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Viruses are ubiquitous and abundant in the oceans, and viral metagenomes (viromes) have been investigated extensively via several large-scale ocean sequencing projects. However, there have not been any systematic viromic studies in estuaries. Here, we investigated the viromes of the Delaware Bay and Chesapeake Bay, two Mid-Atlantic estuaries. Deep sequencing generated a total of 48,190 assembled viral sequences (>5 kb) and 26,487 viral populations (9,204 virus clusters and 17,845 singletons), including 319 circular viral contigs between 7.5 kb and 161.8 kb. Unknown viruses represented the vast majority of the dominant populations, while the composition of known viruses, such as pelagiphage and cyanophage, appeared to be relatively consistent across a wide range of salinity gradients and in different seasons. A difference between estuarine and ocean viromes was reflected by the proportions of Myoviridae, Podoviridae, Siphoviridae, Phycodnaviridae, and a few well-studied virus representatives. The difference in viral community between the Delaware Bay and Chesapeake Bay is significantly more pronounced than the difference caused by temperature or salinity, indicating strong local profiles caused by the unique ecology of each estuary. Interestingly, a viral contig similar to phages infecting Acinetobacter baumannii (“Iraqibacter”) was found to be highly abundant in the Delaware Bay but not in the Chesapeake Bay, the source of which is yet to be identified. Highly abundant viruses in both estuaries have close hits to viral sequences derived from the marine single-cell genomes or long-read single-molecule sequencing, suggesting that important viruses are still waiting to be discovered in the estuarine environment. IMPORTANCE This is the first systematic study about spatial and temporal variation of virioplankton communities in estuaries using deep metagenomics sequencing. It is among the highest-quality viromic data sets to date, showing remarkably consistent sequencing depth and quality across samples. Our results indicate that there exists a large pool of abundant and diverse viruses in estuaries that have not yet been cultivated, their genomes only available thanks to single-cell genomics or single-molecule sequencing, demonstrating the importance of these methods for viral discovery. The spatiotemporal pattern of these abundant uncultivated viruses is more variable than that of cultured viruses. Despite strong environmental gradients, season and location had surprisingly little impact on the viral community within an estuary, but we saw a significant distinction between the two estuaries and also between estuarine and open ocean viromes.

International audience; The impact of fish and oysters on components of the pelagic microbial food web (MFW) was studied in a 10 d mesocosm experiment using Mediterranean coastal waters. Two mesocosms contained natural water only (‘Controls’), 2 contained natural water with Crassostrea gigas (‘Oyster’), and 2 contained natural water with Atherina spp. (‘Fish’). Abundances and biomasses of microorganisms (viruses, bacteria, phytoplankton, heterotrophic flagellates, and ciliates) were measured to estimate their contribution to the total microbial carbon biomass. Two MFW indices, the microbial autotroph:heterotroph C biomass ratio (A:H) structural index and the gross primary production:respiration ratio (GPP:R) functional index, were defined. In the Fish mesocosms, selective predation on zooplankton led to a trophic cascade with 51% higher phyto- plankton C biomass and consequently higher A:H and GPP:R than in the Controls. By the end of the experiment, the Oyster mesocosms had a bacterial C biomass 87% higher and phytoplankton C biomass 93% lower than the Controls, giving significantly lower A:H and GPP:R (1), whereas oyster activities made the MFW more hetero- trophic (both indices

Temporal and spatial changes of virioplankton abundance (VLP), chlorophyll-a (Chl-a) concentration, and some environmental variables, were assessed from October 2004 to April 2005 at four sampling sites in a high-mountain reservoir in the Colombian Andes. VLP ranged from 2.4-10.5×10 7 and 3.6- 6.5×10 7 VLP mL −1 in the samples from the photic zone and hypolimnion, respectively. Surface VLP showed a progressive increase from November to April in the limnetic zone, and until February in the littoral stations. This trend coincided with the gradual increase of the water column stratification, as well as the augment of the reservoir hydraulic volume. Principal components analysis showed a grouping of environmental (dissolved oxygen, pH, water temperature) and biological variables (VLP, Chl-a, bacterioplankton abundance and biomass) within the first component (26.4% of explained variance), associated to an increase of photosynthetic activity, as a potential supplement of organic substrates to heterotrophic viral hosts. High values of virus-to-prokaryote ratio (VPR), and a negative relationship between VLP and average biomass per prokaryotic cell, indicate strong viral control which is probably more intense on active and larger cells. These results provide a framework for understand the virioplankton responses to regional hydrological conditions and hydraulic behavior of this reservoir. The spatiotemporal scale of this study does not allow to confirm that viral dynamics is significantly affected by human activities causing potential alterations on nutrient cycling.

Here we present a comprehensive review of the diversity revealed by research in limnology and microbial ecology conducted in Byers Peninsula (Livingston Island, South Shetland Islands, Antarctica) during the last two decades. The site constitutes one of the largest ice-free areas within the Antarctic Peninsula region. Since it has a high level of environmental protection, it is less human-impacted compared to other sites within the South Shetland archipelago. The main investigations in Byers Peninsula focused on the physical and chemical limnology of the lakes, ponds, rivers, and wetlands, as well as on the structure of their planktonic and benthic microbial communities, and on the functional ecology of the microbial food webs. Lakes and ponds in Byers range along a productivity gradient that extends from the less productive lakes located upland to the eutrophic coastal lakes. Their planktonic assemblages include viruses, bacteria, a metabolically diverse community of protists (i.e., autotrophs, heterotrophs, and mixotrophs), and a few metazooplankton species. Most of the studies conducted in the site demonstrate the strong influence of the physical environment (i.e., temperature, availability of light, and water) and nutrient availability in structuring these microbial communities. However, top-down biotic processes may occur in summer, when predation by zooplankton can exert a strong influence on the abundance of protists, including flagellates and ciliated protozoa. As a consequence, bacterioplankton could be partly released from the grazing pressure exerted by these protists, and proliferates fueled by external nutrient subsidies from the lake’s catchment. As summer temperatures in this region are slightly above the melting point of water, biotic processes, such as those related to the productivity of lakes during ice-free periods, could become even more relevant as warming induced by climate change progresses. The limnological research carried out at the site proves that Byers Peninsula deserves special attention in the framework of the research in extreme environments. Together with nearby sites, such as Signy Island, Byers Peninsula comprises a featuring element of the Maritime Antarctic region that represents a benchmark area relative to the global distribution and diversity of aquatic microorganisms.

The microbial community inhabiting Sfax solar salterns on the east coast of Tunisia has been studied by means of different molecular and culture-dependent tools that have unveiled the presence of novel microbial groups as well as a community structure different from that of other coastal hypersaline environments. We have focused on the study of the viral assemblages of these salterns and their changes along the salinity gradient and over time. Viruses from three ponds (C4, M1, and TS) encompassing salinities from moderately hypersaline to saturated (around 14, 19, and 35%, respectively) were sampled in May and October 2009 and analyzed by transmission electron microscopy (TEM) and pulsed-field gel electrophoresis (PFGE). Additionally, for all three October samples and the May TS sample, viral metagenomic DNA was cloned in fosmids, end sequenced, and analyzed. Viral concentration, as well as virus-to-cell ratios, increased along the salinity gradient, with around 1010 virus-like particles (VLPs)/ml in close-to-saturation ponds, which represents the highest viral concentration reported so far for aquatic systems. Four distinct morphologies could be observed with TEM (spherical, tailed, spindled, and filamentous) but with various proportions in the different samples. Metagenomic analyses indicated that every pond harbored a distinct viral assemblage whose G+C content could be roughly correlated with that of the active part of the microbial community that may have constituted the putative hosts. As previously reported for hypersaline metaviromes, most sequences did not have matches in the databases, although some were conserved among the Sfax metaviromes. BLASTx, BLASTp, and dinucleotide frequency analyses indicated that (i) factors additional to salinity could be structuring viral communities and (ii) every metavirome had unique gene contents and dinucleotide frequencies. Comparison with hypersaline metaviromes available in the databases indicated that the viral assemblages present in close-to-saturation environments located thousands of kilometers apart presented some common traits among them in spite of their differences regarding the putative hosts. A small core metavirome for close-to-saturation systems was found that contained 7 sequences of around 100 nucleotides (nt) whose function was not hinted at by in silico search results, although it most likely represents properties essential for hyperhalophilic viruses. This work was funded by projects A/018342/08 from the Spanish Agency for International Cooperation (AECI) and CGL2009-12651-C02-01 from the Spanish Ministry of Science and Innovation (MICINN), including FEDER funds.

International audience; This study presents an original depth-related survey of virioplankton lytic activity in relation to prokaryotic production and potential protistan bacterivory in the deep (Zmax092 m) meromictic volcanic Lake Pavin (Massif Central, France). The sampling strategy was designed to be representative of the physico-chemical gradients of the water column of the lake, and of the seasonal variability as well, i.e. 12 different depths sampled in triplicates from April to December 2005. In the space, viral lytic activity estimated from the frequency of visibly infected prokaryotic cells and from burst size over the study period generally decreased with depth. This was viewed as a paradox compared to the abundances of viruses and prokaryotes and to the prokaryotic production which increased with depth. The seasonal variability in viral lytic activity was correlated with prokaryotic variables (abundance and production) in the deepest waters, i.e. from the hypolimnion downwards. Compared to previous studies known from the mixolimnion, we conclude that the deep waters in Lake Pavin represent an exclusive environment for heterotrophic prokaryotes whose seasonal activity offers an optimal and unique resource for thriving viral communities, some of which may be typical, endemic to the ambient dark, cold and stable deep water masses. Overall, the main findings in the present study get well around a previous statement that the ecology of the deepest waters of Lake Pavin is essentially driven by the dark viral loop (dissolved organic matter-prokaryotes- viruses) processes, which can sequester organic matters and nutrients for a long-lived turnover time. This is in agreement with recent demonstrations from marine systems that meso- and bathypelagic waters are optimal environments for viral survival and proliferation.

International audience; This study presents a depth-related survey of virioplankton abundance in Lake Pavin (Massif Central, France), in relation to the abundances of heterotrophic prokaryotes, picocyanobacteria (Pcy), autotrophic picoeukaryotes (Peu), and of autotrophic (ANF) and heterotrophic (HNF) nanoflagellates. The sampling strategy was designed to be representative of the physico-chemical gradients of the whole water column of the lake, and the seasonal variability as well. In mixolimnic surface waters, all communities were present and viral abundance peaked in summer and autumn. Viral abundance was significantly correlated (p < 0.001) with Pcy, Peu, and ANF, indicating that cyanophages and perhaps other phytoplankton viruses represent a significant pool of viral standing stocks in the mixolimnion of Lake Pavin. Microautotrophs were absent in the deep monimolimnic water masses, where viruses and heterotrophic prokaryotes exhibited highest seasonal abundances in summer and/or autumn and were significantly correlated (p < 0.001) to each other. This indicates that the anoxic monimolimnion of Lake Pavin is an exclusive habitat for viruses and heterotrophic prokaryotes. We conclude that in this habitat, host availability is prevalent over other factors (temperature, oxygen, nutrients, grazers) in favoring viral proliferation.

Bathypelagic ecosystems depend on prokaryotic heterotrophic biomass fuelled by vertical particulate organic matter (POM) fluxes, but very little information is available on the interactions among viruses, prokaryotes and nanoflagellates in deep waters. We simultaneously investigated the relative importance of the viral and heterotrophic nanoflagellate (HNF) grazing-induced prokaryotic mortality in bathypelagic waters by means of dilution experiments performed on samples collected at 1500 m depth from the Atlantic Ocean to the Eastern Mediterranean Sea. Prokaryotic abundance (range: 1.4 to 8.9 × 104 cells ml–1), although different from one station to another, was on average not significantly different among biogeographic regions. The potential predators of prokaryotes (viruses, HNF and microzooplankton) followed a similar spatial pattern. Viruses were responsible for an important fraction of prokaryotic mortality (on average 13.4% d–1). Dilution experiments carried out to estimate the potential predation of HNF suggested a high effect on prokaryotic abundance. However, since the latter experiments also include the effect of viruses on prokaryotes, when this factor was disentangled from the overall mortality, the potential rates of HNF predation on prokaryotes (on average 49.5%) were ca. 4 times higher than the effect due to viral infections. Conversely to patterns of distribution, the relative importance of virus-mediated mortality vs. HNF predation changed significantly among different regions. Results should be treated with caution due to the intrinsic difficulty in reproducing experimentally natural deep-sea conditions, but they permit disentangling of the relative effect of viruses and HNF on prokaryotes and compare the potential predatory control in different biogeographic regions.

International audience; Samples were collected at four depths every 6 h over a 42-h period during two contrasting seasons (June vs. December) from Lake Bourget, France, for evidence of circadian fluctuations in the concentrations and potential activities of viruses, prokaryotes and protists in relation to environmental conditions: temperature, chlorophyll a and dissolved organic carbon (DOC) concentrations. Considerable vertical and temporal fluctuations were observed for all variables. Circadian variations were noted for DOC and chlorophyll a concentrations. Despite the external abiotic forcing (light, water movements), the fluctuations of microbial variables (including viruses) in most cases were apparently linked to biotic factors and interactions. Standing stocks and activities, as well as the number and levels of correlations among the microbial components, were, surprisingly, higher in winter than in summer. We speculate that this was because trophic interactions prevailed over the seasonal forcing (i.e. temperature) in shaping the observed differences.

International audience; The study of lysogeny in aquatic systems is an often overlooked aspect of microbial ecology, especially in tropical environments. Herein, the fraction of lysogenized cells (FLC) was detected in the surface waters of 20 coastal stations distributed from the eutrophicated shoreline to seaward waters of Hann Bay (Senegal). Concurrently, viral lytic infection rates were extrapolated from the frequency of visibly infected bacterial cells (FVIC), as determined from transmission electron microscopy observations. The experimental induction of prophage was observed in less than 3% of indigenous marine bacteria, suggesting that lysogenic stages of infection are rare in Hann Bay. Similarly, only 0.5-4.7% of bacteria showed visible signs of lytic infection. However, the positive correlation between the fraction of lysogenic and lytic cells (r = 0.67, p < 0.05, n = 20) may actually indicate that the coexistence of both lifestyles may be due to the massive and rapid induction of lysogens, potentially from the high levels of local UV radiation. Overall, we suggest that the determination of FVIC and FLC to examine the predominance of one type of cycle versus the other may be a source of misinterpretation in some particular aquatic environments.

Temporal and spatial changes of viral and bacterial abundance were examined in relation to environmental factors and hydrography at five stations between May 2000 and July 2001 in the brackish lake of Ganzirri (Sicily, Italy). Virioplankton abundance ranged from 5.26 × 104 to 7.54 × 108VLP ml−1 (on average 1.38 × 108particles ml−1) and was significantly higher at the three eutrophic stations located in the lake of Ganzirri (Stations 1, 2, and 3) than in the channel connecting the lake with the Straits of Messina. The virus-to-bacterium abundance ratio (VBR range, 0.4–117; average:14) showed the highest values in channel connecting the lake of Ganzirri with the meromictic lake of Faro. VBR values