Small Heterotrophic Flagellates Research Articles

Feeding by heterotrophic flagellates on marine archaea

Archaea live in diverse habitats including extreme environments. Although they are sometimes the dominant heterotrophic organisms in microbial communities, relatively little is known about their ecological roles in microbial food webs. To explore predator–prey relationships between archaea and heterotrophic protists, we determined feeding occurrence by the heterotrophic nanoflagellates Bodo sp., Goniomonas sp., and Pedospumella sp. the common heterotrophic dinoflagellates Cryptoperidiniopsis brodyi, Gyrodinium dominans, Oblea rotunda, Oxyrrhis marina, and Polykrikos kofoidii on ammonia-oxidizing archaea (AOA) by examining the food vacuoles of these heterotrophic flagellates under an epifluorescence microscope using fluorescently labeled AOA. In addition, we measured the ingestion rates of Goniomonas sp., C. brodyi, G.dominans, and O. marina on AOA as a function of prey concentration. Cells of Bodo sp., Goniomonas sp., Pedospumella sp., C. brodyi, G.dominans, and O. marina were able to feed on AOA, whereas O. rotunda and P. kofoidii were not. With increasing initial prey concentration up to 1.7 × 106 – 9.3 × 106 cells mL−1, the ingestion rates of Goniomonas sp., C. brodyi, G.dominans, and O. marina on AOA increased rapidly but increased slowly or became saturated at higher concentrations. Although the maximum ingestion rates of the smaller flagellate Goniomonas sp. (23.1 cells predator−1 h−1) on AOA was much lower than that of the larger flagellate O. marina (296.4 cells predator−1 h−1), AOA supported the positive growth of only Goniomonas sp. The results of the present study suggest that these small heterotrophic flagellates could be effective heterotrophic protist grazers of AOA in marine ecosystems.

Notable predominant morphology of the smallest most abundant protozoa of the open ocean revealed by electron microscopy.

In the microbe-driven ecosystems of the open ocean, the small heterotrophic flagellates (sHF) are the chief microbial predators and recyclers of essential nutrients to phototrophic microbes. Even with intensive molecular phylogenetic studies of the sHF, the origins of their feeding success remain obscure because of limited understanding of their morphological adaptations to feeding. Here, we examined the sHF morphologies in the largest, most oligotrophic South Pacific and Atlantic (sub)tropical gyres and adjacent mesotrophic waters. On four research cruises, the sHF cells were flow cytometrically sorted from bacterioplankton and phytoplankton for electron microscopy. The sorted sHF comprised chiefly heterokont (HK) biflagellates and unikont choanoflagellates numerically at around 10-to-1 ratio. Of the four differentiated morphological types of HK omnipresent in the open ocean, the short-tinsel heterokont (stHK), whose tinsel flagellum is too short to propagate a complete wave, is predominant and a likely candidate to be the most abundant predator on Earth. Modeling shows that the described stHK propulsion is effective in feeding on bacterioplankton cells at low concentrations; however, owing to general prey scarcity in the oligotrophic ocean, selective feeding is unsustainable and omnivory is equally obligatory for the seven examined sHF types irrespective of their mode of propulsion.

Changes in planktonic microbial components in interaction with juvenile oysters during a mortality episode in the Thau lagoon (France)

Oysters modify the planktonic microbial community structure by their filtration and NH4 excretion activities. While many studies have been conducted on this subject with adult oysters, none had been carried out in situ with juveniles. Pacific oyster juveniles (Magallana gigas, previously Crassostrea gigas) died massively all over the world since 2008 in relation with OsHV-1 infection. During mortality episodes, sick and dead oysters are not separated from healthy live ones, and left to decay in the surrounding environment, with unknown consequences for the nutrient cycle and planktonic microbial components (PMC). The present study aimed to elucidate for the first time the interactions between oyster juveniles and PMC during a mortality episode. Innovative 425-L pelagic chambers were deployed weekly in situ around oyster lanterns along a stocking-density gradient in the Thau Mediterranean lagoon (France) before, during and after an oyster mortality episode, from April to May 2015. This study reveals (i) significant changes of planktonic microbial community structure during mortality episodes, with a proliferation of picoplankton (<3 μm) and ciliates (Balanion sp., Uronema sp.) within 2 weeks when mortality rates and numbers of moribund juvenile oysters were highest. These changes were probably induced by oyster tissue leaching, decomposition and mineralization, which probably began during the moribund period, as suggested by an increase of PO4 concentration and N:P ratio decrease, (ii) oyster juveniles mainly retained 3–20 μm plankton. In contrast to adults, picophytoplankton and small heterotrophic flagellates (<3 μm) were significantly depleted in the presence of oyster juveniles. Depletion of picoplankton occurred only at the starting of the mortality episode and during the moribund phase. (iii) Oyster juvenile filtration and mortality shifted the planktonic microbial structure toward a heterotrophic microbial system, where ciliates and heterotrophic flagellates acted as a trophic link between picoplankton and oysters. The next stage of our investigation is to examine the effect of a mortality episode on pathogen fluxes in the water column, exploring their relationships with planktonic components and dead oyster flesh.

Combined morphological and phylogenomic re-examination of malawimonads, a critical taxon for inferring the evolutionary history of eukaryotes.

Modern syntheses of eukaryote diversity assign almost all taxa to one of three groups: Amorphea, Diaphoretickes and Excavata (comprising Discoba and Metamonada). The most glaring exception is Malawimonadidae, a group of small heterotrophic flagellates that resemble Excavata by morphology, but branch with Amorphea in most phylogenomic analyses. However, just one malawimonad, Malawimonas jakobiformis, has been studied with both morphological and molecular-phylogenetic approaches, raising the spectre of interpretation errors and phylogenetic artefacts from low taxon sampling. We report a morphological and phylogenomic study of a new deep-branching malawimonad, Gefionella okellyi n. gen. n. sp. Electron microscopy revealed all canonical features of ‘typical excavates’, including flagellar vanes (as an opposed pair, unlike M. jakobiformis but like many metamonads) and a composite fibre. Initial phylogenomic analyses grouped malawimonads with the Amorphea-related orphan lineage Collodictyon, separate from a Metamonada+Discoba clade. However, support for this topology weakened when more sophisticated evolutionary models were used, and/or fast-evolving sites and long-branching taxa (FS/LB) were excluded. Analyses of ‘–FS/LB’ datasets instead suggested a relationship between malawimonads and metamonads. The ‘malawimonad+metamonad signal’ in morphological and molecular data argues against a strict Metamonada+Discoba clade (i.e. the predominant concept of Excavata). A Metamonad+Discoba clade should therefore not be assumed when inferring deep-level evolutionary history in eukaryotes.

Phytoplankton growth and microzooplankton grazing in the Homa Lagoon (İzmir Bay, Turkey)

Phytoplankton growth and microzooplankton grazing were investigated at one station in the Homa Lagoon from February to January in 2006-2007. Our results showed significant seasonal variations in phytoplankton dynamics. Microzooplankton was mainly composed of dinoflagellates and tintinnid ciliates and nauplii. Microzooplankton grazing increased with increasing of temperature. Grazing rate was maximum levels in spring and summer. Microphytoplankton, which dominated the total algal biomass and production, were characterized by the proliferation of several chain-forming diatoms. Small heterotrophic flagellates and aloricate ciliates were the main controllers of phytoplankton. Phytoplankton represented a significant for micrograzers, which grazing represented 20–120% of diatom and cynabacteria algal production during 2006-2007. Microzooplankton has, however, a relatively high impact on microphytoplankton, as > 45% of microalgal production was consumed throughout the year. These results suggest that the low grazing was one of the factors contributing to the development of the spring bloom. All of seasonal bases, the phytoplankton production were grazed by microzooplankton in summer, autumn and winter– spring seasons have been changing between 20%-120%. The seasonal variation in the microzooplankton grazing pressure seems to result from the dominant size class of the phytoplankton community of this lagoon diatom and cynabacteria.

A story of famine in the pelagic realm: temporal and spatial patterns of food limitation in rotifers from an oligotrophic tropical lake

Food limitation is one of the most important extrinsic factors influencing population dynamics in aquatic ecosystems, but the spatial patterns of resource limitation and their influence on zooplankton species have not been investigated. We tested the hypothesis that the occurrence and intensity of food limitation may vary both spatially and temporally, and that the irregular population dynamics of the rotifers Brachionus group plicatilis and Hexarthra jenkinae in a saline, warm-monomictic, oligotrophic lake (Alchichica, Central Mexico) can be attributed to resource limitation. We estimated plankton abundance and composition and performed 19 laboratory experiments to assess the intensity of food limitation (Dr) at three selected depths in the water column. Most of the phytoplankton biomass consisted of algae inedible to rotifers, and good-quality food ( phytoplankton � 20 mm þ small heterotrophic flagellates) was frequently below the threshold food concentration for Brachionus species. The vertical distribution and abundance of rotifers were correlated with the availability of food resources. Food limitation was observed during mixing and late stratification; however, the intensity of limitation was rather similar throughout the water column owing to the overall scarcity of resources in the lake. Hexarthra seldom appeared. We conclude that the irregular pattern of rotifer abundance we observed is a consequence of the interplay between the harsh conditions of scarce food availability, resulting in intense resource competition and opportunities for rapid exploitation of opportunities when food outbursts occur.

Hidden diversity among aquatic heterotrophic flagellates: ecological potentials of zoosporic fungi

Since the emergence of the ‘microbial loop’ concept, heterotrophic flagellates have received particular attention as grazers in aquatic ecosystems. These microbes have historically been regarded incorrectly as a homogeneous group of bacterivorous protists in aquatic systems. More recently, environmental rDNA surveys of small heterotrophic flagellates in the pelagic zone of freshwater ecosystems have provided new insights. (i) The dominant phyla found by molecular studies differed significantly from those known from morphological studies with the light microscope, (ii) the retrieved phylotypes generally belong to well-established eukaryotic clades, but there is a very large diversity within these clades and (iii) a substantial part of the retrieved sequences cannot be assigned to bacterivorous but can be assigned instead to parasitic and saprophytic organisms, such as zoosporic true fungi (chytrids), fungus-like organisms (stramenopiles), or virulent alveolate parasites (Perkinsozoa and Amoebophrya sp.). All these microorganisms are able to produce small zoospores to assure dispersal in water during their life-cycles. Based on the existing literature on true fungi and fungus-like organisms, and on the more recently published eukaryotic rDNA environmental studies and morphological observations, we conclude that previously overlooked microbial diversity and related ecological potentials require intensive investigation (i) for an improved understanding of the roles of heterotrophic flagellates in pelagic ecosystems and (ii) to properly integrate the concept of ‘the microbial loop’ into modern pelagic microbial ecology.

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.

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 planktonic food web of the Bizerte lagoon (south-western Mediterranean) during summer: I. Spatial distribution under different anthropogenic pressures

The structure and the trophic interactions of the planktonic food web were investigated during summer 2004 in a coastal lagoon of south-western Mediterranean Sea. Biomasses of planktonic components as well as bacterial and phytoplankton production and grazing by microzooplankton were quantified at four stations (MA, MB, MJ and R) inside the lagoon. Station MA was impacted by urban discharge, station MB was influenced by industrial activity, station MJ was located in a shellfish farming sector, while station R represented the lagoon central area. Biomasses and production rates of bacteria (7–33 mg C m −3; 17.5–35 mg C m −3 d −1) and phytoplankton (80–299 mg C m −3; 34–210 mg C m −3 d −1) showed high values at station MJ, where substantial concentrations of nutrients (NO 3 − and Si(OH) 4) were found. Microphytoplankton, which dominated the total algal biomass and production (>82%), were characterized by the proliferation of several chain-forming diatoms. Microzooplankton was mainly composed of dinoflagellates ( Torodinium, Protoperidinium and Dinophysis) and aloricate ( Lohmaniellea and Strombidium) and tintinnid ( Tintinnopsis, Tintinnus, Favella and Eutintinnus) ciliates. Higher biomass of these protozoa (359 mg C m −3) was observed at station MB, where large tintinnids were encountered. Mesozooplankton mainly represented by Calanoida ( Acartia, Temora, Calanus, Eucalanus, Paracalanus and Centropages) and Cyclopoida ( Oithona) copepods, exhibited higher and lower biomasses at stations MA/MJ and MB, respectively. Bacterivory represented only 35% of bacterial production at stations MB and R, but higher fractions (65–70%) were observed at stations MA and MJ. Small heterotrophic flagellates and aloricate ciliates seemed to be the main controllers of bacteria. Pico- and nanophytoplankton represented a significant alternative carbon pool for micrograzers, which grazing represented 67–90% of pico- and nano-algal production in all stations. Microzooplankton has, however, a relatively low impact on microphytoplankton, as ≤45% of microalgal production was consumed in all stations. This implies that an important fraction of diatom production would be channelled by herbivorous meso-grazers to higher consumers at stations MA and MJ where copepods were numerous. Most of the microalgal production would, however, sink particularly at station MB where copepods were scare. These different trophic interactions suggest different food web structures between stations. A multivorous food web seemed to prevail in stations MJ and MA, whereas microbial web was dominant in the other stations.

Predation limitation in the pelagic microbial food web: an in situ study in an oligotrophic aquatic system

Temporal and spatial variations of pelagic microorganisms in the northern Baltic Sea were studied, as well as factors influencing their abundance and growth rates. Three main questions were asked 1) How does increased productivity influence the structure of the microbial food web? 2) Does predation limitation vary between trophic levels? 3) What is the relative importance of resource and predation limitation at different trophic levels?A field study in the northern Baltic Sea showed that dominating protozoa, flagellates and ciliates, increased with increasing primary productivity from north to south. Furthermore, relatively small protozoan cells dominated in the low productive north, while larger cells became more dominant in the south. The relationship between plankton size structure and productivity was further studied in an experimental system. In agreement with present theories regarding nutrient status of pelagic food webs, increased productivity caused a lengthening of the food chain as well as a change in plankton size structure. While microplankton dominated in nutrient rich treatments pico- and nanoplankton dominated during nutrient poor treament. The flagellate community was dominated by a potentially mixotroph, Chrysochromulina sp., at low nutrient concentrations. To our knowledge this is the first experimental study showing that Chrysochromulina sp. in resemblance with other mixotrophs is favoured by nutrient poor conditions compared to strict autotrophs and heterotrophs.During a stratified summer period autotrophic microorganisms in the northern Baltic Sea did not respond to removal of potential predators, indicating that they were primarily limited by inorganic nutrients. An exception was small eucaryotic picoplankton that showed a large response to predator removal. Among the heterotrophic microorganisms direct effect of predation seemed to increase from ciliates, heterotrophic bacteria, small heterotrophic flagellates, medium flagellates to large flagellates. No quick indirect effect was observed, but after four days trophic cascades were detected.The relative importance of resource and predation limitation was studied among heterotrophic bacteria, flagellates and ciliates in the northern Baltic Sea. For all these groups, resource limitation seemed to prevail during the summer period. The results also indicated that the relative importance of predation increased with the productivity of the system. To our knowledge there are no earlier measurements on the relative importance of resource and predation limitation for micoorganisms in the pelagic environment.

Structural changes in an aquatic microbial food web caused by inorganic nutrient addition

Temporal and spatial variations of pelagic microorganisms in the northern Baltic Sea were studied, as well as factors influencing their abundance and growth rates. Three main questions were asked 1) How does increased productivity influence the structure of the microbial food web? 2) Does predation limitation vary between trophic levels? 3) What is the relative importance of resource and predation limitation at different trophic levels?A field study in the northern Baltic Sea showed that dominating protozoa, flagellates and ciliates, increased with increasing primary productivity from north to south. Furthermore, relatively small protozoan cells dominated in the low productive north, while larger cells became more dominant in the south. The relationship between plankton size structure and productivity was further studied in an experimental system. In agreement with present theories regarding nutrient status of pelagic food webs, increased productivity caused a lengthening of the food chain as well as a change in plankton size structure. While microplankton dominated in nutrient rich treatments pico- and nanoplankton dominated during nutrient poor treament. The flagellate community was dominated by a potentially mixotroph, Chrysochromulina sp., at low nutrient concentrations. To our knowledge this is the first experimental study showing that Chrysochromulina sp. in resemblance with other mixotrophs is favoured by nutrient poor conditions compared to strict autotrophs and heterotrophs.During a stratified summer period autotrophic microorganisms in the northern Baltic Sea did not respond to removal of potential predators, indicating that they were primarily limited by inorganic nutrients. An exception was small eucaryotic picoplankton that showed a large response to predator removal. Among the heterotrophic microorganisms direct effect of predation seemed to increase from ciliates, heterotrophic bacteria, small heterotrophic flagellates, medium flagellates to large flagellates. No quick indirect effect was observed, but after four days trophic cascades were detected.The relative importance of resource and predation limitation was studied among heterotrophic bacteria, flagellates and ciliates in the northern Baltic Sea. For all these groups, resource limitation seemed to prevail during the summer period. The results also indicated that the relative importance of predation increased with the productivity of the system. To our knowledge there are no earlier measurements on the relative importance of resource and predation limitation for micoorganisms in the pelagic environment.

Succession of Protists on Estuarine Aggregates

Colonization by and succession of bacteria and bacterivorous protists on laboratory-made aggregates were determined over a period of 14 days during winter and spring in 1997. Aggregates were generated from natural water from the limnetic zone of the Elbe Estuary using a tilting tube roller system. Within 1 h after the beginning of the experiments, macroaggregates started to form. Aggregates reached a maximum size of 1 mm with a tendency toward large sizes at the end of the experiment after the 10th day. On the first day, high bacterial densities of more than 10(9) cells ml(-1) were detected within the aggregates. The abundances of flagellates and ciliates within aggregates were also two or three orders of magnitude higher than in the surrounding water. Densities of aggregate associated organisms are comparable to those occuring in sediments. The first protistan colonizers on the aggregates were small heterotrophic flagellates, such as choanoflagellates and small euglenids. Later, beginning on the 4th day, small sarcodines and ciliates became abundant. The most abundant ciliates associated with aggregates were small species of the Hypotrichia, Cyrtophorida, and Hymenostomata. After 9 days, large omnivorous and carnivorous ciliates, such as large members of the Hypotrichia and the Pleurostomatida, occurred. In spring, large heterotrophic flagellates and amebae also appeared at this time. These findings indicated the existence of a succession of protists on newly formed aggregates and a microbial food net within the aggregates based on bacterial production. Additionally, most of the species observed during this study were adapted for living on surfaces. In natural environments they are more common in benthic than in pelagic environments. For them, aggregates are havens in the water column comparable to sediment communities.

Periphyton and sphagnicolous protists of dystrophic bog lakes (Brandenburg, Germany): I. Annual cycles, distribution and comparison to other lakes

The periphyton of acid, dystrophic bogs was investigated. Glass-slides were exposed in four different bog lakes about 100 km north of Berlin, Germany, in order to examine the species composition and the annual cycle of the protist community. The annual cycles of the periphyton in bog lakes were comparable to those in other mesotrophic lakes: During the winter months small heterotrophic flagellates (HF) dominated the periphyton community. Besides that bacterivorous ciliates were present as typical pioneer species. At the end of the ice covering, almost exclusively unicellular green algae colonized the slides. In May high abundances of ciliates and choanoflagellates were observed. At the same time, a maximum of species was reached. From August to October peritrich ciliates significantly contributed to the periphyton. Their number decreased when early frosts occurred. The four investigated bog lakes exhibit very similar species compositions, almost all of the species were present in each of the four lakes. Differences were only distinct concerning the dominance of some protistan groups. The largest deflections, however, occurred not between two lakes, but between two habitats within one lake: The upper 30 cm of the Sphagnum mat comprise two very different protistan communities, determined by a strong vertical zoning. Due to the different light and nutrient conditions, the slides of the upper region exhibited a denser colonization, mainly consisting of autotrophic cryptomonads and vagile ciliates. In contrast to this, the slides of the deeper region were mainly colonized by heterotrophic flagellates and sessile peritrich ciliates.

Primary and bacterial production in sea ice in the northern Baltic Sea

The temporal variation of ice primary and bacterial production along with ice algal, bacterial and heterotrophic flagellate biomass were studied at a coastal station in the northern Baltic Sea throughout the ice-covered period of 1996 (January to April). Ice core samples were taken every week and analyzed for abundance and production of different microorganisms. In addition, physical and chemical parameters were measured. The ice algae were Limited by light during the first 3 mo of the study. The algal production showed a peak in the middle of April, which coincided with a marked increase in light availability. Shortly after that, the system became phosphorus depleted and primary production decreased rapidly. Bacterial biomass and production rates were relatively low and stable before the ice algal bloom. After the ice algal bloom, bacterial production increased rapidly, while the biomass remained low. The growth rate of small heterotrophic flagellates (<l0 pm), calculated from increase in biomass, was more than 1 order of magnitude higher than the bacterial production rate following the ice algal bloom. Thus, small heterotrophic flagellates were using food sources other than bacteria for growth after the ice algal bloom. On an annual basis, the ice algal and bacterial production accounted for < l % and <0.1% respectively of the total production (ice + pelagic) due to a short ice-covered season. During the ice-covered season, however, the ice algae accounted tor 10% of the total algal production, whlle ice bacterial production was 0.2 % of the total bacterial produckon.

Notes on protozoa in agricultural soil with emphasis on heterotrophic flagellates and naked amoebae and their ecology.

Heterotrophic flagellates and naked amoebae are usually very numerous in agricultural soils; with numbers in the magnitude of 10,000 to 100,000 (active+encysted) cells per gram of soil. In 'hotspots' influenced by living roots or by dead organic material, the number may occasionally be as high as several millions per gram of soil. An exact enumeration of these organisms is virtually impossible. As they most often adhere closely to the soil particles, direct counting will underestimate numbers since the organisms will be masked. The method usually applied for enumeration of these organisms, the 'most probable number (MPN) method', is based on the ability of the organisms to grow on particular culture media. This method will in many cases underestimate the total protozoan number (active+encysted). It is uncertain how many of the heterotrophic flagellates and naked amoebae are actively moving and how many are encysted at a particular time; the 'HCl-method' which has usually been used to discriminate between active and encysted has proven to be highly unreliable. Despite the methodological difficulties many investigations of these organisms indicate that they play an important role in agricultural soils as bacterial consumers, and to a minor extent as consumers of fungi. Because of their small size and their flexible body they are able to graze bacteria in small pores in the soil in which larger organisms are precluded from coming. Key factors restricting the number and activity of heterotrophic flagellates and naked amoebae in soils seem to be water potential and soil structure and texture. In micro-cosm experiments, small heterotrophic flagellates and naked amoebae regulate the size and composition of the bacterial community. Bacterial activity seems to be stimulated by these organisms in most cases as well as the mineralization of carbon and nitrogen and possibly other mineral nutrients. In the rhizosphere of living plants the activity of protozoa has proven to stimulate uptake of nitrogen in pot experiments, and it has been hypothesized that organic matter liberated by plants in the root zone will stimulate bacterial and protozoan activity, leading to mineralization of organic soil nitrogen which is subsequently taken up by the plants.

Notes on protozoa in agricultural soil with emphasis on heterotrophic flagellates and naked amoebae and their ecology

Heterotrophic flagellates and naked amoebae are usually very numerous in agricultural soils; with numbers in the magnitude of 10000 to 100000 (active+ encysted) cells per gram of soil. In ‘hotspots’ influenced by living roots or by dead organic material, the number may occasionally be as high as several millions per gram of soil. An exact enumeration of these organisms is virtually impossible. As they most often adhere closely to the soil particles, direct counting will underestimate numbers since the organisms will be masked. The method usually applied for enumeration of these organisms, the ‘most probable umber (MPN) method’, is based on the ability of the organisms to grow on particular culture media. This method will in many cases underestimate the total protozoan number (active+ encysted). It is uncertain how many of the heterotrophic flagellates and naked amoebae are actively moving and how many are encysted at a particular time; the ‘HCI-method’ which has usually been used to discriminate between active and encysted has proven to be highly unreliable. Despite the methodological difficulties many investigations of these organisms indicate that they play an important role in agricultural soils as bacterial consumers, and to a minor extent as consumers of fungi. Because of their small size and their flexible body they are able to graze bacteria in small pores in the soil in which larger organisms are precluded from coming. Key factors restricting the number and activity of heterotrophic flagellates and naked amoebae in soils seem to be water potential and soil structure and texture. In micro-corm experiments, small heterotrophic flagellates and naked amoebae regulate the size and composition of the bacterial community. Bacterial activity seems to be stimulated by these organisms in most cases as well as the mineralization of carbon and nitrogen and possibly other mineral nutrients. In the rhizosphere of living plants the activity of protozoa has proven to stimulate uptake of nitrogen inpot experiments, and it has been hypothesized that organic matter liberated by plants in the root zone will stimulate bacterial and protozoan activity, leading to mineralization of organic soil nitrogen which is subsequently taken up by the plants.

Flow cytometry: a new method for characterization of differential ingestion, digestion and egestion by suspension feeders

Flow cytometry: a new method for characterization of differential ingestion, digestion and egestion by suspension feeders