Posterior Flagellum Research Articles

Flagellar proteomic analysis of the brown alga Mutimo cylindricus revealed a novel calcium-binding protein abundantly localized in the anterior flagellum

Mutimo cylindricus gametes have two flagella with different structures (anterior and posterior flagella). Their flagellar waveforms are regulated by calcium ions through different mechanisms, but very little information is available on the factors involved in their regulation. To elucidate the molecular basis underlying the difference between the two flagella, we performed a flagellar proteomic analysis using male M. cylindricus gametes. We were able to detect 848 proteins that were shared with Ectocarpus siliculosus, including 28 calcium-binding proteins. Among these EF-hand proteins, a protein with a mass of 111 kDa showed predominant localization along the anterior flagellum. Immunogold localization suggested that this protein is associated with outer doublet microtubules. This is the first report to show heterogeneous localization of a calcium-binding protein between two flagellar axonemes and suggests that calcium-binding proteins are involved in the specific regulation of the anterior flagellum.

Characterization of Skoliomonas gen. nov., a haloalkaliphilic anaerobe related to barthelonids (Metamonada).

Metamonads are a large and exclusively anaerobic group of protists. Additionally, they are one of the three clades proposed to ancestrally possess an "excavate" cell morphology, with a conspicuous ventral groove accompanied by a posterior flagellum with a vane. Here, we cultivate and characterize four anaerobic bacterivorous flagellates from hypersaline and alkaline soda lake environments, which represent a novel clade. Small subunit ribosomal RNA (SSU rRNA) gene phylogenies support recent phylogenomic analyses in placing them as the sister of barthelonids, a group that is itself sister to or deeply branching within Fornicata (Metamonada). The new isolates have a distinctive morphology: the hunchbacked cell body is traversed by a narrow ventral groove ending in a large opening to a conspicuous recurrent cytopharynx. The right margin of the groove is defined by a thin "lip." The posterior flagellum bears a wide ventral-facing vane. The narrow ventral groove and elongate cytopharynx are shared with barthelonids. We describe one isolate as Skoliomonas litria, gen. et sp. nov. Further investigation of their mitochondrial-related organelles (MROs) and detailed ultrastructural studies would be important to understanding the adaptation to anaerobic conditions in Metamonads-especially fornicates-as well as the evolution of the "excavate" cell architecture.

Foraging mechanisms in excavate flagellates shed light on the functional ecology of early eukaryotes

The phagotrophic flagellates described as "typical excavates" have been hypothesized to be morphologically similar to the Last Eukaryotic Common Ancestor and understanding the functional ecology of excavates may therefore help shed light on the ecology of these early eukaryotes. Typical excavates are characterized by a posterior flagellum equipped with a vane that beats in a ventral groove. Here, we combined flow visualization and observations of prey capture in representatives of the three clades of excavates with computational fluid dynamic modeling, to understand the functional significance of this cell architecture. We record substantial differences amongst species in the orientation of the vane and the beat plane of the posterior flagellum. Clearance rate magnitudes estimated from flow visualization and modeling are both like that of other similarly sized flagellates. The interaction between a vaned flagellum beating in a confinement is modeled to produce a very efficient feeding current at low energy costs, irrespective of the beat plane and vane orientation and of all other morphological variations. Given this predicted uniformity of function, we suggest that the foraging systems of typical excavates studied here may be good proxies to understand those potentially used by our distant ancestors more than 1 billion years ago.

Distinct regulation of two flagella by calcium during chemotaxis of male gametes in the brown alga Mutimo cylindricus (Cutleriaceae, Tilopteridales).

Brown algal male gametes show chemotaxis to the sex pheromone that is released from female gametes. The chemotactic behavior of the male gametes is controlled by the changes in the beating of two flagella known as the anterior and posterior flagellum. Our previous study using Mutimo cylindricus showed that the sex pheromone induced an increment in both the deflection angle of the anterior flagellum and sustained unilateral bend of the posterior flagellum, but the mechanisms regulating these two flagellar waveforms were not fully revealed. In this study, we analyzed the changes in swimming path and flagellar waveforms with a high-speed recording system under different calcium conditions. The extracellular Ca2+ concentration at 10-3 M caused an increment in the deflection angle of the anterior flagellum only when ionomycin was absent. No sustained unilateral bend of the posterior flagellum was induced either in the absence or presence of ionomycin in extracellular Ca2+ concentrations below 10-2 M. Real-time Ca2+ imaging revealed that there is a spot near the basal part of anterior flagellum showing higher Ca2+ than in the other parts of the cell. The intensity of the spot slightly decreased when male gametes were treated with the sex pheromone. These results suggest that Ca2+-dependent changes in the anterior and posterior flagellum are regulated by distinct mechanisms and that the increase in the anterior flagellar deflection angle and sustained unilateral bend of the posterior flagellum may not be primarily induced by the Ca2+ concentration.

Fine structure and molecular characterization of two new parabasalid species that naturally colonize laboratory mice, Tritrichomonas musculus and Tritrichomonas casperi.

Tritrichomonas muris is a common flagellated protist isolated from the cecum of wild rodents. This commensal protist has been shown previously to alter immune phenotypes in laboratory mice. Other trichomonads, referred to as Tritrichomonas musculis and Tritrichomonas rainier, also naturally colonize laboratory mice and cause immune alterations. This report formally describes two new trichomonads, Tritrichomonas musculus n. sp., and Tritrichomonas casperi n. sp., at the ultrastructural and molecular level. These two protists were isolated from laboratory mice and were differentiated by their size and the structure of their undulating membrane and posterior flagellum. Analysis at the 18S rRNA and trans-ITS genetic loci supported their designation as distinct species, related to T. muris. To assess the true extent of parabasalid diversity infecting laboratory mice, 135 mice bred at the National Institutes of Health (NIH) were screened using pan-parabasalid primers that amplify the trans-ITS region. Forty-four percent of mice were positive for parabasalids, encompassing a total of eight distinct sequence types. Tritrichomonas casperi and Trichomitus-like protists were dominant. T. musculus and T. rainier were also detected, but T. muris was not. Our work establishes a previously underappreciated diversity of commensal trichomonad flagellates that naturally colonize the enteric cavity of laboratory mice.

The Unicellular, Parasitic Fungi, Sanchytriomycota, Possess a DNA Sequence Possibly Encoding a Long Tubulin Polymerization Promoting Protein (TPPP) but Not a Fungal-Type One.

The unicellular, parasitic fungi of the phylum Sanchytriomycota (sanchytrids) were discovered a few years ago. These unusual chytrid-like fungi parasitize algae. The zoospores of the species of the phylum contain an extremely long kinetosome composed of microtubular singlets or doublets and a non-motile pseudocilium (i.e., a reduced posterior flagellum). Fungi provide an ideal opportunity to test and confirm the correlation between the occurrence of flagellar proteins (the ciliome) and that of the eukaryotic cilium/flagellum since the flagellum occurs in the early-branching phyla and not in terrestrial fungi. Tubulin polymerization promoting protein (TPPP)-like proteins, which contain a p25alpha domain, were also suggested to belong to the ciliome and are present in flagellated fungi. Although sanchytrids have lost many of the flagellar proteins, here it is shown that they possess a DNA sequence possibly encoding long (animal-type) TPPP, but not the fungal-type one characteristic of chytrid fungi. Phylogenetic analysis of p25alpha domains placed sanchytrids into a sister position to Blastocladiomycota, similarly to species phylogeny, with maximal support.

Characterisation and Cultivation of New Lineages of Colponemids, a Critical Assemblage for Inferring Alveolate Evolution

There are several alveolate groups outside the well-studied trio – ciliates, dinoflagellates, and apicomplexans – that are crucial for understanding the evolution of this major taxon. One such assemblage is the “colponemids”, which are eukaryotrophic biflagellates, usually with a ventral groove associated with the posterior flagellum. Previous phylogenetic studies show colponemids forming up to three distinct deep branches within alveolates (e.g. sister groups to Myzozoa or all other alveolates). We have developed dieukaryotic (predator–prey) cultures of four colponemid isolates. One represents the first stable culture of the halophile Palustrimonas (feeding on Pharyngomonas), while SSU rDNA phylogenies show the other isolates as two distinct new lineages. Neocolponema saponarium gen. et sp. nov. is a swimming alkaliphile with a large groove, which feeds on a kinetoplastid. Loeffela hirca gen. et sp. nov. is halophilic, has a subtle groove, usually moves along surfaces, and feeds on Pharyngomonas and Percolomonas. Prey capture in both new genera is raptorial, involves a specialized structure/region to the right of the proximal posterior flagellum, and presumed extrusomes. The relationships amongst Myzozoa, ciliates, and the (now) five described colponemid clades are unresolved, signaling that colponemid diversity represents both a challenge and important resource for tracing deep alveolate evolution.

Amphibian mucus triggers a developmental transition in the frog-killing chytrid fungus

The frog-killing chytrid fungus Batrachochytrium dendrobatidis (Bd) is decimating amphibian populations around the world.1-4Bd has a biphasic life cycle, alternating between motile zoospores that disperse within aquatic environments and sessile sporangia that grow within the mucus-coated skin of amphibians.5,6 Zoospores lack cell walls and swim rapidly through aquatic environments using a posterior flagellum and crawl across solid surfaces using actin structures similar to those of human cells.7,8Bd transitions from this motile dispersal form to its reproductive form by absorbing its flagellum, rearranging its actin cytoskeleton, and rapidly building a chitin-based cell wall-a process called "encystation."5-7 The resulting sporangium increases in volume by two or three orders of magnitude while undergoing rounds of mitosis without cytokinesis to form a large ceonocyte. The sporangium then cellurizes by dividing its cytoplasm into dozens of new zoospores. After exiting the sporangium through a discharge tube onto the amphibian skin, daughter zoospores can then reinfect the same individual or find a new host.5 Although encystation is critical to Bd growth, whether and how this developmental transition is triggered by external signals was previously unknown. We discovered that exposure to amphibian mucus triggers rapid and reproducible encystation within minutes. This response can be recapitulated with purified mucin, the bulk component of mucus, but not by similarly viscous methylcellulose or simple sugars. Mucin-induced encystation does not require gene expression but does require surface adhesion, calcium signaling, and modulation of the actin cytoskeleton. Mucus-induced encystation may represent a key mechanism for synchronizing Bd development with the arrival at the host.

Coordination of two opposite flagella allows high-speed swimming and active turning of individual zoospores.

Phytophthora species cause diseases in a large variety of plants and represent a serious agricultural threat, leading, every year, to multibillion dollar losses. Infection occurs when their biflagellated zoospores move across the soil at their characteristic high speed and reach the roots of a host plant. Despite the relevance of zoospore spreading in the epidemics of plant diseases, individual swimming of zoospores have not been fully investigated. It remains unknown about the characteristics of two opposite beating flagella during translation and turning, and the roles of each flagellum on zoospore swimming. Here, combining experiments and modeling, we show how these two flagella contribute to generate thrust when beating together, and identify the mastigonemes-attached anterior flagellum as the main source of thrust. Furthermore, we find that turning involves a complex active process, in which the posterior flagellum temporarily stops, while the anterior flagellum keeps on beating and changes its gait from sinusoidal waves to power and recovery strokes, similar to Chlamydomonas's breaststroke, to reorient its body to a new direction. Our study is a fundamental step toward a better understanding of the spreading of plant pathogens' motile forms, and shows that the motility pattern of these biflagellated zoospores represents a distinct eukaryotic version of the celebrated 'run-and-tumble' motility class exhibited by peritrichous bacteria.

Scanning electron microscopic observation of the in vitro cultured protozoan, Perkinsus olseni, isolated from the Manila clam, Ruditapes philippinarum

BackgroundPerkinsosis is a major disease affecting the commercially important marine mollusk Ruditapes philippinarum (Manila clam) in Asian waters. In this study, we investigated the morphological characteristics of Perkinsus olseni, the causative agent of perkinsosis, cultured under laboratory conditions at different stages of its life cycle using a scanning electron microscope (SEM).ResultsThe prezoosporangia formed after induction with Ray’s fluid thioglycollate medium (RFTM) developed into zoosporangia. During this process, a discharge tube formed a porous sponge-like structure that detached before the zoospores were released; thus, this organelle operated as a bung. Liberated zoospores gradually transformed into immature trophozoites, during which detachment of the anterior flagella occurred, but the loss of the posterior flagella was not clearly observed in the present study. Mature trophozoites underwent schizogony by cleaving the cell forming some merozoites in schizonts, which were released by the rupturing of the cellular membrane of the schizont within a few days.ConclusionsOur morphological and ultrastructural studies contribute new information on the life cycle and propagation of P. olseni.

Sperm ultrastructure in several species of Carabidae beetles (Insecta, Adephaga) and their organization in spermatozeugmata

Sperm structure of seven species from different Carabidae tribes was studied. Carabus preslii and Carabus granulatus interstitialis form sperm bundles (spermatozeugmata) in which the heads of conventional insect sperm cells are embedded in the apical cap (spermatostyle), leaving the posterior flagella free. On the contrary, Pterostichus morio, Pterostichus melas, Pterostichus melanarius and Amara aulica form complex spermatozeugmata bearing conspicuous axial spermatostyles associated with lateral groups of sperm. Individual sperm cells are contained in chambers delimited by laminar extensions of the cortical region of the spermatostyle. Demetrias atricapillus shares the general spermatozeugma structure with the above species, but the anterior spermatostyle region has a different structure and posteriorly, the lateral groups of sperm are separated from the spermatostyle but remain connected to it by peduncles consisting of extensions of the cortical region of the spermatostyle. The sperm of the species examined in the study consist of a short, flat acrosomal cap and, with the exception of the Carabus species, show long nuclei which extend parallel to the axonemes, along the flagella. Two relatively small mitochondrial derivatives and two small accessory bodies flank the axoneme. These bodies become very thick in the posterior flagellar region of D. atricapillus. The study revealed different spermatozeugma models and different sperm organization in the Carabidae family.

Ploeotids Represent Much of the Phylogenetic Diversity of Euglenids

Ploeotids are an assemblage of rigid phagotrophic euglenids that have 10-12 pellicular strips and glide on their posterior flagellum. Molecular phylogenies place them as a poorly resolved, likely paraphyletic assemblage outside the Spirocuta clade of flexible euglenids, which includes the well-known phototrophs and primary osmotrophs. Here, we report SSU rRNA gene sequences from 38 ploeotids, using both single-cell and culture-based methods. Several contain group I or non-canonical introns. Our phylogenetic analyses place ploeotids in 8 distinct clades: Olkasia n. gen., Hemiolia n. gen., Liburna n. gen., Lentomonas, Decastava, Keelungia, Ploeotiidae, and Entosiphon. Ploeotia vitrea, the type of Ploeotia, is closely related to P. oblonga and Serpenomonas costata, but not to Lentomonas. Ploeotia cf. vitrea sensu Lax and Simpson 2013 is not related to P. vitrea and has a different pellicle strip architecture (as imaged by scanning electron microscopy): it instead represents a novel genus and species, Olkasia polycarbonata. We also describe new genera, Hemiolia and Liburna, for the morphospecies Anisonema trepidum and A. glaciale. A recent system proposing 13 suprafamilial taxa that include ploeotids is not supported by our phylogenies. The exact relationships between ploeotid groups remain unresolved and multigene phylogenetics or phylogenomics are needed to address this uncertainty.

Ophirina amphinema n. gen., n. sp., a New Deeply Branching Discobid with Phylogenetic Affinity to Jakobids

We report a novel nanoflagellate, Ophirina amphinema n. gen. n. sp., isolated from a lagoon of the Solomon Islands. The flagellate displays ‘typical excavate’ morphological characteristics, such as the presence of a ventral feeding groove with vanes on the posterior flagellum. The cell is ca. 4 µm in length, bears two flagella, and has a single mitochondrion with flat to discoid cristae. The flagellate exists in two morphotypes: a suspension-feeder, which bears flagella that are about the length of the cell, and a swimmer, which has longer flagella. In a tree based on the analysis of 156 proteins, Ophirina is sister to jakobids, with moderate bootstrap support. Ophirina has some ultrastructural (e.g. B-fibre associated with the posterior basal body) and mtDNA (e.g. rpoA–D) features in common with jakobids. Yet, other morphological features, including the crista morphology and presence of two flagellar vanes, rather connect Ophirina to non-jakobid or non-discobid excavates. Ophirina amphinema has some unique features, such as an unusual segmented core structure within the basal bodies and a rightward-oriented dorsal fan. Thus, Ophirina represents a new deeply-branching member of Discoba, and its mosaic morphological characteristics may illuminate aspects of the ancestral eukaryotic cellular body plan.

Spermatogenesis and sperm structure of Neopanorpa lui and Neopanorpa lipingensis (Mecoptera: Panorpidae) with phylogenetic considerations

Spermatogenesis and sperm ultrastructure were investigated in Neopanorpa lui Chou & Ran and Neopanorpa lipingensis Cai & Hua by light and transmission electron microscopy. Two lateral lamellae run along the lateral grooves of the spermatid nucleus during spermiogenesis, but disappear in the mature spermatozoa. Sperm cells are very similar in ultrastructure between the two species. The spermatozoa are elongated cells, each comprising an apical bilayered acrosome inserted on the anterior nuclear region, an elongate nucleus that occupies the anterior sperm region, a neck region, and a posterior long flagellum. The nucleus has two prominent lateral grooves and assumes a helical appearance. The flagellum is helical and consists of a 9 + 2 axoneme, two mitochondrial derivatives of unequal size, one accessory body, and two extra-axonemal accessory structures. The uniqueness of spermatozoa of Neopanorpa lies in the specifically spoon-shaped larger mitochondrial derivative along most of the flagellum length. A tentative phylogenetic analysis based on sperm structure supports a possible monophyletic clade comprising Panorpidae, Panorpodidae, Bittacidae, and Boreidae of Mecoptera with two non-homoplasious characters: glycocalyx organised in longitudinal ridges and two extra-axonemal accessory structures/rods in flagellum. Closer affinities of Siphonaptera and the mecopteran clade are suggested by the characters of two asymmetric mitochondrial derivatives and the presence of mitochondrial derivative with crystalline structure. The sister group relationship of Panorpidae with Panorpodidae is supported by the presence of nuclear fiber and the irregular shape of the larger mitochondrial derivative.

Chytrid Batrachochytrium dendrobatidis fungal infection in freshwater prawn, Macrobrachium rosenbergii (de Man) - A new report

For the first time the occurrence of a chytrid, Batrachochytrium dendrobatidis (Bd) fungus infection in the fresh water prawn, Macrobrachium rosenbergii in grow out farms is reported. An extensive survey carried out between 2007 and 2011 in 15 farms (F) indicates that B. dendrobatidis is responsible for major pandemics in M. rosenbergii culture ponds as evidenced by the unique characteristics of the presence of fungal hyphae in claws and other features in the infected prawn. The pathogenicity of the fungus, LD50, histopathology, hematology and immunological response of the infected prawn is reported; the clinical symptoms include dull body color, abnormal protrusion associated with a grayish white color, wooly appearance with radiating grayish white fungal filaments on claw. Histopathology studies revealed that the infection resulted in faded eye color, occurrence of dark spots and melanization on the cephalothorax and muscles, translucent musculature, cuticular erosion, tissue atrophy with proliferated and infiltrated cells causing distension and gross enlargement of gill lamellae. An optimum dose of 105 zoospores/prawn (LD50) resulted in 50% mortality from 4 to 6days. At high dose of 108 zoospores/prawn, the infection caused total mortality from 2 to 4days. After 2nd week the total hemocyte counts (THCs) significantly decreased while the phenoloxidase (PO) level, relative superoxide anion production level, and superoxide dismutase (SOD) activity registered a significant increase (p<0.05) when compared to control. The cumulative mortality was 90% when challenged with Bd for 30days. The prevalence of infection was high in cooler or winter season (October, November, and December) and low in hot or summer season (March, April, May, and June) indicating a seasonal influence wielding a significant effect on mortality, hematology, and immunology profiles. Statement of relevanceIn India about 1000ha of shrimp/prawn ponds were affected by the disease, resulting in an annual loss of Rs. 10 million (US$0.23 million) during 1994–95 and Rs. 25 million (US$0.58 million) during 1995–96 (Ayyappan, 2000). Intensive prawn farming favors the rapid deterioration of environmental conditions which increase the incidence of diseases and failure of culture. Therefore augmenting the immune resistance and anti-environmental stress of prawns are of primary concern. Chytridiomycosis is an infectious disease caused by the chytrid, Batrachochytrium dendrobatidis; it is a non-hyphal zoospore fungus belonging to Chytridiomycota (Canter-Lund and Lund, 1995) or Blastocladiomycota (James et al., 2006) phylum. Chytrids are a diverse group of microscopic fungi (Barr, 1980; Barr, 2001; Kendrick, 1992) that disperse in an aqueous environment by motile zoospores with a single posterior whiplash flagellum (Sparrow, 1960). Chytrids have been well documented in amphibians (Longcore et al., 1999; Berger et al., 1999). A number of fungal pathogens were isolated and identified from prawns and shrimps (Krishnika and Ramasamy, 2014; Karthikeyan and Gopalakrishnan, 2014; Hatai, 2012; Cruz da Silva et al., 2011; Ramaiah, 2006). Recently, fungal pathogen like Aspergillus awamori has been isolated from marine shrimp, Liptopenaeus vannamei (Karthikeyan et al., 2015) and Gilbertella persicaria from Penaeus monodon. To our knowledge there is no report of the Batrachochytrium dendrobatidis infection in fish, prawn, and shrimp.

A New Heterotrophic Cryptomonad: Hemiarma marina n. g., n. sp.

We report a new heterotrophic cryptomonad Hemiarma marina n. g., n. sp. that was collected from a seaweed sample from the Republic of Palau. In our molecular phylogenetic analyses using the small subunit ribosomal RNA gene, H.marina formed a clade with two marine environmental sequences, and the clade was placed as a sister lineage of the freshwater cryptomonad environmental clade CRY1. Alternatively, in the concatenated large and small subunit ribosomal RNA gene phylogeny, H.marina was placed as a sister lineage of Goniomonas. Light and electron microscopic observations showed that H.marina shares several ultrastructural features with cryptomonads, such as flattened mitochondrial cristae, a periplast cell covering, and ejectisomes that consist of two coiled ribbon structures. On the other hand, H.marina exhibited unique behaviors, such as attaching to substrates with its posterior flagellum and displaying a jumping motion. H.marina also had unique periplast arrangement and flagellar transitional region. On the basis of both molecular and morphological information, we concluded that H.marina should be treated as new genus and species of cryptomonads.

Chemotaxis in the anisogamous brown alga Mutimo cylindricus (Cutleriaceae, Tilopteridales)

:Locomotion of female and male gametes of Mutimo cylindricus was analysed using a high-speed recording system. Female and male gametes morphologically differed in terms of their cell size and flagellar length. Male gametes swam significantly faster than female gametes; although, the flagellar waveforms were almost the same. Thus, the different swimming velocities of male and female gametes may have been due to differences in their cell size. Chemotactic male gametes treated with the sex pheromone from settled female gametes behaved in a similar manner to the male gametes of the isogamous Ectocarpus siliculosus. The results of the study revealed the following: (1) free-swimming male gametes exhibited thigmotaxis, (2) the beat frequency of the anterior flagellum and the swimming velocity decreased, (3) the deflection angle of the anterior flagellum increased, (4) the unilateral beat of the posterior flagellum was affected by the sex pheromone gradient, and (5) the tracking radius of male gametes around female gametes decreased.

Chemotactic movement in sperm of the oogamous brown algae, Saccharina japonica and Fucus distichus.

In oogamous species of brown algae such as Saccharina japonica and Fucus distichus, the sperm possess an unusual long posterior flagellum, which oscillates actively and produces a propulsive force during swimming. In this study, we quantitatively analyzed the effect of chemotactic responses on sperm swimming and flagellar waveforms by high-speed video recordings. We found that the thigmotactic response to the chemo-attractant was not enhanced during chemotactic swimming and that the swimming velocity of sperm did not decrease. As concentration of the chemo-attractant decreased, the sperm performed drastic U-turn movements, which was caused by a rapid and large bend of the posterior flagellum. Unilateral bending of the posterior flagellum when sensing a decrease in the concentration of the chemo-attractant may be a common response in male gametes during fertilization of brown algae both oogamous and isogamous species.

Flagellar waveforms of gametes in the brown alga Ectocarpus siliculosus

Brown algae are members of the Stramenopiles and their gametes generally have two heterogeneous flagella: a long anterior flagellum (AF) with mastigonemes and a short posterior flagellum (PF). In this study, swimming paths and flagellar waveforms in free-swimming and thigmotactic-swimming male and female gametes and in male gametes during chemotaxis, were quantitatively analysed in the model brown alga Ectocarpus siliculosus. This analysis was performed using a high-speed video camera. It was revealed that the AF plays a role in changing the locomotion of male and female gametes from free-swimming to thigmotactic-swimming and also in changing the swimming path of male gametes from linear to circular during chemotaxis. In the presence of a sex pheromone, male gametes changed their swimming path from linear (swimming path curvature, 0–0.02 µm–1) to middle and small circular path (swimming path curvature, 0.04–0.20 µm–1). The flagellar asymmetry and the deflection angle of the AF became larger, whereas the oscillation pattern of the AF was stable. However, there was no correlation between the flagellar asymmetry and the deflection angle of the AF and the path curvature when the male gametes showed middle to small circular paths. The PF irregularly changed the deflection angle and the oscillation pattern was unstable depending on the gradient of the sex pheromone concentration. AF waveforms were independent of PF locomotion during chemotaxis. This means that the AF has the ability to change the swimming path of male gametes – for example, from a highly linear path to a circular path – while changes in locomotion from a middle circle path to a small circle path is the result of beating of the PF.

Morphology and life cycle of amoeboflagellate Pharyngomonas sp. (Heterolobosea, Excavata) from hypersaline inland Razval Lake

The morphology, life cycle, feeding and multiplication of amoeboflagellate Pharyngomonas sp., strain RL isolated from hypersaline inland Razval Lake was studied. Life cycle includes flagellate, amoeboid, and cyst stages. Flagellated cells have two anterior flagella and two posterior flagella emerging apically from the anterior end of the cell. The nucleus localizes closely to the base of flagella. The longitudinal groove is on the ventral side of the cell. Flagellated cells have a distinct cytopharynx in the anterior part of the ventral groove. The cytopharynx passes into an intracellular channel reaching the posterior end of the cell. Amoeboid cells have two morphotypes—heterolobosean and lobosean. Cysts are spherical, their wall is smooth. Cell division occurs either at flagellated stage or at amoeboid one. Distinctions of Pharyngomonas sp., strain RL, from Pharyngomonas kirbyi were determined. They include longer flagella and long filamentous branching pseudopodia at the posterior end of amoeboid cells. The morphological distinctions are in agreement with differences in structure of the 18S rRNA gene. The revealed features may be used for differentiation of Pharyngomonas species.