The surface film of the hydrosphere covers more than 70% of the world's surface. The sea surface microlayer (SML) or "skin" of the ocean is a sink for natural and anthropogenic material originating from the atmosphere and the water column. Organisms living in this SML are called "neuston." Our knowledge of the biology of the SML is still in its infancy. Research of the sea surface microlayer requires the use of appropriate sampling techniques and strategies, and the question of what is the most suitable device has not yet been answered. In the present study, we have compared the efficiency of the Harvey glass plate (GP) and the Garrett metal screen (MS) to analyze a wide range of microbiological parameters in SML samples collected at two coastal stations in the NW Mediterranean Sea. Two types of membranes (Teflon and polycarbonate) were also used to collect bacterioneuston. The MS was the most appropriate technique for most biological parameters providing higher enrichment factors as compared to the GP and, therefore, the highest enrichment factors compared with underlying waters (UW). Control experiments with UW demonstrated that the enrichment reported for the MS was not biased by any selectivity of the sampler itself. Therefore, we recommend the use of the MS when the aim is to compare different biological parameters. In contrast, there is clear evidence that hydrophobic and hydrophilic membranes have an important drawback and should not be used for quantification purposes. AcknowledgmentsThis work was supported by the European Commission (Research Directorate General-Environment Program-Marine Ecosystems) through the AIRWIN project "Structure and role of biological communities involved in the transport and transformation of persistent pollutants at the marine AIR-Water Interface" (contract EVK3-CT2000-00030). The AIRWIN project is part of the EC IMPACTS cluster. We thank Frédérique François-Carcaillet for helpful comments on statistical analysis and Danielle Boissé for language improvements. We are also grateful to the two anonymous reviewers for valuable comments on a previous version of the manuscript.
In most aquatic environments, at least 2 subpopulations of bacterial cells can be discriminated by flow cytometry based on their nucleic acid content. Recent investigations have shown that the cells with a high nucleic acid (HNA) content have a higher cell-specific activity (CSA) cell than those with a low nucleic acid (LNA) content. In this study, the CSA and biomass-specific activities (BSA) of HNA and LNA cells from different aquatic ecosystems, including marine, brackish and freshwater, were investigated using radioactive leucine incorporation and cell sorting by flow cytometry. The genetic diversity of natural assemblages, HNA and LNA cells was investigated using the SSCP (PCR single-strand conformation polymorphism) method. Data showed that both CSA and BSA of HNA cells were always significantly higher than CSA and BSA of LNA cells. In addition, HNA cells had a dominant contribution to the production of the total community (77 to 98%). For the different samples, the SSCP fingerprints from the natural assemblage and from the 2 sorted fractions were not significantly different. This clearly suggests that HNA and LNA subpopulations were composed by the same dominant species and, thus, confirms an important heterogeneity of physiological states within most natural populations.
Seven blue nucleic acid dyes from Molecular Probes Inc. (SYTO-9, SYTO-11, SYTO-13, SYTO-16, SYTO-BC, SYBR-I and SYBR-II) were compared with the DAPI (4′,6-diamidino-2-phenylindole) method for flow cytometric enumeration of live and fixed bacteria in aquatic systems. It was shown that SYBR-II and SYTO-9 are the most appropriate dyes for bacterial enumeration in nonsaline waters and can be applied to both live and dead bacteria. The fluorescence signal/noise ratio was improved when SYTO-9 was used to stain living bacteria in nonsaline waters. Inversely, SYBR-II is more appropriate than SYTO dyes for bacterial enumeration of unfixed and fixed seawater samples.
Dissolved organic matter (DOM) and heterotrophic bacteria are highly diverse components of the ocean system, and their interactions are key in regulating the biogeochemical cycles of major elements. How chemical and phylogenetic diversity are linked remains largely unexplored to date. To investigate interactions between bacterial diversity and DOM, we followed the response of natural bacterial communities to two sources of phytoplankton-derived DOM over six bacterial generation times in continuous cultures. Analyses of total hydrolysable neutral sugars and amino acids, and ultrahigh resolution mass spectrometry revealed large differences in the chemical composition of the two DOM sources. According to 454 pyrosequences of 16S ribosomal ribonucleic acid genes, diatom-derived DOM sustained higher levels of bacterial richness, evenness and phylogenetic diversity than cyanobacteria-derived DOM. These distinct community structures were, however, not associated with specific taxa. Grazing pressure affected bacterial community composition without changing the overall pattern of bacterial diversity levels set by DOM. Our results demonstrate that resource composition can shape several facets of bacterial diversity without influencing the phylogenetic composition of bacterial communities, suggesting functional redundancy at different taxonomic levels for the degradation of phytoplankton-derived DOM.
The effectiveness of SYTOX Green nucleic acid stain for measuring bacterial viability was tested on starved populations of Escherichia coli and Salmonella typhimurium. This stain underestimates the fraction of dead cells within starved populations containing cells with damaged nucleic acids or membranes. Its application to natural samples should be considered with caution.
The Southern Ocean (SO) hosts plankton communities that impact the biogeochemical cycles of the global ocean. However, weather conditions in the SO restrict mainly in situ observations of plankton communities to spring and summer, preventing the description of biological successions at an annual scale. Here, we use shipboard observations collected in the Indian sector of the SO to develop a multivariate relationship between physical and bio‐optical data, and, the composition and carbon content of the plankton community. Then we apply this multivariate relationship to five biogeochemical Argo (BGC‐Argo) floats deployed within the same bio‐geographical zone as the ship‐board observations to describe spatial and seasonal changes in plankton assemblage. The floats reveal a high contribution of bacteria below the mixed layer, an overall low abundance of picoplankton and a seasonal succession from nano‐ to microplankton during the spring bloom. Both naturally iron‐fertilized waters downstream of the Crozet and Kerguelen Plateaus show elevated phytoplankton biomass in spring and summer but they differ by a nano‐ or microplankton dominance at Crozet and Kerguelen, respectively. The estimated plankton group successions appear consistent with independent estimations of particle diameter based on the optical signals. Furthermore, the comparison of the plankton community composition in the surface layer with the presence of large mesopelagic particles diagnosed by spikes of optical signals provides insight into the nature and temporal changes of ecological vectors that drive particle export. This study emphasizes the power of BGC‐Argo floats for investigating important biogeochemical processes at high temporal and spatial resolution.
Abstract. Microbial food web dynamics were determined during the onset of several spring phytoplankton blooms induced by natural iron fertilization off Kerguelen Island in the Southern Ocean (KEOPS2). The abundances of heterotrophic bacteria and heterotrophic nanoflagellates, bacterial heterotrophic production, bacterial respiration, and bacterial growth efficiency, were consistently higher in surface waters of the iron-fertilized sites than at the reference site in HNLC (high nutrient low chlorophyll) waters. The abundance of virus-like particles remained unchanged, but viral production increased by a factor of 6 in iron-fertilized waters. Bacterial heterotrophic production was significantly related to heterotrophic nanoflagellate abundance and viral production across all sites, with bacterial production explaining about 70 and 85%, respectively, of the variance of each in the mixed layer (ML). Estimated rates of grazing and viral lysis, however, indicated that heterotrophic nanoflagellates accounted for a substantially higher loss of bacterial production (50%) than viruses (11%). Combining these results with rates of primary production and export determined for the study area, a budget for the flow of carbon through the microbial food web and higher trophic levels during the early (KEOPS2) and the late phase (KEOPS1) of the Kerguelen bloom is provided.
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