspirin) are of particuler note, as they showed atypical effects on the physiological processes studied.

International audience; The Wolbachia are symbiotic bacteria vertically transmitted from one host generation to another. However, a growing amount of data shows that horizontal transfers of Wolbachia also frequently occur within and between host species. The consequences of the arrival of new symbionts on host physiology can be studied by their experimental introduction in asymbiotic hosts. After experimental transfers of the eight major isopod Wolbachia strains in the isopod Porcellio dilatatus only two of them (wCon and wDil) were found to (1) have no pathogenic effect on the host and (2) be able to pass vertically to the host offspring. In the present work, we studied the influence of these two strains, able to complete an horizontal transfer, on immunity and reproduction of P. dilatatus at two stages of the transfer: (1) in recipient hosts that encounter the symbionts: to test the influence of symbiont when acquired during host life and (2) in vertically infected offspring: to test the influence of a symbiotic interaction occurring all lifelong. The impact of Wolbachia varied depending on the stage: there were clearer effects in vertically infected individuals than in those that acquired the symbionts during their lives. Moreover, the two Wolbachia strains showed contrasted effects: the strain wCon tended to reduce the reproductive investment but to maintain or increase immune parameters whilst wDil had positive effects on reproductive investment but decreased the investment in some immune parameters. These results suggest that horizontally acquisition of Wolbachia can influence the balance between host immune and reproductive traits.

International audience; Seeds of Lactuca sativa L. 'Romaine' were subjected to priming treatments with UV-C radiation at 0.85 or 3.42 kJ m À2. Seedlings obtained from both primed (Pr) and non-primed (NPr) seeds were grown in an hydroponic culture system supplemented with 0 (control) or 100 mM NaCl. After 21 days of NaCl treatment, root and leaf biomass, root lengths, leaf numbers, and leaf surface area were measured. Ions (Na þ and K þ) accumulation was determined in roots and leaves. Total phenolic compound and flavonoid concentrations, as well as antioxidant and antiradical activities were measured in L. sativa leaves. Salt stress resulted in a lower increase in fresh weight of roots and leaves, which was more pronounced in roots than in leaves, due to reduced root elongation, leaf number and leaf expansion, as well as leaf thickness. The lower increase in fresh weight was accompanied by a restriction in tissue hydration and K þ ion uptake, as well as an increase in Na þ ion concentrations in all organs. These effects were mitigated in plants from the UV-C primed seeds. The mitigating effect of UV-C was more pronounced at 0.85 than at 3.42 kJ m À2. Salt stress also resulted in an increase in total phenolic compounds and flavonoid concentrations and in the total antioxidant capacity in leaves. The highest diphenylpicrylhydrazyl radical (DPPH) scavenging activity was found in the leaves of plants from both Pr seeds. Our results suggest that plants grown from seed primed by exposure to moderate UV-C radiation exhibited a higher tolerance to salinity stress.

International audience; Aphanomyces astaci is an invasive pathogenic oomycete responsible for the crayfish plague, a disease that has devastated European freshwater crayfish. So far, five genotype groups of this pathogen have been identified by applying random amplified polymorphic DNA analysis on axenic cultures. To allow genotyping of A. astaci in host tissue samples, we have developed co-dominant microsatellite markers for this pathogen, tested them on pure cultures of all genotype groups, and subsequently evaluated their use on tissues of (1) natural A. astaci carriers, i.e., North American crayfish species, and (2) A. astaci-infected indigenous European species from crayfish plague outbreaks. Out of over 200 potential loci containing simple sequence repeat (SSR) motifs identified by 454 pyrosequencing of SSR-enriched library, we tested 25 loci with highest number of repeats, and finally selected nine that allow unambiguous separation of all known RAPD-defined genotype groups of A. astaci from axenic cultures. Using these markers, we were able to characterize A. astaci strains from DNA isolates from infected crayfish tissues when crayfish had a moderate to high agent level according to quantitative PCR analyses. The results support the hypothesis that different North American crayfish hosts carry different genotype groups of the pathogen, and confirm that multiple genotype groups, including the one originally introduced to Europe in the 19th century, cause crayfish plague outbreaks in Central Europe. So far undocumented A. astaci genotype seems to have caused one of the analysed outbreaks from the Czech Republic. The newly developed culture-independent approach allowing direct genotyping of this pathogen in both axenic cultures and mixed genome samples opens new possibilities in studies of crayfish plague pathogen distribution, diversity and epidemiology.

International audience; Sexual selection predicts that mate choice increases individual fitness. Infection by parasites (from eukaryotes to bacteria or viruses) can reduce this individual fitness, altering the infected individuals' sexual traits and molecular cues. In this case, one would expect to observe mechanisms for avoiding infection during mate choice. The vast majority of host responses to infection in terms of mate choice are intended to avoid infection, but the costs of mate choice can also hinder infection avoidance. This paper highlights the main limitations in current knowledge and empirical experiments, and summarizes the key factors that should be taken into account to test the hypothesis of infection avoidance in mate choice: the time of host-parasite coevolution in the biological interaction implied, the choosy sex tested (male, female or both) and the genetic background of the individuals tested.

International audience; : In crustaceans, the androgenic gland (AG), thanks to the synthesis of the androgenic gland hormone (AGH), controls the differentiation of the primary and secondary male sexual characters. In this study, we amplified 12 new AGH cDNAs in species belonging to five different families of the infra-order Ligiamorpha of terrestrial isopods. Putative essential amino acids for the production of a functional AGH protein exhibit signatures of negative selection and are strictly conserved including typical proteolytic cleavage motifs, a putative N-linked glycosylation motif on the A chains and the eight Cys positions. An insulin-like growth factor motif was also identified in Armadillidium AGH sequences. The phylogenetic relationships of AGH sequences allowed to distinguish two main clades, corresponding to members of the Armadillidiidae and the Porcellionidae families which is congruent with the narrow specificity of AG heterospecific grafting. An in-depth understanding of the regulation of AGH expression would help deciphering the interaction between Wolbachia, a widespread feminizing endosymbiotic bacteria in isopods, and the sex differentiation of their hosts.

International audience; : Free-living amoebae are naturally present in water. These protozoa could be pathogenic and could also shelter pathogenic bacteria. Thus, they are described as a potential hazard for health. Also, free-living amoebae have been described to be resistant to biocides, especially under their cyst resistant form. There are several studies on amoeba treatments but none of them compare sensitivity of trophozoites and cysts from different genus to various water disinfectants. In our study, we tested chlorine, monochloramine and chlorine dioxide on both cysts and trophozoites from three strains, belonging to the three main genera of free-living amoebae. The results show that, comparing cysts to trophozoites inactivation, only the Acanthamoeba cysts were highly more resistant to treatment than trophozoites. Comparison of the disinfectant efficiency led to conclude that chlorine dioxide was the most efficient treatment in our conditions and was particularly efficient against cysts. In conclusion, our results would help to adapt water treatments in order to target free-living amoebae in water networks.

International audience; In this paper, the salicylic acid (o-hydroxy benzoic acid) (SA) uptake by the pulvinar tissues of Mimosa pudica L. pulvini was shown to be strongly pH-dependent, increasing with acidity of the assay medium. This uptake was performed according to a unique affinity system (K(m) = 5.9 mM, V(m) = 526 pmol mgDW(-1)) in the concentration range of 0.1-5 mM. The uptake rate increased with increasing temperature (5-35 °C) and was inhibited following treatment with sodium azide (NaN3) and carbonyl cyanide m-chlorophenylhydrazone (CCCP), suggesting the involvement of an active component. Treatment with p-chloromercuribenzenesulfonic acid (PCMBS) did not modify the uptake, indicating that external thiol groups were not necessary. KCl, which induced membrane depolarization had no significant effect, and fusicoccin (FC), which hyperpolarized cell membrane, stimulated the uptake, suggesting that the pH component of the proton motive force was likely a driving force. These data suggest that the SA uptake by the pulvinar tissues may be driven by two components: an ion-trap mechanism playing a pivotal role and a putative carrier-mediated mechanism. Unlike other benzoic acid derivatives acting as classical respiration inhibitors (NaN3 and KCN), SA modified the pulvinar cell metabolism by increasing the respiration rate similar to CCCP and 2,4-dinitrophenol (DNP). Furthermore, SA inhibited the osmoregulated seismonastic reaction in a pH dependent manner and induced characteristic damage to the ultrastructural features of the pulvinar motor cells, particularly at the mitochondrial level.

International audience; Free-living amoebae (FLA) are protozoa that can be found in water networks where they prey on bacteria within biofilms. Most bacteria are digested rapidly by phagocytosis, however some are able to survive within amoebae and some are even able to multiply, as it is the case for Legionella pneumophila. These resisting bacteria are a potential health problem as they could also resist to macrophage phagocytosis. Several publications already reported intra-amoebal bacteria but the methods of identification did not allow metagenomic analysis and are partly based on co-culture with one selected amoebal strain. The aim of our study was to conduct a rRNA-targeted metagenomic analysis on amoebae and intra-amoebal bacteria found in drinking water network, to provide the first FLA microbiome in environmental strains. Three sites of a water network were sampled during four months. Culturable FLA were isolated and total DNA was prepared, allowing purification of both amoebal and bacterial DNA. Metagenomic studies were then conducted through 18S or 16S amplicons sequencing. Hartmannella was by far the most represented genus of FLA. Regarding intra-amoebal bacteria, 54 genera were identified, among which 21 were newly described intra-amoebal bacteria, underlying the power of our approach. There were high differences in bacterial diversity between the three sites. Several genera were highly represented and/or found at least in two sites, underlying that these bacteria could be able to multiply within FLA. Our method is therefore useful to identify FLA microbiome and could be applied to other networks to have a more comprehensive view of intra-amoebal diversity.

International audience; This field study aimed at determining the effect of metal-polluted environments on fluctuating asymmetry (FA) level of Porcellio scaber as a measure of developmental stability (DS). For this, woodlice and litter were collected on 8 sites of northern France located either away from metal pollution source or near metallurgical smelters. Physico-chemical parameters such as pH, C/N ratio and CaCO(3) were measured in litter since they could have potential effects on woodlouse population. Moreover, Cd, Pb, and Zn concentrations in litter were determined to evaluate the contamination degree of each site. The metal body burdens were determined in woodlice to evaluate their impregnation degree. Fluctuating asymmetry levels on males and gravid females were performed on 9 traits (the 2nd and the 3rd articles of antennae, the 1st flagellum segment of the antennae, and the merus, carpus and propodus of the 6th and the 7th pereiopods) according (1) to each site and (2) to two habitat types (cf. metal-contaminated and uncontaminated habitats). Results showed close correlations between litter metal concentrations and metal body burdens in woodlice. However, contrary to the initial hypothesis that metal pollution would induce an increase of the FA level, this seemed to be lower for individuals from metal-contaminated habitats than for those from uncontaminated habitats, and particularly for males. The role of antennae and pereiopods on the foraging behavior and reproduction processes on woodlice were discussed and could explain these observations. (C) 2012 Elsevier Ltd. All rights reserved.

International audience; Candida yeasts colonize humans' oral cavities as commensals or opportunistic pathogens. They may be isolated from water circulating in dental unit waterlines mixed with saliva traces mainly because of dysfunction of anti-retraction valves. Free-living amoebae (FLA), like Hartmannella vermiformis, are frequently found in aquatic environments and they have also been already isolated from dental unit waterlines. They can be implicated as reservoir for pathogens or directly in infections. This work deals with the survival of three species of Candida (Candida albicans, Candida glabrata and Candida parapsilosis), in co-cultivation with FLA in tap-water. One strain of each Candida species was studied. Microbiological and microscopic approaches were used; amoebae-yeasts co-cultivation assays were performed at different temperatures of incubation. Results have shown that H. vermiformis were able to internalize Candida yeasts and promote their proliferation in tap-water with or without saliva traces (2% v/v). Amoebae interact differently with Candida depending on the temperature used and the studied species of yeasts. In conclusion, this study emphasizes the survival of yeasts and/or FLA in tap-water. The ability of yeasts and amoebae to interact should be taken into account because it could lead to candidiasis infection for the frailest patients after a dental treatment.

International audience