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To better understand the potential risks of Nipah virus emergence in Cambodia by studying different components of the interface between humans and bats.From 2012 to 2016, we conducted a study at two sites in Kandal and Battambang provinces where fruit bats (Our results confirmed circulation of Nipah virus in fruit bats (28 of 3930 urine samples positive by polymerase chain reaction testing). We identified clear potential routes for virus transmission to humans through local practices, including fruit consumed by bats and harvested by humans when Nipah virus is circulating, and palm juice production. Nevertheless, in the serological survey of 418 potentially exposed people, none of them were seropositive to Nipah virus. Differences in agricultural practices among the regions where Nipah virus has emerged may explain the situation in Cambodia and point to actions to limit the risks of virus transmission to humans.Human practices are key to understanding transmission risks associated with emerging infectious diseases. Social science disciplines such as anthropology need to be integrated in health programmes targeting emerging infectious diseases. As bats are hosts of major zoonotic pathogens, such integrated studies would likely also help to reduce the risk of emergence of other bat-borne diseases.Mieux comprendre les risques potentiels d'émergence du virus Nipah au Cambodge en étudiant différents composants de l'interface entre humains et chauves-souris.De 2012 à 2016, nous avons réalisé une étude à deux endroits dans les provinces de Kandal et Battambang, où nichent les chauves-souris frugivores (Nos résultats ont confirmé la circulation du virus Nipah chez les chauves-souris frugivores (28 échantillons d'urine sur 3930 se sont révélés positifs lors des tests de réaction en chaîne par polymérase). Nous avons clairement identifié plusieurs vecteurs de transmission potentiels aux humains liés aux pratiques locales, dont les fruits consommés par les chauves-souris et récoltés par les humains lorsque le virus Nipah circule, ainsi que la production de jus de palme. Néanmoins, une enquête sérologique menée auprès de 418 personnes potentiellement exposées a montré qu'aucune d'entre elles n'était séropositive au virus Nipah. Les différences au niveau des pratiques agricoles dans les régions où le virus Nipah est apparu pourrait expliquer la situation au Cambodge, et conduire à des actions qui permettront de limiter les risques de transmission du virus aux humains.Les pratiques humaines sont la clé d'une meilleure compréhension des risques de transmission associés aux maladies infectieuses émergentes. Certaines disciplines de sciences sociales telles que l'anthropologie doivent être intégrées dans les programmes de santé qui ciblent les maladies infectieuses émergentes. Étant donné que les chauves-souris sont porteuses d'agents pathogènes zoonotiques majeurs, des études intégrées de ce type pourraient contribuer à réduire le risque d'émergence d'autres maladies propagées par les chauves-souris.Entender mejor los riesgos potenciales de la aparición del virus Nipah en Camboya mediante el estudio de los diversos componentes de la interfaz entre los seres humanos y los murciélagos.De 2012 a 2016, se realizó un estudio en dos lugares de las provincias de Kandal y Battambang en donde habitan los murciélagos fruteros (Los resultados confirmaron la circulación del virus Nipah en murciélagos fruteros (28 de 3930 muestras de orina positivas en la prueba de reacción en cadena de la polimerasa). Se identificaron las rutas claras de transmisión del virus a los seres humanos a través de las prácticas locales, incluidas las frutas que consumen los murciélagos y que los seres humanos recolectan cuando el virus Nipah está en circulación, y la producción de zumo de palma. Sin embargo, según el estudio serológico de 418 personas potencialmente expuestas, ninguna de ellas fue seropositiva al virus Nipah. Es posible que las diferencias en las prácticas agrícolas entre las regiones en donde el virus Nipah se ha presentado expliquen la situación en Camboya y señalen las medidas para limitar los riesgos de transmisión del virus a los seres humanos.Las prácticas de los seres humanos son esenciales para entender los riesgos de transmisión asociados a las enfermedades infecciosas emergentes. Las disciplinas de las ciencias sociales, como la antropología, se deben integrar en los programas de salud que se ocupan de esas enfermedades. Teniendo en cuenta que los murciélagos son huéspedes de los principales patógenos zoonóticos, es probable que esos estudios integrados también ayuden a reducir el riesgo de aparición de otras enfermedades que se transmiten por los murciélagos.الغرض تحقيق فهم أفضل للمخاطر المحتملة لظهور فيروس نيباه في كمبوديا عن طريق دراسة المكونات المختلفة للتفاعل بين البشر والخفافيش. الطريقة من عام 2012 حتى عام 2016، قمنا بإجراء دراسة في موقعين في مقاطعتي كاندال وباتامبانج حيث تستوطن خفافيش الفاكهة (旨在通过研究人与蝙蝠之间相互接触的不同部分，更好地了解柬埔寨境内的尼帕病毒潜在风险。.从 2012 年至 2016 年，我们在坎达尔和马德望省的两个地点进行了研究，这些地方都栖息着果蝠（我们的结果证实了尼帕病毒在果蝠中的传播（通过聚合酶链反应测试发现 3930 个尿液样本中有 28 个呈阳性）。我们通过当地实践确定了将病毒传播给人类的明确潜在途径，包括传播尼帕病毒时蝙蝠食用的和人类收获的果实，以及棕榈汁的生产。尽管如此，在对 418 名潜在接触者进行的血清学调查中，没有人对尼帕病毒呈血清反应阳性。已经出现尼帕病毒的地区中在农业实践方面的差异可能说明了柬埔寨的情况以及限制病毒传播给人类的风险的行动指向。.人类实践是了解与新发传染病相关的传播风险的关键。需要将人类学等社会科学学科纳入针对新发传染病的卫生方案。由于蝙蝠是主要的人畜共患病病原体的宿主，因此此类综合研究还可能有助于降低其他蝙蝠传播疾病出现的风险。.Добиться лучшего понимания потенциальных рисков появления вируса Нипах в Камбодже за счет изучения разного рода компонентов взаимодействия людей и летучих мышей.В период с 2012 по 2016 год авторы провели исследование в двух районах провинций Кандаль и Баттамбанг со значительными колониями крыланов (Было подтверждено, что в организмах крыланов циркулирует вирус Нипах (28 из 3930 образцов мочи были положительными по результатам тестирования методом полимеразной цепной реакции). Авторы выявили четкие потенциальные пути передачи вируса людям через принятые в этих местностях практикуемые виды деятельности, включая сбор людьми фруктов, которыми питаются крыланы, во время циркуляции вируса Нипах и добычу пальмового сока. Тем не менее в серологическом исследовании 418 потенциально зараженных людей ни один человек не продемонстрировал положительного результата при серологической диагностике вируса Нипах. Различия в сельскохозяйственных практиках, существующие между регионами Камбоджи, где отмечается появление вируса Нипах, могут объяснять сложившуюся ситуацию и указывать на необходимые мероприятия с целью ограничения риска передачи вируса людям.Практикуемые людьми виды деятельности являются ключом к пониманию рисков, связанных с передачей новых инфекционных заболеваний. Социальные научные дисциплины, такие как антропология, должны быть включены в программы медицинского обслуживания, направленные на борьбу с новыми инфекционными заболеваниями. Поскольку летучие мыши являются переносчиками серьезных зоонозных патогенов, такие объединенные исследования могут способствовать снижению риска появления других заболеваний, передающихся через летучих мышей.

Marine sediments are one of the largest carbon reservoir on Earth, yet the microbial communities, especially the eukaryotes, that drive these ecosystems are poorly characterised. Here, we report implementation of a sampling system that enables injection of reagents into sediments at depth, allowing for preservation of RNA in situ. Using the RNA templates recovered, we investigate the ‘ribosomally active’ eukaryotic diversity present in sediments close to the water/sediment interface. We demonstrate that in situ preservation leads to recovery of a significantly altered community profile. Using SSU rRNA amplicon sequencing, we investigated the community structure in these environments, demonstrating a wide diversity and high relative abundance of stramenopiles and alveolates, specifically: Bacillariophyta (diatoms), labyrinthulomycetes and ciliates. The identification of abundant diatom rRNA molecules is consistent with microscopy-based studies, but demonstrates that these algae can also be exported to the sediment as active cells as opposed to dead forms. We also observe many groups that include, or branch close to, osmotrophic–saprotrophic protists (e.g. labyrinthulomycetes and Pseudofungi), microbes likely to be important for detrital decomposition. The sequence data also included a diversity of abundant amplicon-types that branch close to the Fonticula slime moulds. Taken together, our data identifies additional roles for eukaryotic microbes in the marine carbon cycle; where putative osmotrophic–saprotrophic protists represent a significant active microbial-constituent of the upper sediment layer.

Aims: We aimed to assess whether soil salinity changes the microbial community in the rhizosphere of Hibiscus hamabo, and whether these changes in the microbiome feedback on the growth of H. hamabo. Methods: To test effects of salinity on the rhizosphere microbiome, we first did a greenhouse experiment in which H. hamabo was grown in pots with a sand-soil mixture at different salt concentrations (0, 15, 40 and 90 mM NaCl). Then in another two experiments, we tested effects of the rhizosphere microbiomes on performance of H. hamabo plants by sowing and growing them in pots with a peat-sand-vermiculite mixture inoculated with either soil or root fragments collected from the different salinity treatments (0, 40 and 90 mM NaCl) of the first experiment and crossed with a salinity treatment (0, 40 and 90 mM NaCl). Results: The bacterial rhizosphere community of H. hamabo was less affected by soil salinities than the fungal community was. Germination and biomass of H. hamabo were highest at a salinity of 40 mM NaCl, and higher in the presence than in the absence of microbial inoculums. Moreover, H. hamabo performed best when the microbial inocula came from the same salinity level, particularly at a salinity of 40 mM NaCl. Conclusions: Our study provides evidence that salinity-induced changes in rhizosphere microbial communities tend to promote germination and growth of H. hamabo at the respective salinities.

Microsporidia are obligate parasites that are closely related to Fungi. While the widely known “long-branch” Microsporidia infect mostly metazoans, the hosts of “short-branch” Microsporidia are only partially characterized or not known at all. Here, we used network analyses from Neotropical rainforest soil metabarcoding data, to infer co-occurrences between environmental lineages of short-branch microsporidians and their potential hosts. We found significant co-occurrences with several taxa, especially with Apicomplexa, Cercozoa, and Fungi, as well as some Metazoa. Our results are the first step to identify potential hosts of the environmental lineages of short-branch microsporidians, which can be targeted in future molecular and microscopic studies. Supplementary Information The online version contains supplementary material available at 10.1007/s00248-020-01657-9.

Although it is well established that land use and management affects soil microbial biodiversity, with consequences for soil-mediated ecosystem services, knowledge on human-induced variability in soil bacterial and fungal communities is more abundant for cropping than for livestock systems. In particular, grazing management affects vegetation cover and light availability to lower vegetation layers, particularly in wooded grazing ecosystems. Our objective was to evaluate the use of T-RFLP analysis to characterise soil microbial community patterns. We worked in a grazed temperate savannah in Entre Ríos (Argentina) and focused on two farms with different grazing management, resulting in different vegetation cover. At each farm, we sampled soils from four conditions forming a two shrub cover conditions (open and close) by two light availability conditions (high and low) factorial set with three replicates. T-RFLP data for bacteria and fungi were used to characterise biodiversity patterns (diversity indexes, relative abundance of operational taxonomic units and community structure). Analyses were performed, between and within farms, using nonmetric multidimensional scaling analysis, non-parametric multivariate analysis of variance and network analysis. Variation patterns of soil microbial communities were detected using T-RFLP analysis in savannah soils from these two livestock farms. Farm, light and shrub conditions were statistically significant factors shaping variation patterns. For bacterial communities, light availability and shrub cover affected biodiversity and community structure respectively, but this impact was detected only for the farm with lower grazing intensity. For fungal communities, light availability was the factor shaping community structure, also with slightly higher impact on the farm with lower grazing intensity. These results suggest that in temperate woodlands lower grazing intensity, associated with higher vegetation cover, increases bacterial biodiversity while it decreases fungal community structure. Soil bacterial and fungal communities, showed contrasting patterns, and appear as sensitive indicators to monitor grazing management effects. We showed that T-RFLP analysis is a cost-effective methodology (compared to Illumina sequencing) that would be useful to disentangle impact of grazing intensity and vegetation cover in grazed temperate savannah.

International audience; Mediterranean forests own distinct characteristics resulting from climate, soil, and vegetation that affect soil microbial communities’ assembly and their associated functions. We initiated a multi-scalar analysis of environmental drivers of soil functioning to (1) identify pertinent factorial scales and (2) determine the relative importance of soil, vegetation, and geoclimate influences in shaping soil microbial functions across the French Mediterranean forests. Soil samples (0–15 cm) were collected from 60 forest sites and soil physicochemical and microbiological properties were assessed across different factorial scales i.e., bioclimates, slope exposures, and forest stands. Patterns in microbial catabolic potential (i.e., extracellular enzymes and microbial respiration) and carbon (C) source utilization (i.e., catabolic-level physiological profiling) were partitioned between vegetation cover, soil characteristics, and geoclimate components. Our results reveal that the catabolic potential of soil microbes was strongly influenced by the forest stands and mainly relied on ammonium and nitrate contents. In contrast, variation in C source utilization was mainly explained by vegetation cover. Soil metabolic capacities of microorganisms and resulting C dynamics were largely constrained by climate parameters, which suggests potentially important consequences for soil C storage. Our study revealed diverse structuration patterns between the catabolic potential and the carbon source utilization of soil microbial communities, and gives insights into the underlying mechanisms of soil microbial functioning in Mediterranean forests.

International audience; The strict relationships between bacteria and plants represent one of the major facets of terrestrial ecology. Depending on the type of interaction and amount of metabolic advantage one organism can obtain from such relationships, these are classified as mutualistic, commensal or parasitic interactions. Within this context, Pseudomonas and Xanthomonas are bacterial genera with a worldwide spread, capable of establishing all of the above mentioned interactions with plants. Therefore, they represent good models for studying different lifestyles and, accordingly, deciphering distinct evolutionary trajectories followed by different lineages of a single genus to infect and/or to establish a mutualistic relationships with the plant. Some members of these two genera are regulated pests that are recognized as economically major threats for their host crop(s) both in temperate and tropical environments. Some Pseudomonas and Xanthomonas are key examples of different lifestyles (i.e., mesophyll or vessel-colonizing pathogens, epiphytic pathogens, plant growth-promoting rhizobacteria, non-pathogenic strains of recognized pathogenic species, etc). Refining our knowledge on the ecology and epidemiology of these bacterial groups, as well as deciphering their evolutionary dynamics are keys for understanding their contrasting lifestyles and consequently improving plant disease control. At the same time, insights on the activation of different plant defense mechanisms as challenged by the different repertoires of virulence factors displayed by pseudomonads and xanthomonads, would yield new achievements to reduce the threats they pose to cultivated and wild plant species. This Research Topic focuses on microbial and evolutionary ecology of plant associated Pseudomonas and Xanthomonas, as well as the genomic and molecular diversity of lineages and the virulence and fitness features involved in the interaction with the host-plant. Most of the literature available for this Research Topic has been performed for strains isolated in temperate zones. In line with the long-recognized high social and environmental impact of pests and pathogens in tropical countries, we have welcomed submissions of studies covering such situations for these areas. This Research Topic gathers high-quality contributions (Original Research, Methods, Protocols, Hypothesis & Theory, Reviews, Mini Reviews, Focused Reviews) and in order to promote complementary and original research approaches to improve our knowledge on pseudomonads and xanthomonads-host interactions and their control, it benefited from the scientific communities currently working on Pseudomonas and Xanthomonas such as the teams dealing with the Pseudomonas syringae species complex and the French Network on Xanthomonads (FNX).

International audience; In the ocean, Bacillariophyta are one of the most successful protistan groups. Due to their considerable biogeochemical implications, diatom diversity, development, and seasonality have been at the center of research, specifically large-sized species. In comparison, nanoplanktonic diatoms are mostly disregarded from routine monitoring and are often underrepresented in genetic reference databases. Here, we identified and investigated the temporal dynamics of relevant nanodiatoms occurring in the Western English Channel (SOMLIT-Astan station). Coupling in situ and laboratory approaches, we revealed that nano-species from the genera Minidiscus and Thalassiosira are key components of the phytoplankton community that thrive in these coastal waters, but they display different seasonal patterns. Some species formed recurrent blooms whilst others were persistent year round. These results raise questions about their regulation in the natural environment. Over a full seasonal cycle at the monitoring station, we succeeded in isolating viruses which infect these minute diatoms, suggesting that these mortality agents may contribute to their control. Overall, our study points out the importance of considering nanodiatom communities within time-series surveys to further understand their role and fate in marine systems.

International audience; Microbes in the atmosphere have broad ecological impacts, including the potential to trigger precipitation through species and strains that act as ice nucleation particles. To characterize spatiotemporal trends of microbial assemblages in precipitation we sequenced 16S (bacterial) and 18S (fungal) rRNA gene amplicon libraries collected from 72 precipitation events in three US states (Idaho, Louisiana, and Virginia) over four seasons. We considered these data from the perspective of a novel metacommunity framework. In agreement with our heuristic, we found evidence for distinct mechanisms underlying the composition and diversity of bacterial and fungal assemblages in precipitation. Specifically, we determined that: 1) bacterial operational taxonomic unit (OTU) composition of precipitation was strongly associated with macroscale drivers including season and high altitude characteristics of storms; 2) fungal OTU composition was strongly correlated with mesoscale drivers including particular spatial locations; 3) β‐diversity (heterogeneity of taxa among samples) for both bacteria and fungi was largely maintained by turnover of taxa; however, 4) bacterial assemblages had higher contributions to total β‐diversity from nestedness (i.e., lower richness assemblages were largely taxonomic subsets of richer assemblages), due to losses of taxa during dispersal, particularly among potential ice nucleation active bacteria; and 5) fungal assemblages had higher contributions to total β‐diversity from turnover due to OTU replacement. Spatiotemporal trends in precipitation‐borne metacommunities allowed delineation of a large number of statistically significant indicator taxa for particular sites and seasons, including trends for bacteria that are potentially ice nucleation active. Our findings advance understanding regarding the dispersion of aerosolized microbes via wet deposition, and the development of theory concerning potential assembly rules for bioaerosol assemblages.

At a human/livestock/wildlife interface, Escherichia coli populations were used to assess the risk of bacterial and antibiotic resistance dissemination between hosts. We used phenotypic and genotypic characterization techniques to describe the structure and the level of antibiotic resistance of E. coli commensal populations and the resistant Enterobacteriaceae carriage of sympatric African buffalo ( Syncerus caffer caffer ) and cattle populations characterized by their contact patterns in the southern part of Hwange ecosystem in Zimbabwe. Our results (i) confirmed our assumption that buffalo and cattle share similar phylogroup profiles, dominated by B1 (44.5%) and E (29.0%) phylogroups, with some variability in A phylogroup presence (from 1.9 to 12%); (ii) identified a significant gradient of antibiotic resistance from isolated buffalo to buffalo in contact with cattle and cattle populations expressed as the Murray score among Enterobacteriaceae (0.146, 0.258, and 0.340, respectively) and as the presence of tetracycline-, trimethoprim-, and amoxicillin-resistant subdominant E. coli strains (0, 5.7, and 38%, respectively); (iii) evidenced the dissemination of tetracycline, trimethoprim, and amoxicillin resistance genes ( tet , dfrA , and bla TEM-1 ) in 26 isolated subdominant E. coli strains between nearby buffalo and cattle populations, that led us (iv) to hypothesize the role of the human/animal interface in the dissemination of genetic material from human to cattle and toward wildlife. The study of antibiotic resistance dissemination in multihost systems and at anthropized/natural interface is necessary to better understand and mitigate its multiple threats. These results also contribute to attempts aiming at using E. coli as a tool for the identification of pathogen transmission pathway in multihost systems.

To guarantee food safety, a better deciphering of ecology and adaptation strategies of bacterial pathogens such as Salmonella in food environments is crucial. The role of food processing conditions such as cleaning and disinfection procedures on antimicrobial resistance emergence should especially be investigated. In this work, the prevalence and antimicrobial resistance of Salmonella and the microbial ecology of associated surfaces communities were investigated in a pig slaughterhouse before and after cleaning and disinfection procedures. Salmonella were detected in 67% of samples and isolates characterization revealed the presence of 15 PFGE-patterns belonging to five serotypes: S.4,5,12:i:-, Rissen, Typhimurium, Infantis and Derby. Resistance to ampicillin, sulfamethoxazole, tetracycline and/or chloramphenicol was detected depending on serotypes. 16S rRNA-based bacterial diversity analyses showed that Salmonella surface associated communities were highly dominated by the Moraxellaceae family with a clear site-specific composition suggesting a persistent colonization of the pig slaughterhouse. Cleaning and disinfection procedures did not lead to a modification of Salmonella susceptibility to antimicrobials in this short-term study but they tended to significantly reduce bacterial diversity and favored some genera such as Rothia and Psychrobacter. Such data participate to the construction of a comprehensive view of Salmonella ecology and antimicrobial resistance emergence in food environments in relation with cleaning and disinfection procedures.

Island biogeography theory is one of the most influential paradigms in ecology. That island characteristics, including remoteness, can profoundly modulate biological diversity has been borne out by studies of animals and plants. By contrast, the processes influencing microbial diversity in island systems remain largely undetermined. We sequenced arbuscular mycorrhizal (AM) fungal DNA from plant roots collected on 13 islands worldwide and compared AM fungal diversity on islands with existing data from mainland sites. AM fungal communities on islands (even those >6000 km from the closest mainland) comprised few endemic taxa and were as diverse as mainland communities. Thus, in contrast to patterns recorded among macro-organisms, efficient dispersal appears to outweigh the effects of taxogenesis and extinction in regulating AM fungal diversity on islands. Nonetheless, AM fungal communities on more distant islands comprised a higher proportion of previously cultured and large-spored taxa, indicating that dispersal may be human-mediated or require tolerance of significant environmental stress, such as exposure to sunlight or high salinity. The processes driving large-scale patterns of microbial diversity are a key consideration for attempts to conserve and restore functioning ecosystems in this era of rapid global change.

Investigations into life history of microorganisms that cause plant diseases have been limited mostly to contexts where they are in interaction with plants, and with cropped or otherwise managed vegetation. Therefore, knowledge about the diversity of plant pathogens, about potential reservoirs of inoculum and about the processes that contribute to their survival and adaptation is limited to these contexts. The agro-centric perspective of plant pathogen life histories is incoherent with respect to the capacity of many of them to persist as saprophytes on various substrates. In this context we have investigated the ubiquity of the broad host range necrotrophic fungus Botrytis cinerea, outside of agricultural settings and have determined if the populations in these natural habitats can be distinguished phenotypically and phylogenetically from populations isolated from diseased crops. Over a period of 5 years, we isolated B. cinerea from 235 samples of various substrates collected in France including rainfall, snowpack, river, and lake water, epilithic biofilms in mountain streams, leaf litter and plant debris, rock surfaces, bird feathers and healthy wild plants from outside of agricultural fields. All substrates except rock surfaces harbored B. cinerea leading us to establish a collection of purified strains that were compared to B. cinerea from diseased tomato, grapes and various other crops in France. Phylogenetic comparisons of 321 strains from crop plants and 100 strains from environmental substrates based on sequences of 9 microsatellite markers revealed that strains from crops and the environment could not be distinguished. Furthermore, the genetic diversity of strains outside of agriculture was just as broad as within agriculture. In tests to determine the aggressiveness of strains on tomato stems, the mean disease severity caused by strains from environmental substrates was statistically identical to the severity of disease caused by strains from tomato, but was significantly greater than the severity caused by strains from grape or other crops. Our results suggest that highly diverse populations of this plant pathogen persist outside of agriculture in association with substrates other than plants and that this part of their life history is compatible with its capacity to maintain its potential as plant pathogen.

Traceability of foods is mainly done at the administrative level, and the use of analytical tools is rare. Previous studies have demonstrated that microbial ecology analyses at the molecular level (such as PCR-DGGE) could be used to provide food with a unique biological signature that could be linked to the geographical origin of food. The present study aimed at testing this approach to differentiate farming types by analyzing organic and conventional food products. To this end, the microbial ecology of organic and conventional nectarines was analyzed and statistically compared. Our results show that yeast and bacterial communities were specific of the farming type allowing organic fruits to be discriminated from conventional ones. Several microbial species were identified as potential, biological markers which detection could be used to certify the origin as well as the mode of production of foodstuff. We proposed this analytical tool as a first step to control and authentify organic foods.

Understanding how soil pedogenesis affects microbial communities and their in situ activities according to ecosystem functioning is a central issue in soil microbial ecology, as soils represent essential nutrient reservoirs and habitats for the biosphere. To address this question, soil chronosequences developed from a single, shared mineralogical parent material and having the same climate conditions are particularly useful, as they isolate the factor of time from other factors controlling the character of soils. In our study, we considered a natural succession of uplifted marine terraces in Mendocino, CA, ranging from highly fertile in the younger terrace (about 100,000 years old) to infertile in the older terraces (about 300,000 years old). Using ITS amplicon pyrosequencing, we analysed and compared the diversity and composition of the soil fungal communities across the first terraces (T1 to T3), with a specific focus in the forested terraces (T2 and T3) on soil samples collected below trees of the same species (Pinus muricata) and of the same age. While diversity and richness indices were highest in the grassland (youngest) terrace (T1), they were higher in the older forested terrace (T3) compared to the younger forested terrace (T2). Interestingly, the most abundant ectomycorrhizal (ECM) taxa that we found within these fungal communities showed high homology with ITS Sanger sequences obtained previously directly from ECM root tips from trees in the same study site, revealing a relative conservation of ECM diversity over time. Altogether, our results provide new information about the diversity and composition of the fungal communities as well as on the dominant ECM species in the soil chronosequence of Mendocino in relation to soil age and ecosystem development.

The microbial composition of different types,in ecosystems (including agro-ecosystems), has been investigated in a rapidly growing number of studies in the past few years. The importance of microorganisms, regarding the maintenance and stability of nutrients in agroecosystems, is a key to maintain the sustainability of a crop. Molecular tools to study microbial communities are possible through many methods such as RISA, DGGE, TGGE, clone libraries, T-RFLP, RAPD, SSCP and more recently NGS (Next-Generation Sequencing). DGGE is widely employed to characterize the diversity and the community dynamics of microorganisms in the environment, making possible to find out specific groups through functional genes, allowing access to data that cannot be obtained by cultural methods. The aim of this paper is to review the functional groups related to agroecosystems and to indicate the critical choice of DNA primers pairs and targeted DNA regions that may be used in PCR-based methods such as the DGGE technique in order to evaluate the microbial communities in a variety of environments. (Resume d'auteur)

BGPI : équipe 5; International audience; Recent advancements in sequencing technology allowed researchers to better address the patterns and mechanisms involved in microbial environmental adaptation at large spatial scales. Here we investigated the genomic basis of adaptation to climate at the continental scale in Suillus brevipes, an ectomycorrhizal fungus symbiotically associated with the roots of pine trees. We used genomic data from 55 individuals in seven locations across North America to perform genome scans to detect signatures of positive selection and assess whether temperature and precipitation were associated with genetic differentiation. We found that S. brevipes exhibited overall strong population differentiation, with potential admixture in Canadian populations. This species also displayed genomic signatures of positive selection as well as genomic sites significantly associated with distinct climatic regimes and abiotic environmental parameters. These genomic regions included genes involved in transmembrane transport of substances and helicase activity potentially involved in cold stress response. Our study sheds light on large-scale environmental adaptation in fungi by identifying putative adaptive genes and providing a framework to further investigate the genetic basis of fungal adaptation

International audience; Pollution-induced community tolerance (PICT) approaches involve comparing tolerance levels of natural communities to a particular contaminant or a contaminant mixture using short-term toxicity tests performed under controlled conditions. However, results from toxicity tests can be modulated by various environmental and experimental conditions, raising questions about their reproducibility and comparability. In this context, the present study aimed to determine the influence of exposure duration, periphyton suspension concentration and periphyton maturation stage on the measurement of short-term effects of copper on phototrophic periphyton communities. Our results showed the very weak influence of exposure duration in the tested range (2-6 h) on toxicity level, whereas periphyton biomass in the tested suspension (in terms of both chlorophyll a concentrations and dry weight), proved a crucial determinant in toxicity assessment. Results also highlighted the potential tolerance increase with the periphyton maturation stage. This parameter conditioned the positive linear relationship between tolerance level and periphyton suspension concentration, leading to an increase in the linear regression slope with the maturation stage. This suggests that such a relationship is probably highly periphyton-dependent. Consequently, to enable data toxicity comparisons, an a priori normalization of the periphyton suspension biomass is necessary, and PICT approaches require the use, as much of possible, of periphyton with similar maturation stage. Finally, the present study clearly shows that a better standardization of PICT approaches could help to improve reproducibility. It could thus facilitate the comparison of tolerance levels measured in the same study (e.g. spatial and/or temporal and/or inter-treatment comparison) as well as the comparison obtained from different experimental and in situ research.

International audience; The determination of geographical origin is part of the demand of the traceability system of food products. A hypothesis of tracing the source of a product is to analyse in a global way the bacterial communities of the food samples after their production. For this purpose, molecular techniques employing 16S rDNA profiles generated by PCR–DGGE were used to detect the variation in bacterial community structures of salts from four French regions. When the 16S rDNA profiles were analysed by multivariate analysis, distinct microbial communities were detected. The band profiles of the salt bacteria from different producing areas were different and were specific for each location and could be used as a bar code to certify the origin of salts. These band profiles can be used as specific markers for a specific location. This method is proposed as a new traceability tool which provides salts with a unique bar code that permits to trace back salts from store shelves to their original location.

Methods to ensure the health of crops owe their efficacy to the extent to which we understand the ecology and biology of environmental microorganisms and the conditions under which their interactions with plants lead to losses in crop quality or yield. However, in the pursuit of this knowledge, notions of the ecology of plant-pathogenic microorganisms have been reduced to a plant-centric and agro-centric focus. With increasing global change, i.e. changes that encompass not only climate, but also biodiversity, the geographical distribution of biomes, human demographic and socio-economic adaptations and land use, new plant health problems will emerge via a range of processes influenced by these changes. Hence, knowledge of the ecology of plant pathogens will play an increasingly important role in the anticipation and response to disease emergence. Here, we present our opinion on the major challenges facing the study of the ecology of plant-pathogenic bacteria. We argue that the discovery of markedly novel insights into the ecology of plant-pathogenic bacteria is most likely to happen within a framework of more extensive scales of space, time and biotic interactions than those that currently guide much of the research on these bacteria. This will set a context that is more propitious for the discovery of unsuspected drivers of the survival and diversification of plant-pathogenic bacteria and of the factors most critical for disease emergence, and will set the foundation for new approaches to the sustainable management of plant health. We describe the contextual background of, justification for and specific research questions with regard to the following challenges: Development of terminology to describe plant-bacterial relationships in terms of bacterial fitness. Definition of the full scope of the environments in which plant-pathogenic bacteria reside or survive. Delineation of pertinent phylogenetic contours of plant-pathogenic bacteria and naming of strains independent of their presumed life style. Assessment of how traits of plant-pathogenic bacteria evolve within the overall framework of their life history. Exploration of possible beneficial ecosystem services contributed to by plant-pathogenic bacteria.

BGPI : équipe 5; The structure of pathogen populations is an important driver of epidemics affecting crops and natural plant communities. Comparing the composition of two pathogen populations consisting of assemblages of genotypes or phenotypes is a crucial, recurrent question encountered in many studies in plant disease epidemiology. Determining if there is a significant difference between two sets of proportions is also a generic question for numerous biological fields. When samples are small and data are sparse, it is not straightforward to provide an accurate answer to this simple question because routine statistical tests may not be exactly calibrated. To tackle this issue, we built a computationally-intensive testing procedure, namely the Generalized Monte Carlo Plug-In test with Calibration (GMCPIC test), which is implemented in an R package available at http://dx.doi.org/10.5281/zenodo.53996. A simulation study was carried out to assess the performance of the proposed methodology and to make a comparison with standard statistical tests. This study allows us to give advice on how to apply the proposed method, depending on the sample sizes. The proposed methodology was then applied to real datasets and the results of the analyses were discussed from an epidemiological perspective. The applications to real data sets deal with three topics in plant pathology: the reproduction of Magnaporthe oryzae, the spatial structure of Pseudomonas syringae, and the temporal recurrence of Puccinia triticina.

Here, we investigated the patterns of microbial nitrogen cycling communities along a chronosequence of soil development in a salt marsh. The focus was on the abundance and structure of genes involved in N fixation (nifH), bacterial and archaeal ammonium oxidation (amoA; AOB and AOA), and the abundances of genes involved in denitrification (nirS, nirK, nosZ). Potential nitrification and denitrification activities were also measured, and increases in nitrification were found in soils towards the end of succession, whereas denitrification became maximal in soils at the intermediate stages. The nifH, nirK and nirS gene markers revealed increases in the sizes of the respective functional groups towards the intermediate stage (35 years), remaining either constant (for nifH) or slightly declining towards the latest stage of succession (for nirK and nirS). Moreover, whereas the AOB abundance peaked in soils at the intermediate stage, that of AOA increased linearly along the chronosequence. The abundance of nosZ was roughly constant, with no significant regression. The drivers of changes in abundance and structure were identified using path analysis; whereas the ammonia oxidizers (AOA and AOB) showed patterns that followed mainly N availability, those of the nitrogen fixers followed plant diversity and soil structure. The patterns of denitrifiers were group-dependent, following the patterns of plant diversity (nirK and nirS) and belowground shifts (nosZ). The variation observed for the microbial groups associated with the same function highlights their differential contribution at different stages of soil development, revealing an interplay of changes in terms of niche complementarity and adaptation to the local environment.

BGPI : équipe 5; In addition to being one of the most acute problems impeding chemical control of fungal diseases, the evolution of fungicide resistance is an emblematic case of local adaptation to spatially heterogeneous and temporally variable selection pressures. Here we dissected the adaptation of Botrytis cinerea (the causal agent of grey mould) populations on grapes to several fungicides. We carried out a two-year survey (four collection dates) on three treated/untreated pairs of plots from vineyards in Champagne (France), and monitored the frequency of four resistant phenotypes that are unambiguously associated to four distinct genotypes. For two loci under selection by currently-used fungicides (MDR1 and MDR2), the frequencies of resistant mutations at vintage were greater in treated plots compared to untreated plots, showing that the effect of selection is detectable even at the plot scale. This effect was not detectable for two other loci under selection by previously-used fungicides (BenR1 and ImiR1). We also found that treatment with currently-used fungicides reduced B. cinerea effective population size, leading to a significant decrease in genic diversity and allelic richness in treated versus untreated plots. We further highlight that even under ample drift and migration, fungal populations can present an efficient response to selection. Finally, for the four studied loci, the costs of fungicide resistance were estimated by modelling the decrease in the frequency of resistant mutations in the absence of treatment. We discuss the importance of these estimates for defining strategies for limiting or counteracting the local adaptation of pests to fungicides. This article is protected by copyright. All rights reserved.

One of the most important biotechnological challenges is to develop environment friendly technologies to produce new sources of energy. Microbial production of biohydrogen through dark fermentation, by conversion of residual biomass, is an attractive solution for short-term development of bioH2 producing processes. Efficient biohydrogen production relies on complex mixed communities working in tight interaction. Species composition and functional traits are of crucial importance to maintain the ecosystem service. The analysis of microbial community revealed a wide phylogenetic diversity that contributes in different—and still mostly unclear—ways to hydrogen production. Bridging this gap of knowledge between microbial ecology features and ecosystem functionality is essential to optimize the bioprocess and develop strategies toward a maximization of the efficiency and stability of substrate conversion. The aim of this review is to provide a comprehensive overview of the most up-to-date biodata available and discuss the main microbial community features of biohydrogen engineered ecosystems, with a special emphasis on the crucial role of interactions and the relationships between species composition and ecosystem service. The elucidation of intricate relationships between community structure and ecosystem function would make possible to drive ecosystems toward an improved functionality on the basis of microbial ecology principles.

The conceptual framework considering Anseriformes and Charadriiformes as the main maintenance hosts for influenza A viruses (IAV) in wild birds has shaped IAV research and surveillance over the last decades. We challenge this framework by reviewing the world-wide surveillance data on non-Anseriformes and non-Charadriiformes (NANC) species, generally considered as playing little role in IAV maintenance, available in literature and online data bases (close to 200 sources). Globally, we found an IAV infection rate of 1·51% (95% CI, 1·44–1·59%) for c. 101 000 birds tested from NANC species. If Anseriformes have, as expected, a higher infection rate than any other bird orders, eight bird orders have an infection rate higher or close to the Charadriiformes infection rate, challenging the status of Charadriiformes. We interpret the attention paid in favour of Charadriiformes by an extrapolation bias from data collected in hotspots of IAV infection in Charadriiformes (e.g. Delaware Bay, USA). The growing data on IAV in wild birds world-wide, summarised here, support two non-exclusive hypotheses: (i) the quality of the diagnostic tools and techniques used explain the patterns observed; (ii) IAV maintenance is determined by complex multi-host systems composed of multiple bird species, dependent on the ecosystem and its bird diversity and composition. Synthesis and applications. Our results have two main implications. First, new research and surveillance should be designed in order to understand influenza A viruses ecology in wild birds across the world, along with appropriate diagnostic tools and new hypotheses and dedicated protocols. This should be done in line with our new conceptual framework that conveys less a priori than its predecessor. Second, our results call for more bridging between biological and epidemiological sciences in order to tackle disease ecology in multi-host systems. (Resume d'auteur)

To get insights into the temporal pattern of commensal Escherichia coli populations, we sampled the feces of four healthy cows from the same herd in the Hwange District of Zimbabwe daily over 25 days. The cows had not received antibiotic treatment during the previous 3 months. We performed viable E. coli counts and characterized the 326 isolates originating from the 98 stool samples at a clonal level, screened them for stx and eae genes, and tested them for their antibiotic susceptibilities. We observed that E. coli counts and dominant clones were different among cows, and very few clones were shared. No clone was shared by three or four cows. Clone richness and evenness were not different between cows. Within each host, the variability in the E. coli count was evidenced between days, and no clone was found to be dominant during the entire sampling period, suggesting the existence of clonal interference. Dominant clones tended to persist longer than subdominant ones and were mainly from phylogenetic groups A and B1. Five E. coli clones were found to contain both the stx 1 and stx 2 genes, representing 6.3% of the studied isolates. All cows harbored at least one Shiga toxin-producing E. coli (STEC) strain. Resistance to tetracycline, penicillins, trimethoprim, and sulfonamides was rare and observed in three clones that were shed at low levels in two cows. This study highlights the fact that the commensal E. coli population, including the STEC population, is host specific, is highly dynamic over a short time frame, and rarely carries antibiotic resistance determinants in the absence of antibiotic treatment. IMPORTANCE The literature about the dynamics of commensal Escherichia coli populations is very scarce. Over 25 days, we followed the total E. coli counts daily and characterized the sampled clones in the feces of four cows from the same herd living in the Hwange District of Zimbabwe. This study deals with the day-to-day dynamics of both quantitative and qualitative aspects of E. coli commensal populations, with a focus on both Shiga toxin-producing E. coli and antibiotic-resistant E. coli strains. We show that the structure of these commensal populations was highly specific to the host, even though the cows ate and roamed together, and was highly dynamic between days. Such data are of importance to understand the ecological forces that drive the dynamics of the emergence of E. coli clones of particular interest within the gastrointestinal tract and their transmission between hosts.

International audience; Soilless cultures were originally developed to control soilborne diseases. Soilless cultures provide several advantages for growers such as greater production of crops, reduced energy consumption, better control of growth and independence of soil quality. However, diseases specific to hydroponics have been reported. For instance, zoospore-producing microorganisms such as Pythium and Phytophthora spp. are particularly well adapted to aquatic environments. Their growth in soilless substrates is favoured by the recirculation of the nutrient solution. These pathogenic microorganisms are usually controlled by disinfection methods but such methods are only effective as a preventive measure. Contrary to biofiltration, active treatments such as UV, heat and ozonisation have the disadvantage of eliminating not only the harmful microorganisms but also the beneficial indigenous microorganisms. Here, we review microbial populations that colonise ecological niches of hydroponic greenhouse systems. Three topics are discussed: (1) the general microflora; (2) the pathogenic microflora that are typical to hydroponic systems; and (3) the non-pathogenic and possibly beneficial microflora, and their use in the control of plant diseases in soilless greenhouse systems. Technical, economic and environmental concerns are forcing the adoption of new sustainable methods such as the use of microbial antagonists. Thus, increased attention is now focused on the role of natural microflora in suppressing certain diseases. Managing disease suppression in hydroponics represents a promising way of controlling pathogens. Three main strategies can be used: (1) increasing the level of suppressiveness by the addition of antagonistic microorganisms; (2) using a mix of microorganisms with complementary ecological traits and antagonistic abilities, combined with disinfection techniques; and (3) amending substrates to favour the development of a suppressive microflora. Increasing our knowledge on beneficial microflora, their ecology and treatments that influence their composition will help to commercialise new, ready-to-use substrates microbiologically optimised to protect plants in sustainable management systems.

[Departement_IRSTEA]Ecotechnologies [TR1_IRSTEA]TED [TR2_IRSTEA]INSPIRE; International audience; Although mathematical modelling has reached a degree of maturity in the last decades, microbial ecology is still developing, albeit at a rapid pace thanks to new insights provided by modern molecular tools. However, whilst microbiologists have long enjoyed the perspectives that particular mathematical frameworks can provide, there remains a reluctance to fully embrace the potential of models, which appear too complex, esoteric or distant from the “real-world”. Nevertheless there is a strong case for pursuing the development of mathematical models to describe microbial behaviour and interactions, dynamically, spatially and across scales. Here we put forward perspectives on the current state of mathematical modelling in microbial ecology, looking back at the developments that have defined the synergies between the disciplines, and outline some of the existing challenges that motivate us to provide practical models in the hope that greater engagement with empiricists and practitioners in the microbiological domain may be achieved. We also indicate recent advances in modelling that have had impact in both the fundamental understanding of microbial ecology and its practical application in engineered biological systems. In this way, it is anticipated that interest can be garnered from across the microbiological spectrum resulting in a broader uptake of mathematical concepts in lecture theatres, laboratories and industrial systems.

International audience; Many microbial ecosystems can be seen as microbial 'food chains' where the different reaction steps can be seen as such: the waste products of the organisms at a given reaction step are consumed by organisms at the next reaction step. In the present paper we study a model of a two-step biological reaction with feedback inhibition, which was recently presented as a reduced and simplified version of the anaerobic digestion model ADM1 of the International Water Association (IWA). It is known that in the absence of maintenance (or decay) the microbial 'food chain' is stable. In a previous study, using a purely numerical approach and ADM1 consensus parameter values, it was shown that the model remains stable when decay terms are added. However, the authors could not prove in full generality that it remains true for other parameter values. In this paper we prove that introducing decay in the model preserves stability whatever its parameters values are and for a wide range of kinetics.

Two main types of processing are applied to coffee: the dry and wet processes. The aim of this study was to measure the impact of post-harvest processing on coffee microbial ecology and search for microbial markers of process discrimination. Coffee samples were collected in three farms in two locations of West Cameroon (Bafoussam and Dschang). Samples of Coffea arabica were collected along wet and dry methods and Coffea canephora samples were collected only during treatment using the dry method. Bacterial and fungal communities present on coffee beans were determined by using a molecular method PCR-DGGE (Denaturing Gradient Gel Electrophoresis) generating 16S rDNA and 26S rDNA profiles for bacteria and fungi. Results showed that microbial ecology structures were different depending on three main parameters: i) the method applied, ii) the coffee species and iii) their geographical origin. Statistical analysis applied allowed to measure the impact of all parameters and showed that the method was the major parameter influencing microbial communities' structure. Microbial DNA markers that appear specific of coffee origin, species or treatment were identified, such as lactic acid bacteria (LAB) which were detected only during wet process. They are specific of the fermentation step. DNA from Wallemia sp. was only detected in Robusta coffee beans (Coffea canephora). The study of microbial communities at the molecular level appears as a promising approach to discriminate between coffee processing types.

Introduction. Bissap (Hibiscus sabdariffa L.) is a common plant in the tropics. In Senegal, the calyces are used to make a popular juice. In the food industry, small and medium-sized enterprises (SMEs) are responsible for the transformation of bissap calyces into drinks, concentrates, jam, etc. In spite of the very low pH of the juice (pH < 3), problems of contamination and fermentation are often observed in the final products post-pasteurization. They are mainly due to Pseudomonas spp., E. coli, Klebsiella spp. and Pichia opuntiae. To solve this issue, monitoring of the microbial ecology was performed during the full process of bissap products. Methods and results. Fresh calyces and dried mixed calyces of the two varieties of Hibiscus sabdariffa ('Koor' and 'Vimto'), as well as juice samples, were collected at every stage of the processing of a bissap drink and syrup in a Senegalese SME. The monitoring of microbial flora was performed by using molecular fingerprinting. The molecular technique PCR-DGGE was employed to evaluate the microbial dynamics using bacterial 16S rDNA, yeast 26S rDNA and 28S rDNA mold profiles at each critical stage of the process. Results and discussion. The genetic profiles generated contributed to identifying the critical points in the manufacturing processes. A multivariate analysis based on the presence or absence of spots in the denaturing gradient electrophoresis gels (DGGE) showed that the microbial flora (bacteria, yeasts, molds) of bissap evolved during the following phases: harvest (fresh flower), drying (dried calyces) and processing (before filtration, after pasteurization and before packaging). Conclusion. Our work contributed to determining the microorganisms responsible for the microbial contamination of the final products, and highlighted the origin of these contaminants. The most important critical point was identified as the pasteurization step. (Resume d'auteur)

Many bacterial pathogens are well characterized but, in some cases, little is known about the populations from which they emerged. This limits understanding of the molecular mechanisms underlying disease. The crop pathogen Pseudomonas syringae sensu lato has been widely isolated from the environment, including wild plants and components of the water cycle, and causes disease in several economically important crops. Here, we compared genome sequences of 45 P. syringae crop pathogen outbreak strains with 69 closely related environmental isolates. Phylogenetic reconstruction revealed that crop pathogens emerged many times independently from environmental populations. Unexpectedly, differences in gene content between environmental populations and outbreak strains were minimal with most virulence genes present in both. However, a genome-wide association study identified a small number of genes, including the type III effector genes hopQ1 and hopD1, to be associated with crop pathogens, but not with environmental populations, suggesting that this small group of genes may play an important role in crop disease emergence. Intriguingly, genome-wide analysis of homologous recombination revealed that the locus Psyr 0346, predicted to encode a protein that confers antibiotic resistance, has been frequently exchanged among lineages and thus may contribute to pathogen fitness. Finally, we found that isolates from diseased crops and from components of the water cycle, collected during the same crop disease epidemic, form a single population. This provides the strongest evidence yet that precipitation and irrigation water are an overlooked inoculum source for disease epidemics caused by P. syringae.

International audience; The Anopheles midgut hosts diverse bacterial communities and represents a complex ecosystem. Several evidences indicate that mosquito midgut microbiota interferes with malaria parasite transmission. However, the bacterial composition of salivary glands and ovaries, two other biologically important tissues, has not been described so far. In this study, we investigated the dynamics of the bacterial communities in the mosquito tissues from emerging mosquitoes until 8 days after a blood meal containing Plasmodium falciparum gametocytes and described the temporal colonization of the mosquito epithelia. Bacterial communities were identified in the midgut, ovaries, and salivary glands of individual mosquitoes using pyrosequencing of the 16S rRNA gene. We found that the mosquito epithelia share a core microbiota, but some bacteria taxa were more associated with one or another tissue at a particular time point. The bacterial composition in the tissues of emerging mosquitoes varied according to the breeding site, indicating that some bacteria are acquired from the environment. Our results revealed temporal variations in the bacterial community structure, possibly as a result of the mosquito physiological changes. The abundance of Serratia significantly correlated with P. falciparum infection both in the midgut and salivary glands of malaria challenged mosquitoes, which suggests that interactions occur between microbes and parasites. These bacteria may represent promising targets for vector control strategies. Overall, this study points out the importance of characterizing bacterial communities in malaria mosquito vectors.

This chapter discusses the advances in biological control of cacao diseases over the last 15 years. Most attention has been focused on biological control of frosty pod rot (Moniliophthora roreri), witches’ broom (Moniliophthora perniciosa) and black pod disease (Phytophthora spp.). Research on biocontrol of other diseases in the cacao phyllosphere or rhizosphere is scarce or in its infancy. There is, however, a steady increase in information regarding the factors influencing and the mechanisms underlying biological control of cacao diseases as well as practical aspects such as inoculum production, formulation and application. There has been a clear shift away from inundative approaches using epiphytic BCAs towards more classical biocontrol approaches using bacterial and fungal endophytes as well as vesicular arbuscular mycorrhiza. These have the advantage that they can permanently establish themselves in the cacao tree. Moreover, besides direct competition for space and nutrients, antibiosis and mycoparasitism, through induced resistance and growth promotion, endophytes have a larger arsenal of mechanisms through which they can help protect their host. Endophytic BCAs could thus provide more effective and sustainable disease control. Recent advances in our understanding of the mechanisms through which endophytic biocontrol agents can reduce pest and disease impact provide possibilities for innovative disease control strategies, including combination therapies together with natural or chemical products. Continued work on production, formulation and application is also necessary in order for biocontrol to become economically interesting. However, biological control will not become a stand-alone solution for disease control but should become part of integrated pest management strategies, with cultural management as a central and reinforcing pillar.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., Microorganisms are vital in mediating the earth's biogeochemical cycles; yet, despite our rapidly increasing ability to explore complex environmental microbial communities, the relationship between microbial community structure and ecosystem processes remains poorly understood. Here, we address a fundamental and unanswered question in microbial ecology: 'When do we need to understand microbial community structure to accurately predict function?' We present a statistical analysis investigating the value of environmental data and microbial community structure independently and in combination for explaining rates of carbon and nitrogen cycling processes within 82 global datasets. Environmental variables were the strongest predictors of process rates but left 44% of variation unexplained on average, suggesting the potential for microbial data to increase model accuracy. Although only 29% of our datasets were significantly improved by adding information on microbial community structure, we observed improvement in models of processes mediated by narrow phylogenetic guilds via functional gene data, and conversely, improvement in models of facultative microbial processes via community diversity metrics. Our results also suggest that microbial diversity can strengthen predictions of respiration rates beyond microbial biomass parameters, as 53% of models were improved by incorporating both sets of predictors compared to 35% by microbial biomass alone. Our analysis represents the first comprehensive analysis of research examining links between microbial community structure and ecosystem function. Taken together, our results indicate that a greater understanding of microbial communities informed by ecological principles may enhance our ability to predict ecosystem process rates relative to assessments based on environmental variables and microbial physiology.

International audience; Fermentation is a traditional way of food preservation and is of great importance for human food consumption as it enables the development of nutritional and organoleptic qualities of food. This key traditional process is used for the conservation and transformation of a wide variety of food products of different origins (animal or vegetal) and nature (liquid to solid). For instance, starchy cereal-based food, meat, fish and sea food, vegetables and fruits, dairy products, cocoa, coffee and many others are transformed by fermentation. Food fermentation is utilized in many different geographical areas and, most of the time, occurs spontaneously. In Africa and Asia, traditional fermented foods represent a large part of local population diet. They often require process optimization to extend their production and commercialization (Aidoo et al. 2006, Sanni 1993). Fermentation is either alcoholic, resulting in alcoholic beverages or bread, lactic as is usually seen for dairy foods and vegetables, or acetic (vinegar). But food products may also result from a combination of these. Moreover, frequently for traditional foods, fermentative agents are undetermined and may involve bacterial or mould species.

International audience; How pathogens coevolve with and adapt to their hosts are critical to understanding how host jumps and/or acquisition of novel traits can lead to new disease emergences. The Xanthomonas genus includes Gram-negative plant-pathogenic bacteria that collectively infect a broad range of crops and wild plant species. However, individual Xanthomonas strains usually cause disease on only a few plant species and are highly adapted to their hosts, making them pertinent models to study host specificity. This review summarizes our current understanding of the molecular basis of host specificity in the Xanthomonas genus, with a particular focus on the ecology, physiology, and pathogenicity of the bacterium. Despite our limited understanding of the basis of host specificity, type III effectors, microbe-associated molecular patterns, lipopolysaccharides, transcriptional regulators, and chemotactic sensors emerge as key determinants for shaping host specificity.

Microbial species richness and assemblages across ultramafic ecosystems were investigated to assess the relationship between their distributional patterns and environmental traits. The structure of microorganism communities in the Koniambo massif, New Caledonia, was investigated using a metagenetic approach correlated with edaphic and floristic factors. Vegetation cover and soil properties significantly shaped the large phylogenetic distribution of operational taxonomic unit within microbial populations, with a mean per habitat of 3,477 (± 317) for bacteria and 712 (± 43) for fungi. Using variance partitioning, we showed that the effect of aboveground vegetation was the most significant descriptor for both bacterial and fungal communities. The floristic significant predictors explained 43% of the variation both for the bacterial and fungal community structures, while the edaphic significant predictors explained only 32% and 31% of these variations, respectively. These results confirm the previous hypothesis, that the distribution of microorganisms was more structured by the vegetation cover rather than the edaphic characteristics and that microbial diversity is not limited in ultramafic ecosystems.

Whole Genome Shotgun (WGS) metagenomics is increasingly used to study the structure and functions of complex microbial ecosystems, both from the taxonomic and functional point of view. Gene inventories of otherwise uncultured microbial communities make the direct functional profiling of microbial communities possible. The concept of community aggregated trait has been adapted from environmental and plant functional ecology to the framework of microbial ecology. Community aggregated traits are quantified from WGS data by computing the abundance of relevant marker genes. They can be used to study key processes at the ecosystem level and correlate environmental factors and ecosystem functions. In this paper we propose a novel model based approach to infer combinations of aggregated traits characterizing specific ecosystemic metabolic processes. We formulate a model of these Combined Aggregated Functional Traits (CAFTs) accounting for a hierarchical structure of genes, which are associated on microbial genomes, further linked at the ecosystem level by complex co-occurrences or interactions. The model is completed with constraints specifically designed to exploit available genomic information, in order to favor biologically relevant CAFTs. The CAFTs structure, as well as their intensity in the ecosystem, is obtained by solving a constrained Non-negative Matrix Factorization (NMF) problem. We developed a multicriteria selection procedure for the number of CAFTs. We illustrated our method on the modelling of ecosystemic functional traits of fiber degradation by the human gut microbiota. We used 1408 samples of gene abundances from several high-throughput sequencing projects and found that four CAFTs only were needed to represent the fiber degradation potential. This data reduction highlighted biologically consistent functional patterns while providing a high quality preservation of the original data. Our method is generic and can be applied to other metabolic processes in the gut or in other ecosystems., Author Summary Microbial communities are highly complex ecosystems, harboring a high taxonomic diversity, rapidly varying in time. Focusing on the functional traits of microorganisms instead of species offers an interesting and complementary insight since quite often, owing to environmental selection pressure, traits are less variable in time and shared by many species. By providing gene inventories of otherwise uncultured microbial communities, Whole Genome Shotgun (WGS) metagenomics made the functional profiling of microbial communities possible, since community aggregated traits can be quantified by the abundance of relevant marker genes. Considering a global metabolic process in an ecosystem, we propose a method that exploits gene abundance data to determine combinations of functional traits characterizing this process. When applied to fiber degradation in the human gut microbiota, the method shows that only four complex functional traits are needed to characterize this process. The approach is generic and could be applied to other processes or ecosystems. It allows summarizing complex datasets by a limited number of biologically interpretable functional patterns.

Background The Bacillus cereus Group consists of closely-related bacteria, including pathogenic or harmless strains, and whose species can be positioned along the seven phylogenetic groups of Guinebretière et al. (I–VII). They exhibit different growth-temperature ranges, through thermotolerant to psychrotolerant thermotypes. Among these, B. cytotoxicus is an atypical thermotolerant and food-poisoning agent affiliated to group VII whose thermotolerance contrasts with the mesophilic and psychrotolerant thermotypes associated to the remaining groups I–VI. To understand the role of fatty acid (FA) composition in these variable thermotypes (i.e. growth behavior vs temperatures), we report specific features differentiating the FA pattern of B. cytotoxicus (group VII) from its counterparts (groups I–VI). Findings The FA pattern of thermotolerant group VII (B. cytotoxicus) displayed several specific features. Most notably, we identified a high ratio of the branched-chain FAs iso-C15/iso-C13 (i15/i13) and the absence of the unsaturated FA (UFA) C16:1(5) consistent with the absence of ∆5 desaturase DesA. Conversely, phylogenetic groups II–VI were characterized by lower i15/i13 ratios and variable proportions of C16:1(5) depending on thermotype, and presence of the DesA desaturase. In mesophilic group I, thermotype seemed to be related to an atypically high amount of C16:1(10) that may involve ∆10 desaturase DesB. Conclusion The levels of i15/i13 ratio, C16:1(5) and C16:1(10) UFAs were related to growth temperature variations recorded between thermotypes and/or phylogenetic groups. These FA are likely to play a role in membrane fluidity and may account for the differences in temperature tolerance observed in B. cereus Group strains. Electronic supplementary material The online version of this article (doi:10.1186/s13104-015-1288-4) contains supplementary material, which is available to authorized users.

The behaviour of the sporulating soil-dwellingBacillus cereus sensu lato(B. cereus sl) which includes foodborne pathogenic strains has been extensively studied in relation to its various animal hosts. The aim of this environmental study was to investigate the water compartments (rain and soil water, as well as groundwater) closely linked to the primaryB. cereus slreservoir, for which available data are limited.B. cereus slwas present, primarily as spores, in all of the tested compartments of an agricultural site, including water from rain to groundwater through soil. During rain events, leachates collected after transfer through the soil eventually reached the groundwater and were loaded withB. cereus sl. In groundwater samples, newly introduced spores of aB. cereusmodel strain were able to germinate, and vegetative cells arising from this event were detected for up to 50 days. This firstB. cereus slinvestigation in the various types of interrelated environments suggests that the consideration of the aquatic compartment linked to soil and to climatic events should provide a better understanding ofB. cereus slecology and thus be relevant for a more accurate risk assessment of food poisoning caused byB. cereus slpathogenic strains.

Pas de clé UT; The large bacterial genus Bacillus is widely distributed in the environment and is able to colonize highly diverse niches. Some Bacillus species harbor pathogenic characteristics. The fatty acid (FA) composition is among the essential criteria used to define Bacillus species. Some elements of the FA pattern composition are common to Bacillus species, whereas others are specific and can be categorized in relation to the ecological niches of the species. Bacillus species are able to modify their FA patterns to adapt to a wide range of environmental changes, including changes in the growth medium, temperature, food processing conditions, and pH. Like many other Gram-positive bacteria, Bacillus strains display a well-defined FA synthesis II system that is equilibrated with a FA degradation pathway and regulated to efficiently respond to the needs of the cell. Like endogenous FAs, exogenous FAs may positively or negatively affect the survival of Bacillus vegetative cells and the spore germination ability in a given environment. Some of these exogenous FAs may provide a powerful strategy for preserving food against contamination by the Bacillus pathogenic strains responsible for foodborne illness.

This study aimed to evaluate molecular microbial markers associated with cocoa fermentation and drying steps. The dynamics of microbial populations were analyzed during postharvest processing (fermentation and sun drying) of cocoa from Ivory Coast. The variations in bacterial and yeast communities structure were monitored by a molecular technique (PCR-DGGE) using 16S rRNA and 26S rRNA profiles. Dominant microbial species were determined by sequencing. Our results showed that treatments of cocoa generated great variations in the microbial ecology in terms of number of species detected. The bacterial ecology quickly underwent dramatical changes during the fermentation step after two days. The dominance of Enterobacteria was first observed and then accompanied by Lactic acid bacteria and at the latest stages by acetic acid bacteria. Furthermore, Hanseniaspora opuntiae yeast species predominated during cocoa bean fermentation and drying processes. In addition, two bacterial species were only identified during the drying step of fermented cocoa beans and are potential candidates for biological markers of this process. (Resume d'auteur)

International audience; The Pseudomonas syringae complex is composed of numerous genetic lineages of strains from both agricultural and environmental habitats including habitats closely linked to the water cycle. The new insights from the discovery of this bacterial species in habitats outside of agricultural contexts per se have led to the revelation of a wide diversity of strains in this complex beyond what was known from agricultural contexts. Here, through Multi Locus Sequence Typing (MLST) of 216 strains, we identified 23 clades within 13 phylogroups among which the seven previously described P. syringae phylogroups were included. The phylogeny of the core genome of 29 strains representing nine phylogroups was similar to the phylogeny obtained with MLST thereby confirming the robustness of MLST-phylogroups. We show that phenotypic traits rarely provide a satisfactory means for classification of strains even if some combinations are highly probable in some phylogroups. We demonstrate that the citrate synthase (cts) housekeeping gene can accurately predict the phylogenetic affiliation for more than 97% of strains tested. We propose a list of cts sequences to be used as a simple tool for quickly and precisely classifying new strains. Finally, our analysis leads to predictions about the diversity of P. syringae that is yet to be discovered. We present here an expandable framework mainly based on cts genetic analysis into which more diversity can be integrated.

Diseases transmitted between wildlife and livestock may have significant impacts on local farmers’ health, livestock health and productivity, overall national economies, and conservation initiatives, such as Transfrontier Conservation Areas in Southern Africa. However, little is known on local farmers’ awareness of the potential risks, and how they perceive the role played by wildlife in the epidemiology of these diseases. We investigated the knowledge base regarding livestock diseases of local cattle owners living at the periphery of conservation areas within the Great Limpopo TFCA and the Kavango-Zambezi TFCA in Zimbabwe, using free-listing and semi-structured questionnaires during dipping sessions. The results suggest that information related to cattle diseases circulates widely between cattle farmers, including between different socio-cultural groups, using English and vernacular languages. Most respondents had an accurate perception of the epidemiology of diseases affecting their livestock, and their perception of the potential role played by wildlife species was usually in agreement with current state of veterinary knowledge. However, we found significant variations in the cultural importance of livestock diseases between sites, and owners’ perceptions were not directly related with the local abundance of wildlife. As the establishment of TFCAs will potentially increase the risk of Transboundary Animal Diseases, we recommend an increased participation of communities at a local level in the prioritisation of livestock diseases control and surveillance, including zoonoses.

National audience; Human, animal and plant viral diseases have greatly benefited from recent metagenomics developments. Viral metagenomics is a culture-independent approach used to investigate the complete viral genetic populations of a sample. During the last decade, metagenomics concepts and techniques that were first used by ecologists progressively spread into the scientific field of viral pathology. The sample, which was first for ecologists a fraction of ecosystem, became for pathologists an organism that hosts millions of microbes and viruses. This new approach, providing without a priori high resolution qualitative and quantitative data on the viral diversity, is now revolutionizing the way pathologists decipher viral diseases. This review describes the very last improvements of the high throughput next generation sequencing methods and discusses the applications of viral metagenomics in viral pathology, including discovery of novel viruses, viral surveillance and diagnostic, large-scale molecular epidemiology, and viral evolution.; L’étude des maladies virales humaines, animales et végétales a récemment bénéficié du développement de la métagénomique en écologie. Cette approche, dite sans a priori, consiste à décrire le génome viral total, ou virome, d’une fraction d’un écosystème sans tenir compte de cibles moléculaires déjà connues. Les premiers résultats de la métagénomique en pathologie virale ont été obtenus très localement au niveau d’un tissu ou d’un organe. Ils ont débouché sur la découverte de nouveaux virus et ont permis des avancées sur le plan du diagnostic, de l’épidémiologie moléculaire et de l’évolution virale. Ces travaux sont d’une grande pertinence pour réévaluer des concepts en pathologie et, notamment, le rôle biologique des virus dans un organisme. Ils révèlent la nécessité d’actualiser les méthodes de diagnostic.

In order to identify the function of uncultured microorganisms in their environment, the SIMSISH method, combining in situ hybridization (ISH) and nanoscale secondary ion mass spectrometry (nanoSIMS) imaging, has been proposed to determine the quantitative uptake of specific labelled substrates by uncultured microbes at the single cell level. This technique requires the hybridization of rRNA targeted halogenated DNA probes on fixed and permeabilized microorganisms. Exogenous atoms are introduced into cells and endogenous atoms removed during the experimental procedures. Consequently differences between the original and the apparent isotopic composition of cells may occur. In the present study, the influence of the experimental procedures of SIMSISH on the isotopic composition of carbon in E. coli cells was evaluated with nanoSIMS and compared to elemental analyser-isotopic ratio mass spectrometer (EA-IRMS) measurements. Our results show that fixation and hybridization have a very limited, reproducible and homogeneous influence on the isotopic composition of cells. Thereby, the SIMSISH procedure minimizes the contamination of the sample by exogenous atoms, thus providing a means to detect the phylogenetic identity and to measure precisely the carbon isotopic composition at the single cell level. This technique was successfully applied to a complex sample with double bromine - iodine labelling targeting a large group of bacteria and a specific archaea to evaluate their specific (13)C uptake during labelled methanol anaerobic degradation.

A real-time quantitative PCR method was developed to detect and quantify phenlylurea hydrolase genes’ (puhA and puhB) sequences from environmental DNA samples to assess diuron-degrading genetic potential in some soil and sediment microbial communities. In the soil communities, mineralization rates (determined with [ring-14C]-labeled diuron) were linked to diuron-degrading genetic potentials estimated from puhB number copies, which increased following repeated diuron treatments. In the sediment communities, mineralization potential did not depend solely on the quantity of puhB copies, underlining the need to assess gene expression. In the sediment samples, both puhB copy numbers and mineralization capacities were highly conditioned by whether or not diuron-treated soil was added. This points to transfers of degradative potential from soils to sediments. No puhA gene was detected in soil and sediment DNA extracts. Moreover, some sediments exhibited high diuron mineralization potential even though puhB genes were not detected, suggesting the existence of alternative diuron degradation pathways.

project SEST 009; International audience; Listeria monocytogenes is a ubiquitous opportunistic pathogen responsible for listeriosis. In order to study the processes underlying its ability to adapt to the soil environment, whole-genome arrays were used to analyse transcriptome modifications 15 minutes, 30 minutes and 18 h after inoculation of L. monocytogenes EGD-e in soil extracts. Growth was observed within the first day of incubation and large numbers were still detected in soil extract and soil microcosms one year after the start of the experiment. Major transcriptional reprofiling was observed. Nutrient acquisition mechanisms (phosphoenolpyruvate-dependent phosphotransferase systems and ABC transporters) and enzymes involved in catabolism of specific carbohydrates (b-glucosidases; chitinases) were prevalent. This is consistent with the overrepresentation of the CodY regulon that suggests that in a nutrient depleted environment, L. monocytogenes recruits its extensive repertoire of transporters to acquire a range of substrates for energy production.