Latitudinal distribution of prokaryotic picoplankton populations in the Atlantic Ocean

Members of the prokaryotic picoplankton are the main drivers of the biogeochemical cycles over large areas of the world's oceans. In order to ascertain changes in picoplankton composition in the euphotic and twilight zones at an ocean basin scale we determined the distribution of 11 marine bacterial and archaeal phyla in three different water layers along a transect across the Atlantic Ocean from South Africa (32.9°S) to the UK (46.4°N) during boreal spring. Depth profiles down to 500 m at 65 stations were analysed by catalysed reporter deposition fluorescence in situ hybridization (CARD‐FISH) and automated epifluorescence microscopy. There was no obvious overall difference in microbial community composition between the surface water layer and the deep chlorophyll maximum (DCM) layer. There were, however, significant differences between the two photic water layers and the mesopelagic zone. SAR11 (35 ± 9%) and Prochlorococcus (12 ± 8%) together dominated the surface waters, whereas SAR11 and Crenarchaeota of the marine group I formed equal proportions of the picoplankton community below the DCM (both ～15%). However, due to their small cell sizes Crenarchaeota contributed distinctly less to total microbial biomass than SAR11 in this mesopelagic water layer. Bacteria from the uncultured Chloroflexi‐related clade SAR202 occurred preferentially below the DCM (4–6%). Distinct latitudinal distribution patterns were found both in the photic zone and in the mesopelagic waters: in the photic zone, SAR11 was more abundant in the Northern Atlantic Ocean (up to 45%) than in the Southern Atlantic gyre (～25%), the biomass of Prochlorococcus peaked in the tropical Atlantic Ocean, and Bacteroidetes and Gammaproteobacteria bloomed in the nutrient‐rich northern temperate waters and in the Benguela upwelling. In mesopelagic waters, higher proportions of SAR202 were present in both central gyre regions, whereas Crenarchaeota were clearly more abundant in the upwelling regions and in higher latitudes. Other phylogenetic groups such as the Planctomycetes, marine group II Euryarchaeota and the uncultured clades SAR406, SAR324 and SAR86 rarely exceeded more than 5% of relative abundance.     

Nutrient uptake and alkaline phosphatase (ec 3:1:3:1) activity of emiliania huxleyi (PRYMNESIOPHYCEAE) during growth under n and p limitation in continuous cultures

Emiliania huxleyi (strain L) expressed an exceptional P assimilation capability. Under P limitation, the minimum cell P content was 2.6 fmol P·cell^?1, and cell N remained constant at all growth rates at 100 fmol N·cell^?1. Both, calcification of cells and the induction of the phosphate uptake system were inversely correlated with growth rate. The highest (cellular P based) maximum phosphate uptake rate (V_max^P) was 1400 times (i.e. 8.9 h^?1) higher than the actual uptake rate. The affinity of the P‐uptake system (dV/dS) was 19.8 L·μmol^?1·h^?1 at μ = 0.14 d^?1. This is the highest value ever reported for a phytoplankton species. V_max and dV/dS for phosphate uptake were 48% and 15% lower in the dark than in the light at the lowest growth rates. The half‐saturation constant for growth was 1.1 nM. The coefficient for luxury phosphate uptake (Q_maxt/Q_min) was 31. Under P limitation, E. huxleyi expressed two different types of alkaline phosphatase (APase) enzyme kinetics. One type was synthesized constitutively and possessed a V_max and half‐saturation constant of 43 fmol MFP·cell^?1·h^?1 and 1.9 μM, respectively. The other, inducible type of APase expressed its highest activity at the lowest growth rates, with a V_max and half‐saturation constant of 190 fmol MFP·cell^?1·h^?1 and 12.2 μM, respectively. Both APase systems were located in a lipid membrane close to the cell wall. Under N‐limiting growth conditions, the minimum N quotum was 43 fmol N·cell^?1. The highest value for the cell N‐specific maximum nitrate uptake rate (V_max^N) was 0.075 h^?1; for the affinity of nitrate uptake, 0.37 L·μmol^?1·h^?1. The uptake rate of nitrate in the dark was 70% lower than in the light. N‐limited cells were smaller than P‐limited cells and contained 50% less organic and inorganic carbon. In comparison with other algae, E. huxleyi is a poor competitor for nitrate under N limitation. As a consequence of its high affinity for inorganic phosphate, and the presence of two different types of APase in terms of kinetics, E. huxleyi is expected to perform well in P‐controlled ecosystems.     

Functional Characterization of the Microbial Community in Geothermally Heated Marine Sediments

The microbial population of geothermally heated sediments in a shallow bay of Vulcano Island (Italy) was characterized with respect to metabolic activities and the putatively catalyzing hyperthermophiles. Site-specific anoxic culturing media, most of which were amended with combinations of electron donors (glucose or carboxylic acids) and acceptors (sulfate), were used for selective enrichment of metabolically defined subpopulations. The mostly archaeal chemoautotrophs produced formate at rates of 3.25 and 0.46 fmol cell⁻¹ day⁻¹ with and without sulfate, respectively. The glucose fermenting heterotrophs produced acetate (18 fmol cell⁻¹ day⁻¹) and lactate (2.6 fmol cell⁻¹ day⁻¹) and were identified as predominantly Thermus sp. and coccoid archaea. These archaeal cells also metabolized lactate (5.6 fmol cell⁻¹ day⁻¹), but neither formate nor acetate. The heterotrophic culture enriched on formate/acetate/propionate/sulfate utilized mainly formate (27 fmol cell⁻¹ day⁻¹) and lactate (89–195 fmol cell⁻¹ day⁻¹), and consumed sulfate (38–68 fmol cell⁻¹ day⁻¹). These formate or lactate consuming sulfate reducers were dominated by Archaeoglobales (7% in situ) and unidentified Archaea. The in situ benthic community comprised 15% Crenarchaeota, a significant group only in the autotrophic cultures, and 3% Thermus sp., the putatively predominant group involved in fermentative metabolism. The role of Thermoccales (4% in situ) remained undisclosed in our experiments. This first comprehensive data set established plausible links between several groups of hyperthermophiles in shallow marine hydrothermal systems, their metabolic function within the benthic microbial community, and biogeochemical turnover rates.     

Vertical Distribution of Archaea and Bacteria in a Meromictic Lake as Determined by Fluorescent In Situ Hybridization

The prokaryotic cells distribution in the water column of the coastal saline meromictic Lake Faro (Messina, Italy) was investigated by microscopic counting techniques. Water samples were collected at a central station from the surface to the bottom, when waters were characterized by a marked stratification. A “red-water” layer, caused by a dense growth of photosynthetic sulfur bacteria, was present at a depth of 15 m, defining a transition area between oxic (mixolimnion) and anoxic (monimolimnion) layers. Fluorescently labeled 16S rRNA oligonucleotide, group-specific probes were used to determine the abundance of Bacteria and Archaea, and their subgroups, Green Sulfur Bacteria (GSB), Sulfate Reducing Bacteria (SRB), Cyanobacteria and Chromatium okenii, and Crenarchaeota and Euryarchaeota, as key elements of the microbial community. Bacteria decreased from surface to bottom, while Archaea increased with depth and reached the maximum value at 30 m, where they outnumbered the Bacteria. Bacteria and picophytoplankton prevailed in the mixolimnion. At the chemocline high numbers of prokaryotic cells were present, mainly represented by Cyanobacteria, Chromatium okenii and Euryarchaeota. GSB, SRB, and Crenarchaeota prevailed below the chemocline. Although Archaea constitute a minor fraction of microbial community, they could represent active contributors to the meromictic Lake Faro ecosystem.     

Inoculum effects on community composition and nitritation performance of autotrophic nitrifying biofilm reactors with counter‐diffusion geometry

The link between nitritation success in a membrane‐aerated biofilm reactor (MABR) and the composition of the initial ammonia‐ and nitrite‐oxidizing bacterial (AOB and NOB) population was investigated. Four identically operated flat‐sheet type MABRs were initiated with two different inocula: from an autotrophic nitrifying bioreactor (Inoculum A) or from a municipal wastewater treatment plant (Inoculum B). Higher nitritation efficiencies (NO₂^‐‐N/NH₄⁺‐N) were obtained in the Inoculum B‐ (55.2–56.4%) versus the Inoculum A‐ (20.2–22.1%) initiated reactors. The biofilms had similar oxygen penetration depths (100–150 µm), but the AOB profiles [based on 16S rRNA gene targeted real‐time quantitative PCR (qPCR)] revealed different peak densities at or distant from the membrane surface in the Inoculum B‐ versus A‐initiated reactors, respectively. Quantitative fluorescence in situ hybridization (FISH) revealed that the predominant AOB in the Inoculum A‐ and B‐initiated reactors were Nitrosospira spp. (48.9–61.2%) versus halophilic and halotolerant Nitrosomonas spp. (54.8–63.7%), respectively. The latter biofilm displayed a higher specific AOB activity than the former biofilm (1.65 fmol cell^?1 h^?1 versus 0.79 fmol cell^?1 h^?1). These observations suggest that the AOB and NOB population compositions of the inoculum may determine dominant AOB in the MABR biofilm, which in turn affects the degree of attainable nitritation in an MABR.     

Co-occurrence of denitrification and nitrogen fixation in a meromictic lake, Lake Cadagno (Switzerland)

The nitrogen cycling of Lake Cadagno was investigated by using a combination of biogeochemical and molecular ecological techniques. In the upper oxic freshwater zone inorganic nitrogen concentrations were low (up to ～3.4 μM nitrate at the base of the oxic zone), while in the lower anoxic zone there were high concentrations of ammonium (up to 40 μM). Between these zones, a narrow zone was characterized by no measurable inorganic nitrogen, but high microbial biomass (up to 4 × 10⁷ cells ml^?1). Incubation experiments with ¹⁵N‐nitrite revealed nitrogen loss occurring in the chemocline through denitrification (～3 nM N h^?1). At the same depth, incubations experiments with ¹⁵N₂‐ and ¹³C_DIC‐labelled bicarbonate, indicated substantial N₂ fixation (31.7–42.1 pM h^?1) and inorganic carbon assimilation (40–85 nM h^?1). Catalysed reporter deposition fluorescence in situ hybridization (CARD‐FISH) and sequencing of 16S rRNA genes showed that the microbial community at the chemocline was dominated by the phototrophic green sulfur bacterium Chlorobium clathratiforme. Phylogenetic analyses of the nifH genes expressed as mRNA revealed a high diversity of N₂ fixers, with the highest expression levels right at the chemocline. The majority of N₂ fixers were related to Chlorobium tepidum/C. phaeobacteroides. By using Halogen In Situ Hybridization‐Secondary Ion Mass Spectroscopy (HISH‐SIMS), we could for the first time directly link Chlorobium to N₂ fixation in the environment. Moreover, our results show that N₂ fixation could partly compensate for the N loss and that both processes occur at the same locale at the same time as suggested for the ancient Ocean.     

CELL‐SPECIFIC EXTRACELLULAR PHOSPHATASE ACTIVITY OF DINOFLAGELLATE POPULATIONS IN ACIDIFIED MOUNTAIN LAKES1

High bulk extracellular phosphatase activity (PA) suggested severe phosphorus (P) deficiency in plankton of three acidified mountain lakes in the Bohemian Forest. Bioavailability of P substantially differed among the lakes due to differences in their P loading, as well as in concentrations of aluminum (Al) and its species, and was accompanied by species‐specific responses of phytoplankton. We combined the fluorescently labeled enzyme activity (FLEA) assay with image cytometry to measure cell‐specific PA in natural populations of three dinophyte species, occurring in all the lakes throughout May–September 2007. The mean cell‐specific PA varied among the lakes within one order of magnitude: 188–1,831 fmol · cell^?1 · h^?1 for Gymnodinium uberrimum (G. F. Allman) Kof. et Swezy, 21–150 fmol · cell^?1 · h^?1 for Gymnodinium sp., and 22–365 fmol · cell^?1 · h^?1 for Peridinium umbonatum F. Stein. To better compare cell‐specific PA among the species of different size, the values were normalized per unit of cell biovolume (amol · μm^?3 · h^?1) for further statistical analysis. A step‐forward selection identified concentrations of total and ionic Al together with pH as significant factors (P < 0.05, Monte Carlo permutation test), explaining cumulatively 57% of the total variability in cell‐specific PA. However, this cell‐specific PA showed an unexpected reverse trend compared to an overall gradient in P deficiency of the lake plankton. The autecological insight into dinophyte cell‐specific PA therefore suggested other factors, such as light availability, mixotrophy, and/or zooplankton grazing, causing further PA variations among the acidified lakes.     

Contribution of Archaea to Total Prokaryotic Production in the Deep Atlantic Ocean

Fluorescence in situ hybridization (FISH) in combination with polynucleotide probes revealed that the two major groups of planktonic Archaea (Crenarchaeota and Euryarchaeota) exhibit a different distribution pattern in the water column of the Pacific subtropical gyre and in the Antarctic Circumpolar Current system. While Euryarchaeota were found to be more dominant in nearsurface waters, Crenarchaeota were relatively more abundant in the mesopelagic and bathypelagic waters. We determined the abundance of archaea in the mesopelagic and bathypelagic North Atlantic along a south-north transect of more than 4,000 km. Using an improved catalyzed reporter deposition-FISH (CARD-FISH) method and specific oligonucleotide probes, we found that archaea were consistently more abundant than bacteria below a 100-m depth. Combining microautoradiography with CARD-FISH revealed a high fraction of metabolically active cells in the deep ocean. Even at a 3,000-m depth, about 16% of the bacteria were taking up leucine. The percentage of Euryarchaeota and Crenarchaeaota taking up leucine did not follow a specific trend, with depths ranging from 6 to 35% and 3 to 18%, respectively. The fraction of Crenarchaeota taking up inorganic carbon increased with depth, while Euryarchaeota taking up inorganic carbon decreased from 200 m to 3,000 m in depth. The ability of archaea to take up inorganic carbon was used as a proxy to estimate archaeal cell production and to compare this archaeal production with total prokaryotic production measured via leucine incorporation. We estimate that archaeal production in the mesopelagic and bathypelagic North Atlantic contributes between 13 to 27% to the total prokaryotic production in the oxygen minimum layer and 41 to 84% in the Labrador Sea Water, declining to 10 to 20% in the North Atlantic Deep Water. Thus, planktonic archaea are actively growing in the dark ocean although at lower growth rates than bacteria and might play a significant role in the oceanic carbon cycle.     

Growth and uptake kinetics of phosphate by benthic microalga Nitzschia sp. isolated from Hiroshima Bay,Japan

In the present study, we experimentally investigated the phosphate uptake kinetics of benthic microalga Nitzschia sp. isolated from Hiroshima Bay, Japan. The maximum uptake rate (ρ_max) obtained by short‐term experiments was 6.84 pmol cell^?1 h^?1 for phosphate. The half‐saturation constant for uptake (K_S) was 61.2 µmol cell^?1 h^?1. Both the ρ_max and Ks of this species were extremely high, suggesting that Nitzschia sp. is adapted to benthic environments, where nutrient concentrations are much higher than in the water column. The specific maximum growth rate (µ'_max) and minimum cell quota (Q₀) for the P‐limited condition, obtained by a semi‐continuous growth experiment, were 0.48 day^?1 and 0.045 pmol cell^?1, respectively. It is concluded that Nitzschia sp. could be a ‘storage strategist’ species, meaning it adapts so as to minimize the influence of fluctuations in phosphate conditions resulting from the change in redox conditions of sediment due to bioturbation.     

High rates of net primary production and turnover of floating grasses on the Amazon floodplain: implications for aquatic respiration and regional CO2 flux

We investigated whether rates of net primary production (NPP) and biomass turnover of floating grasses in a central Amazon floodplain lake (Lake Calado) are consistent with published evidence that CO₂ emissions from Amazon rivers and floodplains are largely supplied by carbon from C4 plants. Ground‐based measurements of species composition, plant growth rates, plant densities, and areal biomass were combined with low altitude videography to estimate community NPP and compare expected versus observed biomass at monthly intervals during the aquatic growth phase (January–August). Principal species at the site were Oryza perennis (a C3 grass), Echinochloa polystachya, and Paspalum repens (both C4 grasses). Monthly mean daily NPP of the mixed species community varied from 50 to 96 g dry mass m^?2 day^?1, with a seasonal average (±1SD) of 64±12 g dry mass m^?2 day^?1. Mean daily NPP (±1SE) for P. repens and E. polystachya was 77±3 and 34±2 g dry mass m^?2 day^?1, respectively. Monthly loss rates of combined above‐ and below‐water biomass ranged from 31% to 75%, and averaged 49%. Organic carbon losses from aquatic grasses ranged from 30 to 34 g C m^?2 day^?1 from February to August. A regional extrapolation indicated that respiration of this carbon potentially accounts for about half (46%) of annual CO₂ emissions from surface waters in the central Amazon, or about 44% of gaseous carbon emissions, if methane flux is included.     

Effects of diurnal fluctuations in light intensity on the efficiency of growth of Skeletonema costatum (Grev.) Cleve (Bacillariophyceae) in a cyclostat

The effects of diurnal variations in light intensity on the biomass characteristics and the efficiency of daily growth of Skeletonema costatum (Grev.) Cleve were evaluated. The relative importance of changes in carbon specific rates of respiration and organic release to the efficiency of growth was determined. Light intensity was either constant at 130 μE · m^?2 · s^?1 during the light period or fluctuated throughout the light period from 500 to 10 μE · m^?2 · s^?1 at rates of either 1 or 12 cycles · day^?1. Total daily light was equivalent for all light regimes at 5.6 E · m^?2 · day^?1.Daily rates of growth remained comparable at ≈ 1 · day^?1 under constant and fluctuating light regimes. Cell size as daily mean carbon · cell^?1, nitrogen · cell^?1 and cellular volume was decreased under diurnally varying light whereas daily mean chlorophyll a · cell^?1 was unaffected.Rates of respiration, organic release and gross production were elevated several fold under diurnally varying light in comparison to constant light. Net growth efficiency decreased from 0.69 under constant light to values of 0.50 and 0.38 under 1 and 12 cycles · day^?1, respectively. Decreased efficiency of growth under diurnally fluctuating light resulted mostly from greater respiratory activity while organic release remained < 10% of gross production. Increased rates of gross production reflected enhancement in the efficiency of carbon fixation with fluctuating light.     

Fluctuating water levels control water chemistry and metabolism of a charophyte‐dominated pond

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Abundance and diversity of heterotrophic bacterial cells assimilating phosphate in the subtropical North Atlantic Ocean

Microorganisms play key roles in the cycles of carbon and nutrients in the ocean, and identifying the extent to which specific taxa contribute to these cycles will establish their ecological function. We examined the use of ³³P‐phosphate to identify heterotrophic bacteria actively involved in the cycling of phosphate, an essential inorganic nutrient. Seawater from the sub‐tropical North Atlantic Ocean was incubated with ³³P‐phosphate and analysed by microautoradiography to determine the proportion and diversity of the bacterial community‐assimilating phosphate. Complementary incubations using ³H‐leucine and ³H‐thymidine were also conducted. We found that a higher proportion of total heterotrophic bacterial cells in surface water samples assimilated phosphate compared with leucine or thymidine. Bacteria from all of the phylogenetic groups we identified by CARD‐FISH were able to assimilate phosphate, although the abundances of cells within each group did not scale directly with the number found to assimilate phosphate. Furthermore, a significantly higher proportion of Alphaproteobacteria, Gammaproteobacteria and Cytophaga‐like cells assimilated phosphate compared with leucine or thymidine. Our results suggest that a greater proportion of bacterial cells in surface waters are actively participating in the biogeochemical cycling of phosphorus, and possibly other elements, than is currently estimated through the use of ³H‐leucine or ³H‐thymidine.     

OLISTHODISCUS LUTEUS (CHRYSOPHYCEAE). III. UPTAKE AND UTILIZATION OF NITROGEN AND PHOSPHORUS1

Uptake and assimilation of nitrogen and phosphorus were studied in Olisthodiscus luteus Carter. A diel periodicity in nitrate reductase activity was observed in log and stationary phase cultures; there was a 10-fold difference in magnitude between maximum and minimum rates, but other cellular features such as chlorophyll a, carbon, nitrogen, C:N ratio (atoms) · cell^?1 were less variable. K_s values (~2 μM) for uptake of nitrate-N and ammonium-N were observed. Phosphorus assimilated · cell^?1· day^?1 varied with declining external phosphorus concentrations; growth rates <0.5 divisions · day^?1 were common at <0.5 μM PO_4-P. Phosphate uptake rates (K_s= 1.0–1.98 μM) varied with culture age and showed multiphasic kinetic features. Alkaline phosphatase activity was not detected. Comparisons of the nutrient dynamics of O. luteus to other phytoplankton species and the ecological implications as related to the phytoplankton community of Narragansett Bay (Rhode Island) are discussed.     

Abundance and production of bacteria, and relationship to phytoplankton production, in a large tropical lake (Lake Tanganyika)

1. Abundance and bacterial production (BP) of heterotrophic bacteria (HBact) were measured in the north and south basins of Lake Tanganyika, East Africa, during seasonal sampling series between 2002 and 2007. The major objective of the study was to assess whether BP can supplement phytoplankton particulate primary production (particulate PP) in the pelagic waters, and whether BP and particulate PP are related in this large lake. HBact were enumerated in the 0–100 m surface layer by epifluorescence microscopy and flow cytometry; BP was quantified using ³H‐thymidine incorporation, usually in three mixolimnion layers (0–40, 40–60 and 60–100 m). 2. Flow cytometry allowed three subpopulations to be distinguished: low nucleic acid content bacteria (LNA), high nucleic acid content bacteria (HNA) and Synechococcus‐like picocyanobacteria (PCya). The proportion of HNA was on average 67% of total bacterial abundance, and tended to increase with depth. HBact abundance was between 1.2 × 10⁵ and 4.8 × 10⁶ cells mL⁻¹, and was maximal in the 0–40 m layer (i.e. roughly, the euphotic layer). Using a single conversion factor of 15 fg C cell⁻¹, estimated from biovolume measurements, average HBact biomass (integrated over a 100‐m water column depth) was 1.89 ± 1.05 g C m⁻². 3. Significant differences in BP appeared between seasons, especially in the south basin. The range of BP integrated over the 0–100 m layer was 93–735 mg C m⁻² day⁻¹, and overlapped with the range of particulate PP (150–1687 mg C m⁻² day⁻¹) measured in the same period of time at the same sites. 4. Depth‐integrated BP was significantly correlated to particulate PP and chlorophyll‐a, and BP in the euphotic layer was on average 25% of PP. 5. These results suggest that HBact contribute substantially to the particulate organic carbon available to consumers in Lake Tanganyika, and that BP may be sustained by phytoplankton‐derived organic carbon in the pelagic waters.     

Concentrations of dimethylsulphoniopropionate and activities of dimethylsulphide-producing enzymes in batch cultures of nine dinoflagellate species

Dinoflagellates are recognised as one of the major phytoplankton groups that produce dimethylsulphoniopropionate (DMSP), the precursor of the marine trace gas dimethylsulphide (DMS) which has climate-cooling potential. To improve the prospects for including dinoflagellates in global climate models that include DMSP-related processes, we increased the data base for this group by measuring DMSP, DMS-producing enzyme activity (DPEA), carbon, nitrogen and Chl a in nine clonal dinoflagellate cultures (1 heterotrophic and 8 phototrophic strains). Growth rates ranged from 0.11 to 1.92?day^?1 with the highest value being for the heterotroph Crypthecodinium cohnii. Overall, we observed two orders of magnitude variability in DMSP content (11–364?mM) and detected DPEA in five of the nine strains (0.61–59.73?fmol?cell^?1?h^?1). Cell volume varied between 454 and 18,439?μm³ and whilst C and N content were proportional to the cell volume, DMSP content was not. The first DMSP measurements for a dinoflagellate from Antarctic waters and a species with diatom-like plastids are included. Lower DMSP concentrations were found in three small athecate species and a dinoflagellate with haptophyte-like plastids. The highest concentrations and production rates tended to be in globally distributed dinoflagellates and the heterotroph. Photosynthetic species that are distributed in temperate to tropical waters showed low DMSP concentrations and production rates and the polar representative showed moderate concentration and a low production rate. Estuarine species had the lowest concentrations and production rates. These data should help refine the inclusion of dinoflagellates as a functional group in future global climate models.     

Flagellate nutritional versatility as a key to survival in two contrasting Antarctic saline lakes

1. Seasonal patterns of grazing and photosynthesis were investigated in two saline Antarctic lakes (Highway and Ace) in the Vestfold Hills (68°S). The phototrophic nanoflagellate (PNAN) community was dominated by Pyramimonas gelidicola and two morphological forms of a cryptophyte species that occurred throughout the year. Both species were mixotrophic on bacteria, and in Highway Lake they also exploited dissolved organic carbon as determined by the uptake of fluorescently labelled dextrans. 2. Clearance rates ranged between 0.02 and 0.21 nL h^?1 cell^?1 in Ace Lake and 0.004–1.05 nL h^?1 cell^?1 in Highway Lake. On occasion cryptophyte grazing equalled that of the heterotrophic nanoflagellates (HNAN). 3. Photosynthetic rates showed similar trends in both lakes, but there were differences in chlorophyll a specific rates and photosynthetic efficiency, probably related to the meromictic characteristic of Ace Lake. Primary production was measurable in winter and peaked in summer following the maxima of mixotroph grazing. 4. The HNAN community of Highway Lake achieved clearance rates of 0.02–1.80 nL h^?1 cell^?1 and removing between 50 and 693 ng bacterial carbon L^?1 day^?1, with highest impact in winter when HNAN were most abundant. The HNAN also ingested fluorescently labelled dextrans showing a preference for 4 and 500 kDa molecules. The more diverse HNAN community of Ace Lake had lower clearance rates (0.04–0.37 nL h^?1 cell^?1) and exerted a lower grazing pressure on bacterioplankton. In Highway Lake, where the HNAN community was dominated by the choanoflagellate Diaphanoeca grandis, there was a significant correlation between mean cell volume and clearance rate. 5. The major feature was that the microbial plankton functioned throughout the year by employing nutritional versatility.     

FATTY ACIDS IN PHOTOTROPHIC AND MIXOTROPHIC GYRODINIUM GALATHE‐ANUM (DINOPHYCEAE)

Fatty acids were measured in G. galatheanum grown either phototrophically, or mixotrophically with Storeatula major (Cryptophyceae) as prey. G. galatheanum, like many photosynthetic dinoflagellates, contains high amounts of n‐3 long‐chain‐polyunsaturated fatty acids (LC‐PUFA) such as docosahexaenoic acid (DHA, 22:6n‐3) and the hemolytic toxic fatty acid 18:5n‐3. We hypothesize that a benefit of phagotrophy in G. galatheanum is the acquisition of precursor linolenic acid (18:3n‐3) that fuels LC‐PUFA synthesis. Phototrophs grew at 0.37 d^?1, while mixotrophs grew at 0.40 d^?1 with a feeding rate of 0.62 d^?1. Photosynthesis was lower in mixotrophs (3.7 pg C cell^?1 h^?1) than phototrophs (4.9 pg C cell^?1 h^?1). DHA levels were higher in mixotrophs [3.7 (+/? 0.11) pg cell^?1] than phototrophs [3.0 (+/? 0.16) pg cell^?1] and prey [0.4 (+/? 0.01) pg cell^?1]. 18:5n–3 levels [1.7 (+/? 0.03) pg cell^?1] were similar in phototrophs and mixotrophs. An intermediate in n‐3 LC‐PUFA synthesis, 20:4n‐3, accumulated in mixotrophs [0.6 (+/? 0.27) pg cell^?1] relative to phototrophs (not detected) and prey [0.03 (+/? 0.002) pg cell^?1]. Low ratios of linolenic acid to DHA in phototrophic G. galatheanum (0.14) relative to mixotrophic G. galatheanum (0.29) and prey (2.14) are consistent with substrate limitation of LC‐PUFA synthesis in phototrophs. Accumulation of 20:4n‐3 suggests incomplete conversion of linolenic acid to DHA, possibly due to conditions in batch culture. We conclude that precursors for n‐3 LC‐PUFA biosynthesis in G. galatheanum may be acquired through ingestion of S. major, and may partially control feeding/photosynthesis in mixotrophic populations.     

Low‐temperature (9°C) AMD treatment in a sulfidogenic bioreactor dominated by a mesophilic Desulfomicrobium species

The possibilities for the treatment of low‐temperature mine waste waters have not been widely studied. The amenability of low‐temperature sulfate reduction for mine waste water treatment at 9°C was studied in a bench‐scale fluidized‐bed bioreactor (FBR). Formate was used as the electron and carbon source. The first influent for the FBR was acidic, synthetic waste water containing iron, nutrients, and sulfate, followed by diluted barren bioleaching solution (DBBS). The average sulfate reduction rates were 8 mmol L^?1 day^?1 and 6 mmol L^?1 day^?1 with synthetic waste water and DBBS, respectively. The corresponding specific activities were 2.4 and 1.6 mmol SO g VSS^?1 day^?1, respectively. The composition of the microbial community and the active species of the FBR was analyzed by extracting the DNA and RNA, followed by PCR‐DGGE with the universal bacterial 16S rRNA gene primers and dsrB‐primers specific for sulfate‐reducing bacteria. The FBR microbial community was simple and stable and the dominant and active species belonged to the genus Desulfomicrobium. In summary, long‐term operation of a low‐temperature bioreactor resulted in enrichment of formate‐utilizing, psychrotolerant mesophilic sulfate reducing bacteria. Biotechnol. Bioeng. 2009; 104: 740–751 © 2009 Wiley Periodicals, Inc.