Phytoplankton composition and biomass during leg 7 of Galathea 3 cruise across the southern Indian Ocean

Phytoplankton composition and biomass was investigated across the southern Indian Ocean. Phytoplankton composition was determined from pigment analysis with subsequent calculations of group contributions to total chlorophyll a (Chl a) using CHEMTAX and, in addition, by examination in the microscope. The different plankton communities detected reflected the different water masses along a transect from Cape Town, South Africa, to Broome, Australia. The first station was influenced by the Agulhas Current with a very deep mixed surface layer. Based on pigment analysis this station was dominated by haptophytes, pelagophytes, cyanobacteria, and prasinophytes. Sub-Antarctic waters of the Southern Ocean were encountered at the next station, where new nutrients were intruded to the surface layer and the total Chl a concentration reached high concentrations of 1.7 µg Chl a/L with increased proportions of diatoms and dinoflagellates. The third station was also influenced by Southern Ocean waters, but located in a transition area on the boundary to subtropical water. Prochlorophytes appeared in the samples and Chl a was low, i.e., 0.3 µg/L in the surface with prevalence of haptophytes, pelagophytes, and cyanobacteria. The next two stations were located in the subtropical gyre with little mixing and general oligotrophic conditions where prochlorophytes, haptophytes and pelagophytes dominated. The last two stations were located in tropical waters influenced by down-welling of the Leeuwin Current and particularly prochlorophytes dominated at these two stations, but also pelagophytes, haptophytes and cyanobacteria were abundant. Haptophytes Type 6 (sensu Zapata et al., 2004), most likely Emiliania huxleyi, and pelagophytes were the dominating eucaryotes in the southern Indian Ocean. Prochlorophytes dominated in the subtrophic and oligotrophic eastern Indian Ocean where Chl a was low, i.e., 0.043-0.086 µg total Chl a/L in the surface, and up to 0.4 µg Chl a/L at deep Chl a maximum. From the pigment analyses it was found that the dinoflagellates of unknown trophy enumerated in the microscope at the oligotrophic stations were possibly heterotrophic or mixotrophic. Presence of zeaxanthin containing heterotrophic bacteria may have increased the abundance of cyanobacteria determined by CHEMTAX.

Physical oceanography and hydrochemistry measured on water bottle samples during METEOR cruise M10/1

During a R.V. Meteor JGOFS-NABE cruise to a tropical site in the northeast Atlantic in spring 1989, three different vertical regimes with respect to nitrate distribution and availability within the euphotic zone were observed. Besides dramatic variations in the depth of the nitracline, a previously undescribed nose-like nitrate maximum within the euphotic zone was the most prominent feature during this study. Both the vertical structure of phytoplankton biomass and the degree of absolute and relative new production were related to the depth of the nitracline, which in turn was dependent on the occurrence/non-occurrence of the subsurface subtropical salinity maximum (Smax). The mesoscale variability of the nitracline depth, as indicated from a pre-survey grid, and published data on the frequent occurrence of the Smax in tropical waters suggest higher variability of new production and F-ratio than usually expected for oligotrophic oceans. The importance of salt fingering and double diffusion for nitrate transport into the euphotic zone is discussed.

Seawater carbonate chemistry and phytoplankton and eubacterial community compositions in the northwest subarctic Pacific

On-deck CO2-Fe-manipulated incubation experiments were conducted using surface seawater collected from the Western Subarctic Gyre of the NW Pacific in the summer of 2008 to elucidate the impacts of ocean acidification and Fe enrichment on the abundance and community composition of phytoplankton and eubacteria in the study area. During the incubation, excluding the initial period, the mean partial pressures of CO2 in non-Fe-added bottles were 230, 419, 843, and 1124 µatm, whereas those in Fe-added treatments were 152, 394, 791, and 1008 µatm. Changes in the abundance and community composition of phytoplankton were estimated using HPLC pigment signatures with the program CHEMTAX and flow cytometry. A DGGE fingerprint technique targeting 16S rRNA gene fragments was also used to estimate changes in eubacterial phylotypes during incubation. The Fe addition induced diatom blooms, and subsequently stimulated the growth of heterotrophic bacteria such as Roseobacter, Phaeobacter, and Alteromonas in the post-bloom phase. In both the Fe-limited and Fe-replete treatments, concentrations of 19'-hexanoyloxyfucoxanthin, a haptophyte marker, and the cell abundance of coccolithophores decreased at higher CO2 levels (750 and 1000 ppm), whereas diatoms exhibited little response to the changes in CO2 availability. The abundances of Synechococcus and small eukaryotic phytoplankton (<10 µm) increased at the higher CO2 levels. DGGE band positions revealed that Methylobacterium of Alphaproteobacteria occurred solely at lower CO2 levels (180 and 380 ppm) during the post-bloom phase. These results suggest that increases in CO2 level could affect not only the community composition of phytoplankton but also that of eubacteria. As these microorganisms play critical roles in the biological carbon pump and microbial loop, our results indicate that the progression of ocean acidification can alter the biogeochemical processes in the study area.

Seawater carbonate chemistry and processes during experiments with amphipod Gammarus locusta, 2009

We report an investigation of the effects of increases in pCO2 on the survival, growth and molecular physiology of the neritic amphipod Gammarus locusta which has a cosmopolitan distribution in estuaries. Amphipods were reared from juvenile to mature adult in laboratory microcosms at three different levels of pH in nominal range 8.1-7.6. Growth rate was estimated from weekly measures of body length. At sexual maturity the amphipods were sacrificed and assayed for changes in the expression of genes coding for a heat shock protein (hsp70 gene) and the metabolic enzyme glyceraldehyde-3-phosphate dehydrogenase (gapdh gene). The data show that the growth and survival of this species is not significantly impacted by a decrease in sea water pH of up to 0.5 units. Quantitative real-time PCR analysis indicated that there was no significant effect of growth in acidified sea water on the sustained expression of the hsp70 gene. There was a consistent and significant increase in the expression of the gapdh gene at a pH of ~7.5 which, when combined with observations from other workers, suggests that metabolic changes may occur in response to acidification. It is concluded that sensitive assays of tissue physiology and molecular biology should be routinely employed in future studies of the impacts of sea water acidification as subtle effects on the physiology and metabolism of coastal marine species may be overlooked in conventional gross "end-point" studies of organism growth or mortality.

Villefranche sur Mer: multistressors experiment March 2012

The effect of ocean warming and acidification was investigated on a natural plankton assemblage from an oligotrophic area, the bay of Villefranche (NW Mediterranean Sea). The assemblage was sampled in March 2012 and exposed to the following four treatments for 12 days: control ( 360 µatm, 14°C), elevated pCO2 ( 610 µatm, 14°C), elevated temperature ( 410 µatm, 17°C), and elevated pCO2 and temperature ( 690 µatm, 17°C). Nutrients were already depleted at the beginning of the experiment and the concentrations of chlorophyll a (chl a), heterotrophic prokaryotes and viruses decreased, under all treatments, throughout the experiment. There were no statistically significant effects of ocean warming and acidification, whether in isolation or combined, on the concentrations of nutrients, particulate organic matter, chl a and most of the photosynthetic pigments. Furthermore, 13C labelling showed that the carbon transfer rates from 13C-sodium bicarbonate into particulate organic carbon were not affected by seawater warming nor acidification. Rates of gross primary production followed the general decreasing trend of chl a concentrations and were significantly higher under elevated temperature, an effect exacerbated when combined to elevated pCO2 level. In contrast to the other algal groups, the picophytoplankton population (cyanobacteria, mostly Synechococcus) increased throughout the experiment and was more abundant in the warmer treatment though to a lesser extent when combined to high pCO2 level. These results suggest that under nutrient-depleted conditions in the Mediterranean Sea, ocean acidification has a very limited impact on the plankton community and that small species will benefit from warming with a potential decrease of the export and energy transfer to higher trophic levels.

Micro- and picoplankton in waters of the Saint Paul Island, Pribilof Islands, Bering Sea

On the basis of materials collected in June-August 1994 characteristic data on microplankton were gathered in three biotopes of the eastern shelf of the Bering Sea: open shelf (coastal zone), the harbor, and the salt lagoon of Saint Paul Island (Pribiof Islands). The following parameters of microplanktonic communities were analyzed: abundance, biomass, and production of autotrophic picoplankton (picoalgae and cyanobacteria); abundance, biomass, growth rate constant, and production of bacterioplankton; role of filiform bacteria in bacterioplankton; species composition of heterotrophic flagellates and ciliates, their abundance, and biomass. Growth rates and consumption rates of picoplankton and bacterioplankton by heterotrophic nano- and microplankton were estimated in the experiments using the dilution method. Temporal dynamics of all structural and functional parameters of microplankton were analyzed. The minor role of autotrophic picoplankton and significant role of bacterioplankton as well as heterotrophic nano- and microplankton in planktonic communities of studied biotopes during summer months was shown. During certain periods, bacterial biomass was as high as 50-65% of phytoplankton biomass, and production of bacteria was as high as 20-40% of primary production. In the middle of the season biomass of nano- and microheterotrophic organisms in different biotopes exceeded biomass of mesozooplankton 2-10 times. Average consumption of bacterial production by nano- and microplankton during the period of observations was 85-94%.

Abundance and Biomass from heterotrophic bacteria, Synechococcus, Prochlorococcus and Virus in the eastern Mediteranean Sea in October 2008 during SES_GR2

The HCMR_SES_LAGRANGIAN_GR2_ MICROBIAL PARAMETERS dataset is based on samples collected in the framework of the project SESAME, in the North Aegean Sea during October 2008. The objectives were to measure the standing stocks and calculate the production of the microbial compartment of the food web, describe the vertical distribution pattern and characterize its structure and function through the water column as influenced by the BSW. Heterotrophic bacteria, Synechococcus, Prochlorococcus and Virus abundance: Subsamples for virus, heterotrophic bacteria and cyanobacteria (Synechococcus spp. and Prochlorococcus spp.) counting were analyzed using a FACSCalibur (Becton Dickinson) flow cytometer equipped with a standard laser (488 nm) and filter set and using deionized water as sheath fluid. Fluorescent beads with a diameter of 0.97 µm (Polysciences) were added to each sample as an internal standard, and all parameters were normalized to the beads and expressed as relative units. SYBRGreen I stain (Molecular Probe) was used to stain viral and heterotrophic bacterial DNA. Viruses were counted according to (Brussaard 1984). In order to avoid bulk consentrations of viruses samples we dilluted to Tris-EDTA (pH=8,0) buffer to a final sollution of 1/5 to 1/100. Total abundance and nucleid content classes were calculated using the Paint-A-Gate software (Becton Dickinson). Heterotrophic Nanoflagellate abundance: Subsamples (30-150 ml) were concentrated on 25mm black polycarbonate filters of porosity 0.6?m and stained with DAPI for 10 min (Porter and Feig 1980). Under epifluorescence microscopy heterotrophic nanoflagellates (HNAN) were distinguished using UV and blue excitation and enumerated. Nanoflagellates were classified in size categories and the biovolume was calculated. Ciliate abundance: For ciliate identification and enumeration, 100-3000 ml samples were left for 24h-4d for sedimentation and then observed under an inverted microscope. Ciliates were counted, distinguished into size-classes and major taxonomic groups and identified down to genus or species level where possible (Pitta et al. 2005). Heterotrophic bacteria, Synechococcus, Prochlorococcus bacteria: Subsamples for virus, heterotrophic bacteria and cyanobacteria (Synechococcus spp. and Prochlorococcus spp.) counting were analyzed using a FACSCalibur (Becton Dickinson) flow cytometer equipped with a standard laser (488 nm) and filter set and using deionized water as sheath fluid. Fluorescent beads with a diameter of 0.97 µm (Polysciences) were added to each sample as an internal standard, and all parameters were normalized to the beads and expressed as relative units. SYBRGreen I stain (Molecular Probe) was used to stain viral and heterotrophic bacterial DNA. Viruses were counted according to (Brussaard 1984). In order to avoid bulk consentrations of viruses samples we dilluted to Tris-EDTA (pH=8,0) buffer to a final sollution of 1/5 to 1/100. Total abundance and nucleid content classes were calculated using the Paint-A-Gate software (Becton Dickinson). Abundance data were converted into C biomass using 250 fgC cell-1 (Kana & Glibert 1987) for Synechococcus, 50 fgC cell-1 (Campbell et al. 1994) for Prochlorococcus and 20fgC cell-1 (Lee & Fuhrman 1987) for heterotrophic bacteria. Heterotrophic Nanoflagellate biomass: Subsamples (30-150 ml) were concentrated on 25mm black polycarbonate filters of porosity 0.6µm and stained with DAPI for 10 min (Porter and Feig 1980). Under epifluorescence microscopy heterotrophic nanoflagellates (HNAN) were distinguished using UV and blue excitation and enumerated. Nanoflagellates were classified in size categories and the biovolume was calculated. Abundance data were converted into C biomass using 183 fgC µm**3 (Caron et al. 1995). Ciliate biomass: For ciliate identification and enumeration, 100-3000 ml samples were left for 24h-4d for sedimentation and then observed under an inverted microscope. Ciliates were counted, distinguished into size-classes and major taxonomic groups and identified down to genus or species level where possible (Pitta et al. 2005). Ciliate cell sizes were measured and converted into cell volumes using appropriate geometric formulae using image analysis. For biomass estimation, the conversion factor 190 fgC µm**3 was used (Putt and Stoecker 1989).