Trophic interactions between viruses, bacteria and nanoflagellates under various nutrient conditions and simulated climate change

Population dynamics in the microbial food web are influenced by resource availability and predator/parasitism activities. Climatic changes, such as an increase in temperature and/or UV radiation, can also modify ecological systems in many ways. A series of enclosure experiments was conducted using natural microbial communities from a Mediterranean lagoon to assess the response of microbial communities to top-down control [grazing by heterotrophic nanoflagellates (HNF), viral lysis] and bottom-up control (nutrients) under various simulated climatic conditions (temperature and UV-B radiations). Different biological assemblages were obtained by separating bacteria and viruses from HNF by size fractionation which were then incubated in whirl-Pak bags exposed to an increase of 3°C and 20% UV-B above the control conditions for 96 h. The assemblages were also provided with an inorganic and organic nutrient supply. The data show (i) a clear nutrient limitation of bacterial growth under all simulated climatic conditions in the absence of HNF, (ii) a great impact of HNF grazing on bacteria irrespective of the nutrient conditions and the simulated climatic conditions, (iii) a significant decrease in burst size (BS) (number of intracellular lytic viruses per bacterium) and a significant increase of VBR (virus to bacterium ratio) in the presence of HNF, and (iv) a much larger temperature effect than UV-B radiation effect on the bacterial dynamics. These results show that top-down factors, essentially HNF grazing, control the dynamics of the lagoon bacterioplankton assemblage and that short-term simulated climate changes are only a secondary effect controlling microbial processes.     

Ecology of phytoplankton communities dominated by Aureococcus anophagefferens: the role of viruses, nutrients, and microzooplankton grazing

We investigated the impact of viruses, nutrient loading, and microzooplankon grazing on phytoplankton communities in two New York estuaries that hosted blooms of the brown tide alga Aureococcus anophagefferens during 2000 and 2002. The absence of a bloom at one location during 2002 allowed for the fortuitous comparison of a bloom and non-bloom year at the same location as well as a comparison of two sites experiencing bloom and non-bloom conditions during the same year. During the study, blooms were found at locations with high levels of dissolved organic nitrogen and lower nitrate concentrations compared to a non-bloom location. Experimental additions of inorganic nitrogen and phosphorus yielded growth rates within the total phytoplankton community which significantly exceeded control treatments in 83% of experiments, while A. anophagefferens experienced significantly increased growth during only 20% of experimental inorganic nutrient additions. Consistent with prior research, these results suggest brown tides are not caused by eutrophication, but instead are more likely to occur when sources of labile DOM are readily available. Microzooplankton grazing rates on the total phytoplankton community during a bloom were lower than grazing rates at a non-bloom site, and grazing rates on A. anophagefferens were lower than grazing rates on the total community on some dates, suggesting that reduced grazing mortality may also promote brown tides. Mean densities of viruses during blooms (3 × 10⁸ ml⁻¹) were elevated compared to most estuarine environments and were twice the levels found at a non-bloom site. Experimental enrichment of the natural viral densities yielded a significant increase in A. anophagefferens growth rates relative to control treatments when background levels of viruses were low (<1.7 × 10⁸ ml⁻¹), suggesting that viruses may promote bloom occurrence by regenerating DOM or altering the composition of microbial communities.     

Response of Alteromonadaceae and Rhodobacteriaceae to glucose and phosphorus manipulation in marine mesocosms

Microbial successions were studied in experimental mesocosms of marine water in the presence of additional organic carbon (glucose), phosphorus (P) or both. P addition lead to pronounced blooms of phytoplankton and to significantly enhanced bacterial production. Characteristic succession patterns were observed for two phylogenetic groups of bacteria that both transiently formed > 50% of total cells. An initial bloom of bacteria affiliated to the Alteromonadaceae could not be assigned to any specific treatment and was interpreted as a response to the manipulations during mesocosm set-up. These bacteria rapidly declined with the appearance of heterotrophic nanoflagellates, suggesting a negative effect of selective grazing. The persistence of Alteromonadaceae in the microbial assemblages was significantly favored by the presence of additional glucose. During the second half of the experiment, bacteria affiliated to Rhodobacteriaceae formed a dominant component of the experimental assemblages in treatments with addition of P. The community contribution of Rhodobacteriaceae was significantly correlated with chlorophyll a concentrations only in the P-amended mesocosms (r(2) = 0.58). This was more pronounced in the absence of glucose (r(2) = 0.85). The phylogenetic and morphological diversity among Rhodobacteriaceae was high, and treatment-specific temporal successions of genotypes related to Rhodobacteriaceae were observed. We suggest that the observed succession patterns reflect different niche preferences: Alteromonadaceae rapidly responded to disturbance and profited from allochthonous glucose input, whereas Rhodobacteriaceae benefited from the phytoplankton bloom.     

Microbial Community Response during the Iron Fertilization Experiment LOHAFEX

Iron fertilization experiments in high-nutrient, low-chlorophyll areas are known to induce phytoplankton blooms. However, little is known about the response of the microbial community upon iron fertilization. As part of the LOHAFEX experiment in the southern Atlantic Ocean, Bacteria and Archaea were monitored within and outside an induced bloom, dominated by Phaeocystis-like nanoplankton, during the 38 days of the experiment. The microbial production increased 1.6-fold (thymidine uptake) and 2.1-fold (leucine uptake), while total cell numbers increased only slightly over the course of the experiment. 454 tag pyrosequencing of partial 16S rRNA genes and catalyzed reporter deposition fluorescence in situ hybridization (CARD FISH) showed that the composition and abundance of the bacterial and archaeal community in the iron-fertilized water body were remarkably constant without development of typical bloom-related succession patterns. Members of groups usually found in phytoplankton blooms, such as Roseobacter and Gammaproteobacteria, showed no response or only a minor response to the bloom. However, sequence numbers and total cell numbers of the SAR11 and SAR86 clades increased slightly but significantly toward the end of the experiment. It seems that although microbial productivity was enhanced within the fertilized area, a succession-like response of the microbial community upon the algal bloom was averted by highly effective grazing. Only small-celled members like the SAR11 and SAR86 clades could possibly escape the grazing pressure, explaining a net increase of those clades in numbers.     

The annual cycle of heterotrophic freshwater nanoflagellates: role of bottom-up versus top-down control

The population dynamics of heterotrophic nanoflagellates (HNF)were analyzed in pre-alpine Lake Constance over three consecutiveyears. A recurrent seasonal pattern led to the identificationof five seasonal phases: winter, spring, clear-water, summerand autumn. HNF biomass was lowest in winter and highest m latespring several weeks after the phytoplankton spring bloom. Theaverage biomass of HNF was 5–12% of bacterial biomassand 13–34% of ciliate biomass respectively. The largestHNF cells were recorded during the spring phase, whereas theaverage cell size was reduced to one-third during the subsequentclear-water phase. The pronounced differences in the mean cellsize were attributed mainly to varying grazing impact on HNFThroughout most of the year, HNF production was balanced bygrazing of microzooplankton, namely ciliates, within the microbialloop. During the dear-water phase, however, the grazing impactwas mainly due to rotifers and daphnids. Changing grazing impactwas primarily responsible for the observed 2-fold interannualdifference m the mean biomass of HNF Overall, top-down controlby grazing was more important in governing the population dynamicsof HNF than bottom-up control by bacterial food supply.     

8 Grazing by heterotrophic nanoflagellates on virus- and bacteria-sized particles in two lakes of different trophy

Although viruses are now widely recognized to infect aquatic bacteria as well as prokaryotic and eukaryotic phytoplankton, our understanding of how they fit into aquatic foodwebs remains still deficient. The present study examined the potential nutritional food source that viral particles represent for natural assemblages of heterotrophic nanoflagellates (HNF). We determined the seasonal and depth-related variability in the grazing activity of HNF on virus- and bacteria-sized particles in the oligomesotrophic Lake Pavin and the eutrophic Lake Aydat. Ingestion rates were determined using 50 and 500 nm diameter fluorescent microspheres. Estimated ingestion rates ranged from 0.01 to 1.7 viruses cell⁻¹·h⁻¹ (Aydat) and from 0.01 to 2 viruses cell⁻¹·h⁻¹ (Pavin). Derived clearance rates for viruses represented 3–5% (Aydat) and 10–12% (Pavin) of those for bacteria. In general, HNF grazing on both viruses and bacteria showed a similar pattern characterized by an apparent decrease in summer followed by autumnal peaks, with lowest values always being recorded in the hypolimnion. Viral production, estimated from the frequency of visibly infected cells (using transmission electronic microscopy), was consumed by HNF at rates averaging 2% (Aydat) and 10% (Pavin). This study suggests that, viruses, which are assumed to correspond to 1×10⁻⁷ ng C/particle, are likely to be of low nutritive value for HNF in both lakes. However, these results imply that HNF grazing may contribute significantly to the removal of viruses in freshwaters, especially in low productive systems.     

Seasonal and vertical distribution of planktonic bacteria and heterotrophic nanoflagellates in the middle Adriatic Sea

Temporal and spatial patterns of bacteria and heterotrophic nanoflagellates (HNF) were studied monthly from January 1997 to December 1998 in the middle Adriatic Sea. Bacterial and HNF relationships with phytoplankton biomass and temperature were analyzed to examine how the relative importance of bottom-up and top-down factors may shift over seasons and locations. For the coastal area, an inconsistent relationship between bacterial abundance and chlorophyll a and a stronger relationship between bacterial abundance and bacterial production suggest that other substrates than those of phytoplankton origin are important for bacteria. The analysis of simultaneous effects of temperature and bacterial production on bacterial abundance showed that the effect of temperature obscured the effects of bacterial production, suggesting that bacterial growth itself is highly temperature-dependent. The relationship between HNF abundance and bacterial abundance was slightly improved by the inclusion of in situ temperature, bacterial production or both parameters, as additional independent variables. About 60% of the variability in HNF abundance can be explained by bacterial abundance, bacterial production and temperature. In the open sea, tight coupling of bacterial abundance with chlorophyll a concentrations implied that phytoplankton-derived substrates have a dominant role in controlling bacterial abundance. During the colder months, bacterial abundance was high enough to support higher HNF abundance than observed, suggesting that predation exerted a minor depressing influence on bacterial abundance during that period. During the spring-summer period, HNF controlled bacterial standing stocks by direct cropping of bacterial production.Communicated by: H.-D. Franke     

Shifts in bacterial community composition associated with increased carbon cycling in a mosaic of phytoplankton blooms

Marine microbes have a pivotal role in the marine biogeochemical cycle of carbon, because they regulate the turnover of dissolved organic matter (DOM), one of the largest carbon reservoirs on Earth. Microbial communities and DOM are both highly diverse components of the ocean system, yet the role of microbial diversity for carbon processing remains thus far poorly understood. We report here results from an exploration of a mosaic of phytoplankton blooms induced by large-scale natural iron fertilization in the Southern Ocean. We show that in this unique ecosystem where concentrations of DOM are lowest in the global ocean, a patchwork of blooms is associated with diverse and distinct bacterial communities. By using on-board continuous cultures, we identify preferences in the degradation of DOM of different reactivity for taxa associated with contrasting blooms. We used the spatial and temporal variability provided by this natural laboratory to demonstrate that the magnitude of bacterial production is linked to the extent of compositional changes. Our results suggest that partitioning of the DOM resource could be a mechanism that structures bacterial communities with a positive feedback on carbon cycling. Our study, focused on bacterial carbon processing, highlights the potential role of diversity as a driving force for the cycling of biogeochemical elements.     

A cautionary note: Examples of possible microbial community dynamics in dilution grazing experiments

Dilution experiments are used commonly to provide estimates of grazing pressure exerted on phytoplankton and bacterioplankton as well as estimate their growth rates. However, very little attention has been given to the dynamics of grazers, especially heterotrophic nanoflagellates (HNF), in such experiments. We found temporal changes in concentrations of ciliates and HNF in a dilution experiment using water from the oligotrophic N.W. Mediterranean Sea. Ciliates decreased markedly over 24 h when held in seawater diluted with particle-free water (60% and 20% final conc whole seawater) while HNF increased in concentration in the same treatments. Using a time-course approach in a second experiment, we monitored changes in HNF and bacterioplankton concentrations in 20% whole seawater (80% particle-free seawater). Both HNF and heterotrophic bacteria displayed stable concentrations for the first 12 h and then grew rapidly, especially HNF, from 12 to 24 h. Examination of bacterial community composition using denaturing gel gradient electrophoresis (DGGE) showed a change in community composition over the 24 h incubation period. Dilution can have differential effects on the distinct components of the marine microbial food web.     

Uptake of Dissolved Organic Carbon by Gammaproteobacterial Subgroups in Coastal Waters of the West Antarctic Peninsula

Heterotrophic bacteria are well known to be key players in the turnover of dissolved organic material (DOM) in the oceans, but the relationship between DOM uptake and bacterial clades is still not well understood. Here we explore the turnover and single-cell use of glucose, an amino acid mixture, N-acetylglucosamine (NAG), and protein by gammaproteobacterial clades in coastal waters of the West Antarctic Peninsula in summer and fall. More than 60% of the cells within two closely related gammaproteobacterial clades, Ant4D3 and Arctic96B-16, were active in using the amino acid mixture, protein, and NAG. In contrast, an average of only 7% of all SAR86 cells used amino acids and protein even in summer when DOM use was high. In addition to DOM uptake within a group, we explored the contribution of the three gammaproteobacterial groups to total community uptake of a compound. SAR86 contributed 5- to 10-fold less than the other gammaproteobacterial subgroups to the uptake of all compounds. We found that the overall contribution of the Ant4D3 clade to DOM uptake was highest, whereas the SAR86 clade contributed the least to DOM turnover in West Antarctic Peninsula waters. Our results suggest that the low growth activity of a bacterial clade leads to low abundance, fewer active cells and a low contribution to the turnover of DOM components.     

北冰洋海域微食物环研究进展

海洋微食物环在海洋生态系统中起着重要作用.北冰洋因常年为海冰所覆盖,对微食物环的研究较为有限.现有研究表明,微食物环在北冰洋生态系统中的作用与海域和季节相关.近年来环境的快速变化、特别是夏季海冰覆盖面积的迅速减少,会对微食物环的结构和功能产生重大影响,已有研究显示其生态作用有望进一步提高.综合近年来已有的研究成果,对北冰洋微食物环的主要类群:原核生物、真核浮游植物、原生动物和浮游病毒等的基本生态特征进行了概述,讨论了各类群间的相互关系,并对未来的研究重点进行了展望.     

Viral lysis and grazing loss of bacteria in nutrient- and carbon-manipulated brackish water enclosures

A 3 week enclosure experiment was carried out at the Gulf of Finland, the Baltic Sea. After additions of inorganic nutrients [nitrogen (N) + phosphorus (P)] and a carbon source (sucrose), we followed bacterial, viral and heterotrophic nanoflagellate (HNF) abundances, as well as bacterial production and the frequency of bacteria visibly infected with viruses. Furthermore, the decay rate of virus particles was measured three times during the enclosure experiment from the KCN-treated water samples. Bacterial mortality caused by viral lysis was estimated using the decay rates and the fraction of bacteria infected. Nutrient (N + P) additions stimulated phytoplankton growth [the chlorophyll (Chl) a concentration increased from <5 g l^-1 up to 19 g l^-1], while sucrose additions increased bacterial production (from 4-6 x 10⁷ l^-1 h^-1). The phytoplankton blooms affected bacterial production only slightly. Bacterial mortality that was explained by viruses ranged from <2% to 13% when estimated from the visibly infected cells, and from 8% to 808% when the decay rates (range 0.052-0.765 h^-1) were used. Assuming a clearance rate of 5 nl flagellate^-1 h^-1, the HNF community could graze 16-135% of total bacterial loss.      

Effects of Small-Scale Turbulence on Bacteria: A Matter of Size

We examined the influence of small-scale turbulence and its associated shear on bacterioplankton abundance and cell size. We incubated natural microbial assemblages and bacteria-only fractions and subjected them to treatments with turbulence and additions of mineral nutrients and/or organic carbon. Bacterial abundance was not affected directly by turbulence in bacteria-only incubations. In natural microbial assemblage incubations, bacterial concentrations were higher under turbulence than in still-water controls when nutrients were added. In general, in the turbulence treatments bacteria increased significantly in size, mainly due to elongation of cells. The addition of inorganic nutrients had a negative effect on bacterial size, but a significantly positive effect on abundance independently of other factors such as turbulence and the presence of predators. Flagellate grazing did not trigger an increase in bacterial size as a grazing resistance response in unmixed containers. With the addition of organic carbon, bacteria elongated and partly settled to the bottom of the containers, in both the turbulent and still treatment, but bacterial abundance did not further increase. Furthermore, bacteria aggregated in the turbulence treatments after the second day of incubation even in the absence of other components of the microbial community. We found that turbulence and the associated shear increase bacterial size and change bacterial morphology, at least under certain nutrient conditions. This might be due to a physiological response (enhanced growth rate and/or unbalanced growth) or due to the selection of opportunistic strains when organic carbon is in excess compared to mineral nutrients. We suggest that shear associated with turbulent flow enhances the DOM flux to bacteria directly as well as indirectly through enhanced grazing activity and photosynthetic release. The formation of bacterial aggregates and filaments under turbulence might give selective advantage to bacteria in terms of nutrient uptake and grazing resistance.     

Net production and consumption of fluorescent colored dissolved organic matter by natural bacterial assemblages growing on marine phytoplankton exudates

An understanding of the distribution of colored dissolved organic matter (CDOM) in the oceans and its role in the global carbon cycle requires a better knowledge of the colored materials produced and consumed by marine phytoplankton and bacteria. In this work, we examined the net uptake and release of CDOM by a natural bacterial community growing on DOM derived from four phytoplankton species cultured under axenic conditions. Fluorescent humic-like substances exuded by phytoplankton (excitation/emission [Ex/Em] wavelength, 310 nm/392 nm; Coble's peak M) were utilized by bacteria in different proportions depending on the phytoplankton species of origin. Furthermore, bacteria produced humic-like substances that fluoresce at an Ex/Em wavelength of 340 nm/440 nm (Coble's peak C). Differences were also observed in the Ex/Em wavelengths of the protein-like materials (Coble's peak T) produced by phytoplankton and bacteria. The induced fluorescent emission of CDOM produced by prokaryotes was an order of magnitude higher than that of CDOM produced by eukaryotes. We have also examined the final compositions of the bacterial communities growing on the exudates, which differed markedly depending on the phytoplankton species of origin. Alteromonas and Roseobacter were dominant during all the incubations on Chaetoceros sp. and Prorocentrum minimum exudates, respectively. Alteromonas was the dominant group growing on Skeletonema costatum exudates during the exponential growth phase, but it was replaced by Roseobacter afterwards. On Micromonas pusilla exudates, Roseobacter was replaced by Bacteroidetes after the exponential growth phase. Our work shows that fluorescence excitation-emission matrices of CDOM can be a helpful tool for the identification of microbial sources of DOM in the marine environment, but further studies are necessary to explore the association of particular bacterial groups with specific fluorophores.     

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.     

DECOMPOSITION IN PELAGIC MARINE ECOSYSTEMS

SUMMARY

During the decomposition of plant detritus, complex microbial successions develop which are dominated in the early stages by a number of distinct bacterial morphotypes. The microheterotrophic community rapidly becomes heterogenous and may include cyanobacteria, fungi, yeasts and bactivorous protozoans.

Microheterotrophs in the marine environment may have a biomass comparable to that of all other heterotrophs and their significance as a resource to higher trophic orders, and in the regeneration of nutrients, particularly nitrogen, that support ‘regenerated’ primary production, has aroused both attention and controversy.

Soluble low molecular weight substrates (dissolved organic matter, or DOM) are for the most part rapidly turned over and readily taken up with a high growth efficiency by bacteria although detrital particulate organic material (POM) is turned over slowly and utilized with a low growth efficiency, owing to the structural complexity of the detritus. The presence of appropriate substrate-specific strains of bacteria Is important in the decomposition of both DOM and POM.

Estimates of the transfer of photosynthetically fixed carbon and nitrogen through the pelagic microbial community have recently become widespread. However, the quantification of C and N fluxes through bacteria and microzooplankton is very sensitive to accurate measures of microbial biomass, production, net growth yield, bacterial activity and bactivory by microzooplankton. These processes also vary significantly in both spatial and temporal dimensions during the development and decay of phytoplankton blooms.

Recent attempts to model decomposition processes and C and N fluxes In pelagic marine ecosystems are described. This review examines the most sensitive components and predictions of the models with particular reference to estimates of bacterial production, net growth yield and predictions of N cycling determined by ¹⁵N methodology.     

Bottom-up excitable models of phytoplankton blooms

A simple nutrient-phytoplankton model is used to explore the dynamics of phytoplankton blooms. The model exhibits excitable behaviour in the sense that a large scale outbreak can only be triggered when a critical nutrient threshold is exceeded. The model takes into account several features often neglected but whose combined effect proves very important: (i) rapid nutrient recycling associated with the microbial loop and patch formation; (ii) self-shading; and (iii) a bottom-up approach, whereby nutrient levels are responsible for both the triggering and the demise of the bloom. Although the literature is replete with studies of ‘top-down’ models in which zooplankton grazing control the triggering and demise of the bloom, bottom-up models are nevertheless appropriate in many circumstances. We provide a full mathematical investigation of the effects of these three different features in an excitable system framework.     

Pseudomonas aeruginosa Biofilms in Disease

Analysis of the composition of the marine-dissolved organic matter has highlighted the importance of d-amino acids, whose origin is attributed mainly to the remains of bacterial peptidoglycan released as a result of grazing or viral lysis. However, very few studies have focused on the active release of d-amino acids by bacteria. With this purpose, we measured the concentration of dissolved amino acids in both enantiomeric forms with two levels of complexity: axenic cultures of Vibrio furnissii and Vibrio alginolyticus and microcosms created from marine microbial assemblages (Biscay Bay, Cantabrian Sea) with and without heterotrophic nanoflagellates (HNFs). Axenic cultures showed that only d-Ala was significantly released and accumulated in the medium up to a concentration of 120 nM, probably as a consequence of the rearrangement of peptidoglycan. The marine microbial assemblages showed that only two d-amino acids significantly accumulated in the environment, d-Ala and d-aspartic acid (Asp), in both the absence and presence of HNFs. The d/l ratio increased during the incubation and reached maximum values of 3.0 to 4.3 for Ala and 0.4 to 10.6 for Asp and correlated with prokaryotic and HNF abundance as well as the rate of prokaryotic thymidine and leucine incorporation. Prokaryotes preferentially consumed l-amino acids, but the relative uptake rates of d-Ala significantly increased in the growth phase. These results demonstrate that bacteria can release and consume d-amino acids at high rates during growth, even in the absence of viruses and grazers, highlighting the importance of bacteria as producers of dissolved organic matter (DOM) in the sea.     

Phytoplankton exudation: exploitation of the microbial loop as a defence against algal viruses

Healthy phytoplankton cells exude dissolved organic matter (DOM).In a model, DOM exudation is demonstrated to be a cost-effective,indirect, means of reducing virus infection, which can be amajor cause of phytoplankton mortality. Diffusion theoly showsthat, for a given biomass, small particles will have a muchhigher rate of wlsorption of solutes than will large ones. Thuscolloidal viruses are far more likely to come into contact withbacteria than with phytoplankton if the same biomasses are present.B can destroy viruses in a large proportion of contacts. Althoughflagellate protozoa have lower contact rates with viruses, theymay consume more of the viruses that they do encounter and theydeal with larger viruses particularly effectively. The exudedDOM supports bacterial growth, which in turn may support flagellates.Even fairly low levels of exudation can maintain the biomassof bacteria or small flagellates required to remove >50%of viruses before they have a chance to infect their host, atleast for larger phytoplankton. High rates of virus removalmay occur at the high exudation rates that are typical of lateblooms, It is concluded that healthy phytoplankton cells exudeDOM in order to remain healthy.     

Relationship between bacteria, phytoplankton and heterotrophic nanoflagellates along the trophic gradient

Bacterial and heterotrophic nanoflagellates (HNF) abundance, as well as bacterial production and chlorophylla levels, were measured at five sites extending from the coastal zone toward the open Adriatic in the period from March to October 1995. The investigated areas were grouped into trophic categories according to concentrations of chlorophylla. All the biotic-para-meters increased along the trophic gradient, leading to eutrophy, but they did not increase at the same rate. The bacterial biomass: phytoplankton biomass (BB: chla) ratio decreased from about 10 in the very oligotrophic area to 0.8 at the eutrophic site. In contrast, the bacterial abundance: HNF abundance ratio (B: HNF) increased from 1000 bacteria per 1 flagellate in the oligotrophic system to 1700 bacteria flagellate⁴ in the eutrophic area. Decreasing BB: chla and increasing B: HNF ratios along the trophic gradient might reflect the different structures of the microbial food web. Relationships between bacterial abundance and production, and chla and HNF showed that bacterial abundance along the trophic gradient was regulated by the interplay between nutrient supply and grazing pressure. But in the oligotrophic system, bacterial abundance was more closely related to bacterial production and chla than in the eutrophic system, suggesting stronger control of bacterial abundance by substrate supply. On the other hand, the coupling between bacteria and HNF, and uncoupling between bacterial abundance and production in the eutrophic system, showed that the importance of bacteriovory increased in richer systems.