A new method for evaluating defense of microalgae against protozoan grazing

Body-size spectrum has proved to be a highly informative indicator to summarize the functional structure of a community at taxon-free resolution. In this study, an approach based on body-size spectrum of protozoan communities was used to detect the defense of microalgae against protozoan grazing. The biofilm-dwelling protozoan communities were used as a test predator system, and two algal species, Chlorella sp. and Nannochloropsis oceanica, were employed as test microalgae. A nine-day bioassay test was carried out by exposing biofilm-dwelling protozoan communities to a gradient of concentrations 10⁰ (control), 10⁴, 10⁵, 10⁶, and 10⁷ cell ml⁻¹ of both microalgae, respectively. Results showed that both algal species represented strong defense effects on the test predator system at different levels of concentration. The body-size distinctness of the protozoan assemblages showed a sharp decrease at high concentration level more than 10⁶ cell ml⁻¹ in both algal treatments. Based on the paired body-size distinctness indices of the protozoa, ellipse tests demonstrated that the body-size spectrum showed an increasing trend of departure from the expected pattern with increasing concentrations of both test algae. Thus, it is suggested that the body-size spectrum of protozoa may be used as a useful indicator to identify the defense of microalgae against protozoan grazing.     

Size-selective grazing of coastal bacterioplankton by natural assemblages of pigmented flagellates,colorless flagellates,and ciliates

Fluorescently-labelled bacteria (FLB) were used to study the feeding strategies of a natural assemblage of estuarine protozoans and to examine whether the protozoan grazing could account for the in situ size structure of the bacterioplankton. The FLB, DTAF-stained enterococci, ranging in volume from 0.01 to 0.30 × 10^–1 µm³, were added to a natural planktonic assemblage at a density of 5.5% of the natural bacterioplankton. Initial densities (individuals ml^–1) were as follows: total natural bacteria, 2.2 × 10⁶; FLB, 1.2 × 10⁵; pigmented flagellates, 300; colorless flagellates, 250; and ciliates, 30. FLB consumption rates were determined by examining the contents of protozoan food vacuoles, and the long-term effect of grazing (over a period of 100 hours) was determined by monitoring the decline in the FLB density in experimental vessels. The average consumption rates of FLB by pigmented flagellates were similar to those by flagellates that lacked chloroplasts (0.9 and 0.6 FLB protozoan^–1 hour^–1, respectively). The ciliates consumed bacteria at an average rate that was 17-fold higher (per cell) than flagellates, and they displayed a greater preference for larger bacteria than did the flagellates. FLB of the mid-size classes (0.025–0.100 µm³) were heavily grazed by the entire protozoan assemblage; the smallest (<0.025 µm³) and the largest (>0.100 µm³) FLB escaped protozoan grazing. This had a profound effect on the resulting size distribution of FLB. At the end of a 100-hour incubation, the percentage of mid-size FLB (0.025 to 0.100 µm³) decreased 2.0–2.2-fold, while the percentage of the smallest and the largest FLB increased 2.0–2.5-fold. Resultant densities of FLB were consistent with initial clearance rates determined for the protozoan groups. The grazing rates of protozoans on FLB were species-specific; whereas some species consumed FLB, others did not demonstrate bacterivory. The results suggest that protozoan grazing has a major effect on the size distribution of coastal bacterioplankton. By selectively feeding on a particular size-class of bacteria, planktonic ciliates may consume 15–90% day^–1 of the standing stock of largest size classes of bacterioplankton. Thus, ciliates, which were present in low abundance in the field, could not balance the production of the entire bacterial community, but they may strongly influence the portion of the bacterial community represented by the largest bacterial class. The direct effect of flagellates (e.g., grazing) was limited to smaller bacteria.Offprint requests to: M. P. Shiaris.     

Possible food chain relationships between bacterioplankton,protozoans, and cladocerans in a reservoir

Ingestion of fluorescent particles by natural protozoan assemblage was studied in the Řimov Reservoir (Southern Bohemia) from April to October, 1987. Attached and free-living bacterial abundance, proportion of active bacteria, density of suspended particles and biomass of cladocerans were also monitored. Heterotrophic nanoflagellates (HNF; 5–12.8 10²ml⁻¹) were the dominant bacterial micrograzers during the spring period and consumed 3 to 9% of the total bacteria per day. After the spring phytoplankton bloom maximum densities of suspended particles and attached bacteria (up to 28% of the total counts) were found. Development of cladocerans in May sharply decreased the proportion of attached bacteria and kept them below 5% of the total counts. All the studied components of plankton except Cladocera decreased during the clearwater phase. The most significant drop was observed in the numbers of protozoans, and they were negligible for bacterial elimination. Bacterial losses during that time apparently were due to cladoceran grazing. During the summer period, ciliates (15–142 ml⁻¹) were mostly dominant micrograzers, and protozoan community grazing increased up to 21% of bacterial standing stock per day. The proportion of active bacteria was strongly correlated with protozoan grazing (r=0.83).     

Viral abundance, production, decay rates and life strategies (lysogeny versus lysis) in Lake Bourget (France)

We have investigated the ecology of viruses in Lake Bourget (France) from January to August 2008. Data were analysed for viral and bacterial abundance and production, viral decay, frequency of lysogenic cells, the contribution of bacteriophages to prokaryotic mortality and their potential influence on nutrient dynamics. Analyses and experiments were conducted on samples from the epilimnion (2 m) and the hypolimnion (50 m), taken at the reference site of the lake. The abundance of virus‐like particles (VLP) varied from 3.4 × 10⁷ to 8.2 × 10⁷ VLP ml^?1; with the highest numbers and virus‐to‐bacterium ratio (VBR = 69) recorded in winter. Viral production varied from 3.2 × 10⁴ VLP ml^?1 h^?1 (July) to 2 × 10⁶ VLP ml^?1 h^?1 (February and April), and production was lower in the hypolimnion. Viral decay rate reached 0.12–0.15 day^?1, and this parameter varied greatly with sampling date and methodology (i.e. KCN versus filtration). Using transmission electron microscopy (TEM) analysis, viral lysis was responsible for 0% (January) to 71% (February) of bacterial mortality, while viral lysis varied between 0% (April) and 53% (January) per day when using a modified dilution approach. Calculated from viral production and burst size, the virus‐induced bacterial mortality varied between 0% (January) and 68% (August). A weak relationship was found between the two first methods (TEM versus dilution approach). Interestingly, flow cytometry analysis performed on the dilution experiment samples revealed that the viral impact was mostly on high DNA content bacterial cells whereas grazing, varying between 8.3% (June) and 75.4% (April), was reflected in both HDNA and LDNA cells equally. The lysogenic fraction varied between 0% (spring/summer) and 62% (winter) of total bacterial abundance, and increased slightly with increasing amounts of mitomycin C added. High percentages of lysogenic cells were recorded when bacterial abundance and activity were the lowest. The calculated release of carbon and phosphorus from viral lysis reached up to 56.5 µgC l^?1 day^?1 (assuming 20 fgC cell^?1) and 1.4 µgP l^?1 day^?1 (assuming 0.5 fgP cell^?1), respectively, which may represent a significant fraction of bacterioplankton nutrient demand. This study provides new evidence of the quantitative and functional importance of the virioplankton in the functioning of microbial food webs in peri‐alpine lakes. It also highlights methodologically dependent results.     

A Mesocosm Study of the Changes in Marine Flagellate and Ciliate Communities in a Crude Oil Bioremediation Trial

Protozoan grazers play an important role in controlling the density of crude-oil degrading marine communities as has been evidenced in a number of microcosm experiments. However, small bioreactors contain a low initial titre of protozoa and the growth of hydrocarbon-depleting bacteria is accompanied by the fast depletion of mineral nutrients and oxygen, which makes microcosms rather unsuitable for simulating the sequence of events after the oil spill in natural seawater environment. In the present study, the population dynamics of marine protozoan community have been analysed in a 500 l mesocosm experiment involving bioaugmented oil booms that contained oil sorbents and slow-release fertilisers. A significant increase in numbers of marine flagellates and ciliates on biofilms of oil-degrading microbes was microscopically observed as early as 8 days after the start of the experiment, when protozoa exhibited a population density peak making up to 3,000 cells ml⁻¹. Further, the protozoan density varied throughout the experiment, but never dropped below 80 cells ml⁻¹. An 18S rRNA gene-based fingerprinting analysis revealed several changes within the eukaryotic community over the whole course of the experiment. Initial growth of flagellates and small ciliates was followed by a predominance of larger protozoa. According to microscopic observations and SSU rRNA molecular analyses, most predominant were the ciliates belonging to Euplotidae and Scuticociliatia. This is the first study to characterise the eukaryotic communities specifically in a large-scale oil bioremediation trial using both microscopy-based and several molecular techniques.     

Impact of Water Column Acidification on Protozoan Bacterivory at the Lake Sediment-Water Interface

Although the impact of acidification on planktonic grazer food webs has been extensively studied, little is known about microbial food webs either in the water column or in the sediments. Protozoon-bacterium interactions were investigated in a chronically acidified (acid mine drainage) portion of a lake in Virginia. We determined the distribution, abundance, apparent specific grazing rate, and growth rate of protozoa over a pH range of 3.6 to 6.5. Protozoan abundance was lower at the most acidified site, while abundance, in general, was high compared with other systems. Specific grazing rates were uncorrelated with pH and ranged between 0.02 and 0.23 h^-1, values similar to those in unacidified systems. The protozoan community from an acidified station was not better adapted (P = 0.95) to low-pH conditions than a community from an unacidified site (multivariate analysis of variance on growth rates for each community incubated at pHs 4, 5, and 6). Both communities had significantly lower (P < 0.05) growth rates at pHs 4 and 5 than at pH 6. Reduced protozoan growth rates coupled with high grazing rates and relatively higher bacterial yields (ratio of bacterial-protozoan standing stock) at low pH indicate reduced net protozoan growth efficiency and a metabolic cost of acidification to the protozoan community. However, the presence of an abundant, neutrophilic protozoan community and high bacterial grazing rates indicates that acidification of Lake Anna has not inhibited the bacterium-protozoon link of the sediment microbial food web.     

An Under-appreciated Component of Biodiversity in Plankton Communities: The Role of Protozoa in Lake Michigan (A Case Study)

Recent technological advances have led to the discovery that free-living, planktonic protozoa are ubiquitous in nature and appear to be important components of pelagic food webs (e.g., fluorescent straining, flow cytometry). Despite this, limited information exists tying their seasonality to rate processes that drive succession patterns. The abundance, and seasonal growth and grazing loss of an entire protozoan assemblage were evaluated in Lake Michigan. The protozoan assemblage was species-rich (100 taxa) and abundant throughout the year in Lake Michigan. Nano-sized protozoa (Hnano and Pnano, <20 μm in size) ranged in abundance from 10² to 10³ cells ml⁻¹, while micro-protozoa (Hmicro and Pmico, >20 and <200 μm in size) ranged in abundance from 4 to 17 cells ml⁻¹. The biomass of Hnano and Hmicro by itself represented more than 70–80% of crustacean zooplankton biomass, while Pnano and Pmicro constituted nearly 50% of phytoplankton biomass. Protozoa exhibited growth rates comparable to other components of the plankton in Lake Michigan, and some populations grew at rates similar to maximum rates determined in the laboratory (rates of 1–2 day⁻¹). Overall, it appears that macro-zooplankton predation is a major loss factor counter-balancing growth with only small differences between the two rate processes (<0.1 day⁻¹). Discrepancies between growth and grazing loss in the spring were likely attributed to sedimentation losses for larger species of tintinnids and dinoflagellates (Codonella, Tintinnidium, and Gymnodinium) that can account for their occurrence in the deep chlorophyll layer. In the summer, carnivory among similar sized species (Chromulina and small ciliates) may be additional loss factors impinging on the protozoan assemblage.     

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.     

Effects of grazing and excretion by pelagic mysids (Mysis spp.) on the size structure and biomass of the phytoplankton community

The aim of this study was to determine the effects of pelagic mysids (Mysis mixta and M. relicta) on the biomass and size-structure of the phytoplankton community during the period following the spring bloom. Mysids excreted phosphate (4.5 ± 0.7 nmol ind⁻¹ h⁻¹) and ammonium (123.6 ± 31.6 and 45.0 ± 3.2 nmol ind⁻¹ h⁻¹) and increased the total chlorophyll-a concentration of phytoplankton slightly. However, the presence of mysids affected different size-classes of phytoplankton differently. Mysids mainly grazed on large-sized (>10 μm) phytoplankton cells. Small-sized (<10 μm) algal cells avoided grazing, gained a competitive advantage and were able to utilize the nutrients excreted by mysids. According to this study, both top-down and bottom-up mechanisms simultaneously mould the structure of the phytoplankton community. A large zooplankton biomass might promote the increase of small flagellates by a combination of repleting nutrient stores, selective grazing on large algal cells and heavy predation on protozoa which, consequently, might have a cascading effect on the most favoured protozoan food source, small flagellates.

     

Mobility of Protozoa through Narrow Channels

Microbes in the environment are profoundly affected by chemical and physical heterogeneities occurring on a spatial scale of millimeters to micrometers. Physical refuges are critical for maintaining stable bacterial populations in the presence of high predation pressure by protozoa. The effects of microscale heterogeneity, however, are difficult to replicate and observe using conventional experimental techniques. The objective of this research was to investigate the effect of spatial constraints on the mobility of six species of marine protozoa. Microfluidic devices were created with small channels similar in size to pore spaces in soil or sediment systems. Individuals from each species of protozoa tested were able to rapidly discover and move within these channels. The time required for locating the channel entrance from the source well increased with protozoan size and decreased with channel height. Protozoa of every species were able to pass constrictions with dimensions equal to or smaller than the individual's unconstrained cross-sectional area. Channel geometry was also an important factor affecting protozoan mobility. Linear rates of motion for various species of protozoa varied by channel size. In relatively wide channels, typical rates of motion were 300 to 500 μm s⁻¹ (or about 1 m per hour). As the channel dimensions decreased, however, motilities slowed more than an order of magnitude to 20 μm s⁻¹. Protozoa were consistently observed to exhibit several strategies for successfully traversing channel reductions. The empirical results and qualitative observations resulting from this research help define the physical limitations on protozoan grazing, a critical process affecting microbes in the environment.     

Relative importance of biotic and abiotic factors affecting bacterial abundance in Lake Biwa: an empirical analysis

To determine the relative importance of factors affecting bacterial abundance in Lake Biwa, correlation and multiple regression analyses were performed with relevant biotic and abiotic variables. Data used in the analyses were collected weekly from April 1997 to June 1998 at a pelagic site in the north basin. The bacterial abundance ranged from 1 to 7 × 10⁶ cells ml⁻¹, and its spatio-temporal pattern was virtually identical to that in previous studies conducted 12–15 years ago. In the surface layer (0–12.5 m), bacterial abundance was significantly correlated with water temperature and with protozoan and metazoan grazers, but not with chlorophyll a and nutrient concentrations. The results suggest that loss factors rather than growth factors are more important in determining bacterial abundance in this lake. However, grazing effects on bacterial abundance differed among zooplankton. Bacterial abundance correlated negatively with phagotrophic nanoflagellates (PNF) and Daphnia, but positively with Eodiaptomus. Thus, PNF and Daphnia act to reduce the bacterial abundance, while Eodiaptomus acts to stimulate. In contrast, these biotic factors did not explain changes in bacterial abundance in the middle (12.5–25 m) and deep (>25 m) layers. Instead, the bacterial abundance in the deep layer was highly correlated with vertical mixing regimes, suggesting that bacterial abundance was directly or indirectly affected by abiotic factors. These results indicate that bacterial abundance in Lake Biwa was regulated by different factors at different depths. Received: February 8, 2000 / Accepted: August 29, 2000     

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.     

Interactions of bacteria and microflagellates in sequencing batch reactors exhibiting enhanced mineralization of toxic organic chemicals

Community level interactions were studied in non-axenic sequencing batch reactors (SBRs) being used to treat 2,4-dinitrophenol (DNP). Increasing the influent DNP concentrations from 1 to 10 μg ml⁻¹ eliminated large predatory organisms such as rotifers and ciliated protozoa from the SBRs. Under steady-state conditions at a DNP concentration of 10 μg ml⁻¹, supplemental additions of glucose enhanced DNP degradation and led to the establishment of a microbial community consisting of five species of bacteria and a variety of microflagellates. The bacteria and flagellates exhibited oscillating population dynamics in this system, possibly indicating predator-prey interactions between these two groups. Only two of the five bacteria isolated from this system could utilize glucose as a growth substrate, and one of these two species was the only organism that could mineralize DNP in the system. The other three bacteria could grow using metabolic by-products of one of the glucose-utilizing strains (Bacillus cereus) found in the reactors. Supplemental glucose additions increased the average size of bacterial floc particles to 172 μm, compared with 41 μm in SBRs not receiving glucose. It is theorized that the enhanced mineralization of DNP in this non-axenic system was attributable to increased community interactions resulting in increased bacterial flocculation in SBRs receiving supplemental glucose additions. Offprint requests to: S. K. Schmidt.     

Heterotrophic bacterial and viral dynamics in Arctic freshwaters: results from a field study and nutrient-temperature manipulation experiments

Heterotrophic bacterial and viral concentrations (range, 0.7 × 10⁴ to 206.2 × 10⁴ ml⁻¹ and 0.05 × 10⁶ to 128.9 × 10⁶ ml⁻¹, respectively) were determined in several Arctic freshwater environments, including lakes and glacial ecosystems (78.55°N, 11.56°E). Our bacteria and virus results mirrored trends seen in temperate lakes, with an average virus-to-bacteria ratio (VBR) of 13 (range, 7.3–25.2) and viral concentrations and DOC positively correlated with bacterial concentrations (R = 0.964, P < 0.01 and R = 0.813, P < 0.05, respectively). Lysogenic bacteria, determined by induction with Mitomycin C, were not detected in any of the investigated Arctic freshwater environments. Nutrient-addition experiments at in situ and at elevated temperatures were performed to elucidate the factors which influenced the bacterial growth and the virus–bacteria interactions in Arctic freshwaters. Our results suggest that multiple limiting factors interacted and constrained bacterial growth. Bacterial concentrations and doubling times increased at elevated temperatures and appeared to be co-stimulated by phosphorus and carbon. However, viral concentrations showed a lack of response to nutrient addition thus indicating an uncoupling between bacteria and viruses in the experiment.     

Experimental demonstration of the roles of bacteria and bacterivorous protozoa in plankton nutrient cycles

We have used a model food chain composed of a natural bacterial assemblage, a pennate diatom and a bacterivorous microflagellate to investigate the factors controlling the relative importance of bacteria and protozoa as sources for regenerated nitrogen in plankton communities. In bacterized diatom cultures in which diatom growth was nitrogen-limited, the carbon:nitrogen (C:N) ratio of the bacterial substrate greatly affected which population was responsible for the uptake of nitrogen. When nitrogen was added as NH ₄ ⁺ and the cultures were supplemented with glucose, the bacteria competed successfully with the algae for NH ₄ ⁺ and prevented the growth of algae by rapidly assimilating all NH ₄ ⁺ in the cultures. Bacterivorous protozoa inoculated into these cultures grazed the bacterial population and remineralized NH ₄ ⁺ , thus relieving the nitrogen limitation of algal growth and allowing an increase in algal biomass. In contrast, bacteria in cultures supplemented with the amino acid glycine (C:N = 2) were major remineralizers of nitrogen, and the influence of protozoan grazing was minimal. We conclude that the relative importance of bacteria and protozoa as nutrient regenerators in the detrital food loop is dependent largely on the overall carbon:nutrient ratio of the bacterial substrate. The role of bacterivorous protozoa as remineralizers of a growth-limiting nutrient is maximal in situations where the carbon:nutrient ratio of the bacterial substrate is high.     

Seasonal dynamics of crustacean zooplankton, heterotrophic nanoflagellates and bacteria in a shallow, eutrophic lake

1. The seasonal development of crustacean zooplankton, heterotrophic nanoflagellates (HNF) and bacteria was examined in Grosser Binnensee, a shallow, eutrophic lake in northern Germany. The grazing impact of Daphnia on bacteria and nanoflagellates was estimated from field data on population abundances and from clearance rates obtained in laboratory experiments. 2. The seasonal succession of zooplankton showed distinct peaks of Daphnia magna, cyclopopid copepods, Bosmina longirostris and Daphnia galeata and D. hynlina. The population dynamics of Dapfinia had the strongest impact on all sestonic components. Daphnia maxima coincided with clearwater phases, and were negatively correlated with particulate organic carbon (POC), HNF and phytoplankton. Bacterial abundance was only slightly affected although daphnids were at times more important as bacterial consumers than HNF, as estimated from measured bacterial clearance rates. Other crustaceans (copepods, Bosmina) were probably of minor importance as grazers of bacteria and nanoplankton. 3. HNF abundance varied from 550 ml^?1 to more than 30000 ml^?1. HNF appeared to be suppressed by daphnids and reached highest densities when copepods dominated the metazooplankton. The variation in HNF abundance was not reflected in the concentration of heterotrophic bacteria, which fluctuated rather irregularly between 5 and 20 ± 10⁶ ml^?1. Long filamentous bacteria which were probably resistant to protozoan grazing, however, appeared parallel to the development of HNF. These bacterial cells, although small in number, could comprise more than 30% of the total bacterial biomass.     

Protozoan Bacterivory and Escherichia coli Survival in Drinking Water Distribution Systems

The development of bacterial communities in drinking water distribution systems leads to a food chain which supports the growth of macroorganisms incompatible with water quality requirements and esthetics. Nevertheless, very few studies have examined the microbial communities in drinking water distribution systems and their trophic relationships. This study was done to quantify the microbial communities (especially bacteria and protozoa) and obtain direct and indirect proof of protozoan feeding on bacteria in two distribution networks, one of GAC water (i.e., water filtered on granular activated carbon) and the other of nanofiltered water. The nanofiltered water-supplied network contained no organisms larger than bacteria, either in the water phase (on average, 5 × 10⁷ bacterial cells liter⁻¹) or in the biofilm (on average, 7 × 10⁶ bacterial cells cm⁻²). No protozoa were detected in the whole nanofiltered water-supplied network (water plus biofilm). In contrast, the GAC water-supplied network contained bacteria (on average, 3 × 10⁸ cells liter⁻¹ in water and 4 × 10⁷ cells cm⁻² in biofilm) and protozoa (on average, 10⁵ cells liter⁻¹ in water and 10³ cells cm⁻² in biofilm). The water contained mostly flagellates (93%), ciliates (1.8%), thecamoebae (1.6%), and naked amoebae (1.1%). The biofilm had only ciliates (52%) and thecamoebae (48%). Only the ciliates at the solid-liquid interface of the GAC water-supplied network had a measurable grazing activity in laboratory test (estimated at 2 bacteria per ciliate per h). Protozoan ingestion of bacteria was indirectly shown by adding Escherichia coli to the experimental distribution systems. Unexpectedly, E. coli was lost from the GAC water-supplied network more rapidly than from the nanofiltered water-supplied network, perhaps because of the grazing activity of protozoa in GAC water but not in nanofiltered water. Thus, the GAC water-supplied network contained a functional ecosystem with well-established and structured microbial communities, while the nanofiltered water-supplied system did not. The presence of protozoa in drinking water distribution systems must not be neglected because these populations may regulate the autochthonous and allochthonous bacterial populations.     

Autotrophic picoplankton community dynamics in a pre-alpine lake in British Columbia,Canada

Autotrophic picoplankton (APP) were studied in Chilko Lake, a large, deep ultra-oligotrophic pre-alpine lake (elevation: 1172 m) in the south central coast mountains of British Columbia. Data from 1985 (untreated) and 1990 (treated) were used to compare and contrast APP community response to a whole-lake fertilization experiment. The APP communities of Chilko Lake were dominated by the coccoid cyanobacteria Synechococcus and its colonial morph which comprised about 99% of the APP community of Chilko Lake. Chlorella-like eukaryotic picoplankters and small cyanobacteria were rare, comprising < 1 % of the APP community. In 1990 autotrophic picoplankters contributed an average of 73% to total chlorophyll, and 54% to total photosynthesis. Average APP abundance ranged from lows of 4,000–5,000 cells ml^-1 in winter and spring to highs of 50000–150000 cells ml^-1 in early August with no apparent autumnal increase. APP populations were uniformly distributed in the epilimnion, but during calm periods in August often formed a peak near the metalimnion/hypolimnion boundary. Seasonal and vertical distribution patterns of APP showed little relation to temperature or to light. When nutrients were added to the lake in 1990, APP populations doubled within 3 wk of addition and average abundance (6.16 × 10⁴ cells · ml^-1) was twice 1985 APP numbers. Bottom-up control by scarce nutrient supplies is considered the primary factor regulating community composition and abundance during the initial population growth phase (June, July) with top-down control by grazing during nutrient colimitation periods when the epilimnion is deplete of both nitrogen and phosphorus (August, September).     

Unusual bloom of star-like prosthecate bacteria and filaments as a consequence of grazing pressure

In seawater used for shrimp aquaculture in French Polynesia, the grazing of small bacteria (rods and coccoids) allowed the growth ofAncalomicrobium cells (to more than 2×10⁶ cells ml^–1) and large filaments > 10m in length (5×10⁶ cells ml^–1). Their contribution to the increase in total bacterial number after grazing was 27.8 and 9.8%, respectively. These large bacteria are not grazed on by microflagellates, but are available for mesoplankton larvae.     

Importance of passive diffusion in the uptake of polychlorinated biphenyls by phagotrophic protozoa

Unicellular protozoan grazers represent a size class of organisms where a transition in the mechanism of chlorobiphenyl (CB) introduction, from diffusion through surface membranes to ingestion of contaminated prey, could occur. This study compares the relative importance of these two processes in the overall uptake of polychlorinated biphenyls by protists. Uptake rates and steady-state concentrations were compared in laboratory cultures of grazing and nongrazing protozoa. These experiments were conducted with a 10-microm marine scuticociliate (Uronema sp.), bacterial prey (Halomonas halodurans), and a suite of 21 CB congeners spanning a range of aqueous solubilities. The dominant pathway of CB uptake by both grazing and nongrazing protozoa was diffusion. Organic-carbon-normalized CB concentrations (in the protozoan cell) were equivalent in grazing and nongrazing protozoa for all congeners studied. Rate constants for uptake into and loss from the protozoan cell were independently determined by using [3,3',4, 4'-(14)C]tetrachlorobiphenyl (IUPAC no. 77), 0.38 +/- 0.03 min(-1) and (1.1 +/- 0.1) x 10(-5) (g of organic carbon)(-1) min(-1), respectively. Magnitudes of the uptake and loss processes were calculated and compared by using a numerical model. The model result was consistent with data from the bioaccumulation experiment and supported the hypothesis that diffusive uptake is faster than ingestive uptake in phagotrophic unicellular protozoa.