Bacterioplankton Secondary Production Estimates for Coastal Waters of British Columbia, Antarctica, and California

The principal objective of this study was to quantify the rate of heterotrophic bacterioplankton production. Production was estimated by two approaches: (i) measurement of increasing bacterial abundance with time in filtered (3-μm pore size) seawater and (ii) estimation of bacterial deoxyribonucleic acid synthesis by tritiated thymidine incorporation in unfractionated seawater. The two approaches yielded comparable results when used at the Controlled Ecosystem Population Experiment (Saanich Inlet, British Columbia, Canada), at McMurdo Sound (Antarctica), and off Scripps Pier (La Jolla, Calif.). Estimated bacterioplankton production was lower in Antarctic samples (ranging from ~0 to 2.9 μg of C liter⁻¹ day⁻¹) than in those from the other two sites (ranging from 0.7 to 71 μg of C liter⁻¹ day⁻¹). In all three regions studied, it appeared that a significant fraction of the total primary production was utilized by the bacterioplankton and that substantial growth could occur in the absence of large particles. These results support the conclusion that bacterioplankton are a quantitatively important component of coastal marine food webs.     

Effect of Protistan Grazing on the Frequency of Dividing Cells in Bacterioplankton Assemblages

Grazing by phagotrophic flagellates and ciliates is a major source of mortality for bacterioplankton in both marine and freshwater systems. Recent studies have demonstrated a positive relationship between clearance rate and prey size for bacterivorous protists. We tested the idea that, by selectively grazing the larger (more actively growing or dividing) cells in a bacterial assemblage, protists control bacterial standing stock abundances by directly cropping bacterial production. Samples of estuarine water were passed through 0.8-μm-pore-size filters (bacteria only) or 20-μm-mesh screens (bacteria and bacterivorous protists) and placed in dialysis tubing suspended in 7 liters of unfiltered water. Changes in total bacterial biovolume per milliliter (bacterial biomass), frequency of dividing cells (FDC), and average per cell biovolume were followed over a period of 24 h. In three experiments, the FDC increased more rapidly and attained higher values in water passed through 0.8-μm-pore-size filters (average, 5.1 to 8.9%; maximum, 15.5%) compared with FDC values in water passed through 20-μm-mesh screens (average, 2.7 to 5.3%; maximum, 6.7%). Increases in bacterial biomass per milliliter lagged behind increases in FDC by about 4 to 6 h. Grazed bacterial assemblages were characterized by lower total biomasses and smaller average cell sizes compared with those of cells in nongrazed assemblages. We conclude that bacterivorous protists control bacterial standing stock abundances partly by preferentially removing dividing cells. Selective grazing of the more actively growing cells may also explain, in part, the ability of slow-growing cells to persist in bacterioplankton assemblages.     

Spatial Distribution, Structure, Biomass, and Physiology of Microbial Assemblages across the Southern Ocean Frontal Zones during the Late Austral Winter

We examined the spatial distributions of picoplankton, nanoplankton, and microplankton biomass and physiological state relative to the hydrography of the Southern Ocean along 90 degrees W longitude and across the Drake Passage in the late austral winter. The eastern South Pacific Ocean showed some large-scale biogeographical differences and size class variability. Microbial ATP biomass was greatest in euphotic surface waters. The horizontal distributions of microbial biomass and physiological state (adenylate energy charge ratio) coincided with internal currents (fronts) of the Antarctic Circumpolar Current. In the Drake Passage, the biological scales in the euphotic and aphotic zones were complex, and ATP, total adenylate, and adenylate energy charge ratio isopleths were compressed due to the extension of the sea ice from Antarctica and constriction of the Circumpolar Current through the narrow passage. The physiological state of microbial assemblages and biomass were much higher in the Drake Passage than in the eastern South Pacific Ocean. The temperature of Antarctic waters, not dissolved organic carbon, was the major variable controlling picoplankton growth. Estimates of picoplankton production based on ATP increments with time suggest that production under reduced predation pressure was 1 to 10 mug of carbon per liter per day. Our results demonstrate the influence of large-scale hydrographic processes on the distribution and structure of microplankton, nanoplankton, and picoplankton across the Southern Ocean.     

Assessing Phytoplankton and Bacterioplankton Production During Early Spring in Lake Erken, Sweden

The spring development of both phytoplankton and bacterioplankton was investigated between 18 April and 7 May 1983 in mesotrophic Lake Erken, Sweden. By using the lake as a batch culture, our aim was to estimate, via different methods, the production of phytoplankton and bacterioplankton in the lake and to compare these production estimates with the actual increase in phytoplankton and bacterioplankton biomass. The average water temperature was 3.5°C. Of the phytoplankton biomass, >90% was the diatom Stephanodiscus hantzchii var. pusillus, by the peak of the bloom. The ¹⁴C and O₂ methods of estimating primary production gave equivalent results (r = 0.999) with a photosynthetic quotient of 1.63. The theoretical photosynthetic quotient predicted from the C/NO₃ N assimilation ratio was 1.57. The total integrated incorporation of [¹⁴C]bicarbonate into particulate material (>1 μm) was similar to the increase in phytoplankton carbon determined from cell counts. Bacterioplankton increased from 0.5 × 10⁹ to 1.52 × 10⁹ cells liter⁻¹ (~0.5 μg of C liter⁻¹ day⁻¹). Estimates of bacterioplankton production from rates of [³H]thymidine incorporation were ca. 1.2 to 1.7 μg of C liter⁻¹ day⁻¹. Bacterial respiration, measured by a high-precision Winkler technique, was estimated as 4.8 μg of C liter⁻¹ day⁻¹, indicating a bacterial growth yield of 25%. The bulk of the bacterioplankton production was accounted for by algal extracellular products. Gross bacterioplankton production (production plus respiration) was 20% of gross primary production, per square meter of surface area. We found no indication that bacterioplankton production was underestimated by the [³H]thymidine incorporation method.     

Uncoupling of Bacterioplankton and Phytoplankton Production in Fresh Waters Is Affected by Inorganic Nutrient Limitation

Pelagic bacterial production is often positively correlated, or coupled, with primary production through utilization of autotrophically produced dissolved organic carbon. Recent studies indicate that inorganic N or P can directly limit both bacterial and phytoplanktonic growth. Our mesocosm experiments, with whole communities from mesotrophic Calder Lake, test whether this apparent bacterial-algal coupling may be the result of independent responses to limiting inorganic nutrients. In systems without N additions, numbers of bacteria but not phytoplankton increased 2- to 2.5-fold in response to P fertilization (0 to 2.0 μmol of P per liter); this resulted in uncoupled production patterns. In systems supplemented with 10 μmol of NH₄NO₃ per liter, P addition resulted in up to threefold increases in bacteria and two- to fivefold increases in total phytoplankton biomass (close coupling). P limitation of pelagic bacteria occurred independently of phytoplankton dynamics, and regressions between bacterial abundance and phytoplankton chlorophyll a were nonsignificant in all systems without added N. We describe a useful and simple coupling index which predicts that shifts in phytoplankton and bacterioplankton growth will be unrelated (Δ bacteria/Δ phytoplankton → either + ∞ or - ∞) in systems with inorganic N/P (molar) ratios of <~40. In systems with higher N/P ratios (>40), the coupling index will approach 1.0 and close coupling between bacteria and phytoplankton is predicted to occur.     

Respiratory Succession and Community Succession of Bacterioplankton in Seasonally Anoxic Estuarine Waters

Anoxia occurs in bottom waters of stratified estuaries when respiratory consumption of oxygen, primarily by bacteria, outpaces atmospheric and photosynthetic reoxygenation. Once water becomes anoxic, bacterioplankton must change their metabolism to some form of anaerobic respiration. Analysis of redox chemistry in water samples spanning the oxycline of Chesapeake Bay during the summer of 2004 suggested that there was a succession of respiratory metabolism following the loss of oxygen. Bacterial community doubling time, calculated from bacterial abundance (direct counts) and production (anaerobic leucine incorporation), ranged from 0.36 to 0.75 day and was always much shorter than estimates of the time that the bottom water was anoxic (18 to 44 days), indicating that there was adequate time for bacterial community composition to shift in response to changing redox conditions. However, community composition (as determined by PCR-denaturing gradient gel electrophoresis analysis of 16S rRNA genes) in anoxic waters was very similar to that in surface waters in June when nitrate respiration was apparent in the water column and only partially shifted away from the composition of the surface community after nitrate was depleted. Anoxic water communities did not change dramatically until August, when sulfate respiration appeared to dominate. Surface water populations that remained dominant in anoxic waters were Synechococcus sp., Gammaproteobacteria in the SAR86 clade, and Alphaproteobacteria relatives of Pelagibacter ubique, including a putative estuarine-specific Pelagibacter cluster. Populations that developed in anoxic water were most similar (<92% similarity) to uncultivated Firmicutes, uncultivated Bacteroidetes, Gammaproteobacteria in the genus Thioalcalovibrio, and the uncultivated SAR406 cluster. These results indicate that typical estuarine bacterioplankton switch to anaerobic metabolism under anoxic conditions but are ultimately replaced by different organisms under sulfidic conditions.     

Frequency of Dividing Cells, a New Approach to the Determination of Bacterial Growth Rates in Aquatic Environments

Frequency of dividing cells is suggested to be an indirect measure of the mean growth rate of an aquatic bacterial community. Seasonal changes in frequency of dividing cells were found which covariated with the bacterial uptake of ¹⁴C-labeled phytoplankton exudates. Batch and continuous culture growth experiments, using brackish water bacteria in pure and mixed enrichment cultures, were performed to establish a relationship between frequency of dividing cells and growth rate. An improved technique for bacterial direct counts, using fluorescent staining and epifluorescence microscopy, is presented. Based on a 6-month survey in a coastal area of the Baltic Sea, the bacterial production in the photic zone is estimated. Compared to the total primary production in the area, the bacterial population during this period utilized approximately 25% of the amount of carbon originally fixed by the primary producers.     

Relationship between Bacterioplankton Richness, Respiration, and Production in the Southern North Sea

We investigated the relationship between bacterioplankton production (BP), respiration (BR), and community composition measured by terminal restriction fragment length polymorphism in the southern North Sea over a seasonal cycle. Major changes in bacterioplankton richness were apparent from April to December. While cell-specific BP decreased highly significantly with increasing bacterioplankton richness, cell-specific BR was found to be variable along the richness gradient, suggesting that bacterioplankton respiration is rather independent from shifts in the bacterial community composition. As a consequence, the bacterial growth efficiency [BGE = BP/(BP + BR)] was negatively related to bacterioplankton richness, explaining ~43% of the variation in BGE. Our results indicate that despite the observed shifts in the community composition, the main function of the bacterioplankton, the remineralization of dissolved organic carbon to CO₂, is rather stable.     

Double-Staining Epifluorescence Technique to Assess Frequency of Dividing Cells and Bacteriovory in Natural Populations of Heterotrophic Microprotozoa

We have developed a double-staining procedure for use with epifluorescence microscopy which allows the detection both of dividing cells and of ingested bacteria in food vacuoles of heterotrophic microprotozoa. Microprotozoan cells are stained sequentially with the DNA-specific fluorochrome DAPI (4′,6-diami-dino-2-phenylindole) and the nonspecific protein stain fluorescein isothiocyanate. During microscopic examination, heterotrophic microprotozoan cells are first located with fluorescein isothiocyanate fluorescence and then epifluorescence filter sets are switched to permit inspection under DAPI fluorescence of the cell nuclei and of the contents of food vacuoles. Among in situ populations of estuarine microprotozoa sampled over a tidal cycle, we found from 2.2 to 5.2% of the heterotrophic cells in a recognizable stage of division (nuclei elongated or double). Batch culture growth experiments were also carried out both with natural populations and with two isolated species of estuarine microprotozoa. In these experiments, the frequency of dividing cells ranged from 1.2 to 3.8% and appeared to be negatively correlated with growth rate. Microprotozoan populations sampled in continental shelf waters off Savannah, Ga., had mean frequencies of dividing cells ranging from 2.0 to 5.0%. A large fraction of cells in heterotrophic microprotozoan populations (an average of 27.4 ± 1.0% in estuarine water and of 30.1 ± 4.8% in shelf water) had DAPI-stained inclusions, presumably recently ingested bacteria, in their food vacuoles.     

Abundance of Viruses in Marine Waters: Assessment by Epifluorescence and Transmission Electron Microscopy

Abundance of bacteria and tiny DNA-associated particles in the upper layer of Japanese coastal and offshore waters was evaluated by epifluorescence microscopy with 0.015-μm-pore-size Nuclepore filters. The number of tiny DNA-associated particles was compared with the abundance of virus particles estimated by transmission electron microscopy. Although a large variation in virus abundance (1.2 × 10⁶ to 35 × 10⁶ ml⁻¹) was obtained with the transmission electron microscopy method, the ratio of 4′,6-diamidino-2-phenylindole-reactive tiny particles to viruses was in a rather narrow range (1.0 to 1.6), indicating that the majority of the tiny DNA-associated particles identified by epifluorescence microscopy were actually virus particles. This result implies the possibility of using epifluorescence microscopy for the evaluation of virus abundance in marine environments.     

Viral Abundance, Decay, and Diversity in the Meso- and Bathypelagic Waters of the North Atlantic

To elucidate the potential importance of deep-water viruses in controlling the meso- and bathypelagic picoplankton community, the abundance, decay rate, and diversity of the virioplankton community were determined in the meso- and bathypelagic water masses of the eastern part of the subtropical North Atlantic. Viral abundance averaged 1.4 × 10⁶ ml⁻¹ at around 100 m of depth and decreased only by a factor of 2 at 3,000 to 4,000 m of depth. In contrast, picoplankton abundance decreased by 1 order of magnitude to the Lower Deep Water (LDW; 3,500- to 5,000-m depth). The virus-to-picoplankton ratio increased from 9 at about 100 m of depth to 110 in the LDW. Mean viral decay rates were 3.5 × 10⁻³ h⁻¹ between 900 m and 2,750 m and 1.1 × 10⁻³ h⁻¹ at 4,000 m of depth, corresponding to viral turnover times of 11 and 39 days, respectively. Pulsed-field gel electrophoresis fingerprints obtained from the viral community between 2,400 m and 4,000 m of depth revealed a maximum of only four bands from 4,000 m of depth. Based on the high viral abundance and the low picoplankton production determined via leucine incorporation, we conclude that the viral production calculated from the viral decay is insufficient to maintain the high viral abundance in the deep North Atlantic. Rather, we propose that substantial allochthonous viral input or lysogenic or pseudolysogenic production is required to maintain the high viral abundance detected in the meso- and bathypelagic North Atlantic. Consequently, deep-water prokaryotes are apparently far less controlled in their abundance and taxon richness by lytic prokaryotic phages than the high viral abundance and the virus-to-picoplankton ratio would suggest.     

Patterns of Species Number and Abundance in Macroalgal Communities in Coastal Waters

Predictable patterns of species number have been observed in relation to habitat size, habitat heterogeneity and environmental conditions, while patterns in relative abundance of species have been examined for few communities and no assembly rules have been established. We studied communities of attached macroalgae in 61 individual sites located in four different areas; the inner, middle and outer parts of three neighbouring low-tidal estuaries and the adjacent open waters of the Kattegat, Denmark. The objectives were to determine (1) the relationships of species number and rank-abundance to the environmental conditions, and (2) the importance of scale and the consistency of species rank number at the sites for these relationships. We found that species number increased significantly from the inner estuaries to the open coastal waters along with decreasing nutrient concentrations. Turnover (β) diversity was lowest in the open waters suggesting that species composition was more similar among samples there than in the estuaries. Rank-abundance curves did not differ between depth intervals and individual sites across the environmental gradients. However, the summed rank-abundance patterns for two sites showed significantly steeper initial slopes and dominance of few species (i.e., low evenness) in the inner estuaries than in open waters. This pattern was due to high rank consistency of dominant species among sites in the inner estuaries. In open waters rank consistency was low, and the summed abundance across sites showed an even abundance of species. The results imply that the scale of the study and the community variability observed at that particular scale, is the main determinant of abundance patterns.     

Abundance and function of bacteria in the Southern Ocean.

The very low water temperatures existing in polar oceans that experience seasonal advance and retreat of pack ice do not inhibit the presence of large bacterial populations. Bacteria may contribute significantly to the energy transfers within the Southern Ocean. In the last decades, notable progress has been made in the knowledge of the role of marine bacteria in the Southern Ocean. A short overview of the abundance and function ofAntarctic marine bacteria is given, with respect to metabolic activity. The importance of spatial and temporal variability is described. The ecological function of Antarctic marine bacterioplankton is discussed. Depending on food web structure, bacteria may be either a link in food webs supporting metazoan production, or a sink where bacterial production is metabolised by microorganisms. In the more oligotrophic areas and during certain periods of the year bacterial biomass dominates phytoplankton. The microbial food web is therefore the dominant pathway for carbon and energy flow in Antarctic seawater.     

Abundance and breeding of the Antarctic Tern Sterna vittata at the James Ross and Seymour Islands, NE Antarctic Peninsula

The Antarctic Peninsula (AP) is experiencing rapid environmental change associated with warming and sea ice retreat, which is likely to affect locally breeding birds. Yet, contrary to the knowledge of bird biology along the maritime West coasts of the AP, there is a remarkable lack of data from the more continental East coast. We report on the distribution, abundance, and breeding of the Antarctic Tern at the James Ross and Seymour islands, the two largest snow-free areas in the NE part of the AP, where this species breeds under harsh climate conditions probably close to its limits. Terns were found breeding in most ice-free areas, with nests located up to 2.9 km from coastlines at altitudes up to 180 m a.s.l. While the large-scale density was relatively low (c. 450 pairs per 127 km² of surveyed ice-free area), the local density (total colony area: 3 nests per ha; nest clusters: 100–140 nests per ha) was as high as elsewhere. Mean clutch size (1.21, n = 196) was smaller than in the west AP or in the maritime Antarctic. Daily nest survival rate during incubation varied between years and locations (mean = 0.977; 95 % CI: 0.966–0.985). While both predation and weather-caused mortality were locally important, the impact of skua predation might be lower in areas with alternative prey (penguin colonies). We suggest that the Antarctic Tern deserves attention as a species potentially suitable for monitoring of environmental impacts upon bird populations around the AP.     

Diel Fluctuations in the Abundance and Community Diversity of Coastal Bacterioplankton Assemblages over a Tidal Cycle

The diel change in abundance and community diversity of the bacterioplankton assemblages within the Pacific Ocean at a fixed location in Monterey Bay, California (USA) were examined with several culture-independent (i.e., nucleic acid staining, fluorescence in situ hybridization {FISH}, and 16S ribosomal RNA gene libraries) approaches over a tidal cycle. FISH analyses revealed the quantitative predominance of bacterial members belonging to the Cytophaga-Flavobacterium cluster as well as two Proteobacteria (α- and γ-) subclasses within the bacterioplankton assemblages, especially during high tide (HT) and outgoing tide (OT) than the other tidal events. While the clone libraries showed that majority of the sequences were similar to the 16S rRNA gene sequences of unknown bacteria (32% to 73%), however, the operational taxonomic units from members of the α-Proteobacteria, Bacteroidetes, Firmicutes, and Cyanobacteria were also well represented during the four tidal events examined. Comparatively, sequence diversity was highest in OT, lowest in low tide, and very similar between HT and incoming tide. The results indicate that the dynamics of bacterial occurrence and diversity appeared to be more pronounced during HT and OT, further indicative of the ecological importance of several environmental variables including temperature, light intensity, and nutrient availability that are also concurrently fluctuating during these tidal events in marine systems.     

Generation and Fates of Supernumerary Centrioles in Dividing Cells

The centrosome is a subcellular organelle from which a cilium assembles. Since centrosomes function as spindle poles during mitosis, they have to be present as a pair in a cell. How the correct number of centrosomes is maintained in a cell has been a major issue in the fields of cell cycle and cancer biology. Centrioles, the core of centrosomes, assemble and segregate in close connection to the cell cycle. Abnormalities in centriole numbers are attributed to decoupling from cell cycle regulation. Interestingly, supernumerary centrioles are commonly observed in cancer cells. In this review, we discuss how supernumerary centrioles are generated in diverse cellular conditions. We also discuss how the cells cope with supernumerary centrioles during the cell cycle.     

DNA Synthesis, Mitosis and Ploidy of Dividing Cells in Early Crown Gall

DNA synthesis, mitosis and ploidy of dividing cells were studied during 30 h after wounding around wounds inoculated with Agrobacterium tumefaciens, around sterile wounds and in control stems of Vicia faba. DNA synthesis was examined by counting nuclei labelled with ³H-thymidine in slides in which mitoses had been counted and analyzed before autoradiography. The main results were that around inoculated wounds, DNA synthesis and the number of mitoses showed a peak between 12 and 22.5 h. Both types of wounding induced mitoses, many of them polyploid (DNA content higher than 4C), both in the pith and the cortex, whereas in the control stems only diploid mitoses, mostly in the stelar area, were seen. The first polyploid (8C) mitoses around the inoculated wounds took place at 12 h and at 15.5 h 32C mitoses were seen; around the sterile wounds the first 8C divisions occurred at 26 h. The frequency of polyploid mitoses and their degree of ploidy continued to be considerably higher around the crown gall than around the control wounds. When a cell with a higher than 4C content is induced to divide, the 12 chromosomes, as a rule, consist of four, eight, 16 or 32 chromatids, instead of the normal two. The early division of highly polyploid cells around the inoculated wounds is obviously caused by growth factors which are known to be produced by the bacteria. It appears possible that this ability to synthesize excessive amounts of growth factors is subsequently transferred to the host cells through bacterial DNA.     

Regulation of Bacterioplankton Production and Cell Volume in a Eutrophic Estuary

During three periods of 16 to 25 days, bacterioplankton production, bacterial cell volume, chlorophyll a, CO₂ assimilation, and particulate organic carbon were measured in enclosures situated in the eutrophic estuary Roskilde Fjord, Denmark. The enclosures were manipulated with respect to sediment contact and contents of inorganic nutrients, planktivorous fish, and suspension-feeding bivalves. Nutrient enrichment, the presence of suspension feeders, and sediment contact induced pronounced changes in bacterial production, as well as minor changes in bacterial cell volume; however, these effects seemed to be indirect, transmitted via phytoplankton. Bacterial production, measured as [³H]thymidine incorporation, closely followed changes in phytoplankton biomass and production, with time lags of 5 to 10 days. Good correlations of mean bacterioplankton production to chlorophyll a concentration and CO₂ assimilation suggested phytoplankton to be the dominating source of bacterial substrate, apparently independent of nutrient stress. Zooplankton >140 μm, bivalves, and sediment seemed to provide insignificant, if any, substrate for bacterioplankton, and benthic suspension feeders seemed not to act as direct competitors for dissolved organic carbon. The bacterioplankton mean cell volume, measured by image analysis, changed seasonally, with the smallest cells during the summer. Within each period, the bacterial cell volume correlated positively to growth rate and negatively to temperature.