Search

International audience; Metagenomics studies have revealed tremendous viral diversity in aquatic environments. Yet, while the genomic data they have provided is extensive, it is unannotated. For example, most phage sequences lack accurate information about their bacterial host, which prevents reliable phage identification and the investigation of phage–host interactions. This study aimed to take this knowledge further, using a viral metagenomic framework to decipher the composition and diversity of phage communities and to predict their bacterial hosts. To this end, we used water and sediment samples collected from seven sites with varying contamination levels in the Ebrié Lagoon in Abidjan, Ivory Coast. The bacterial communities were characterized using the 16S rRNA metabarcoding approach, and a framework was developed to investigate the virome datasets that: (1) identified phage contigs with VirSorter and VIBRANT; (2) classified these contigs with MetaPhinder using the phage database (taxonomic annotation); and (3) predicted the phages’ bacterial hosts with a machine learning-based tool: the Prokaryotic Virus-Host Predictor. The findings showed that the taxonomic profiles of phages and bacteria were specific to sediment or water samples. Phage sequences assigned to the Microviridae family were widespread in sediment samples, whereas phage sequences assigned to the Siphoviridae, Myoviridae and Podoviridae families were predominant in water samples. In terms of bacterial communities, the phyla Latescibacteria, Zixibacteria, Bacteroidetes, Acidobacteria, Calditrichaeota, Gemmatimonadetes, Cyanobacteria and Patescibacteria were most widespread in sediment samples, while the phyla Epsilonbacteraeota, Tenericutes, Margulisbacteria, Proteobacteria, Actinobacteria, Planctomycetes and Marinimicrobia were most prevalent in water samples. Significantly, the relative abundance of bacterial communities (at major phylum level) estimated by 16S rRNA metabarcoding and phage-host prediction were significantly similar. These results demonstrate the reliability of this novel approach for predicting the bacterial hosts of phages from shotgun metagenomic sequencing data.

Like other seafood products, tuna is highly perishable and sensitive to microbial spoilage. Its consumption, whether fresh or canned, can lead to severe food poisoning due to the activity of specific microorganisms, including histamine-producing bacteria. Yet, many grey areas persist regarding their ecology, conditions of emergence, and proliferation in fish. In this study, we used 16S rRNA barcoding to investigate postmortem changes in the bacteriome of fresh and brine-frozen yellowfin tuna (Thunnus albacares), until late stages of decomposition (i.e. 120 h). The results revealed that despite standard refrigeration storage conditions (i.e. 4°C), a diverse and complex spoilage bacteriome developed in the gut and liver. The relative abundance of spoilage bacterial taxa increased rapidly in both organs, representing 82% of the bacterial communities in fresh yellowfin tuna, and less than 30% in brine-frozen tuna. Photobacterium was identified as one of the dominant bacterial genera, and its temporal dynamics were positively correlated with histamine concentration in both gut and liver samples, which ultimately exceeded the recommended sanitary threshold of 50 ppm in edible parts of tuna. The results from this study show that the sanitary risks associated with the consumption of this widely eaten fish are strongly influenced by postcapture storage conditions.

Ecological traits of aquatic microorganisms have been poorly investigated in tropical latitudes, especially in lagoons, which are often subjected to strong anthropogenic influence, conducive to microbial development. In this study, we examined the abundance of both viral and bacterial communities, as well as their interactions (lytic and lysogenic infections) in the water and sediment of seven main stations of the Ebrié Lagoon (Ivory Coast) with contrasting levels of eutrophication. The highest bacterial and viral concentrations in both planktonic and benthic samples were found in the most eutrophicated stations, where viral lytic infections also exhibited their highest values. Conversely, the highest fractions of inducible lysogens were measured in the most oligotrophic stations, suggesting that these two main viral life strategies are mutually exclusive in this lagoon. Our findings also revealed the importance that nutrients (especially ammonium) play as drivers of the interactions between viruses and their bacterial hosts in tropical lagoons.

Although tunas represent a significant part of the global fish economy and a major nutritional resource worldwide, their microbiome still remains poorly documented. Here, we conducted an analysis of the taxonomic composition of the bacterial communities inhabiting the gut, skin, and liver of two most consumed tropical tuna species (skipjack and yellowfin), from individuals caught in the Atlantic and Indian oceans. We hypothesized that each organ harbors a specific microbial assemblage whose composition might vary according to different biotic (sex, species) and/or abiotic (environmental) factors. Our results revealed that the composition of the tuna microbiome was totally independent of fish sex, regardless of the species and ocean considered. Instead, the main determinants of observed diversity were (i) tuna species for the gut and (ii) sampling site for the skin mucus layer and (iii) a combination of both parameters for the liver. Interestingly, 4.5% of all amplicon sequence variants (ASV) were shared by the three organs, highlighting the presence of a core-microbiota whose most abundant representatives belonged to the genera Mycoplasma, Cutibacterium, and Photobacterium. Our study also revealed the presence of a unique and diversified bacterial assemblage within the tuna liver, comprising a substantial proportion of potential histamine-producing bacteria, well known for their pathogenicity and their contribution to fish poisoning cases. These results indicate that this organ is an unexplored microbial niche whose role in the health of both the host and consumers remains to be elucidated.

Off the coast of Vietnam, two recent events seem to have contributed to the death of coral reefs: the rapid invasion of a small marine snail called Drupella, and the emergence of a disease that spreads over corals. We decided to investigate whether there is a link between the corals, the snails, and the disease. We analyzed the mucus covering the surface of corals, both in healthy corals and those infested by Drupella snails. Mucus produced by corals is usually full of helpful microbes, protecting the corals from harmful microbes that can make them ill or kill them. Our analyses showed that, when Drupella eats corals, the snail also removes one of the corals’ primary defenses by eliminating most of the tiny protectors in their mucus. This allows harmful microbes to cause additional damage to the corals.

Background : Although tunas represent a significant part of the global fish economy and a major nutritional resource worldwide, their consumption poses a risk of food poisoning through the development of particular bacterial pathogens. However, their microbiome still remains poorly documented. Here, we conducted a multi-compartmental analysis of the taxonomic composition of the bacterial communities inhabiting the gut, skin and liver of two most consumed tropical tuna species (skipjack and yellowfin), from individuals caught in the Atlantic and Indian oceans. Results : Our results revealed that the composition of the microbiome was independent of fish sex, regardless of the species and ocean considered. Instead, the main determinants were (i) tuna species for the gut and(ii) sampling site for the skin mucus layer, and (iii) a combination of both parameters for the liver. Interestingly, only 4.5% of all ASVs were shared by the three compartments, raising numerous questions about the circulation of microorganisms within the tuna body. Our results also revealed the presence of a unique and diversified bacterial assemblage within the liver, comprising a substantial proportion of histamine-producing bacteria, well known for their potential pathogenicity and their contribution to fish poisoning cases. Conclusions : These results indicate that the tuna liver is an unexplored microbial niche whose role in the health of both the host and consumers remains to be elucidated.

WOS:000667043200001; International audience; Cortisol is recognized as a physiological indicator of stress in fish. However, this hormone is typically measured in plasma samples. In this study, cortisol content was evaluated for the first time in the scales of tropical tuna. The sample included 20 skipjack tuna (Katsuwonus pelamis) and 25 yellowfin tuna (Thunnus albacares) collected in the Atlantic (the Gulf of Guinea off the Ivory Coast) and the Indian Ocean (off Reunion Island), and their scales were analyzed using an ELISA test. The results indicated that on average, cortisol concentration (1) did not show any geographical pattern, (2) was independent of fish size, and (3) was significantly higher in skipjack (mean 4.75 ± 6.56 ng g−1) than in yellowfin (mean 1.65 ± 1.85 ng g−1), although the difference was mainly due to four skipjack individuals. Larger datasets would be needed to confirm any species difference in cortisol concentration. The particularly high cortisol concentration observed in four individuals may be due to fish from different schools, suggesting the need for future sampling from free-swimming schools to investigate intra- and inter-school variability in cortisol concentration. This study opens the door for future research, including the collection of scales on tagged individuals, to investigate the links between chronic stress and behavior in these species.

International audience; In recent years, a growing number of studies sought to examine the composition and the determinants of the gut microflora in marine animals, including fish. For tropical tuna, which are among the most consumed fish worldwide, there is scarce information on their enteric bacterial communities and how they evolve during fish growth. In this study, we used metabarcoding of the 16S rDNA gene to (1) describe the diversity and composition of the gut bacteriome in the three most fished tuna species (skipjack, yellowfin and bigeye), and (2) to examine its intra-specific variability from juveniles to larger adults. Although there was a remarkable convergence of taxonomic richness and bacterial composition between yellowfin and bigeyes tuna, the gut bacteriome of skipjack tuna was distinct from the other two species. Throughout fish growth, the enteric bacteriome of yellowfin and bigeyes also showed significant modifications, while that of skipjack tuna remained relatively homogeneous. Finally, our results suggest that the gut bacteriome of marine fish may not always be subject to structural modifications during their growth, especially in species that maintain a steady feeding behavior during their lifetime.

The basic bacterial ecology and diversity was investigated in five running water systems of Southern New Caledonia. These running waters were characterized by potential P-limitation and high concentrations of Ni, Fe, Mn, Cr and Co. The low concentrations of dissolved organic carbon, bacterial and viral abundance, bacterial production and growth efficiency support the characterization of the running waters as oligotroph to ultraoligotroph. Despite these similarities, there were strong differences (Betaproteobacteria is typical for freshwater systems, however, we found also a strong representation of Gammaproteobacteria. Indeed the three bands found at all stations were related to Limnohabitans (Comamonadaceae) and Alteromonadaceae. Strong differences were also found between the free-living and the attached bacterial fraction with Gammaproteobacteria dominating in two systems. A higher representation of Gammaproteobacteria seems typical for metal-rich freshwater habitats. Consistent with fresh water habitats, majority of phylotypes detected in the sediment was affiliated to proteobacteria. Also, none of the sequences showed a 100% identity with data bases, and 10 of the 22 and 2 of the 23 sequences had similarities higher than 97% in the freshwater and sediment. This could indicate specific adaptations of the community composition either due to the high metal concentrations or due to the geographical isolation of the New Caledonia.

International audience; Coral reefs host hundreds of thousands of animal species that are increasingly threatened by anthropogenic disturbances. These animals host microbial communities at their surface, playing crucial roles for their fitness. However, the diversity of such microbiomes is mostly described in a few coral species and still poorly defined in other invertebrates and vertebrates. Given the diversity of animal microbiomes, and the diversity of host species inhabiting coral reefs, the contribution of such microbiomes to the total microbial diversity of coral reefs could be important, yet potentially vulnerable to the loss of animal species. Analysis of the surface microbiome from 74 taxa, including teleost fishes, hard and soft corals, crustaceans, echinoderms, bivalves and sponges, revealed that more than 90% of their prokaryotic phylogenetic richness was specific and not recovered in surrounding plankton. Estimate of the total richness associated with coral reef animal surface microbiomes reached up to 2.5% of current estimates of Earth prokaryotic diversity. Therefore, coral reef animal surfaces should be recognized as a hotspot of marine microbial diversity. Loss of the most vulnerable reef animals expected under present-day scenarios of reef degradation would induce an erosion of 28% of the prokaryotic richness, with unknown consequences on coral reef ecosystem functioning.

International audience; The main objective of this pilot study, conducted in June 2015 in the Can Gio Mangrove Biosphere Reserve (Can Gio MBR, Viet Nam), was to develop an interdisciplinary approach to assess some key services provided by reforested mangroves subject to external pressures and varying management policies. We focused on the abundance of viruses, bacteria, endo-and epi-and macrofauna and the diversity of crabs in the mangrove and the exploitation of its resources. The main social finding was that the local inhabitants are aware of the levels of protection of the different zones within the Can Gio MBR and respect them. The core and the buffer zones seem to present a similar ecological status. Genotyping showed a low level of crab diversity although there were many different morphotypes. In the future, we need to understand the stakeholders' general perception of the biodiversity and environment changes by developing an integrated, multi-scale approach. (C) 2017 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved.

Microbial communities from hypersaline ponds, dominated by halophilic archaea, are considered specific of such extreme conditions. The associated viral communities have accordingly been shown to display specific features, such as similar morphologies among different sites. However, little is known about the genetic diversity of these halophilic viral communities across the Earth. Here, we studied viral communities in hypersaline ponds sampled on the coast of Senegal (8-36% of salinity) using metagenomics approach, and compared them with hypersaline viromes from Australia and Spain. The specificity of hyperhalophilic viruses could first be demonstrated at a community scale, salinity being a strong discriminating factor between communities. For the major viral group detected in all samples (Caudovirales), only a limited number of halophilic Caudovirales clades were highlighted. These clades gather viruses from different continents and display consistent genetic composition, indicating that they represent related lineages with a worldwide distribution. Non-tailed hyperhalophilic viruses display a greater rate of gene transfer and recombination, with uncharacterized genes conserved across different kind of viruses and plasmids. Thus, hypersaline viral communities around the world appear to form a genetically consistent community that are likely to harbour new genes coding for enzymes specifically adapted to these environments.

The factors controlling estuarine phytoplankton diversity and production are relatively well known in temperate systems. Less however is known about the factors affecting phytoplankton community distribution in tropical estuaries. This is surprising given the economic and ecological importance of these large, deltaic ecosystems, such as are found in South East Asia. Here we present the results from an investigation into the factors controlling phytoplankton distribution and phytoplankton-bacterial coupling in the Bach Dang Estuary, a sub-estuary of the Red River system, in Northern Vietnam. Phytoplankton diversity and primary and bacterial production, nutrients and metallic contaminants (mercury and organotin) were measured during two seasons: wet (July 2008) and dry (March 2009). Phytoplankton community composition differed between the two seasons with only a 2% similarity between July and March. The large spatial extent and complexity of defining the freshwater sources meant that simple mixing diagrams could not be used in this system. We therefore employed multivariate analyses to determine the factors influencing phytoplankton community structure. Salinity and suspended particulate matter were important factors in determining phytoplankton distribution, particularly during the wet season. We also show that phytoplankton community structure is probably influenced by the concentrations of mercury species (inorganic mercury and methyl mercury in both the particulate and dissolved phases) and of tri-, di, and mono-butyl tin species found in this system. Freshwater phytoplankton community composition was associated with dissolved methyl mercury and particulate inorganic mercury concentrations during the wet season, whereas, during the dry season, dissolved methyl mercury and particulate butyl tin species were important factors for the discrimination of the phytoplankton community structure. Phytoplankton-bacterioplankton coupling was also investigated during both seasons. In the inshore, riverine stations the ratio between bacterial production and dissolved primary production was high supporting the hypothesis that bacterial carbon demand is supported by allochthonous riverine carbon sources. The inverse was true in the offshore stations, where BP:DPP values were less than 1, potentially reflecting differences in primary production due to shifting phytoplankton community diversity.

International audience; The grazing activity by specific marine organisms represents a growing threat to the survival of many scleractinian species. For example, the recent proliferation of the corallivorous gastropod Drupella now constitutes a critical case in all South-East Asian waters. If the damaging effects caused by this marine snail on coral polyps are relatively well known, the indirect incidence of predation on coral microbial associates is still obscure and might also potentially impair coral health. In this study, we compared the main ecological traits of coral-associated bacterial and viral communities living in the mucus layer of Acropora formosa and Acropora millepora, of healthy and predated individuals (i.e., colonized by Drupella rugosa), in the Bay of Van Phong (Vietnam). Our results show a substantial impact of the gastropod on a variety of microbiological markers. Colonized corals harbored much more abundant and active epibiotic bacteria whose community composition shifted toward more pathogenic taxa (belonging to the Vibrionales, Clostridiales, Campylobacterales, and Alteromonadales orders), together with their specific phages. Viral epibionts were also greatly influenced by Drupella corallivory with spectacular modifications in their concentrations, life strategies, genotype richness, and diversity. Novel and abundant circular Rep-encoding ssDNA viruses (CRESS-DNA viruses) were detected and characterized in grazed corals and we propose that their occurrence may serve as indicator of the coral health status. Finally, our results reveal that corallivory can cause severe dysbiosis by altering virus-bacteria interactions in the mucus layer, and ultimately favoring the development of local opportunistic infections.

A recent hypothesis considers that many coral pathologies are the result of a sudden structural alteration of the epibiotic bacterial communities in response to environmental disturbances. However, the ecological mechanisms that lead to shifts in their composition are still unclear. In the ocean, viruses represent a major bactericidal agent but little is known on their occurrence within the coral holobiont. Recent reports have revealed that viruses are abundant and diversified within the coral mucus and therefore could be decisive for coral health. However, their mode of action is still unknown, and there is now an urgent need to shed light on the nature of the relationships they might have with the other prokaryotic and eukaryotic members of the holobiont. In this opinion letter, we are putting forward the hypothesis that coral-associated viruses (mostly bacterial and algal viruses), depending on the environmental conditions might either reinforce coral stability or conversely fasten their decline. We propose that these processes are presumably based on an environmentally driven shift in infection strategies allowing viruses to regulate, circumstantially, both coral symbionts (bacteria or Symbiodinium) and surrounding pathogens.

Summary Viruses attract increasing interest from environmental microbiologists seeking to understand their function and role in coral health. However, little is known about their main ecological traits within the coral holobiont. In this study, a quantitative and qualitative characterization of viral and bacterial communities was conducted on the mucus of seven different coral species of the Van Phong Bay (Vietnam). On average, the concentrations of viruses and bacteria were, respectively, 17- and twofold higher in the mucus than in the surrounding water. The examination of bacterial community composition also showed remarkable differences between mucus and water samples. The percentage of active respiring cells was nearly threefold higher in mucus (m = 24.8%) than in water (m = 8.6%). Interestingly, a positive and highly significant correlation was observed between the proportion of active cells and viral abundance in the mucus, suggesting that the metabolism of the bacterial associates is probably a strong determinant of the distribution of viruses within the coral holobiont. Overall, coral mucus, given its unique physicochemical characteristics and sticking properties, can be regarded as a highly selective biotope for abundant, diversified and specialized symbiotic microbial and viral organisms.

International audience; Large amounts of soot are continuously deposited on the global ocean. Even though significant concentrations of soot particles are found in marine waters, the effects of these aerosols on ocean ecosystems are currently unknown. Using a combination of in situ and experimental data, and results from an atmospheric transport model, we show that the deposition of soot particles from an oil-fired power plant impacted biogeochemical properties and the functioning of the pelagic ecosystem in tropical oligotrophic oceanic waters off New Caledonia. Deposition was followed by a major increase in the volume concentration of suspended particles, a change in the particle size spectra that resulted from a stimulation of aggregation processes, a 5% decrease in the concentration of dissolved organic carbon (DOC), a decreases of 33 and 23% in viral and free bacterial abundances, respectively, and a factor~2 increase in the activity of particle-attached bacteria suggesting that soot introduced in the system favored bacterial growth. These patterns were confirmed by experiments with natural seawater conducted with both soot aerosols collected in the study area and standard diesel soot. The data suggest a strong impact of soot deposition on ocean surface particles, DOC, and microbial processes, at least near emission hot spots.

International audience; The direct counts of bacteria inhabiting coral mucus were performed by flow cytometry testing four fluorescent dyes (SYBR®Green I, HCS, TOPRO®3, SYTO®62) with three different scleractinian species. Results obtained with SYTO62 were the most reliable based on the comparison with standardized epifluorescence counts and the resolution of cytograms.

Since the discovery, 2–3 decades ago, that viruses of microbes are abundant in marine ecosystems, aquatic viral ecology has grown increasingly to reach the status of a full scientific discipline in environmental sciences. A dedicated society, the International Society for Viruses of Microorganisms (ISVM) (http://www.isvm.org/), was recently launched. Increasing studies in aquatic viral ecology are a source of novel knowledge related to the biodiversity, the functioning of ecosystems and the evolution of the cellular world. This is because viruses are perhaps the most diverse, abundant, and ubiquitous biological entities in the world’s aquatic ecosystems. They exhibit various lifestyles that intimately depend on living cell metabolism, and are ultimately replicated by members of all three domains of life (Bacteria, Eukarya, Archaea). This establishes viruses as microbial killers and manipulators in the hydrosphere. Lake Pavin is one of the pioneer sites where original ecological data were first provided on the qualitative, quantitative and functional significance of both lytic and temperate viruses of prokaryotes in temperate freshwater lakes, taking into account both seasonal and depth-related variability in the water column and in the sediments. These data were acquired by means of original protocols we developed. In addition to these protocols, we herein provide a synthesis of Lake Pavin studies on viral ecology, focusing on: (i) spatio-temporal dynamics of the diversity of viral communities, (ii) the significance of seasonal and depth-related variations of viral abundance and lytic and lysogenic activities, and (iii) the relative importance of lytic viruses and grazers for bacterial mortality and the biogeochemical implications for the food web dynamics. Unexpected and novel putative viruses were discovered in the deep-aged, dark, and permanently anoxic monimolimnic waters and sediments of Lake Pavin, highlighting the possible endemicity of these habitats. Some of these original viruses resembled dsDNA viruses of hyperthermophilic and hyperhalophilic Archaea. Unusual types of spherical and cubic virus-like particles (VLPs) were also observed for the first time. Infected prokaryotic cells were detected in deep sediment cores, and their vertical distribution correlated with both viral and prokaryotic abundances. Pleomorphic ellipsoid VLPs were visible in filamentous cells tentatively identified as representatives of the archaeal genus Methanosaeta, a major group of methane producers on Earth.

We describe a new method to produce marine aggregates from natural organic material based on the sticking properties of transparent exopolymeric particles. Seawater samples were prescreened and ultrafiltered to concentrate the 30 kDa to 10 µm size fraction. First, we produced small magnetizable aggregates by combining glass microfibers and paramagnetic 1-µm beads with the organic matter present in the concentrated solution. The second step involved clustering the small aggregates into a single macro-aggregate, using a small ringshaped magnet as an aggregation nucleus. Viral and bacterial densities, determined after dissolution of the newly formed aggregates with methanol, averaged 13.8 × 106 ± 3.6 × 106 vir. agg.−1 and 4.1 × 106 ± 1.1 × 106 bact. agg.−1. Bacterial respiration and production measurements of single aggregates averaged 8.47 ± 1.72 nmol O2 agg.−1 h−1 and 1.54 ± 0.45 ng C agg.−1 h−1, respectively. Particulate organic carbon and nitrogen content of the newly formed macro-aggregates averaged 31.92 ± 2.67 µg C agg.−1 and 3.44 ± 0.43 µg N agg.−1, respectively. This approach allows the concentration and isolation of the organic matter precursors that compose natural aggregates and provides a simple protocol for recombining those precursors into single newly formed macro-aggregates, which can then be easily manipulated for further investigation. This method is a new tool for investigations into the interactions between microorganisms and marine aggregates and their implications at the ecosystem level, but also into the interactions between aggregates and dissolved organic or inorganic substances.

International audience; Size fractionation was performed using water from the Great Reef of Toliara (Madagascar) taken from two different habitats (ocean and lagoon) during the dry and wet seasons, to study the growth and mortality rates of bacterioplankton. Experiments were conducted with 1 and 100% of heterotrophic nanoflagellate (HNF) concentrations and virus-free water was obtained by tangential filtration (10 kDa). During the dry season, in both environments, bacterial abundance and production were significantly lower than values recorded during the wet season. Bacterial growth rates without grazers were 0.88 day–1 in the lagoon and 0.58 day–1 in the ocean. However, growth rates were statistically higher without grazers and viruses (1.58 day–1 and 1.27 day–1). An estimate of virus-induced bacterial mortality revealed the important role played by viruses in the lagoon (0.70 day–1) and the ocean (0.69 day–1). During the wet season, bacterial growth rates without grazers were significantly higher in both environments than were values obtained in the dry season. However, the bacterial growth rates were paradoxally lower in the absence of viruses than with viruses in both environments. Our results suggest that changes in nutrient concentrations can play an important role in the balance between viral lysis and HNF grazing in the bacterial mortality. However, virus-mediated bacterial mortality is likely to act simultaneously with nanoflagellates pressure in their effects on bacterial communities.

Population dynamics in the microbial food web are influenced by resource availability and predator/parasitism activities. Climatic changes, such as an increase in temperature and/or UV radiation, can also modify ecological systems in many ways. A series of enclosure experiments was conducted using natural microbial communities from a Mediterranean lagoon to assess the response of microbial communities to top-down control [grazing by heterotrophic nanoflagellates (HNF), viral lysis] and bottom-up control (nutrients) under various simulated climatic conditions (temperature and UV-B radiations). Different biological assemblages were obtained by separating bacteria and viruses from HNF by size fractionation which were then incubated in whirl-Pak bags exposed to an increase of 3°C and 20% UV-B above the control conditions for 96 h. The assemblages were also provided with an inorganic and organic nutrient supply. The data show (i) a clear nutrient limitation of bacterial growth under all simulated climatic conditions in the absence of HNF, (ii) a great impact of HNF grazing on bacteria irrespective of the nutrient conditions and the simulated climatic conditions, (iii) a significant decrease in burst size (BS) (number of intracellular lytic viruses per bacterium) and a significant increase of VBR (virus to bacterium ratio) in the presence of HNF, and (iv) a much larger temperature effect than UV-B radiation effect on the bacterial dynamics. These results show that top-down factors, essentially HNF grazing, control the dynamics of the lagoon bacterioplankton assemblage and that short-term simulated climate changes are only a secondary effect controlling microbial processes.

Virus-prokaryote interactions were investigated in four natural sites in Senegal (West Africa) covering a salinity gradient ranging from brackish (10‰) to near salt saturation (360‰). Both the viral and the prokaryote communities exhibited remarkable differences in their physiological, ecological and morphological traits along the gradient. Above 240‰ salinity, viral and prokaryotic abundance increased considerably with the emergence of (1) highly active square haloarchaea and of (2) viral particles with pleiomorphic morphologies (predominantly spindle, spherical and linear shaped). Viral life strategies also showed some salinity-driven dependence, switching from a prevalence of lytic to lysogenic modes of infection at the highest salinities. Interestingly, the fraction of lysogenized cells was positively correlated with the proportion of square cells. Overall, the extraordinary abundance of viruses in hypersaline systems (up to 6.8 × 10(8) virus-like particles per milliliter) appears to be partly explained by their high stability and specific ability to persist and proliferate in these apparently restrictive habitats.

12 pages, 4 figures, 3 tables, Previous studies indicate that lysogeny is preponderant when environmental conditions are challenging for the bacterial communities and when their metabolism is reduced. Furthermore, it appears that lysogeny is more frequent within certain bacterial phylogenetic groups. In this comparative study from 10 freshwater reservoirs and 10 coastal lagoons, we aim to disentangle the influence of these different factors. In eight reservoirs and four lagoons, lysogeny was detected by induction assays with mitomycin C, and induction significantly modified the bacterial community composition (BCC), whereas community composition remained constant in ecosystems in which lysogeny was not observed. Among the phylogenetic groups studied, the most abundant ones were Bacteroidetes and α-proteobacteria in lagoons, and β-proteobacteria and Bacteroidetes in reservoirs. These dominant groups comprised the highest proportions of inducible lysogens. In order to unravel the effects of bacterial metabolism from phylogeny on lysogeny, we measured bacterial community physiology and the specific activities of selected phylogenetic groups. The proportion of inducible lysogens within the α- and the β-proteobacteria decreased with increasing group-specific metabolism in lagoons and reservoirs, respectively. In contrast, this relationship was not observed for the other lysogen-containing groups. Hence, both host physiology and phylogeny are critical for the establishment of lysogeny. This study illustrates the importance of lysogeny among the most abundant phylogenetic groups, and further suggests its strong structuring impact on BCC, This work was supported by the French ANR Aquaphage 07 BDIV 015–02 (to TB and YB). Work of HS was supported by the Spanish MEC (SB2006–0060), MCyI (Juan de la Cierva Fellowship JCI-2008–2727) and Portuguese FCT do Ministério da Ciência, Tecnologia e Ensino Superior’ (Grant SFRH/BPD/34041/2006)

Summary Remarkable morphological diversity of virus-like par- ticles was observed by transmission electron micros- copy in a hypersaline water sample from Lake Retba, Senegal. The majority of particles morphologically resembled hyperthermophilic archaeal DNA viruses isolated from extreme geothermal environments. Some hypersaline viral morphotypes have not been previously observed in nature, and less than 1% of observed particles had a head-and-tail morphology, which is typical for bacterial DNA viruses. Culture- independent analysis of the microbial diversity in the sample suggested the dominance of extremely halophilic archaea. Few of the 16S sequences corre- sponded to known archeal genera (Haloquadratum, Halorubrum and Natronomonas), whereas the major- ity represented novel archaeal clades. Three sequences corresponded to a new basal lineage of the haloarchaea. Bacteria belonged to four major phyla, consistent with the known diversity in saline environments. Metagenomic sequencing of DNA from the purified virus-like particles revealed very few similarities to the NCBI non-redundant database at either the nucleotide or amino acid level. Some of the identifiable virus sequences were most similar to previously described haloarchaeal viruses, but no sequence similarities were found to archaeal viruses from extreme geothermal environments. A large pro- portion of the sequences had similarity to previously sequenced viral metagenomes from solar salterns.

Little is known about the ability of phages to successfully colonize contrasting aquatic niches. We conducted experimental cross-infections between viruses and prokaryotes from three tropical sites of West Africa, with distinct salinities: a freshwater reservoir, a marine coastal station and a hypersaline lake. A cellular poison-based method (potassium cyanide) revealed that the addition of native viruses (regardless of the water type) consistently stimulated viral production. Conversely, in all incubations conducted with allochtonous (non-native) viruses, their overall production was not promoted, which suggests a lytic failure. Prokaryotic heterotrophic production increased in fresh and marine water supplemented with native viruses, but not in the hypersaline water. These results point to the role of the viral shunt in low-salinity environments, where the release of bioavailable lysis products might be of high nutritional value for the noninfected prokaryotes. In contrast, in hypersaline water where glycerol is a major carbon and energy source for the heterotrophic community, dissolved organic matter (DOM) of lytic origin may represent a less important DOM source for prokaryotes. Finally, our results suggest that cosmopolitan phages capable of moving between biomes are probably rare in aquatic habitats, supporting the common idea that most wild phages are relatively limited in their host range.

International audience; Viral abundance and processes in the water column and sediments are well studied for some systems; however, we know relatively little about virus–host interactions on particles and how particles influence these interactions. Here we review virus–prokaryote interactions on inorganic and organic particles in the water column. Profiting from recent methodological progress, we show that confocal laser scanning microscopy in combination with lectin and nucleic acid staining is one of the most powerful methods to visualize the distribution of viruses and their hosts on particles such as organic aggregates. Viral abundance on suspended matter ranges from 105 to 1011 ml–1. The main factors controlling viral abundance are the quality, size and age of aggregates and the exposure time of viruses to aggregates. Other factors such as water residence time likely act indirectly. Overall, aggregates appear to play a role of viral scavengers or reservoirs rather than viral factories. Adsorption of viruses to organic aggregates or inorganic particles can stimulate growth of the free-living prokaryotic community, e.g. by reducing viral lysis. Such mechanisms can affect microbial diversity, food web structure and biogeochemical cycles. Viral lysis of bacterio- and phytoplankton influences the formation and fate of aggregates and can, for example, result in a higher stability of algal flocs. Thus, viruses also influence carbon export; however, it is still not clear whether they short-circuit or prime the biological pump. Throughout this review, emphasis has been placed on defining general problems and knowledge gaps in virus–particle interactions and on providing avenues for further research, particularly those linked to global change.

Virucidal properties of sunlight and temperature have been identified for a long time. However, it is less well established if virioplankton communities are evenly susceptible or geographically adapted to these environmental factors. Transplant experiments were conducted between a tropical (Senegal) and temperate (France) site to examine the effect of natural daylight conditions and temperature on the persistence of free viruses in water. Fresh- and seawater viruses from both countries were simultaneously subjected to 12 h of full sunlight and dark exposure under their native and "transplanted" latitudes. Consistently, viruses decayed much faster when incubated abroad, regardless of origin (latitude and/or water type). However, viral losses, in most cases, were not significantly different between sunlight exposed and dark samples, implying that ambient radiation does not exert strong negative effects on virioplankton particles. Rather, temperature clearly emerged as the principal determinant Of viral survival in all samples. This suggests that viruses, under local temperature, can adapt through an evolutionary process to survive for longer periods of time thus increasing their chances to encounter their hosts.

There is increasing suspicion that viral communities play a pivotal role in maintaining coral health, yet their main ecological traits still remain poorly characterized. In this study, we examined the seasonal distribution and reproduction pathways of viruses inhabiting the mucus of the scleractinians Fungia repanda and Acropora formosa collected in Nha Trang Bay (Vietnam) during an 11-month survey. The strong coupling between epibiotic viral and bacterial abundance suggested that phages are dominant among coral-associated viral communities. Mucosal viruses also exhibited significant differences in their main features between the two coral species and were also remarkably contrasted with their planktonic counterparts. For example, their abundance (inferred from epifluorescence counts), lytic production rates (KCN incubations), and the proportion of lysogenic cells (mitomycin C inductions) were, respectively, 2.6-, 9.5-, and 2.2-fold higher in mucus than in the surrounding water. Both lytic and lysogenic indicators were tightly coupled with temperature and salinity, suggesting that the life strategy of viral epibionts is strongly dependent upon environmental circumstances. Finally, our results suggest that coral mucus may represent a highly favorable habitat for viral proliferation, promoting the development of both temperate and virulent phages. Here, we discuss how such an optimized viral arsenal could be crucial for coral viability by presumably forging complex links with both symbiotic and adjacent nonsymbiotic microorganisms.

International audience; Animal-associated microbiota of animals form complex communities, which are suspected to play crucial functions for their host fitness. However, the biodiversity of these communities, including their differences between host species and individuals, have been scarcely studied, especially in case of skin-associated communities. In addition, the intra-individual variability (i.e. between body parts) has never been assessed to date. The objective of this study was to characterize skin bacterial communities of two teleostean fish species, namely the European seabass (Dicentrarchus labrax) and gilthead seabream (Sparus aurata), using a high-throughput DNA sequencing method. In order to focus on intrinsic factors of host-associated bacterial community variability, individuals of the two species were raised in controlled conditions. Bacterial diversity was assessed using a set of four complementary indices, describing the taxonomic and phylogenetic facets of biodiversity and their respective composition (based on presence/absence data) and structure (based on species relative abundances) components. Variability of bacterial diversity was quantified at the interspecific, inter- and intra-individual scales. We demonstrated that fish surfaces host highly diverse bacterial communities, whose composition was very different from that of surrounding bacterioplankton. This high total biodiversity of skin-associated communities was supported by the important variability, between host species, individuals and the different body parts (dorsal, anal, pectoral and caudal fins).

Viruses attract increasing interest from environmental microbiologists seeking to understand their function and role in coral health. However, little is known about their main ecological traits within the coral holobiont. In this study, a quantitative and qualitative characterization of viral and bacterial communities was conducted on the mucus of seven different coral species of the Van Phong Bay (Vietnam). On average, the concentrations of viruses and bacteria were, respectively, 17- and twofold higher in the mucus than in the surrounding water. The examination of bacterial community composition also showed remarkable differences between mucus and water samples. The percentage of active respiring cells was nearly threefold higher in mucus (m = 24.8%) than in water (m = 8.6%). Interestingly, a positive and highly significant correlation was observed between the proportion of active cells and viral abundance in the mucus, suggesting that the metabolism of the bacterial associates is probably a strong determinant of the distribution of viruses within the coral holobiont. Overall, coral mucus, given its unique physicochemical characteristics and sticking properties, can be regarded as a highly selective biotope for abundant, diversified and specialized symbiotic microbial and viral organisms.

International audience; The periodic rise in seawater temperature is one of the main environmental determinants of coral bleaching. However, the direct incidence of these episodic thermal anomalies on coral-associated microbiota and their subsequent effects on coral health are still not completely understood. In this study, we investigated the dynamics of three main microbial communities of the coral holobiont (e.g., Symbiodinium, bacteria and viruses), during an experimental thermal stress (+4 °C) conducted on the scleractinian Fungia repanda. The heat-treatment induced coral bleaching after 11 days and resulted in a final elevation of ca. 9, 130 and 250-fold in the abundance of mucosal viruses, bacteria, and Symbiodinium, respectively. On the contrary, the proportion of actively respiring bacterial cells declined by 95% in heat-stressed corals. The community composition of epibiotic bacteria in healthy corals also greatly differed from bleached ones, which also exhibited much higher production rates of viral epibionts. Overall, our results suggest that the shift in temperature induced a series of microbial changes, including the expulsion and transfer of Symbiodinium cells from the coral polyps to the mucus, the collapse of the physiological state of the native bacterial associates, a substantial alteration in their community structure, and accompanied by the development of a cortege of highly active virulent phages. Finally, this study provides new insights into the environmentally-driven microbial and viral processes responsible for the dislocation of the coral holobiont.

International audience; Yeast cells transformed with high-copy number plasmids comprising a green fluorescent protein (GFP)-encoding gene optimized for yeast under the control of the new DIN7 or PLM2 and the established RNR2 and RAD54 promoters were used to assess the genotoxic potential of chemical compounds. The activity of potential DNA-damaging agents was investigated by genotoxicity assays and by OxoPlate assay in the presence of various test compounds. The fluorescence signal generated by GFP in response to DNA damage was related to the different concentrations of analytes and the analyte-dependent GFP synthesis. The use of distinct DNA damage-inducible promoters presents alternative genotoxicity testing strategies by selective induction of promoters in response to DNA damage. The new DIN7 and PLM2 systems show higher sensitivity than the RNR2 and RAD54 systems in detecting 4-nitroquinoline-N-oxide and actinomycin D. Both DIN7 and PLM2 systems are able to detect camptothecin while RNR2 and RAD54 systems are not. Automated laboratory systems with assay performance on 384-well microplates provide for cost-effective high-throughput screening of DNA-damaging agents, reducing compound consumption to about 53% as compared with existing eukaryotic genotoxicity bioassays.

Phytoplankton diversity and abundance in estuarine systems are controlled by many factors. Salinity, turbidity, and inorganic nutrient concentrations and their respective ratios have all been proposed as principal factors that structure phytoplankton diversity and influence the emergence of potentially toxic species. Although much work has been conducted on temperate estuaries, less is known about how phytoplankton diversity is controlled in tropical, monsoonal systems that are subject to large, seasonal shifts in hydrology and to rapidly changing land use. Here, we present the results of an investigation into the factors controlling phytoplankton species composition and distribution in a tropical, monsoonal estuary (Bach Dang estuary, North Vietnam). A total of 245 taxa, 89 genera from six algal divisions were observed. Bacillariophyceae were the most diverse group contributing to 51.4 % of the microalgal assemblage, followed by Dinophyceae (29.8 %), Chlorophyceae (10.2 %), Cyanophyceae (3.7 %), Euglenophyceae (3.7 %) and Dictyochophyceae (1.2 %). The phytoplankton community was structured by inorganic nutrient ratios (DSi:DIP and DIN:DIP) as well as by salinity and turbidity. Evidence of a decrease in phytoplankton diversity concomitant with an increase in abundance and dominance of certain species (e.g., Skeletonema costatum) and the appearance of some potentially toxic species over the last two decades was also found. These changes in phytoplankton diversity are probably due to a combination of land use change resulting in changes in nutrient ratios and concentrations and global change as both rainfall and temperature have increased over the last two decades. It is therefore probable in the future that phytoplankton diversity will continue to change, potentially favoring the emergence of toxic species in this system.

A novel virus (Chaetoceros nuclear inclusion virus: CspNIV) causing lysis of a culture of the diatom Chaetoceros cf. gracilis was isolated from the Chesapeake Bay, USA, in April 2003. Trans- mission electron microscopy of ultrathin sections of infected C. cf. gracilis revealed that CspNIV pro- liferates within the nucleus and forms paracrystalline arrays corresponding to the alignment of icosahedral viral particles of about 25 nm diameter. CspNIV shows some strong similarities with Heterosigma akashiwo nuclear inclusion virus (HaNIV) (cf. Lawrence et al. 2001; J Phycol 37: 216-222). The latent period of CspNIV is

1. The seasonal and vertical distribution of the abundance of virus-like particles (VLPs), together with the abundances of other microbial organisms (bacteria, unpigmented and pigmented nanoflagellates and ciliates), were determined in an oligomesotrophic lake (Pavin, France) and in a eutrophic lake (Aydat, France) between March and December 2000. 2. The abundance of the viral plankton and those of other microbial taxa were significantly higher in the more productive system. The same was for the virus-to-bacteria quotient (VBQ), which averaged seven in Lake Pavin and nine in Lake Aydat. 3. The abundance of viruses increased during the period of thermal stratification in both lakes, with the highest values being recorded at the end of summer/early autumn in the epi- and the metalimnion. The seasonal pattern of abundance of viruses in both lakes in the surface layer was similar, indicating that the dynamics of viruses may be controlled by environmental factors such as light conditions. 4. There was no correlation between the abundance of viruses and protists. We found correlations between viruses and heterotrophic bacteria in the whole water column in Lake Pavin, but only in the dark bottom waters in Lake Aydat. 5. Overall, the empirical findings in this study lead us to speculate that the weaker correlation between bacteria and viruses in Lake Aydat than in Lake Pavin, as well as the higher VBQ in the former, is a consequence of the increasing relative abundance of non-bacteriophage VLPs along the trophic gradient of aquatic systems.