Microbial processes in a high-latitude fjord (Kongsfjorden, Svalbard): II. Ciliates and dinoflagellates

The composition and ecological role of ciliates and dinoflagellates were investigated at one station in Kongsfjorden, Svalbard, during six consecutive field campaigns between March and December 2006. Total ciliate and dinoflagellate abundance mirrored the seasonal progression of phytoplankton, peaking with 5.8 × 10⁴ cells l⁻¹ in April at an average chlorophyll a concentration of 10 μg l⁻¹. Dinoflagellates were more abundant than ciliates, dominated by small athecates. Among ciliates, aloricate oligotrichs dominated the assemblage. A large fraction (>60%) of ciliates and dinoflagellates contained chloroplasts in spring and summer. The biomass of the purely heterotrophic fraction of the ciliate and dinoflagellate community (protozooplankton) was with 14 μg C l⁻¹ highest in conjunction with the phytoplankton spring bloom in April. Growth experiments revealed similar specific growth rates for heterotrophic ciliates and dinoflagellates (<0–0.8 d⁻¹). Food availability may have controlled the protozooplankton assemblage in winter, while copepods may have exerted a strong control during the post-bloom period. Calculations of the potential grazing rates of the protozooplankton indicated its ability to control or heavily impact the phytoplankton stocks at most times. The results show that ciliates and dinoflagellates were an important component of the pelagic food web in Kongsfjorden and need to be taken into account when discussing the fate of phytoplankton and biogeochemical cycling in Arctic marine ecosystems.     

Changes in bacterial and ciliate densities with trophic status in Mediterranean shallow lakes

Ciliate and bacterial densities and their link with eutrophication were studied in fourteen shallow lakes in northwest Spain. Total phosphorus (TP) in these lakes varied between 30 μg l⁻¹ and 925 μg l⁻¹ and chlorophyll a concentration (chla) between 0.5 μg l⁻¹ and 107 μg l⁻¹. Bacterial abundance ranged from 1 × 10⁶ to 14 × 10⁶ cells ml⁻¹, while ciliate abundance ranged from 0.6 cells ml⁻¹ to 229 cells ml⁻¹. Lakes were classified into three trophic types from their TP and chla concentrations. Bacterial abundance was significantly correlated with trophic type, as well as with TP and with chla separately, whereas ciliate abundance was only correlated with chla. No significant relationship could be established between bacterial and ciliate abundance across the trophic gradient. A general pattern was observed in the ratios of bacterial abundance to TP and chla concentrations, of decreasing ratios with increases in the nutrient loading. This pattern was not found for ciliates. The dominant zooplankton group in 13 of the 14 lakes studied was Rotifera, which accounted for a mean of 71% of total zooplankton abundance (41% of zooplankton biomass). The positive correlation between bacteria and ciliates with this group, and the absence of any relationship with Cladocera suggest that top down control by cladocerans was weaker in our lakes than previously shown in northern European shallow lakes. Rotifers could be important predators of bacteria in the high-nutrient lakes of our study. Higher slopes of regressions on bacterial abundance towards the hypertrophic range indicate that top-down control was weaker in our lakes than in northern European shallow lakes.     

Annual and interannual variability in phytoplankton at a permanent station off Kerguelen Islands, Southern Ocean

From November 1992 to February 1995 a quantitative and qualitative phytoplankton study was conducted at a permanent station (Kerfix) southwest off the Kerguelen Islands, in the vicinity of the Polar Front (50°40′S–68°25′E). Phytoplankton populations are low in this area both during summers and winters. They consist, in order of decreasing cell abundance, of pico- and nanoflagellates (1.5–20 μm), coccolithophorids (<10 μm), diatoms (5–80 μm) and dinoflagellates (6–60 μm). Flagellates form the dominant group throughout the year and attain the highest summer average of 3.0 × 10⁵ cells l⁻¹. Next in abundance year-round are coccolithophorids with the dominant Emiliania huxleyi (highest summer 1992 average 1.9 × 10⁵ cells l⁻¹), diatoms (summer 1992 average 1.0 × 10⁵ cells l⁻¹) and dinoflagellates (average 3.8 × 10⁴ cells l⁻¹). Winter mean numbers of flagellates and picoplankton do not exceed 8.4 × 10⁴ cells l⁻¹; those of the three remaining algal groups together attain 2 × 10⁴ cells l⁻¹. Summer peaks of diatoms and dinoflagellates are mainly due to the larger size species (>20 μm). The latter group contributes most to the total cell carbon biomass throughout the year. Dominant diatoms during summer seasons include: Fragilariopsis kerguelensis, Thalassionema nitzschioides, Chaetoceros dichaeta, C. atlanticus, Pseudonitzschia heimii, and P. barkleyi/lineola. This diatom dominance structure changes from summer to summer with only F. kerguelensis and T. nitzschioides retaining their first and second positions. Any one of the co-dominant species might be absent during some summer period. The variable diatom community structure may be due to southward meandering of the Polar Front bringing “warmer” species from the north, and to the mixing of the water masses in this area. The entire community structure characterized both during summer and winters by the dominance of flagellates can be related to deep mixing (ca. 40–200 m) of the water column as the probable controlling factor. Received: 13 November 1997 / Accepted: 11 May 1998     

A comparison of phytoplankton size-fractions in Mondsee,an alpine lake in Austria: distribution,pigment composition and primary production rates

Production rates, abundance, chlorophyll a (Chl a) concentrations and pigment composition were measured for three size classes (<2 μm, 2–11 μm and >11 μm) of phytoplankton from May to December 2000 in deep, mesotrophic, alpine lake Mondsee in Austria. The study focuses on differences among phytoplankton size fractions characterised by their surface area to volume ratio ([mm² l⁻¹: mm³l⁻¹]), pigment distribution patterns and photosynthetic rates. Particular attention was paid to autotrophic picophytoplankton (APP, fraction <2 μm) since this size fraction differed significantly from the two larger size fractions. Among the three fractions, APP showed the highest surface area to volume ratios and a high persistence in the pattern of lipophilic pigments between temporarily and spatially successive samples (about 80% similarity of pigment composition between samples over seasons and depths). The epilimnetic abundance of APP varied seasonally with an annual maximum of 180 × 10³ cells ml⁻¹ in June (at 4–9 m). The minimum (October at 12 m) was more than an order of magnitude lower (4.9 × 10³ ml⁻¹). APP peaked during autumn and contributed between 24% and 42% to the total area-integrated Chl a (10–23 mg m⁻²) and between 16% and 58% to total area-integrated production (5–64 mg m⁻²  h⁻¹) throughout seasons.     

Relevance of bacterioplankton abundance and production in the oligotrophic equatorial Indian Ocean

Bacterioplankton abundance and production, chlorophyll a (Chl a) concentrations and primary production (PP) were measured from the equatorial Indian Ocean (EIO) during northeast (NEM), southwest (SWM) and spring intermonsoon (SpIM) seasons from 1°N to 5°S along 83°E. The average bacterial abundance was 0.52 ± 0.29, 0.62 ± 0.33 and 0.46 ± 0.19 (× 10⁸ cells l⁻¹), respectively during NEM, SWM and SpIM in the top 100 m. In the deep waters (200 m and below), the bacterial counts averaged ∼0.35 ± 0.14 × 10⁸ cells l⁻¹ in SWM and 0.39 ± 0.16 × 10⁸ cells l⁻¹ in SpIM. The 0–120 m column integrated bacterial production (BP) ranged from 19 to 115 and from 10 to 51 mg C m⁻² d⁻¹ during NEM and SWM, respectively. Compared with many open ocean locations, bacterial abundance and production in this region are lower. The bacterial carbon production, however, is notably higher than that of phytoplankton PP (BP:PP ratio 102% in SWM and 188% in NEM). With perpetually low PP (NEM: 20, SWM: 18 and SpIM: 12 mg C m⁻² d⁻¹) and Chl a concentration (NEM: 16.5, SWM: 15.0 and SpIM: 20.9 mg m⁻²), the observed bacterial abundance and production are pivotal in the trophodynamics of the EIO. Efficient assimilation and mineralization of available organics by bacteria in the euphotic zone might serve a dual role in the ultra-oligotrophic regions including EIO. Thus, bacteria probably sustain microheterotrophs (micro- and meso-zooplankton) through microbial loop. Further, rapid mineralization by bacteria will make essential nutrients available to autotrophs.     

Partitioning of the food ration of marine ciliates between pico- and nanoplankton

Food size-range for 13 species of Tintinnina and 18 species of Oligotrichina were studied using electronic particle counting and in situ observation of food vacuole contents. Tintinnids consume nanoplankton in the size range 2–20 μm. Oligotrichous naked ciliates consume particles in the size range 0.5–10 μm. Ciliates smaller than 30 μm take 72% picoplankton and 28% nanoplankton. For ciliates between 30 μm and 50 μm the proportions are reversed (30% pico- and 70% nanoplankton), while the larger ciliates (> 50 μm) take nanoplankton almost exclusively (95% nano- and 5% picoplankton). A seasonal study of total Oligotrichida grazing showed that natural particles were consumed at rates that varied from 1 to 20 μg C 1⁻¹ day⁻¹. This included between 1 and 38% of the bacterioplankton production and 9 to 52% of the nanoplankton production. In the N-W Mediterranean the total ciliate production varied from 0.4 to 8.2 μg C 1⁻¹ day⁻¹. Research supported by CNRS-PIROCEAN AIP-RTM-953146-GRECO P4 and by CNRS-UA 716 (FR), and by the University of Nice and by the CNRS-GRECO 88 (MLP)     

Small nanoplankton and bacteria in the Western Ross Sea during sea-ice retreat (spring 1994)

Spatial changes of small nanoplankton (2–10 μm) were investigated in relation to sea-ice conditions, hydrography and receding ice processes in the Ross Sea (Antarctica) during spring 1994. Abundance and biomass of heterotrophic and autotrophic nanoplankton, as well as bacterioplankton, were determined along a south-north transect from the open waters polynya towards the pack ice. Autotrophic and heterotrophic nanoplankton biomass ranged from 758 to 4570 mgC m⁻² and from 3 to 387 mgC m⁻², respectively. Heterotrophic nanoplankton accounted, on average, for about 9% of the total (i.e. autotrophic plus heterotrophic) nanoplankton biomass. The size structure of both auto- and heterotrophic nanoplankton in the Ross Sea continental shelf receding ice edge was different from that of nanoplankton associated with the shelf break and open Antarctic ice-edge area. Generally, the highest heterotrophic biomass was found in the pack-ice zone on the continental shelf, while the highest heterotrophic contribution to the total nanoplankton biomass (up to 25%) was encountered at the shelf break where phytoplankton was largely dominated by 2- to 3-μm-size cells. Accepted: 2 May 1999     

Relationships Between Coastal Bacterioplankton Growth Rates and Biomass Production: Comparison of Leucine and Thymidine Uptake with Single-Cell Physiological Characteristics

Specific growth rates of heterotrophic bacterioplankton have been frequently estimated from in situ bacterial production (BP) to biomass (BB) ratios, using a series of assumptions that may result in serious discrepancies with values obtained from predator-free cultures. Here, we used both types of approaches together with a comprehensive assessment of single-cell physiological characteristics (membrane integrity, nucleic acid content, and active respiration) of coastal bacterioplankton during a complete annual cycle (February 2007–January 2008) in the southern Bay of Biscay off Xixón, Spain. Both leucine and thymidine incorporation rates were used in conjunction with empirical tracer to carbon or cells conversion factors (eCFs) to accurately derive BP. Leu and TdR incorporation rates covaried year-round, as did the corresponding eCFs at 0 and 50 m depth. eCFs peaked in autumn, with mean annual values close to the theoretical ones (3.4 kg C mol Leu⁻¹ and 2.0 × 10¹⁸ cells mol TdR⁻¹). Bacterial abundance (0.2–1.5 × 10⁶ cells L⁻¹) showed a bimodal distribution with maxima in May and October and minima in March. Live (membrane-intact) cells dominated year-round (79–97%), with high nucleic acid cells (42–88%) and actively respiring bacteria (CTC+, 1–16%) showing distinct surface maxima in April and July, respectively. BB (557–1,558 mg C m⁻²) and BP (7–139 mg C m⁻² day⁻¹) presented two distinct peaks in spring and autumn, both of similar size due to a strong upwelling event observed in September. Specific growth rates (0.35–3.8 day⁻¹) were one order of magnitude higher in predator-free incubations than bacterial turnover rates derived from integrated BP:BB ratios (0.01–0.16 and 0.01–0.09 day⁻¹, for Leu and TdR, respectively) and were not correlated, probably due to a significant contribution of low activity cells to total standing stocks. The Leu:TdR molar ratio averaged for the water column (6.6–25.5) decreased significantly with higher integrated BB, indicating that low standing stocks tend to present unbalanced growth. Discrepancies about the true magnitude of specific growth rates must be solved before fully appreciating the role of bacteria in the ocean carbon cycle.     

Seasonal variations of microphytoplankton assemblages and environmental variables in the coastal zone of Bozcaada Island in the Aegean Sea (NE Mediterranean Sea)

Seasonal changes in the microphytoplankton assemblages were examined in the coastal zone of Bozcaada Island with regard to some major physical and chemical variables. Samples were collected from May 2000 to December 2001 at four stations. A total of 108 dinoflagellates, 102 diatoms, 1 chrysophycean, 3 dictyochophycean, and 1 prasinophycean species were identified and quantified during the study period. Diatoms and dinoflagellates were the most important in terms of species number and abundance. The maximum values of total microphytoplankton were observed at 0.5 m depth (46.2 × 10³ cells l⁻¹ at st. 3) in May as this was the month when the diatom Pseudo-nitzschia pungens bloomed. Chlorophyll (chl) a concentration ranged between 0.08 (August) and 0.78 μg l⁻¹ (February). May was another important month in which chlorophyll a increased (0.41–0.47 μg l⁻¹). Species diversity values (H′_log2) ranged from 1.66 bits (June, 20 m) to 4.11 bits (November, 0.5 m). The increase was attributed to a more balanced distribution of abundance among species. The amounts of nitrate + nitrite (0.6−3.7 μg-at N l⁻¹), phosphate (0.2−0.6 μg-at P l⁻¹) and silicate (0.7−2.5 μg-at Si l⁻¹) were recorded on each sampling occasion. Nutrient concentrations and chl a values of the research area were found to be poorer than those of the many other coastal areas in the northeastern Mediterranean. The mean atomic ratio of nitrogen to phosphorus varied from 1.3 (June) to 12.9 (February). This ratio was lower than the Redfield ratio of 16 for ocean phytoplankton, and phytoplankton was potentially limited by nitrogen for most of the months. The result of this study confirms and emphasizes the oligotrophic nature of the eastern Mediterranean.     

The effect of <Emphasis Type="Italic">Poterioochromonas</Emphasis> abundance on production of intra- and extracellular microcystin-LR concentration

Due to its capability for producing various microcystins, Microcystis aeruginosa is recognized as one of the most toxic, bloom-forming cyanobacteria. In this study, the fates of intra- and extracellular microcystin-LR (MC-LR) were investigated when the mixotrophic golden alga Poterioochromonas sp. (ZX1) was grazing on M. aeruginosa cells. In the control groups, the total MC-LR concentration increased with the growth of M. aeruginosa with an MC-LR content per cell of 0.5–1.5 × 10⁻⁸ μg cell⁻¹. In the treatment with ZX1, the total MC-LR decreased linearly throughout the incubation period. In particular, intracellular MC-LR disappeared with a loss of M. aeruginosa cells in the first few days. Part of the intracellular MC-LR was released to the medium under the grazing stress, resulting in an increase of extracellular MC-LR. The degradation rate of MC-LR was positively related to the initial abundance of ZX1 and negatively related to that of M. aeruginosa. The inhibition ratio of MC-LR production dropped sharply from 98 to 67% when the initial abundance of M. aeruginosa increased from 10⁶ to 10⁷ cells ml⁻¹. However, it increased from 84 to 99% when the initial ZX1 abundance increased from 10⁴ to 10⁵ cells ml⁻¹. The effective removal of both M. aeruginosa cells and MC-LR was observed under lower M. aeruginosa abundance (<10⁶ cells ml⁻¹) and higher ZX1 abundance (>1% of M. aeruginosa abundance). Light had little impact on MC-LR degradation, but MC-LR degradation decreased due to the loss of ZX1 after 10 days of darkness. This study showed that the interactions between M. aeruginosa and ZX1 were strongly influenced by their initial abundances.     

The importance and distinctiveness of small-sized phytoplankton in the Magellan Straits

The distribution of summer phytoplankton across the Straits of Magellan (SOM) was studied with the aims of tracing differences among the distinct subregions of the area and contributing to the knowledge of its biodiversity. Samples collected at 25 stations were observed and counted in light microscopy. Selected samples were observed with transmission electron microscopy. The main unifying feature of the phytoplankton in the SOM was the high abundance and numerical dominance of small-sized (<10 μm) eukaryotic species, among which coccoid cells of <3 μm size were predominant (56.2 ± 30.6 of the total phytoplankton abundance). They mostly belonged to the prasinophyte Pycnococcus provasolii, which was abundant (0.8–6,834 cells × 10³ ml⁻¹) at all stations with the exception of those in proximity to the Atlantic entrances, where it was not recorded. Small-sized (<3 and 3–5 μm) diatoms (Minidiscus trioculatus, Lennoxia faveolata and other undetermined centric species) attained high densities (<3,757 cells 10³ ml⁻¹) especially at stations of the Patagonian sectors, whereas microplanktonic diatoms were only found at the two entrances of the Straits. Dinoflagellates were constituted mainly by >10 μm forms in the Andean subregion and <10 μm naked species in the Patagonian subregion, contributing up to 75.9 and 41.8% of the total carbon in these two areas, respectively. In the Patagonian subregion, flagellates mainly constituted by <5 μm forms and by cryptomonads <10 μm comprised up to 53.9% of the total biomass. Several species identified in this study have never been reported in other investigations in the SOM, while others, including Pycnococcus provasolii and Lennoxia faveolata, have rarely been recorded elsewhere. Overall, the summer phytoplankton of the Straits does not resemble that of any other region of the world’s seas. Although some of the predominant species might have been overlooked elsewhere, their abundance and relative importance apparently constitute a distinctive feature of the SOM.     

Abundance and composition of the summer phytoplankton community along a transect from the Barguzin River to the central basin of Lake Baikal

The abundance and composition of phytoplankton were investigated at six stations along a transect from the Barguzin River inflow to the central basin of Lake Baikal in August 2002 to clarify the effect of the river inflow on the phytoplankton community in the lake. The water temperature in the epilimnion was high near the shore at Station 1 (17.3°C), probably due to the higher temperature of the river water, and gradually decreased offshore at Station 6 (14.5°C). Thermal stratification developed at Stations 2–6, and a thermocline was observed at a 17–22-m depth at Stations 2–4 and an 8–12-m depth at Stations 5 and 6. The concentrations of nitrogen and phosphorus nutrients in the epilimnion at all stations were <1.0 μmol N l⁻¹ and <0.16 μmol P l⁻¹, respectively. Relatively high concentrations of nutrients (0.56–7.38 μmol N l⁻¹ and 0.03–0.28 μmol P l⁻¹) were detected in the deeper parts of the euphotic zone. Silicate was not exhausted at all stations (>20 μmol Si l⁻¹). The chlorophyll a (chl. a) concentration was high (>10 μg l⁻¹) near the shore at Station 1 and low (<3 μg l⁻¹) at five other stations. The <2 μm fraction of chl. a in Stations 2–6 ranged between 0.80 and 1.85 μg l⁻¹, and its contribution to total chl. a was high (>60%). In this fraction, picocyanobacteria were abundant at all stations and ranged between 5 × 10⁴ and 5 × 10⁵ cells ml⁻¹. In contrast, chl. a in the >2 μm fraction varied significantly (0.14–11.17 μg l⁻¹), and the highest value was observed at Station 1. In this fraction, the dominant phytoplankton was Aulacoseira and centric diatoms at Station 1 and Cryptomonas, Ankistrodesmus, Asterionella, and Nitzschia at Stations 2–6. The present study demonstrated the dominance of picophytoplankton in the pelagic zone, while higher abundance of phytoplankton dominated by diatoms was observed in the shallower littoral zone. These larger phytoplankters in the littoral zone probably depend on nutrients from the Barguzin River.     

A study of the diatom-dominated microplankton summer assemblages in coastal waters from Terre Adélie to the Mertz Glacier, East Antarctica (139°E–145°E)

In January 2004 the microplankton community from the coastal waters of Terre Adélie and Georges V Land (139°E–145°E) was studied. Results showed a diatom-dominated bloom with chlorophyll a levels averaging 0.64 μg l⁻¹ at 5 m depth (range 0.21–1.57 μg l⁻¹). Three geographic assemblages of diatoms were identified, based on principal diatom taxa abundances. The stratified waters near the Mertz Glacier presented highest phytoplankton biomasses (0.28–1.57 μg Chl a l⁻¹ at 5 m) and diatom abundances (6,507–70,274 cells l⁻¹ at 5 m), but low diversity, dominated by Fragilariopsis spp. Lower biomasses (0.38–0.94 μg Chl a l⁻¹ at 5 m) and abundances (394–9,058 cells l⁻¹ at 5 m) were observed in the mixed waters around the Astrolabe Glacier with a diverse diatom community characterised by larger species Corethron pennatum and Rhizosolenia spp. Finally an intermediate zone between them over the shallower shelf waters of the Adélie Bank represented by Chaetoceros criophilus, where biomasses (0.21–0.35 μg Chl a l⁻¹ at 5 m) and abundances (1,190–5,431 cells l⁻¹ at 5 m) were lowest, coinciding with the presence of abundant herbivorous zooplankton.     

Ecological balance between ciliate plankton and its prey candidates,pico- and nanoplankton,in the East China Sea

Standing stocks of ciliate plankton and its prey candidates, both picoplankton and nanoplankton, were investigated in spring in the East China Sea. The former was 1.36 × 10⁵–1.54 × 10⁸ μm³ l⁻¹ in biovolume, and the latter was 5.33 × 10⁶–1.11 × 10⁸ μm³ l⁻¹. The biovolume ratio of ciliate plankton to prey candidates ranged from 1.31 × 10⁻² to 2.00 × 10⁰; it was larger in abundant prey conditions and smaller in sparse preys. Making some plausible assumptions about physiological activity on both organisms, every ratio meet the quantitative restriction that prey production should be equal to or larger than ciliate consumption. However, prey candidates would be so sparsely distributed that ciliate plankton could not capture sufficient prey organisms in its random filter-feeding manner. Even though planktonic ciliates must have some extraordinary mechanisms to capture preys efficiently, this quantitative imbalance might be one of the reasons for decreasing ciliate/prey ratio in sparse prey conditions. Handling editor: K. Martens     

Virus impact on heterotrophic bacterioplankton of water reservoirs

The quantitative distribution of viruses and their impact on heterotrophic bacterioplankton were studied in mesotrophic and eutrophic reservoirs of the Volga and Volga-Baltic waterway. The abundance of planktonic virus particles ranged from 9.4 × 10⁶ to 120 × 10⁶ ml⁻¹ and was from 2.5 to 9 times greater than the bacterial numbers. Production of virioplankton varied from 2.1 × 10⁶ to 132 × 10⁶ particles (ml day)⁻¹ and the population turnover time values were between 0.3 and 11.6 days. The maximum values of numbers and production of virio- and bacterioplankton were observed in the eutrophic Ivan’kovo reservoir. Distribution of the viruses in the Volga reservoirs depended to a significant degree on the number and activity of heterotrophic bacterioplankton. The infected bacteria accounted for 5.5–33.5% of the total bacterial abundance. Phages were an important factor of bacterial mortality. During July to September virus-induced bacterial mortality varied between 6.1 and 40.6% (20.2% on average) of daily bacterioplankton production.     

Growth-associated biosynthesis of astaxanthin in heterotrophic Chlorella zofingiensis (Chlorophyta)

The green microalga Chlorella zofingiensis can produce the ketocarotenoid astaxanthin under heterotrophic culture conditions. Here we report the growth-associated biosynthesis of astaxanthin in this biotechnologically important alga. With glucose as sole carbon and energy source, C. zofinginesis grew fast in the dark with rapid exhaustion of nitrogen and carbon sources from media, leading to a high specific growth rate (0.034 h⁻¹). Cultures started at a cell concentration of about 3.4 × 10⁹ cells l⁻¹ reached, after 6 days, standing biomass values of 1.6 × 10¹¹ cells or 8.5 g dry weight l⁻¹. Surprisingly, the biosynthesis of astaxanthin was found to start at early exponential phase, independent of cessation of cell division. A general trend was observed that the culture conditions benefiting cell growth also benefited astaxanthin accumulation, indicating that astaxanthin was a growth-associated product in this alga. The maximum cell dry biomass and astaxanthin yield were 11.75 g l⁻¹ and 11.14 mg l⁻¹ (about 1 mg g⁻¹), simultaneously obtained in the fed-batch culture with a combined glucose–nitrate mixture addition, which were the highest ever reported in dark-heterotrophic algal cultures. The possible reasons why dark-heterotrophic C. zofingiensis could produce astaxanthin during the course of cell growth were discussed.     

Antarctic sea ice viral dynamics over an annual cycle

Viral abundance, burst sizes, lytic production and temperate phage were investigated in land-fast ice at two sites in Prydz Bay Antarctica (68°S, 77°E) between April and November 2008. Both ice cores and brine were collected. There was no seasonal pattern in viral or bacterial numbers. Across the two sites virus abundances ranged between 0.5 × 10⁵ and 5.1 × 10⁵ viruses ml⁻¹ in melted ice cores and 0.6 × 10⁵–3.5 × 10⁵ viruses ml⁻¹ in brine, and bacterial abundances between 2.7 × 10⁴ and 17.3 × 10⁴ cells ml⁻¹ in melted ice cores and 3.9 × 10⁴–32.5 × 10⁴ cells ml⁻¹ in brine. Virus to bacterium ratios (VBR) showed a clear seasonal pattern in ice cores with lowest values in winter (range 1.2–20.8), while VBRs in brine were lower (0.2–4.9). Lytic viral production range from undetectable to 2.0 × 10⁴ viruses ml⁻¹ h⁻¹ in ice cores with maximum rates in September and November. In brine maximum, lytic viral production occurred in November (1.18 × 10⁴ viruses ml⁻¹ h⁻¹). Low burst sizes were typical (3.94–4.03 viruses per bacterium in ice cores and 3.16–4.0 viruses per bacterium in brine) with unusually high levels of visibly infected cells—range 40–50%. This long-term investigation revealed that viral activity was apparent within the sea ice throughout its annual cycle. The findings are discussed within the context of limited data available on viruses in sea ice.     

The viability to a wall shear stress and propagation of <Emphasis Type="Italic">Bifidobacterium longum</Emphasis> in the intensive membrane bioreactor

Bifidobacterium longum grew at 65 L pilot scale of the membrane bioreactor (MBR), externally fitted with ceramic membrane (0.7 m²). Cell mass at the MBR reached 22.18 g L⁻¹ as dry cell weight in 12 h, which is 8.44 times higher than cell mass attained at the vial culture. The growth rate in the vial culture was μ = 0.385 h⁻ and at the batch culture was μ = 1.13 h⁻ in the exponential period and μ = 0.31 h⁻¹ in the stationary period. In the fed-batch mode was μ = 1.102 h⁻¹ for 6 h with inoculation and declined to μ = 0.456 h⁻¹ with feeding of feed medium. The growth rate at the MBR was μ = 0.134 h⁻¹. The number of viable cells was 6.01 × 10¹² cfu L⁻¹ at the batch culture, but increased to 1.15 × 10¹⁴ cfu L⁻¹ at the MBR culture. The specific growth rate of viable cell number (colony-forming units per liter, per hour) improved by 6.01 times from the batch to the MBR culture. The wall shear stress mainly generated by the pump, and the membrane incorporated into the MBR was controlled during the cultivation at the MBR. The viability of B. longum declined to under 10% in the first 2 weeks of the 4-week stability test (40°C) as B. longum was exposed to over wall shear stress 713 Pa, but the viability improved to 30–40% in wall shear stress of 260 Pa or STR culture. The loss in the cell viability can be saved by managing with wall shear stress during the cultivation at the MBR.     

Seasonal changes of Antarctic marine bacterioplankton and sea ice bacterial assemblages

The distributions of bacterial populations in sea ice and underlying seawater were investigated on the continental shelf of the “Terre Adélie” area. A reference station was sampled weekly from January 1991 to January 1992. In winter, the survey included a minimum of six sampling layers: surface and bottom ice, brine, seawater from the interface, and at 0.5 and 2 m depth. In seawater, the total bacterial abundance ranged from 0.5 × 10⁵ cells ml⁻¹ in July to 6.0 × 10⁵ cells ml⁻¹ after ice break. Values reaching 2.5 × 10⁶ cells ml⁻¹ were recorded in the overlying ice cover. Mean cell volumes were twice as high in brine as in seawater. The saprophytic bacterial abundance ranged from 5.0 × 10⁴ CFU (colony-forming units) ml⁻¹ in some winter interface samples to less than 1.0 × 10³ CFU ml⁻¹ in most of the summer seawater samples. In sea ice a clear decreasing gradient for most of the studied bacterial parameters from the surface layers towards the bottom layer was found. The ice cover had a discernible impact on underlying seawater, but its influence was restricted to a limited interface layer.     

Optimization of protoplast yields from the red algae <Emphasis Type="Italic">Gracilaria dura</Emphasis> (C. Agardh) J. Agardh and G. <Emphasis Type="Italic">verrucosa</Emphasis> (Huds.) Papenfuss

This study reports on the optimization of protoplast yield from two important tropical agarophytes Gracilaria dura and Gracilaria verrucosa using different cell-wall-degrading enzymes obtained from commercial sources. The conditions for achieving the highest protoplast yield was investigated by optimizing key parameters such as enzyme combinations and their concentrations, duration of enzyme treatment, enzyme pH, mannitol concentration, and temperature. The significance of each key parameter was also further validated using the statistical central composite design. The enzyme composition with 4% cellulase Onozuka R-10, 2% macerozyme R-10, 0.5% pectolyase, and 100 U agarase, 0.4 M mannitol in seawater (30‰) adjusted to pH 7.5 produced the highest protoplast yields of 3.7 ± 0.7 × 10⁶ cells g⁻¹ fresh wt for G. dura and 1.2 ± 0.78 × 10⁶ cells g⁻¹ fresh wt for G. verrucosa when incubated at 25°C for 4–6 h duration. The young growing tips maximally released the protoplasts having a size of 7–15 μm in G. dura and 15–25 μm in G. verrucosa, mostly from epidermal and upper cortical regions. A few large-size protoplasts of 25–35 μm, presumably from cortical region, were also observed in G. verrucosa.