Heterotrophic Flagellates Research Articles

An SSU rDNA barcoding approach to the diversity of marine interstitial cercozoans, including descriptions of four novel genera and nine novel species

Environmental DNA surveys have revealed a great deal of hidden diversity within the Cercozoa. An investigation into the biodiversity of heterotrophic flagellates in marine benthic habitats of British Columbia, Canada, demonstrated the presence of several undescribed taxa with morphological features that resemble the cercozoan genera Cryothecomonas and Protaspis. Nine novel species of marine interstitial cercozoans are described that are distributed into five genera, four of which are new. Phylogenetic analyses of small subunit rDNA sequences derived from two uncultured isolates of Protaspis obliqua and nine novel cercozoan species (within four novel genera) provided organismal anchors that helped establish the cellular identities of several different environmental sequence clades. These data, however, also showed that the rarity of distinctive morphological features in cryomonads, and other groups of cercozoans, makes the identification and systematics of the group very difficult. Therefore, a DNA barcoding approach was applied as a diagnostic tool for species delimitation that used a 618 bp region at the 5' end of the SSU rDNA sequence. Nucleotide sequence analysis of this region showed high intergeneric sequence divergences of about 7% and very low intraspecific sequence divergences of 0-0.5%; phylogenetic analyses inferred from this barcoding region showed very similar tree topologies to those inferred from the full-length sequence of the gene. Overall, this study indicated that the 618 bp barcoding region of SSU rDNA sequences is a useful molecular signature for understanding the biodiversity and interrelationships of marine benthic cercozoans.

A Decade of Change in the Skidaway River Estuary. III. Plankton

The Skidaway River estuary, GA (USA), a tidally dominated subtropical system surrounded by extensive Spartina salt marshes, is experiencing steady increases in nutrients, chlorophyll, and particulate matter and decline in dissolved oxygen, associated with cultural eutrophication. A long-term study is documenting changes in these parameters: previous papers Verity (Estuaries 25:944–960, 2002a, Estuaries 25:961–975, b) reported on hydrography, nutrients, chlorophyll, and particulate matter during 1986–1996; plankton community responses are reported here. Phytoplankton, bacteria, heterotrophic nanoplankton and dinoflagellates, ciliates, and copepods exhibited strong seasonal cycles in abundance driven by temperature and resource availability, typically with summer maxima and winter minima. However, cultural eutrophication coincided with altered planktonic food webs as autotrophic and heterotrophic communities responded to increasing concentrations and changing ratios of inorganic and organic nutrients, potential prey, and predators. Small (<8 μm) photosynthetic nanoplankton increased in absolute concentration and also relative to larger cells. In contrast, diatoms did not show consistent increases in abundance, despite significant long-term increases in ambient silicate concentrations. Mean annual bacteria concentrations approximately doubled, and eukaryotic organisms in the microbial food web (heterotrophic and mixotrophic flagellates, dinoflagellates, ciliates, and metazoan zooplankton) also increased. All plankton groups except copepods showed trends of increasing annual amplitudes between seasonal high and low values, with higher peak concentrations each year. These observations suggest that the eutrophication signal was gradually becoming uncoupled from regulatory mechanisms. Theory and evidence from other more impacted waters suggest that, if these patterns continue, changes in the structure and function of higher trophic levels will ensue.

Community Structure and Dynamics of Small Eukaryotes Targeted by New Oligonucleotide Probes: New Insight into the Lacustrine Microbial Food Web

The seasonal dynamics of the small eukaryotic fraction (cell diameter, 0.2 to 5 microm) was investigated in a mesotrophic lake by tyramide signal amplification-fluorescence in situ hybridization targeting seven different phylogenetic groups: Chlorophyceae, Chrysophyceae, Cryptophyceae, Cercozoa, LKM11, Perkinsozoa (two clades), and Fungi. The abundance of small eukaryotes ranged from 1,692 to 10,782 cells ml(-1). The dominant groups were the Chrysophyceae and the Chlorophyceae, which represented 19.6% and 17.9% of small eukaryotes, respectively. The results also confirmed the quantitative importance of putative parasites, Fungi and Perkinsozoa, in the small heterotrophic eukaryotic assemblage. The relative abundances recorded for the Perkinsozoa group reached as much as 31.6% of total targeted eukaryotes during the summer. The dynamics of Perkinsozoa clade 1 coincided with abundance variations in Peridinium and Ceratium spp. (Dinoflagellates), while the dynamics of Perkinsozoa clade 2 was linked to the presence of Dinobryon spp. (Chrysophyceae). Fungi, represented by chytrids, reached maximal abundance in December (569 cells ml(-1)) and were mainly correlated with the dynamics of diatoms, especially Melosira varians. A further new finding of this study is the recurrent presence of Cercozoa (6.2%) and LKM11 (4.5%) cells. This quantitative approach based on newly designed probes offers a promising means of in-depth analysis of microbial food webs in lakes, especially by revealing the phylogenetic composition of the small heterotrophic flagellate assemblage, for which an important fraction of cells are generally unidentified by classical microscopy (on average, 96.8% of the small heterotrophic flagellates were identified by the specific probes we used in this study).

Seasonal variations of microbial abundances and virus- versus flagellate-induced mortality of picoplankton in three peri-alpine lakes

Viruses and small heterotrophic flagellates are known to exert an important control on bacterial populations. In parallel with the study of picoplankton dynamics (abundance and distribution) in surface waters (0-50 m) of Lakes Geneva, Bourget and Annecy, we used a dilution technique during different seasonal periods in order to assess flagellate- versus virus-induced mortality of heterotrophic bacteria, picocyanobacteria and small eukaryotic phytoplankton. Although it was not always possible to detect a significant viral effect (typically in winter), viral lysis and protozoan grazing could be responsible for up to 71 % of the bacterial mortality (in summer). Viral impact, considered alone, never equalled or exceeded predation for heterotrophic bacteria, but could for picocyanobacteria, typically in autumn. In addition, during summer complex interactions between grazing- and virus-induced mortality of bacteria (e.g synergism versus antagonism) could be highlighted for instance with bacterial lysis susceptible to enhance picocyanobacterial growth). The temporal variations observed for experimental viral parasitism and flagellate predation were consistent with the in situ dynamics and statistical relationships found between the targeted communities. This study thus provides new evidence on the critical role played by viruses and small flagellates in the functioning of freshwater microbial food webs and also that these mortality processes vary strongly throughout the seasons.

Role of dispersal in shaping communities of ciliates and heterotrophic flagellates within riverine biofilms

We tested the role of immigration from the plankton on the structure of local surface‐associated communities of ciliates and heterotrophic flagellates (HF) in flow cells fed with fresh riverine water (Rhine, Germany). By applying size fractionation, we reduced the immigration potential of HF from the plankton in mature, pregrown biofilms by 93%, and observed no significant effects on the abundance and taxonomic composition of HF. Compared with a treatment with natural plankton density, a 100% reduction in the number of planktonic cells flowing over mature biofilms resulted in a reduced total abundance of ciliates, although only slight effects on the relative composition of present morphospecies were detectable. When starting with sterile flow cells, we found an initial linear increase in HF abundance proportional to their planktonic abundance; additionally supplemented planktonic bacteria showed that this initial colonization rate was resource independent. The initial phase was followed by an exponential increase, which was also independent from the resource level and strongest when initial colonization was low. In contrast to the early succession stages, the final abundances reached were independent of the plankton abundance but strongly dependent upon the local resource level. Immigration is an important factor controlling the initial substrate colonization and early growth, and ciliate and HF communities in biofilms become increasingly independent of immigration with maturation, being then controlled by local factors such as resources.

Light Microscopic Observations, Ultrastructure, and Molecular Phylogeny of Hicanonectes teleskopos n. g., n. sp., a Deep‐Branching Relative of Diplomonads

We describe Hicanonectes teleskopos n. g., n. sp., a heterotrophic flagellate isolated from low-oxygen marine sediment. Hicanonectes teleskopos has a ventral groove and two unequal flagella, and rapidly rotates during swimming. At the ultrastructural level H. teleskopos is a "typical excavate": it displays flagellar vanes, a split right microtubular root, "I,""B," and "C" fibres, a singlet microtubular root, and a possible composite fibre. Small subunit rRNA (SSU rRNA) gene phylogenies and an "arched" B fibre demonstrate that H. teleskopos belongs to Fornicata (i.e. diplomonads, retortamonads, and relatives). It forms a clade with the deep-branching fornicate Carpediemonas, with moderate-to-strong bootstrap support, although their SSU rRNA gene sequences are quite dissimilar. Hicanonectes differs from Carpediemonas in cell shape, swimming behaviour, number of basal bodies (i.e. 4 vs. 3), number of flagellar vanes (i.e. 2 vs. 3), anterior root organization, and by having a cytopharynx. Like Carpediemonas and Dysnectes, Hicanonectes has conspicuous mitochondrion-like organelles that lack cristae and superficially resemble the hydrogenosomes of parabasalids, rather than the mitosomes of their closer relatives the diplomonads (e.g. Giardia).

Direct Effects of UV-B Radiation on the Freshwater Heterotrophic NanoflagellateParaphysomonassp

The formation of DNA photoproducts in organisms exposed to ambient levels of UV-B radiation can lead to death and/or reduced population growth in aquatic systems. Dependence on photoenzymatic repair to reverse DNA damage caused by UV-B radiation is demonstrated for Paraphysomonas sp., a member of a widely distributed genus of heterotrophic nanoflagellates. At 20 degrees C, Paraphysomonas sp. was exposed to a range of UV-B intensities encountered in natural systems. Populations of the flagellate survived and grew in a dose-dependent manner, but only when simultaneously exposed to photorepair radiation (PRR). In contrast, flagellates exposed to UV-B at 15 degrees C suffered 100% mortality except at the lowest UV-B level (with PRR) tested, which suggested a photorepair temperature optimum above 15 degrees C. After acute UV-B exposures, DNA damage (measured as the formation of pyrimidine dimers) was reduced only in organisms that underwent subsequent exposure to PRR. Populations kept in the dark after UV-B exposure maintained the initial levels of pyrimidine dimers. These results are the first to demonstrate the reliance of a heterotrophic flagellate on photoenzymatic DNA repair for survival from UV-B exposure.

Structure and seasonal dynamics of the protozoan community (heterotrophic flagellates, ciliates, amoeboid protozoa) in the plankton of a large river (River Danube, Hungary)

Seasonal dynamics of all major protozoan groups were investigated in the plankton of the River Danube, upstream of Budapest (Hungary), by bi-weekly sampling over a 1-year long period. Sixty-one heterotrophic flagellate, 14 naked amoeba, 50 testate amoeba, 4 heliozoan and 83 ciliate morphospecies were identified. The estimated abundance ranges of major groups throughout the year were as follows: heterotrophic flagellates, 0.27–7.8×10 6 ind. l −1; naked amoebae, max. 3300 ind. l −1; testaceans, max. 1600 ind. l −1; heliozoans, max. 8500 ind. l −1; ciliates, 132–34,000 ind. l −1. In terms of biovolume, heterotrophic flagellates dominated throughout the year (max. 0.58 mm 3 l −1), and ciliates only exceeded their biovolume in summer (max. 0.76 mm 3 l −1). Naked amoeba and heliozoan biovolume was about one, and testacean biovolume 1–3, orders of magnitude lower than that of ciliates. In winter, flagellates, mainly chrysomonads, had the highest biomass, whilst ciliates were dominated by peritrichs. In 2005 from April to July a long spring/summer peak occurred for all protozoan groups. Beside chrysomonads typical flagellates were choanoflagellates, bicosoecids and abundant microflagellates (large chrysomonads and Collodictyon). Most abundant ciliates were oligotrichs, while Phascolodon, Urotricha, Vorticella, haptorids, Suctoria, Climacostomum and Stokesia also contributed significantly to biovolume during rapid succession processes. In October and November a second high protozoan peak occurred, with flagellate dominance, and slightly different taxonomic composition.

The Choanoflagellates: Heterotrophic Nanoflagellates and Sister Group of the Metazoa

INTRODUCTIONChoanoflagellates are the closest living relatives of the metazoa, and the study of their cell biology and genomes promises to provide new insights into metazoan ancestry and origins. These heterotrophic flagellates are a cosmopolitan group of small, colorless protozoa that are present in marine and freshwater environments as well as in hydrated soils. Monosiga brevicollis, a marine species, has emerged as a representative of the group because it is easily grown and manipulated in the laboratory and was the subject of a recently completed genome project. Because of the similarity in morphology and ecology shown across the order Choanoflagellida, the protocols mentioned in this article should be transferable to most choanoflagellate species.

Distribution of the uncultured protist MAST‐4 in the Indian Ocean, Drake Passage and Mediterranean Sea assessed by real‐time quantitative PCR

Molecular surveys of marine picoeukaryotes have revealed a large number of sequences unrelated to cultured organisms, such as those forming the marine stramenopile (MAST)-4 clade. Recent FISH (fluorescent in situ hybridization) data have shown that MAST-4 cells are uncultured heterotrophic flagellates of 2-3 microm in size that have a global distribution in non-polar marine waters. However, FISH is time-consuming and hard to apply to the many samples generated during oceanographic cruises, so we developed a real-time quantitative polymerase chain reaction (Q-PCR) protocol to determine rapidly the abundance of this group using environmental DNA. We designed a primer set targeting the 18S rRNA genes (rDNA) of MAST-4 and optimized and calibrated the Q-PCR protocol using a plasmid with the target sequence as insert. The Q-PCR was then applied to quantify MAST-4 rDNA molecules along three marine transects, longitudinal in the Indian Ocean, latitudinal in the Drake Passage and coastal-offshore in the Mediterranean Sea, and to a temporal study in a Mediterranean Sea coastal station. MAST-4 was detected in all samples processed (averaged abundances between 500 and 1000 rDNA molecules ml(-1)) except in mesopelagic and Antarctic samples, where it was virtually absent. In general, it was more abundant in the coast than offshore and in the deep chlorophyll maximum than at surface. A comparison of Q-PCR and FISH signals in well-controlled microbial incubations indicated that MAST-4 cells have around 30 copies of the rDNA operon. This Q-PCR assay quickly yielded quantitative data of uncultured MAST-4 cells and confirmed their wide distribution and putative ecological importance.

Grazing rates and functional diversity of uncultured heterotrophic flagellates

Aquatic assemblages of heterotrophic protists are very diverse and formed primarily by organisms that remain uncultured. Thus, a critical issue is assigning a functional role to this unknown biota. Here we measured grazing rates of uncultured protists in natural assemblages (detected by fluorescent in situ hybridization (FISH)), and investigated their prey preference over several bacterial tracers in short-term ingestion experiments. These included fluorescently labeled bacteria (FLB) and two strains of the Roseobacter lineage and the family Flavobacteriaceae, of various cell sizes, which were offered alive and detected by catalyzed reporter deposition-FISH after the ingestion. We obtained grazing rates of the globally distributed and uncultured marine stramenopiles groups 4 and 1 (MAST-4 and MAST-1C) flagellates. Using FLB, the grazing rate of MAST-4 was somewhat lower than whole community rates, consistent with its small size. MAST-4 preferred live bacteria, and clearance rates with these tracers were up to 2 nl per predator per h. On the other hand, grazing rates of MAST-1C differed strongly depending on the tracer prey used, and these differences could not be explained by cell viability. Highest rates were obtained using FLB whereas the flavobacteria strain was hardly ingested. Possible explanations would be that the small flavobacteria cells were outside the effective size range of edible prey, or that MAST-1C selects against this particular strain. Our original dual FISH protocol applied to grazing experiments reveals important functional differences between distinct uncultured protists and offers the possibility to disentangle the complexity of microbial food webs.

Responses of Biofilm-Dwelling Ciliate Communities to Planktonic and Benthic Resource Enrichment

Four experiments covering different seasons were performed to test the impact of increased benthic and planktonic resource availability on the structure of biofilm-dwelling ciliate communities which were cultivated in river bypass systems. The growth of benthic bacteria was stimulated by the addition of dissolved organic carbon. The enrichment of the planktonic resource was achieved by supplementation with suspended bacteria. It was shown that both resource enrichments can differentially influence abundance and taxonomic structure of ciliate communities. Furthermore, both resources can influence different stages during biofilm colonization. Increased benthic bacterial growth mainly resulted in both an accumulation of primarily grazing-resistant bacterial filaments and in an increase in the number of vagile heterotrophic flagellates. This can stimulate nanophagous ciliates (feeding on flagellates) in addition to the direct stimulation of bacteriovorous ciliates. The effects of the planktonic bacteria enrichments were twofold: They could have been utilized either directly by suspension-feeding ciliates or indirectly through an enhanced growth of suspension-feeding attached heterotrophic flagellates, which were then in turn grazed upon by ciliates. The magnitude of responses of the total ciliate abundance to the two resource enrichments further depended on the background conditions, thereby showing temporarily variable limitations of these resources. Furthermore, the particular taxonomic groups stimulated by one resource type sometimes differed between the experiments, an observation which demonstrates that the response depends on different environmental factors and is not easily predictable based simply on resource type. Taken together, our results emphasize the need of a differentiated view on the effects of resources on complex biofilm-dwelling consumer communities with respect to both the origin of carbon source as well as the particular environmental conditions.

Responsibility of microzooplankton and parasite pressure for the demise of toxic dinoflagellate blooms

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 53:211-225 (2008) - DOI: https://doi.org/10.3354/ame01245 Responsibility of microzooplankton and parasite pressure for the demise of toxic dinoflagellate blooms David J. S. Montagnes1,*, Aurélie Chambouvet2,3, Laure Guillou2,3, Andy Fenton1 1School of Biological Sciences, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, UK 2Université Pierre et Marie Curie – Paris 6, Station Biologique de Roscoff, Place Georges Teissier, BP 74, 29680 Roscoff, France 3CNRS, UMR 7144, Laboratoire Adaptation et Diversité en Milieu Marin, Place Georges Teissier, BP 74, 29680 Roscoff, France *Email: dmontag@liv.ac.uk ABSTRACT: Two mechanisms proposed to control dinoflagellate blooms—parasitism by eukaryotes (e.g. Amoebophrya sp.) and grazing by microzooplankton—have been explored through previous laboratory and field studies but lack quantitative assessment. We modelled the relative effect of these mechanisms. We used literature values to embed an Anderson-May host-microparasite model into a microbial food web model (nano- and microphytoplankton, nano- and microzooplankton, a toxic dinoflagellate, a dinoflagellate parasite). Three scenarios were examined to simulate the introduction of a toxic dinoflagellate and its parasite into an environment: (1) a food web, including autotrophic nano- and microplankton, heterotrophic flagellates, and ciliates; (2) as for Case 1 but with a toxic dinoflagellate; (3) as for Case 2 but with a dinoparasite. This mimics observations in a French estuary, where a toxic dinoflagellate began blooming in 1988; since 1998, blooms appear to have become regulated, and numerous parasitic infections by Amoebophrya sp. occurred from 2004 to 2006. After supporting parasite control of dinoflagellate blooms, we assessed the effects of observed ranges of key variables associated with the parasite and other components of the food web on parasite control of the dinoflagellate population. Population dynamics were examined over 50 d. In Case 1, all taxa had 10 to 20 d blooms. In Case 2, the toxic dinoflagellate population dynamics mimicked that of the microphytoplankton, and this dinoflagellate was reduced in numbers but not extirpated by microzooplankton grazing. In Case 3 population blooms occurred, and the parasite virtually eliminated the dinoflagellate over ~10 d. Sensitivity analysis indicated that our assessment was robust. We propose that the decline in toxic dinoflagellates in the French estuary may have been due to an introduced dinoparasite. In general, we suggest that parasites may have greater impact on toxic dinoflagellate blooms than microzooplankton grazers; the parasites have the potential to eliminate the toxic dinoflagellates. We recommend that such parasites be incorporated into more complex food web models. KEY WORDS: Amoebophrya · Alexandrium minutum · Dinospore · Harmful algal bloom · HAB · Microbial food web · Model · Plankton · Red tide Full text in pdf format PreviousNextCite this article as: Montagnes DJS, Chambouvet A, Guillou L, Fenton A (2008) Responsibility of microzooplankton and parasite pressure for the demise of toxic dinoflagellate blooms. Aquat Microb Ecol 53:211-225. https://doi.org/10.3354/ame01245 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 53, No. 2. Online publication date: October 31, 2008 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2008 Inter-Research.

Effects of protozoan grazing within river biofilms under semi-natural conditions

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 52:283-296 (2008) - DOI: https://doi.org/10.3354/ame01236 Effects of protozoan grazing within river biofilms under semi-natural conditions Jennifer K. Wey, Anja Scherwass, Helge Norf, Hartmut Arndt, Markus Weitere* Department of General Ecology and Limnology, Zoological Institute, University of Cologne, Weyertal 119, 50931 Cologne, Germany *Corresponding author. Email: markus.weitere@uni-koeln.de ABSTRACT: Biofilms play an important role in the material flux of many aquatic ecosystems, but little is known about the mechanisms controlling their community structure under natural conditions. In the present study, we focused on the effects of ciliates on the quantity and taxonomic composition of heterotrophic flagellates (HF), and the effects of HF on the quantity and life forms (single cells vs. microcolonies) of bacteria in the early phase of biofilm colonization. For this purpose, we established semi-natural biofilms in flow cells connected to the river Rhine at Cologne, Germany. Using filter cartridges, we size-fractionated the potamoplankton, which is the source of the biofilm community, thus establishing biofilms containing (1) only bacteria (1.2 µm filter), (2) HF and bacteria (8 or 5 µm filter), or (3) ciliates, HF and bacteria (20 µm filter). The presence of ciliates negatively influenced the abundance of biofilm-dwelling HF and selectively altered the taxonomic composition of the HF community. The presence of HF resulted in a significant reduction in the abundance of single bacterial cells, but enhanced the abundance of bacterial microcolonies. Furthermore, the presence of ciliates stimulated the abundance of single-cell bacteria (probably due to an HF-mediated trophic cascade), but had no effect on bacterial microcolonies. Taken together, the results of this study show the importance of protozoan grazing in shaping the species composition and morphology of early river biofilms under semi-natural conditions. KEY WORDS: River biofilm · Benthic protozoans · Size fractionation · Trophic cascade · Top-down control · Microbial communities Full text in pdf format PreviousCite this article as: Wey JK, Scherwass A, Norf H, Arndt H, Weitere M (2008) Effects of protozoan grazing within river biofilms under semi-natural conditions. Aquat Microb Ecol 52:283-296. https://doi.org/10.3354/ame01236 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 52, No. 3. Online publication date: September 11, 2008 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2008 Inter-Research.

Microscopic and Molecular Studies of the Diversity of Free-Living Protozoa in Meat-Cutting Plants

The diversity of free-living protozoa in five meat-cutting plants was determined. Light microscopy after enrichment culturing was combined with sequencing of PCR-amplified, denaturing gradient gel electrophoresis (DGGE)-separated 18S rRNA gene fragments, which was used as a fast screening method. The general results of the survey showed that a protozoan community of amoebae, ciliates, and flagellates was present in all of the plants. Protozoa were detected mainly in floor drains, in standing water on the floor, on soiled bars of cutting tables, on plastic pallets, and in out-of-use hot water knife sanitizers, but they were also detected on surfaces which come into direct contact with meat, such as conveyer belts, working surfaces of cutting tables, and needles of a meat tenderizer. After 7 days of incubation at refrigerator temperature, protozoa were detected in about one-half of the enrichment cultures. Based on microscopic observations, 61 morphospecies were found, and Bodo saltans, Bodo spp., Epistylis spp., Glaucoma scintillans, Petalomonas spp., Prodiscophrya collini, and Vannella sp. were the most frequently encountered identified organisms. Sequencing of DGGE bands resulted in identification of a total of 49 phylotypes, including representatives of the Amoebozoa, Chromalveolata, Excavata, Opisthokonta, and Rhizaria. Sequences of small heterotrophic flagellates were affiliated mainly with the Alveolata (Apicomplexa), Stramenopiles (Chrysophyceae), and Rhizaria (Cercozoa). This survey showed that there is high protozoan species richness in meat-cutting plants and that the species included species related to known hosts of food-borne pathogens.

The Molecular Diversity of Freshwater Picoeukaryotes Reveals High Occurrence of Putative Parasitoids in the Plankton

Eukaryotic microorganisms have been undersampled in biodiversity studies in freshwater environments. We present an original 18S rDNA survey of freshwater picoeukaryotes sampled during spring/summer 2005, complementing an earlier study conducted in autumn 2004 in Lake Pavin (France). These studies were designed to detect the small unidentified heterotrophic flagellates (HF, 0.6–5 µm) which are considered the main bacterivores in aquatic systems. Alveolates, Fungi and Stramenopiles represented 65% of the total diversity and differed from the dominant groups known from microscopic studies. Fungi and Telonemia taxa were restricted to the oxic zone which displayed two fold more operational taxonomic units (OTUs) than the oxycline. Temporal forcing also appeared as a driving force in the diversification within targeted organisms. Several sequences were not similar to those in databases and were considered as new or unsampled taxa, some of which may be typical of freshwater environments. Two taxa known from marine systems, the genera Telonema and Amoebophrya, were retrieved for the first time in our freshwater study. The analysis of potential trophic strategies displayed among the targeted HF highlighted the dominance of parasites and saprotrophs, and provided indications that these organisms have probably been wrongfully regarded as bacterivores in previous studies. A theoretical exercise based on a new ‘parasite/saprotroph-dominated HF hypothesis’ demonstrates that the inclusion of parasites and saprotrophs may increase the functional role of the microbial loop as a link for carbon flows in pelagic ecosystems. New interesting perspectives in aquatic microbial ecology are thus opened.

Complex interactions in the microbial world: underexplored key links between viruses, bacteria and protozoan grazers in aquatic environments

Viruses are the most abundant biological entities in aquatic ecosystems, and hetero- trophic bacteria form the major host reservoir for these viruses. Together with small protozoan graz- ers, these biological compartments are now recognized as the most abundant microorganisms on earth. Bacterial loss, which constitutes a key process in pelagic ecosystems, is mainly a result of lysis by viruses and grazing by protists, which may be comparable at times. Both viruses and protozoan grazers are known to be driving forces in shaping bacterial diversity. A significant correlation between viruses and protozoan grazers has been reported in various ecosystems. Some studies have also shown that grazers can feed directly on viruses, although the transfer of carbon to the higher trophic levels via this process seems to be of minor importance. Thus, there are many routes of inter- action between grazers, bacteria and viruses in pelagic systems, but few studies have attempted to characterize them. Interestingly, experimental studies have recently revealed both direct and indi- rect control by viruses and protozoan grazers on bacteria. Interactions between bacteria, between viruses and between heterotrophic flagellates are also likely to occur. From a mathematical point of view, we can argue that there are 9 different possible pathways. Although some of these have been well documented, others have barely been mentioned. This review attempts to cover advances in the field in recent years and concentrates on addressing the underexplored interactions among viruses, bacteria and heterotrophic nanoflagellates.

Easy Visualization of the Protist Oxyrrhis marina Grazing on a Live Fluorescently Labelled Heterotrophic Nanoflagellate

Planktonic heterotrophic flagellates are ubiquitous eukaryotic microorganisms that play a crucial role in carbon and nutrient fluxes through pelagic food webs. Here we illustrate for the first time a grazing model of planktonic dinoflagellate, Oxyrrhis marina, on the heterotrophic nanoflagellate Goniomonas amphinema, using the DNA-binding fluorescent dye Hoechst 33342. A solution of 1 microg/mL of the fluorochrome allowed viability of the prey for at least 48 hours, provided low fluorescence quenching, and labelled the flagellate without masking the cytoplasm. After 2 hours of contact between the fluorescent prey and the predator, O. marina population had preyed on live G. amphinema at an ingestion rate of 2.2 prey Oxyrrhis (-1) h(-1). Results show that this model is a time-effective and inexpensive approach for the direct observation of heterotrophic flagellate grazing. The fact that prey remain alive while predation occurs, as well as the low rate of quenching, could be of help in studying the fate of real-time trophic interactions between protists in microbial webs.

Heterotrophic flagellate biodiversity and community structure in freshwater streams

The species diversity and community structure of heterotrophic flagellates in watercourses from different geographical regions were investigated. Forty-one species have been identified. The species diversity of cercomonads, kinetoplastids, choanoflagellates, and flagellates is the highest. Bodo saltans, B. designis and Goniomonas truncata are the most common species. Planktonic communities in different streams are most similar in terms of species composition. The highest similarity within benthic cenoses was noted between geographically distant communities. Communities of heterotrophic flagellates seem to be highly heterogenous systems; species distribution depends mainly on the type of habitat and not on geographical factors.

Differential grazer‐mediated effects of high summer temperatures on pico‐ and nanoplankton communities

We investigated the role of a macrograzer (the filter feeding mussel Dreissena polymorpha) in mediating effects of high summer temperatures on the dominant components of natural river plankton (i.e., bacteria, algae, and heterotrophic flagellates) in flow channel experiments. Effects of adaptation (by comparing mussels from a southern and a northern population) and thermal acclimation of the mussels were considered. Both heterotrophic flagellates and algae are released from grazing pressure and increase in abundance at temperatures above 20°C. Bacterial abundance, however, decreased with increasing temperature, suggesting a trophic cascade (mussel‐flagellates‐bacteria) that is altered by the temperature response of the mussel ingestion rate. Warm acclimation of the mussels did not change the outcome of the experiments. The dreissenids from the southern population showed a significantly higher ingestion rate than those from the northern population only in July. The general pattern (i.e., decreasing ingestion rates at high temperatures) was found in both populations. Microbial communities controlled by macrofauna can experience substantial changes in warm summers because of differential development of direct and indirect grazing effects with increasing temperature.