Changes in Microbial (Bacteria and Archaea) Plankton Community Structure after Artificial Dispersal in Grazer-Free Microcosms.

Microbes are considered to have a global distribution due to their high dispersal capabilities. However, our knowledge of the way geographically distant microbial communities assemble after dispersal in a new environment is limited. In this study, we examined whether communities would converge because similar taxa would be selected under the same environmental conditions, or would diverge because of initial community composition, after artificial dispersal. To this aim, a microcosm experiment was performed, in which the temporal changes in the composition and diversity of different prokaryoplankton assemblages from three distant geographic coastal areas (Banyuls-sur-Mer in northwest Mediterranean Sea, Pagasitikos Gulf in northeast Mediterranean and Woods Hole, MA, USA in the northwest Atlantic), were studied. Diversity was investigated using amplicon pyrosequencing of the V1-V3 hypervariable regions of the 16S rRNA. The three assemblages were grown separately in particle free and autoclaved Banyuls-sur-mer seawater at 18 °C in the dark. We found that the variability of prokaryoplankton community diversity (expressed as richness, evenness and dominance) as well as the composition were driven by patterns observed in Bacteria. Regarding community composition, similarities were found between treatments at family level. However, at the OTU level microbial communities from the three different original locations diverge rather than converge during incubation. It is suggested that slight differences in the composition of the initial prokaryoplankton communities, resulted in separate clusters the following days even when growth took place under identical abiotic conditions.

Dynamics of phytoplankton communities in eutrophying tropical shrimp ponds affected by vibriosis.

Tropical shrimp aquaculture systems in New Caledonia regularly face major crises resulting from outbreaks of Vibrio infections. Ponds are highly dynamic and challenging environments and display a wide range of trophic conditions. In farms affected by vibriosis, phytoplankton biomass and composition are highly variable. These conditions may promote the development of harmful algae increasing shrimp susceptibility to bacterial infections. Phytoplankton compartment before and during mortality outbreaks was monitored at a shrimp farm that has been regularly and highly impacted by these diseases. Combining information from flow cytometry, microscopy, pigment and phylogenetic analysis, the presence of Picocyanobacteria, Prasinophyceae and Diatomophyceae were detected as dominant phytoplankton groups and Cryptophyceae, Prymnesiophyceae and Dinophyceae as minor components. At the onset of the first shrimp mortalities, Bacillariophyceae increased while Cyanobacteria, Prymnesiophyceae and Dinophyceae decreased in the water column, followed by proliferation of Prasinophyceae. Several taxa were identified as potential harmful algae (Cyanobacteria, dinoflagellates and Phaeocystis).

An efficient and rapid method for the enumeration of heterotrophic prokaryotes in coastal sediments by flow cytometry.

Flow cytometry offers an easy and powerful way to assess multi-parametric data in different domains, notably in the environmental sciences. Because evaluating heterotrophic prokaryotic abundance is crucial to understand an ecosystem's functioning, we propose a quick and efficient protocol for (1) cell's detachment in muddy coastal sediments followed by (2) enumeration of prokaryotes by flow cytometry compared to epifluorescence microscopy and (3) a type of storage adapted for benthic samples. First, sample preparation by incubation in a detergent mix containing sodium pyrophosphate (0.01M final concentration) and Tween 80 (0.1% final concentration) drastically increased cell detachment from sediment particles (+130.40%) compared to extraction with sodium pyrophosphate only. Cell sorting allowed to control the efficiency of the extraction as few cells were found attached to sediment particles in epifluorescence microscopy after sorting. Flow cytometry gave consistent results with strong reliability by counting 1.81 times more cells compared to epifluorescence microscopy. Thirdly, results revealed that sediment samples fixed with formaldehyde and then liquid-N2 frozen and directly stored at -80°C can be analyzed within 3months. In routine, our method of extraction and counting allowed to evaluate 83.67% of the real abundance in a sediment sample. Finally, this optimized technique was applied on sandy and muddy coastal and freshwater sediments and allowed us to prove the high efficiency of this new method. Flow cytometry is a fast, replicable and low-cost method for counting heterotrophic prokaryotes, even for sediment samples. The two-step method that we developed enables high frequency analyses (30 samples in less than 4h).

Spatial and temporal extension of eutrophication associated with shrimp farm wastewater discharges in the New Caledonia lagoon.

Shrimp farming in New Caledonia typically uses a flow-through system with water exchange rates as a tool to maintain optimum hydrological and biological parameters for the crop. Moreover, the effluent shows hydrobiological characteristics (minerals, phytoplankton biomass and organic matter) significantly higher than that of the receiving environment. Separate surveys were carried out in a bay (CH Bay) with a medium-size intensive farm (30 ha) (PO) and in a mangrove-lined creek (TE Creek) near a larger semi-intensive farm (133 ha) (SO). Net loads of nitrogen exported from the semi-intensive farm and the intensive farm amounted to 0.68 and 1.36 kg ha(-1)day(-1), respectively. At CH Bay, discharge effects were spatially limited and clearly restricted to periods of effluent release. The high residence time at site TE favoured the installation of a feedback system in which organic matter was not exported. Mineralization of organic matter led to the release of nutrients, which in turn, caused in an increased eutrophication of this ecosystem. The study of the pico- and nanophytoplankton assemblages showed (i) a shift in composition from picophytoplankton to nanophytoplankton from offshore towards the coast and (ii) a shift within the picophytoplankton with the disappearance of Prochlorococcus and the increase of picoeucaryotes towards the shoreline. These community changes may partially be related to a nitrogen enrichment of the environment by shrimp farm discharges. Thus, in view of the recent addition of the New Caledonian lagoon to the UNESCO World Heritage list, the data presented here could be a first approach to quantify farm discharges and evaluate their impact on the lagoon.

Nutrient and microbial dynamics in eutrophying shrimp ponds affected or unaffected by vibriosis.

A field survey was conducted on two intensive shrimp farms using similar technical practices: one (DF) historically affected by a vibriosis, the other (HC) in which the pathogen has been observed although no mortality event has occurred. Because historical data suggest that eutrophication process may directly or indirectly play a role in the disease outbreak, we focussed our research on its dynamics. A higher variability of the phytoplanktonic compartment linked to an imbalance in the molar N:P ratio was observed in farm DF compared to farm HC, implying a modification on the linkage between the bacteria and phytoplankton compartments at DF. The beginning of the mortality outbreak at DF followed a shift from pico- to nanophytoplankton. The organic matter mineralization process at the water-sediment interface may explain the disturbance observed in the water column during eutrophication. The consequences of this disturbance on shrimps' health status and pathogen ecology are discussed.

Grazing impact of different heterotrophic nanoflagellates on eukaryotic (Ostreococcus tauri) and prokaryotic picoautotrophs (Prochlorococcus and Synechococcus).

Autotrophic picoplankton (<3 microm) composed of both prokaryotes and eukaryotes are the most abundant primary producers on Earth. In this study we examined the ingestion of the picoeukaryote Ostreococcus tauri by different marine heterotrophic nanoflagellates (HNF) with various morphologies, swimming and feeding behaviours. Cultures of specific bacterivorous nanoflagellates (Rhynchomonas nasuta, Jakoba libera, and a culture of Cafeteria sp./Monosiga sp.) and natural nanoflagellate populations were used as grazers. For comparison with Ostreococcus, we used similar-sized prokaryotes as prey, Prochlorococcus and Synechococcus. We observed large species-specific differences in terms of: use of picoautotrophs among nanoflagellates, time lag between prey addition and prey consumption (0-196 h), grazing rate (0-0.12 h(-1)), growth rate (0-0.3 h(-1)) and maximum abundance of HNF reached in experimental bottles (e.g. from 10(4) to 10(5) cells ml(-1), for a natural coastal population and a Cafeteria sp./Monosiga sp. culture feeding Ostreococcus respectively). Overall, this study shows that the nanoflagellate community composition is conclusive for picoautotrophic community structure and, vice versa, the picoautotrophic community structure favours or inhibits the growth of some nanoflagellate groups.