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PubMed2005

Assimilation efficiency of Vibrio bacterial protein biomass by the flagellate Pteridomonas : assessment using flow cytometric sorting.

A flow cytometric sorting technique for direct determination of bacterial biomass assimilation by phagotrophic flagellates was developed and tested in laboratory culture experiments. Living Vibrio bacteria were quantitatively pulse-chase labelled with [(35)S]methionine tracer and fed to Pteridomonas flagellates. Flow sorting revealed that the isotopically labelled material is in either bacterial prey or flagellate predators and the egested bacterial debris contained negligible amounts of tracer. These experimental results confirm an earlier hypothesis that flagellates release metabolised bacterial proteins primarily in a dissolved form. The assimilation efficiency of the Vibrio protein biomass by Pteridomonas was low, only about 20%, independently of the amount of consumed bacterial biomass, confirming our earlier indirect estimates. Additionally, against expectations that cells decrease their metabolic activity whilst preparing for and engaged in division, we found that the precursor uptake rates by flow sorted bacterial cells at the S+G(2) cell cycle stages were constantly 1.5 times higher than those of cells at the G(1) stage.

Animals↗
PubMed2002

Novel marine flagellate fatty acid: structural elucidation by GC-MS analysis of DMOX derivatives and DMDS adducts.

In situ biodegradation experiments of marine particles were performed in deep Atlantic waters. Lipid changes were associated with the colonization of the decaying detritus by marine flagellates smaller than 10 microm in size. Fatty acid methyl esters (FAMEs) of these flagellates showed high proportion of a FAME with a molecular weight (MW) of 320. Its structure could not be unambiguously resolved by retention times on gas chromatography runs using polar and nonpolar columns, nor by routine gas chromatography coupled to mass spectrometry (GC-MS). Complementary GC-MS analysis of two types of derivatives was performed to fully elucidate the structure of this novel acid. GC-MS analysis of 4,4-dimethyloxazoline (DMOX) derivative of the compound enabled localization of a double bond in position Delta17, whereas other double bond locations could not be unambiguously located by spectrum interpretation. DMDS addition on the flagellate biomarker produced monocyclic triadducts. Fragment suites corresponding to gradual losses of thiomethyl substituents indicated the presence of a five-membered thioether cycle, located on the methyl side of the derivative. Fragment suites produced by cleavage of C linked to sulfured substituents revealed various possible structures. However, interpretation of the spectra in relation with the fragmentation of the DMOX derivative yielded a convergent identification of the flagellate biomarker, as a non-methylene-interrupted C20:3Delta7,13,17 FAME.

Animals↗
PubMed1982

Interactions between the intestinal flagellates Giardia muris and Spironucleus muris and the blood parasites Babesia microti, Plasmodium yoelii and Plasmodium berghei in mice.

In mice infected with the intestinal flagellates Giardia muris or Spironucleus muris, together with the blood parasites Babesia microti or Plasmodium yoelii, there is a temporary decrease of flagellate cyst output coincident with the peak of the blood parasite infections, followed by a rapid return to normal levels. This decrease in cyst output is correlated with decreased numbers of trophozoites in the small intestine. The effect on S. muris is more marked than that on G. muris. Neither blood parasites has any effect on the total duration of the flagellate infection and the flagellates do not affect the blood parasites. In mice infected with G. muris or S. muris and P. berghei there is also a decrease in cyst output but this is less apparent than in infections with B. microti or P. yoelii because of the fatal nature of the P. berghei infection. It is suggested that the decrease in cyst output is probably due to changes in the contents of the small intestine or to non-specific immunological factors rather than to specific immunological changes.

Animals↗
PubMed2005

Amperometric biosensing systems based on motility and gravitaxis of flagellate algae for aquatic risk assessment.

Electrochemical biosensing systems for toxic substances were developed on the basis of motility and negative gravitaxis of the unicellular flagellate Chlamydomonas reinhardtii. Changes in the flagellar movement of the flagellates in response to three toxic chemicals, toluene, copper(II) sulfate, and nickel(II) chloride, were monitored as changes in the redox currents for a coexisiting redox marker. The gravitaxis-based flagellate biosensing system was more sensitive to toluene than the motility-based system. A thin-layer flagellate biosensor was also developed. In comparison with the conventional algal biosensors monitoring the photosynthetic activity, the gravitaxis-based thin-layer sensor was more sensitive by more than 1 order of magnitude.

Animals↗
PubMed2001

Rhodopsin receptors of phototaxis in green flagellate algae.

Green flagellate algae are capable of the active adjustment of their swimming path according to the light direction (phototaxis). This direction is detected by a special photoreceptor apparatus consisting of the photoreceptor membrane and eyespot. Receptor photoexcitation in green flagellates triggers a cascade of rapid electrical events in the cell membrane which plays a crucial role in the signal transduction chain of phototaxis and the photophobic response. The photoreceptor current is the earliest so far detectable process in this cascade. Measurement of the photoreceptor current is at present the most suitable approach to investigation of the photoreceptor pigment in green flagellate algae, since a low receptor concentration in the cell makes application of optical and biochemical methods so far impossible. A set of physiological evidences shows that the phototaxis receptor in green flagellate algae is a unique rhodopsin-type protein. It shares common chromophore properties with retinal proteins from archaea. However, the involvement of photoelectric processes in the signal transduction chain relates it to animal visual rhodopsins. The presence of some enzymatic components of the animal visual cascade in isolated eyespot preparations might also point to this relation. A retinal-binding protein has been identified in such preparations, the amino acid sequence of which shows a certain homology to sequences of animal visual rhodopsins. However, potential function of this protein as the phototaxis receptor has been questioned in recent time.

Chlorophyta↗
PubMed2002

Bacterivory by heterotrophic flagellates: community structure and feeding strategies.

Heterotrophic flagellates (HF) are known as most important grazers of bacteria in many aquatic ecosystem. HF cannot be treated as a black box since HF generally contain a diverse community of species significantly differing in their feeding behaviour and other ecological properties. Today it seems that the dominant taxonomic groups among heterotrophic nano- and microflagellate communities within different marine, brackish and limnetic pelagic communities (heterokont taxa, dinoflagellates, choanoflagellates, kathablepharids) and benthic communities (euglenids, bodonids, thaumatomonads, apusomonads, cercomonads) are relatively similar. HF among protista incertae sedis, often neglected in ecological studies, are abundant bacterivores in all investigated habitats. Recent studies of flagellate feeding processes indicated that there are significant species-specific differences and individual variability regarding the food uptake and food selection of bacterivorous flagellates: Variability of bacterivory is discussed regarding the prevailing feeding modes, the energy budgets, the considerable importance of slight deviations in the time budgets of feeding phases, the ingestion rates and the feeding microhabitat, respectively. The significant flexibility of the grazing impact of bacterivorous flagellate communities creates a complex top-down pressure on bacteria which should have lead to the evolution of efficient predator avoidance mechanisms in bacteria and should be at least partly responsible for the diversity of present bacteria.

Animals↗
PubMed2001

The amoeba-to-flagellate transformation test is not reliable for the diagnosis of the genus Naegleria. Description of three new Naegleria spp.

Trophozoites of several isolates from one location in Australia have failed consistently to transform into flagellates, although they display all other characteristics of the genus Naegleria. When changing the standard transformation test, flagellates were produced. In phylogenetic trees derived from partial small subunit ribosomal DNA (SSUrDNA) sequences, one of these strains branches close to a cluster comprising N. clarki, N. australiensis, N. italica and N. jadini. It is proposed that these Australian isolates represent a new species, named N. fultoni (strain NG885). Failing to form flagellates since their isolation, even when different transformation procedures are used, are two Naegleria strains from Chile and Indonesia. In SSUrDNA-based phylogenetic trees the Chilean strain clusters with N. pussardi and the Indonesian strain clusters with N. galeacystis, but the degree of sequence difference from these described species (3.5% and 2.2%, respectively) is sufficient to propose that both of the strains represent new species, named N. chilensis (strain NG946) and N. indonesiensis (strain NG945), respectively. The close relationships between each of the new species and the Naegleria species with which they cluster in SSUrDNA-based trees were confirmed by ribosomal internal transcribed spacer region (ITS) sequence comparisons. In France, several non-flagellating N. fowleri strains were isolated from one location. ITS rDNA sequence comparisons indicated that they correspond to a 'type' of N. fowleri found in both Europe and the USA. A redefinition of the genus Naegleria is proposed as a consequence of these and previous findings.

Amebiasis↗
PubMed2002

The soil flagellate Proleptomonas faecicola: cell organisation and phylogeny suggest that the only described free-living trypanosomatid is not a kinetoplastid but has cercomonad affinities.

The only putative free-living trypanosomatid is Proleptomonas faecicola described first by Woodcock in 1916 as a coprophilic flagellate with striking Leptomonas-like flagellar movement but lacking a kinetoplast. P faecicola was later identified by Sandon in 1927 as a widespread non-phagotrophic inhabitant of soils. No division stages were seen by either observer. An organism conforming to Woodcock's light microscope description has been isolated from tapwater and cultivated axenically in various serum-containing media. Division has been shown to occur in an aflagellate stage enclosed in a thin cyst wall. Electron microscopy of the flagellate stage reveals that, in addition to the long locomotory flagellum, a second non-motile flagellum is present attached to the body along its entire length. The flagellate's ultrastructure lacks all the major features of the Trypanosomatidae. The several mitochondria of Proleptomonas have tubular cristae and lie between intracytoplasmic microtubules originating as a loose cone associated with the flagellar basal bodies. This cytoskeleton is much reduced in the division cyst. A comparable Proleptomonas-like flagellate with similar division cysts has been observed in soil samples from farmland. Phylogenetic analysis based on SSU rRNA gene sequences suggests that the cultured organism identified here as Proleptomonas is unrelated to the Kinetoplastida and has affinities with the Phylum Cercozoa Cavalier-Smith, even though in morphology, life cycle and mode of feeding it bears little resemblance to any member of that diverse grouping.

Animals↗
PubMed1975

Isolation of intracellular symbiotes by immune lysis of flagellate protozoa and characterization of their DNA.

A new method dependent on immune lysis is described for the isolation of intracellular symbiotes from two species of flagellate protozoa Blastocrithidia culicis and Crithidia oncopelti. The symbiote-containing flagellates are exposed to complement and antisera prepared in rabbits against symbiote-free organisms. The immune lysis seems to weaken the plasma membranes of the flagellates so that subsequent application of gentle shearing force liberates the intracellular entities in an undamaged condition. The symbiotes are then separated from other cellular components by DNAse digestion and differential centrifugation. The average recovery of symbiotes isolated by this method is 20%. Light and electron microscopy establishes the structural integrity and numerical abundance of isolated symbiotes in the final fractions. Integrity of symbiotes is further indicated by the high activity of a marker enzyme, uroporphyrinogen I synthetase. The DNA's of symbiote-containing and symbiote-free flagellates, and of isolated symbiotes were purified and compared after isopycnic centrifugation. The comparison establishes the presence of DNA's in symbiotes of both species. The guanine-cytosine (G-C) content of symbiote DNA differs from that of host DNA's in C. oncopelti, but resembles that of kinetoplast DNA in B. culicis. The latter observation was further shown by heat denaturation study. Renaturation kinetics indicate that the genome complexity of symbiote DNA in B. culicis is similar to that of bacteria.

Animals↗
PubMedDate not supplied

Reclinomonas americana N. G., N. Sp., a new freshwater heterotrophic flagellate.

A new heterotrophic flagellate has been discovered from sites in Maryland, Michigan and Wyoming. The flagellate resides within a lorica constructed of a meshwork of intertwined fibrils with the outer surface invested with nail-shaped spines. The organism "reclines" within the lorica with its ventral aspect directed upward, and has two heterodynamic flagella, neither of which bears mastigonemes. One flagellum is directed upward and the other is arched over the ventral aspect of the body. Ingestion of bacteria takes place at the left posterior half of the cell. The organism is anchored to the lorica on the right posterior side by a series of regularly spaced cytoplasmic bridges and at the left anterior of the cell by a cytoplasmic appendage similar to the "languette cytoplasmique" found in some bicosoecids. The right side of the cell is raised into a flattened lip with the outer margin reinforced by a ribbon of microtubules. The new flagellate has mitochondria with tubular cristae and lacks a Golgi. A new genus is created to accommodate both the new flagellate described herein and Histiona campanula Penard. A new family is proposed to include the new genus and Histiona.

Animals↗
PubMedDate not supplied

A survey of flagellate diversity at four deep-sea hydrothermal vents in the Eastern Pacific Ocean using structural and molecular approaches.

Eighteen strains of flagellated protists representing nine species were isolated and cultured from four deep-sea hydrothermal vents: Juan de Fuca Ridge (2,200 m), Guaymas Basin (2,000 m), 21 degrees N (2,550 m) and 9 degrees N (2,000 m). Light and electron microscopy were used to identify flagellates to genus and, when possible, species. The small subunit ribosomal RNA genes of each vent species and related strains from shallow-waters and the American Type Culture Collection were sequenced then used for comparative analysis with database sequences to place taxa in an rDNA tree. The hydrothermal vent flagellates belonged to six different taxonomic orders: the Ancyromonadida, Bicosoecida, Cercomonadida, Choanoflagellida, Chrysomonadida, and Kinetoplastida. Comparative analysis of vent isolate and database sequences resolved systematic placement of some well-known species with previously uncertain taxonomic affinities, such as Ancyromonas sigmoides, Caecitellus parvulus, and Massisteria marina. Many of these vent isolates are ubiquitous members of marine, freshwater, and terrestrial ecosystems worldwide, suggesting a global distribution of these flagellate species.

Animals↗
PubMed1994

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.

Animals↗
PubMed1997

Effect of flagellates on free-living bacterial abundance in an organically contaminated aquifer.

Little is known about the role of protists in the saturated subsurface. Porous media microcosms, containing bacteria and protists, were used to determine whether flagellates from an organically contaminated aquifer could substantively affect the number of free-living bacteria (FLB). When flagellates were present, the 3-40% maximum breakthrough of fluorescently labelled FLB injected into the microcosms was much lower than the 60-130% observed for killed controls. Grazing and clearance rates (3-27 FLB flag-1 h-1 and 12-23 nl flag-1 h-1, respectively) calculated from the data were in the range reported for flagellates in other aqueous environments. The data provide evidence that flagellate bacterivory is an important control on groundwater FLB populations.

Animals↗
PubMed1990

Size-selective grazing on bacteria by natural assemblages of estuarine flagellates and ciliates.

The small average cell size of in situ bacterioplankton, relative to cultured cells, has been suggested to be at least partly a result of selection of larger-sized cells by bacterivorous protozoa. In this study, we determined the relative rates of uptake of fluorescence-labeled bacteria (FLB), of various cell sizes and cell types, by natural assemblages of flagellates and ciliates in estuarine water. Calculated clearance rates of bacterivorous flagellates had a highly significant, positive relationship with size of FLB, over a range of average biovolume of FLB of 0.03 to 0.08 microns3. Bacterial cell type or cell shape per se did not appear to affect flagellate clearance rates. The dominant size classes of flagellates which ingested all types of FLB were 3- to 4-microns cells. Ciliates also showed a general preference for larger-sized bacteria. However, ciliates ingested a gram-positive enteric bacterium and a marine bacterial isolate at higher rates than they did a similarly sized, gram-negative enteric bacterium or natural bacterioplankton, respectively. From the results of an experiment designed to test whether the addition of a preferentially grazed bacterial strain stimulated clearance rates of natural bacterioplankton FLB by the ciliates, we hypothesized that measured differences in rates of FLB uptake were due instead to differences in effective retention of bacteria by the ciliates. In general, clearance rates for different FLB varied by a factor of 2 to 4. Selective grazing by protozoa of larger bacterioplankton cells, which are generally the cells actively growing or dividing, may in part explain the small average cell size, low frequency of dividing cells, and low growth rates generally observed for assemblages of suspended bacteria.

Animals↗
PubMed2002

The bacterivorous soil flagellate Heteromita globosa reduces bacterial clogging under denitrifying conditions in sand-filled aquifer columns.

An exopolymer (slime)-producing soil bacterium Pseudomonas sp. (strain PS+) rapidly clogged sand-filled columns supplied with air-saturated artificial groundwater containing glucose (500 mg liter(-1)) as a sole carbon source and nitrate (300 mg liter(-1)) as an alternative electron acceptor. After 80 days of operation under denitrifying conditions, the effective porosity and saturated hydraulic conductivity (permeability) of sand in these columns had fallen by 2.5- and 26-fold, respectively. Bacterial biofilms appeared to induce clogging by occluding pore spaces with secreted exopolymer, although there may also have been a contribution from biogas generated during denitrification. The bacterivorous soil flagellate Heteromita globosa minimized reductions in effective porosity (1.6-fold) and permeability (13-fold), presumably due to grazing control of biofilms. Grazing may have limited growth of bacterial biomass and hence the rate of exopolymer and biogas secretion into pore spaces. Evidence for reduction in biogas production is suggested by increased nitrite efflux from columns containing flagellates, without a concomitant increase in nitrate consumption. There was no evidence that flagellates could improve flow conditions if added once clogging had occurred (60 days). Presumably, bacterial biofilms and their secretions were well established at that time. Nevertheless, this study provides evidence that bacterivorous flagellates may play a positive role in maintaining permeability in aquifers undergoing remediation treatments.

Animals↗
PubMed2004

Successful predation of filamentous bacteria by a nanoflagellate challenges current models of flagellate bacterivory.

Current models suggest that (i) filamentous bacteria are protected against predation by nanoflagellates, (ii) prey size is positively correlated with prey-predator contact probability, and (iii) contact probability is mainly responsible for size-selective predation by interception-feeding flagellates. We used five strains of filamentous bacteria and one bacterivorous nanoflagellate, Ochromonas sp. strain DS, to test these assumptions. The five strains, including one spirochete and four Betaproteobacteria strains, were isolated by the filtration-acclimatization method. All five strains possess flexible cells, but they differ in average cell length, which ranged from 4.5 to 13.7 micro m. High-resolution video microscopy was used to measure contact, capture, and ingestion rates, as well as selectivity of the flagellate feeding. Growth and feeding experiments with satiating and nonsatiating food conditions, as well as experiments including alternative well-edible prey, were performed. In contrast to predictions by current models, the flagellate successfully consumed all the tested filamentous strains. The ingestion rate was negatively correlated with bacterial length. On the other hand, the lengths of the filamentous bacteria were not positively correlated to the contact rate and capture rate but were negatively correlated to ingestion efficiency. In experiments including alternative nonfilamentous prey, the flagellates showed negative selection for filamentous bacteria, which was independent of food concentration and is interpreted as a passive selection. Our observations indicate that (i) size alone is not sufficient to define a refuge for filamentous bacteria from nanoflagellate predation and (ii) for the investigated filamentous bacteria, prey-predator contact probability could be more influenced by factors other than the prey size.

Animals↗
PubMed1977

Motility of flagellated bacteria in viscous environments.

The lowest viscosity that immobilized flagellated bacteria such as Psedomonas aeruginosa, Spirillum serpens, and Escherichia coli was 60 centipoise (cp). Much higher viscosities (1,000 cp and higher) were required to immobilize two flagellated bacteria selectively isolated from nature by methods based on their ability to migrate through agar gels. The latter finding indicates that certain flagellated bacteria have the ability to swim through environments of relatively high viscosity. It is suggested that these flagellated bacteria possess a specialized type of motility apparatus suited to viscous conditions present in their habitats.

Agar↗
PubMed1980

Polyprenyl p-hydroxybenzoate carboxylase in flagellation of Salmonella typhimurium.

Flagellation of Salmonella typhimurium was found to require a functional pathway for ubiquinone biosynthesis as well as growth in the presence of appropriate carboxylic acids. Induction of flagellation by carboxylic acids was shown to induce incorporation of p-hydroxybenzoic acid into polyprenylphenol. Constitutive flagellation was found to correlate with constitutive incorporation of p-hydroxybenzoic acid into polyprenylphenol. A novel pathway for polyprenyl p-hydroxybenzoic acid decarboxylation to polyprenylphenol was implicated in flagellation of S. typhimurium.

Biological Transport↗