Development of ice biota in a temperate sea area (Gulf of Bothnia)

A study of sea ice biota was carried out in the Gulf of Bothnia (northern Baltic Sea) during the winter of 1989–1990. Samples (ice cores) were taken at a coastal station at regular time intervals during the ice season. Chlorophyll a concentration, algal species distribution, bacterial numbers, and primary and bacterial production were measured. Colonization of the ice began in January when daylight was low. As the available light increased, the algae started to grow exponentially. The vertical chlorophyll a distribution changed and algal species composition and biomass changed during the season. During the initial and middle phase of colonization, ice-specific diatoms, Nitzschia frigida and Navicula pelagica, dominated the algal biomass. Nutrients (PO₄ ^3– and NO_3j) were found to be depleted during the time of algal exponential growth. The maximum algal biomass exceeded 800 g C 1^–1. The primary production supplied food for heterotrophic organisms. The presence of heterotrophic organisms of different trophic levels (bacteria, flagellates, ciliates and rotifers) indicated an active microbial food web.     

Protistan bacterivory in an oligomesotrophic lake: Importance of attached ciliates and flagellates

Seasonal and depth variations of the abundance, biomass, and bacterivory of protozoa (heterotrophic and mixotrophic flagellates and ciliates) were determined during thermal stratification in an oligomesotrophic lake (Lake Pavin, France). Maximal densities of heterotrophic flagellates (1.9 × 10³ cells ml^–1) and ciliates (6.1 cells ml^–1) were found in the metalimnion. Pigmented flagellates dominated the flagellate biomass in the euphotic zone. Community composition of ciliated protists varied greatly with depth, and both the abundance and biomass of ciliates was dominated by oligotrichs. Heterotrophic flagellates dominated grazing, accounting for 84% of total protistan bacterivory. Maximal grazing impact of heterotrophic flagellates was 18.9 × 10⁶ bacteria 1^–1h^–1. On average, 62% of nonpigmented flagellates were found to ingest particles. Ciliates and mixotrophic flagellates averaged 13% and 3% of protistan bacterivory, respectively. Attached protozoa (ciliates and flagellates) were found to colonize the diatom Asterionella formosa. Attached bacterivores had higher ingestion rates than free bacterivorous protozoa and may account for 66% of total protozoa bacterivory. Our results indicated that even in low numbers, epibiotic protozoa may have a major grazing impact on free bacteria. Correspondence: C. Amblard.     

Dynamics of Microbial Communities on Marine Snow Aggregates: Colonization, Growth, Detachment, and Grazing Mortality of Attached Bacteria

We studied the dynamics of microbial communities attached to model aggregates (4-mm-diameter agar spheres) and the component processes of colonization, detachment, growth, and grazing mortality. Agar spheres incubated in raw seawater were rapidly colonized by bacteria, followed by flagellates and ciliates. Colonization can be described as a diffusion process, and encounter volume rates were estimated at about 0.01 and 0.1 cm³ h⁻¹ for bacteria and flagellates, respectively. After initial colonization, the abundances of flagellates and ciliates remained approximately constant at 10³ to 10⁴ and ~10² cells sphere⁻¹, respectively, whereas bacterial populations increased at a declining rate to >10⁷ cells sphere⁻¹. Attached microorganisms initially detached at high specific rates of ~10⁻² min⁻¹, but the bacteria gradually became irreversibly attached to the spheres. Bacterial growth (0 to 2 day⁻¹) was density dependent and declined hyperbolically when cell density exceeded a threshold. Bacterivorous flagellates grazed on the sphere surface at an average saturated rate of 15 bacteria flagellate⁻¹ h⁻¹. At low bacterial densities, the flagellate surface clearance rate was ~5 × 10⁻⁷ cm² min⁻¹, but it declined hyperbolically with increasing bacterial density. Using the experimentally estimated process rates and integrating the component processes in a simple model reproduces the main features of the observed microbial population dynamics. Differences between observed and predicted population dynamics suggest, however, that other factors, e.g., antagonistic interactions between bacteria, are of importance in shaping marine snow microbial communities.     

Massive Regime Shifts and High Activity of Heterotrophic Bacteria in an Ice-Covered Lake

In winter 2009/10, a sudden under-ice bloom of heterotrophic bacteria occurred in the seasonally ice-covered, temperate, deep, oligotrophic Lake Stechlin (Germany). Extraordinarily high bacterial abundance and biomass were fueled by the breakdown of a massive bloom of Aphanizomenon flos-aquae after ice formation. A reduction in light resulting from snow coverage exerted a pronounced physiological stress on the cyanobacteria. Consequently, these were rapidly colonized, leading to a sudden proliferation of attached and subsequently of free-living heterotrophic bacteria. Total bacterial protein production reached 201 µg C L⁻¹ d⁻¹, ca. five times higher than spring-peak values that year. Fluorescence in situ hybridization and denaturing gradient gel electrophoresis at high temporal resolution showed pronounced changes in bacterial community structure coinciding with changes in the physiology of the cyanobacteria. Pyrosequencing of 16S rRNA genes revealed that during breakdown of the cyanobacterial population, the diversity of attached and free-living bacterial communities were reduced to a few dominant families. Some of these were not detectable during the early stages of the cyanobacterial bloom indicating that only specific, well adapted bacterial communities can colonize senescent cyanobacteria. Our study suggests that in winter, unlike commonly postulated, carbon rather than temperature is the limiting factor for bacterial growth. Frequent phytoplankton blooms in ice-covered systems highlight the need for year-round studies of aquatic ecosystems including the winter season to correctly understand element and energy cycling through aquatic food webs, particularly the microbial loop. On a global scale, such knowledge is required to determine climate change induced alterations in carbon budgets in polar and temperate aquatic systems.     

Marine bacterial community structure resilience to changes in protist predation under phytoplankton bloom conditions

To test whether protist grazing selectively affects the composition of aquatic bacterial communities, we combined high-throughput sequencing to determine bacterial community composition with analyses of grazing rates, protist and bacterial abundances and bacterial cell sizes and physiological states in a mesocosm experiment in which nutrients were added to stimulate a phytoplankton bloom. A large variability was observed in the abundances of bacteria (from 0.7 to 2.4 × 10⁶ cells per ml), heterotrophic nanoflagellates (from 0.063 to 2.7 × 10⁴ cells per ml) and ciliates (from 100 to 3000 cells per l) during the experiment (∼3-, 45- and 30-fold, respectively), as well as in bulk grazing rates (from 1 to 13 × 10⁶ bacteria per ml per day) and bacterial production (from 3 to 379 μg per C l per day) (1 and 2 orders of magnitude, respectively). However, these strong changes in predation pressure did not induce comparable responses in bacterial community composition, indicating that bacterial community structure was resilient to changes in protist predation pressure. Overall, our results indicate that peaks in protist predation (at least those associated with phytoplankton blooms) do not necessarily trigger substantial changes in the composition of coastal marine bacterioplankton communities.     

Pico- and nanoplankton dynamics during bloom initiation of Aureococcus in a Long Island, NY bay

The entire microbial plankton community was quantified on a weekly basis April through June of 2000 in Quantuck Bay as part of an ongoing study to identify factors contributing to the initiation of blooms of Aureococcus anophagefferens (brown tide) in Long Island, NY bays. We used flow cytometry, imaging cytometry, fluorescent antibody cell counts, and traditional visual cell counting to quantify the picophytoplankton, heterotrophic bacteria, nanophytoplankton, heterotrophic protists, and microplankton prior to, and during the initiation of a brown tide bloom. Cells passing through a 5 μm mesh dominated the total chlorophyll concentration (>80%) for most of the spring study period. The A. anophagefferens bloom occurred in the context of a larger pico/nanophytoplankton bloom where A. anophagefferens accounted for only 30% of the total cell count when it was at its maximum concentration of 4.8 × 10⁵ mL⁻¹. Levels of dissolved organic nitrogen were enriched during the bloom peak relative to pre-bloom levels and heterotrophic bacteria also bloomed, reaching abundances over 10⁷ mL⁻¹. A trophic cascade within the heterotrophic protist community may have occurred, coinciding with the A. anophagefferens bloom. Before the onset of the bloom, larger grazers increased in abundance, while the next smaller trophic level of grazers were diminished. These smaller grazers were the likely water column predators of A. anophagefferens, and the brown tide bloom initiated when they were depleted. These results suggest that this bloom initiated due to interactions with other pico/nano algae and release from grazing pressure through a trophic cascade.     

The role of phototrophic sulfur bacteria as food for meiobenthic harpacticoid copepods inhabiting eutrophic coastal lagoons

Laboratory experiments were undertaken using Amonardia normani and Schizopera cf. compacta, two meiobenthic harpacticoid copepods commonly found in coastal lagoons. The first experiments were designed to determine if the phototrophic sulfur bacteria Chromatium gracile can be ingested by these copepods and at what concentrations. Egestion rate was used as an index of feeding rate. The response of the egestion rate, expressed in numbers of faecal pellets produced by copepod per day, as a function of bacterial concentration followed the functional model. A. normani attained constant feeding rates from the bacterial concentration of 1 × 10⁷ cells ml^–1 (5 µg C ml^–1) onwards, S. cf. compacta attained constant feeding rates from 2.6 × 10⁷ cells ml^–1 (13 µg C ml^–1) onwards. The faecal pellet volume changed significantly (p<0.05) between food concentrations for A. normani but not for S. cf. compacta (p>0.05). In order to investigate the effect of the phototrophic bacterial diet on the population dynamics of A. normani three groups of nauplii were maintained at 2 × 10⁷ cells ml^–1 and observed every day. The mortality of these nauplii was very high compared to those maintained on a diatom diet (Nitzschia constricta); only in one of the groups did some copepodites develop but no adults were ever observed. Adults fed on bacteria did not have different (p>0.05) survival rates compared to those fed on diatoms, nevertheless, the number of nauplii produced was significantly less (p<0.05) on the bacterial diet. These results lead us to suggest that although the phototrophic sulfur bacteria (Chromatium gracile) can be ingested by both copepod species it cannot sustain the full development of the A. normani population. Thus, a bloom of phototrophic sulfur bacteria does not seem to be a favourable situation for opportunistic benthic copepods to colonize eutrophic coastal lagoons after a dystrophic crisis.     

Effects of Grazing by Flagellates on Competition for Ammonium between Nitrifying and Heterotrophic Bacteria in Chemostats

The enhanced mineralization of organic nitrogen by bacteriophagous protozoa is thought to favor the nitrification process in soils, in which nitrifying bacteria have to compete with heterotrophic bacteria for the available ammonium. To obtain more insight into this process, the influence of grazing by the bacteriovorous flagellate Adriamonas peritocrescens on the competition for limiting amounts of ammonium between the ammonium-oxidizing species Nitrosomonas europaea and the heterotrophic species Arthrobacter globiformis was studied in the presence of Nitrobacter winogradskyi in continuous cultures at dilution rates of 0.004 and 0.01 h^-1. The ammonium concentration in the reservoir was maintained at 2 mM, whereas the glucose concentration was increased stepwise from 0 to 7 mM. A. globiformis won the competition for limiting amounts of ammonium when the glucose concentration in the reservoirs increased, in agreement with previously described experiments in which the flagellates were not included. The numbers of nitrifying bacteria decreased as the numbers of heterotrophic bacteria rose with increasing glucose concentrations. Critical C/N ratios, i.e., ratios between glucose and ammonium in the reservoirs at which no nitrate was found in the culture vessels, of 12.5 and 10.5 were determined at dilution rates of 0.004 and 0.01 h^-1, respectively. Below these critical values, coexistence of the competing species was found. The numbers of nitrifying bacteria decreased more in the presence of flagellates than in their absence, presumably by selective predation on the nitrifying bacteria, either in the liquid culture or on the glass wall of the culture vessels. Despite this, the rate of nitrate production did not decrease more in the presence of flagellates than in their absence. This demonstrates that no correlation has to be expected between numbers of nitrifying bacteria and their activity and that a constant nitrification rate per cell cannot be assumed for nitrifying bacteria. Above the critical C/N ratios, low numbers of nitrifying bacteria were still found in the culture vessels, probably because of attachment of the nitrifying bacteria to the glass wall of the culture vessels. Like the numbers of heterotrophic bacteria, the numbers of flagellates increased when the glucose concentrations in the reservoirs increased. Numbers of 2 × 10⁵ and 12 × 10⁵ flagellates ml^-1 were found at 7 mM glucose at dilution rates of 0.004 and 0.01 h^-1, respectively. It was concluded that the critical C/N ratios were practically unaffected by the presence of protozoa. Although nitrate production rates were equal in the presence and absence of flagellates, the numbers of nitrifying bacteria decreased more strongly in their presence. This indicates a higher activity per nitrifying cell in the presence of flagellates.     

Pelagic food web patterns: do they modulate virus and nanoflagellate effects on picoplankton during the phytoplankton spring bloom?

As agents of mortality, viruses and nanoflagellates impact on picoplankton populations. We examined the differences in interactions between these compartments in two French Atlantic bays. Microbes, considered here as central actors of the planktonic food web, were first monitored seasonally in Arcachon (2005) and Marennes‐Oléron (2006) bays. Their dynamics were evaluated to categorize trophic periods using the models of Legendre and Rassoulzadegan as a reference framework. Microbial interactions were then compared through 48 h batch culture experiments performed during the phytoplankton spring bloom, identified as herbivorous in Marennes and multivorous in Arcachon. Marennes was spatially homogeneous compared with Arcachon. The former was potentially more productive, featuring a large number of heterotrophic pathways, while autotrophic mechanisms dominated in Arcachon. A link was found between viruses and phytoplankton in Marennes, suggesting a role of virus in the regulation of autotroph biomass. Moreover, the virus–bacteria relation was weaker in Marennes, with a bacterial lysis potential of 2.6% compared with 39% in Arcachon. The batch experiments (based on size‐fractionation and viral enrichment) revealed different microbial interactions that corresponded to the spring‐bloom trophic interactions in each bay. In Arcachon, where there is a multivorous web, flagellate predation and viral lysis acted in an opposite way on picophytoplankton. When together they both reduced viral production. Conversely, in Marennes (herbivorous web), flagellates and viruses together increased viral production. Differences in the composition of the bacterial community composition explained the combined flagellate‐virus effects on viral production in the two bays.     

Distribution of Viruses and Dissolved DNA along a Coastal Trophic Gradient in the Northern Adriatic Sea

The distribution of viral and other microbial abundances as well as the concentrations of dissolved DNA (D-DNA) along a trophic gradient in the northern Adriatic Sea were determined. Virus abundances, covering a range of 1.2 × 10⁹ to 8.7 × 10¹⁰ liter^-1 were on average 2.5-fold higher in eutrophic than in mesotrophic stations. A 2.5-fold enrichment was also measured for chlorophyll a concentrations, whereas the densities of bacteria and heterotrophic nanoflagellates were only approximately 1.5-fold higher. The frequency of bacteria containing mature phage increased linearly with bacterial abundance. Assuming that mature phage is only visible during the last 14 to 27% of the latent period (L. M. Proctor, A. Okubo, and J. A. Fuhrman, Microb. Ecol. 25:161-182, 1993), we estimated that between 3.5 and 7.3% of the bacterial population was infected at mesotrophic stations versus between 7.0 and 19.5% at eutrophic stations, indicating that the bacterial mortality due to viral lysis might increase with the degree of eutrophication. The frequency of bacteria with mature phage and the burst size varied significantly with the bacterial morphotype; rod-shape cells, the most abundant morphotype, showed low infection rates but a high burst size. Concentrations of D-DNA varied significantly with season but not with trophic conditions. The estimated percentage of viral DNA on total D-DNA concentrations averaged 17.1% (range, 0.7 to 88.3%). Some kind of interaction between heterotrophic nanoflagellates and viruses is proposed. We conclude (i) that the significance of viruses varies with changing trophic conditions and (ii) that viral activity may play a significant role in food web structure under changing trophic conditions.     

Sea ice microbial communities. III. Seasonal abundance of microalgae and associated bacteria,Mcmurdo Sound,Antarctica

Numbers of bacteria in annual sea ice increased directly with numbers of algae during the 1981 spring ice diatom bloom in McMurdo Sound, Antarctica. Algae and bacteria in a control site grew at rates of 0.10 and 0.05 day^–1, respectively, whereas in an experimentally darkened area neither increased after six weeks. Epiphytic bacteria grew at a rate twice that of the nonattached bacteria and were significantly larger, contributing approximately 30% of the total bacterial biomass after October. The microalgal assemblage was dominated by two species of pennate diatoms, anAmphiprora sp. andNitzschia stellata. Greater than 65% of epiphytic bacteria were associated withAmphiprora sp. after October.N. stellata, however, remained largely uncolonized throughout the study. We hypothesize that microalgae stimulate bacterial growth in sea ice, possibly by providing the bacteria with organic substrates.     

The submicron size fraction of seawater containing high numbers of virus particles as bioactive agent in unicellular plankton community successions

The effect of moderately (2.5 times) increasing the concentrationof the submicron size fraction (2–200 nm) of seawateron unicellular heterotrophic plankton organisms was tested ina series of experiments during algal bloom conditions. Usingelectron microscopy, vims-like particles (VLPs) were by farthe most visible component in this size fraction. We incubatedseawater. which was collected in late winter, in slowly rotatingcylindrical tanks that were exposed to spring temperature andlight conditions. This led to the onset of a mixed diatom bloomwhich was delayed by several days in submicron size-enrichedtreatments. Later, phytoplankton biomass reached values evenslightly higher than in unaltered incubations. The abundanceof heterotrophic unicellular plankton and VLPs exhibited morepronounced oscillation amplitudes during the early phase ofsuccession, thus being stimulated by additional submicron-sizedmaterial (including virions). VLPs are suggested as a possiblesupplementary diet for heterotrophic flagellates. From 190 honwards, toward the end of the incubation experiments (550 h).heterotrophic flagellate density was no longer influenced byinitial enrichment with the submicron size fraction. In contrast,bacteria were consistently repressed in the last two-thirdsof the experimental duration. Our results support the existenceof bioactive substances in the 2-200 nm size fraction, mostprobably virus particles, which are capable of influencing thedynamics and oscillation patterns in plankton community successionaccording to their concentration during algal bloom events.     

Seasonal variations of diatoms and dinoflagellates in a shallow,temperate estuary,with emphasis on neritic assemblages

Seasonal changes in the diatom and dinoflagellate assemblages were examined in the neritic zone of the Urdaibai estuary (north Spain) with regard to some major physical and chemical variables during an annual cycle. A total of 81 diatoms and 38 dinoflagellates were identified and quantified during the study period. Both groups displayed a distinctive pattern of seasonal succession. The seasonal distribution of the Shannon index showed a trend of increasing values from the upper estuary to the lower neritic segment. The diatom diversity maxima were observed in February, April and September, and dinoflagellate maxima in April–May, July and October. Diatoms dominated the assemblages, reaching 1×10⁶ cells l^–1 from April to September. A shift from large diatoms and dinoflagellates to small bloom-forming taxa was observed during winter–early spring. A spring diatom bloom composed of Rhizosolenia spp. was observed in April, while small chain-forming taxa (chiefly Chaetoceros spp.) dominated from June to September. Cell maxima for both groups in late summer were produced by the diatoms Chaetoceros salsugineum and Skeletonema costatum, and by the dinoflagellates Heterocapsa pygmaea and Peridinium quinquecorne. Silicate availability by river supply and strong tidal-mixing of the water column seem to determine the year-round dominance of diatoms over dinoflagellates.     

GROWTH OF VITAMIN B12-REQUIRING MARINE DIATOMS IN MIXED LABORATORY CULTURES WITH VITAMIN B12-PRODUCING MARINE BACTERIA12

The vitamin B₁₂ requirement of several marine diatoms can be satisfied in B₁₂^?limited laboratory cultures by heterotrophic marine bacteria isolated from the same waters and from sediments. The bacteria can utilize diatom excretory products, or the remains of dead diatom cells, in the production of the vitamin. The growth of 12 B₁₂¹? requiring diatoms (7 genera) in mixed cultures with 14 different bacteria (without added B₁₂) was compared to the growth of those same diatoms in axenic cultures with excess added B₁₂. Diatom growth was generally rapid in the first few days, followed by sustained, slower growth. The diatom yields in mixed cultures ranged from 0.8 to 84% of the yields in axenic cultures with added B₁₂. In a detailed study of one mixed culture, increases in diatom densities were paralleled by increases in cell densities of the bacterium during the first few days of exponential diatom growth. During the period of slow diatom growth, when diatom densities oscillated but steadily increased, the decreases in diatom densities were associated with increased bacterial growth. This suggests that death of a fraction of the B₁₂-limited diatom population releases sufficient organic matter to stimulate growth of the bacteria and their subsequent excretion of B₁₂; this B₁₂ in turn stimulates further growth of the diatoms. Diatom-bacteria interactions leading to the production of B₁₂ may be important in maintaining viable populations of B₁₂-requiring diatoms in nutrient-poor waters during periods between blooms, when conditions are unfavorable for rapid growth.     

Seasonal cycle of the microbial plankton in Crooked Lake,Antarctica

Summary Changes in the abundance of the components of the microbial plankton between July 1990 and March 1991 in Crooked Lake, one of the largest and deepest freshwater lakes in Antarctica, are described. Chlorophyll a concentration is low (0.2–0.4g·1^–1) and there is no discernable spring increase. The phytoplankton is largely dominated by flagellates. Bacterioplankton exhibits a seasonal pattern of abundance ranging from 1.0 × 10⁸·1^–1 in July to 3.25 × 10⁸·1^–1 in September. Changes in bacterial abundance probably relate to temperature and grazing by heterotrophic and mixotrophic flagellates. Total flagellated protozoan concentrations ranged between 25–136 × 10²·l^–1. Autotrophic and heterotrophic flagellate abundances were coupled and peaks in their abundance oscillated with peaks in bacterioplankton concentration. Four species of ciliated protozoa, dominated by oligotrichs, particularly the plastidic Strombidium, inhabit the lake. The plankton is characterised by the presence of floes which act as loci for bacteria, flagellates and amoebae and feeding sites for the ciliates and the two sparce metazoan components of the plankton. Crooked Lake is extremely oligotrophic but nonetheless supports a plankton community with a low species diversity and simple trophodynamics.     

Influence of Cyanobacterial Hyperscum on Heterotrophic Activity of Planktonic Bacteria in a Hypertrophic Lake

The response of the planktonic heterotrophic bacterial community to the buildup and breakdown of a semipermanent, crusted, floating cyanobacterial mat, or hyperscum, that covered 1 to 2 ha was studied in a hypertrophic lake (Hartbeespoort Dam, South Africa). The initial response of bacteria in the main basin to the release of dissolved organic carbon (DOC) from the hyperscum 1 km away was an increase in activity per cell from 35 × 10⁻¹² to 153 × 10⁻¹² μg of C cell⁻¹ h⁻¹ for total cell counts, while activity per cell for metabolically active cells increased from 19 × 10⁻¹¹ to 85 × 10⁻¹¹ μg of C cell⁻¹ h⁻¹. No major population growth occurred at this stage. Later, with the continuous supply of DOC from the hyperscum, total bacterial numbers increased from 6.6 × 10⁶ to 20 × 10⁶ cells ml⁻¹, while the activity per cell declined. Metabolically active bacteria followed the same trend. Shorter-term DOC increases caused only increases in bacterial activity per cell. The data from Hartbeespoort Dam demonstrate an interesting and little-documented mechanism by which aquatic bacteria respond to increased DOC concentration and which may be universal for aquatic systems.     

Higher Abundance of Bacteria than of Viruses in Deep Mediterranean Sediments

The interactions between viral abundance and bacterial density, biomass, and production were investigated along a longitudinal transect consisting of nine deep-sea stations encompassing the entire Mediterranean basin. The numbers of viruses were very low (range, 3.6 × 10⁷ to 12.0 × 10⁷ viruses g⁻¹) and decreased eastward. The virus-to-bacterium ratio was always < 1.0, indicating that the deep-sea sediments of the Mediterranean Sea are the first example of a marine ecosystem not numerically dominated by viruses. The lowest virus numbers were found where the lowest bacterial metabolism and turnover rates and the largest cell size were observed, suggesting that bacterial doubling time might play an important role in benthic virus development.     

Viral Lysis and Bacterivory during a Phytoplankton Bloom in a Coastal Water Microcosm

The relative importance of viral lysis and bacterivory as causes of bacterial mortality were estimated. A laboratory experiment was carried out to check the kind of control that viruses could exert over the bacterial assemblage in a non-steady-state situation. Virus-like particles (VLP) were determined by using three methods of counting (DAPI [4′,6-diamidino-2-phenylindole] staining, YOPRO staining, and transmission electron microscopy). Virus counts increased from the beginning until the end of the experiment. However, different methods produced significantly different results. DAPI-stained VLP yielded the lowest numbers, while YOPRO-stained VLP yielded the highest numbers. Bacteria reached the maximal abundance at 122 h (3 × 10⁷ bacteria ml⁻¹), after the peak of chlorophyll a (80 μg liter⁻¹). Phototrophic nanoflagellates followed the same pattern as for chlorophyll a. Heterotrophic nanoflagellates showed oscillations in abundance throughout the experiment. The specific bacterial growth rate increased until 168 h (2.6 day⁻¹). The bacterivory rate reached the maximal value at 96 hours (0.9 day⁻¹). Bacterial mortality due to viral infection was measured by using two approaches: measuring the percentage of visibly infected bacteria (%VIB) and measuring the viral decay rates (VDR), which were estimated with cyanide. The %VIB was always lower than 1% during the experiment. VDR were used to estimate viral production. Viral production increased 1 order of magnitude during the experiment (from 10⁶ to 10⁷ VLP ml⁻¹ h⁻¹). The percentage of heterotrophic bacterial production consumed by bacterivores was higher than 60% during the first 4 days of the experiment; afterwards, this percentage was lower than 10%. The percentage of heterotrophic bacterial production lysed by viruses as assessed by the VDR reached the highest values at the beginning (100%) and at the end (50%) of the experiment. Comparing both sources of mortality at each stage of the bloom, bacterivory was found to be higher than viral lysis at days 2 and 4, and viral lysis was higher than bacterivory at days 7 and 9. A balance between bacterial losses and bacterial production was calculated for each sampling interval. At intervals of 0 to 2 and 2 to 4 days, viral lysis and bacterivory accounted for all the bacterial losses. At intervals of 4 to 7 and 7 to 9 days, bacterial losses were not balanced by the sources of mortality measured. At these time points, bacterial abundance was about 20 times higher than the expected value if viral lysis and bacterivory had been the only factors causing bacterial mortality. In conclusion, mortality caused by viruses can be more important than bacterivory under non-steady-state conditions.     

Enumeration of Bacteriophages and Host Bacteria in Sewage and the Activated-Sludge Treatment Process

Bacteriophage populations in an activated-sludge sewage treatment plant were enumerated. A newly developed assay for quantitation of total phages, employing direct electron microscopic counts, was used in conjunction with the plaque assay. The total concentration of phages was significantly higher in reactor mixed liquor and effluent than in influent sewage, indicating a net production of phages within the reactor. Maximum total phage concentrations in the fluid phase of sewage, activated-sludge mixed liquor, and reactor effluent were 2.2 × 10⁷, 9.5 × 10⁷, and 8.4 × 10⁷/ml, respectively. Conditions were optimized for isolation of predominant heterotrophic aerobic bacteria from sewage and mixed liquor. Blending at ice water temperatures was superior to ultrasound or enzyme treatments for maximum release of viable bacteria from microbial floc. A solidified extract of mixed liquor was superior to standard media for cultivating maximum numbers of heterotrophic bacteria. The highest culture counts for sewage and mixed liquor were 1.4 × 10⁷ and 1.3 × 10⁹/ml, respectively, which represented only 3 and 6.8% of the total microscopic cell counts. Only 3 out of 48 dominant bacterial isolates from either mixed liquor or sewage were hosts for phages present in the system. The sum of phage populations infecting these three hosts accounted for, at best, 3.8% (sewage) and 0.2% (mixed liquor) of the total number of phages present. Generally, specific phage titers were lower in mixed liquor than in sewage, indicating that these hosts were not responsible for the net production of phages in the reactor. This study emphasizes the limitations of the plaque assay for ecological studies of phages, and it suggests that bacteria responsible for phage production in activated-sludge mixed liquor are either minor components of the heterotrophic population, floc-producing strains, or members of other physiological groups.     

Production and Vertical Flux of Attached Bacteria in the Hudson River Plume of the New York Bight as Studied with Floating Sediment Traps

We investigated the growth and vertical flux of attached bacteria with floating sediment traps in the Hudson River Plume of the New York Bight during the spring diatom blooms. Traps were floated at the base of the mixed layer (ca. 10 m) for 1-day periods. After recovery, we measured bacterial abundance and rates of [methyl-³H]thymidine incorporation in the trap samples. The vertical flux of attached bacteria was estimated with a model formulated to distinguish between bacterial accumulation in traps due to in situ growth and that due to vertical flux. Attached bacterial flux ranged from 0.6 × 10¹¹ to 2.0 × 10¹¹ cells m⁻² day⁻¹, and attached bacterial settling rates of 0.1 to 1.0 m day⁻¹ were observed during periods of vertical particulate organic carbon flux ranging from 254 to 1,267 mg of C m⁻² day⁻¹. In situ growth of bacteria in sediment traps was unimportant as a source of bacterial increase when compared with vertical flux during our study. The vertical flux of attached bacteria removed 3 to 67% of the total daily bacterial production from the water column. Particulate organic carbon is not significantly mineralized by attached bacteria during its descent to the sea floor in the plume area during this period, when water temperature and grazing rates are at their annual minima.