Cthulhu Macrofasciculumque n. g., n. sp. and Cthylla Microfasciculumque n. g., n. sp., a Newly Identified Lineage of Parabasalian Termite Symbionts

The parabasalian symbionts of lower termite hindgut communities are well-known for their large size and structural complexity. The most complex forms evolved multiple times independently from smaller and simpler flagellates, but we know little of the diversity of these small flagellates or their phylogenetic relationships to more complex lineages. To understand the true diversity of Parabasalia and how their unique cellular complexity arose, more data from smaller and simpler flagellates are needed. Here, we describe two new genera of small-to-intermediate size and complexity, represented by the type species Cthulhu macrofasciculumque and Cthylla microfasciculumque from Prorhinotermes simplex and Reticulitermes virginicus, respectively (both hosts confirmed by DNA barcoding). Both genera have a single anterior nucleus embeded in a robust protruding axostyle, and an anterior bundle flagella (and likely a single posterior flagellum) that emerge slightly subanteriorly and have a distinctive beat pattern. Cthulhu is relatively large and has a distinctive bundle of over 20 flagella whereas Cthylla is smaller, has only 5 anterior flagella and closely resembles several other parababsalian genera. Molecular phylogenies based on small subunit ribosomal RNA (SSU rRNA) show both genera are related to previously unidentified environmental sequences from other termites (possibly from members of the Tricercomitidae), which all branch as sisters to the Hexamastigitae. Altogether, Cthulhu likely represents another independent origin of relatively high cellular complexity within parabasalia, and points to the need for molecular characterization of other key taxa, such as Tricercomitus.     

Esquamula lacrimiformis n. g., n. sp., a New Member of Thaumatomonads that Lacks Siliceous Scales

We report a new naked cercozoan flagellate, Esquamula lacrimiformis n. g., n. sp., collected from a sandy beach in Japan. Its cells were 4.5–11.3 μm in length and 3.9–8.8 μm in width and possess two unequal flagella. Cells move in a smooth gliding motion and have a trailing long posterior flagellum. Phylogenetic analyses with small and large subunit ribosomal RNA genes revealed that E. lacrimiformis forms a novel lineage within the Thaumatomonadida, the members of which are flagellates with siliceous scales. However, our light and electron microscopic observations indicated that E. lacrimiformis cells do not possess any siliceous structures. Furthermore, other morphological characteristics, such as the shape of the extrusomes and the structural arrangement of the microbody, were clearly different from those of previously described thaumatomonads. On the basis of a combination of these morphological observations and our phylogenetic analyses, we conclude that E. lacrimiformis should be treated as a new species of a new genus and placed into a new family, Esquamulidae n. fam., under Thaumatomonadida.     

Cryptobia udonellae sp. n. (Kinetoplastidea: Cryptobiida) - parasites of the excretory system of Udonella murmanica (Udonellida)

A new cryptobiid flagellates, Cryptobia udonellae sp. n., is described from the excretory channels of Udonella murmanica. The body of flagellates is spindle-shaped. The flagellar pocket is subapical. Two flagella emerge from the pocket. One flagellum turns anterior and is forward-directed; the other flagellum is directed posterior and close to the ventral cell surface. The ventral groove is well developed. The cytostome opens just anterior to the flagellar pocket. The cytostome leads to the short cytopharynx. In the excretory channel of worms the flagellates C. udonellae sp. n. are attached to microvilli of epithelium or lay free in the lumen. Both flagellates have been studied with TEM. The unusual parasite system which involves organisms of four different phylums of animals has been described for the first time.     

Cell morphology and formal description of Ergobibamus cyprinoides n. g., n. sp., another Carpediemonas-like relative of diplomonads

About 20 new isolates of Carpediemonas-like organisms (CLOs) have been reported since 2006. Small subunit rRNA gene phylogenies divide CLOs into six major clades: four contain described exemplars (i.e. Carpediemonas, Dysnectes, Hicanonectes, and Kipferlia), but two include only undescribed organisms. Here we describe a representative of one of these latter clades as Ergobibamus cyprinoides n. g., n. sp., and catalogue its ultrastructure. Ergobibamus cyprinoides is a bean-shaped biflagellated cell, 7-11.5 μm long, with a conspicuous groove. Instead of classical mitochondria there are cristae-lacking rounded organelles 300-400 nm in diameter. The posterior flagellum has a broad ventral vane and small dorsal vane. There are normally four basal bodies, two non-flagellated. There is one anterior root (AR), containing six microtubules. The posterior flagellar apparatus follows the "typical excavate" pattern of a splitting right root supported by fibres "I,"B," and "A," a "composite" fibre, a singlet root, and a left root (LR) with a "C" fibre. The B fibre originates against the LR--a synapomorphy of the taxon Fornicata--supporting the assignation of Ergobibamus to Fornicata, along with diplomonads, retortamonads, and other CLOs. Distinctive features of E. cyprinoides include the complexity of the AR, which is intermediate between Hicanonectes, and Carpediemonas and Dysnectes, and a dorsal extension of the C fibre.     

Flagellum retraction and axoneme depolymerisation during the transformation of flagellates to amoebae inPhysarum

Summary The transformation ofPhysarum polycephalum flagellates to myxamoebae is characterised by disappearance of the flagellum. This transition, from the flagellate to the myxamoeba was observed by phase contrast light microscopy and recorded by time lapse video photography to determine whether flagellates shed their flagella or they are absorbed within the cell. In addition, the kinetics of flagellum disappearance were also studied. Our observations indicate that the flagellum was absorbed within the cell; the process occurred within seconds. Flagellum resorbtion was preceded by typical morphological cell changes. The shape of the nucleus altered and its mobility within the cell decreased. It was not possible to observe the flagellum within the cell with phase contrast video recordings. Thin section electron microscopy was used to study this intracellular phenomenon. Several stages of flagellum dissolution could be identified within the cell. The two most important stages were: an axoneme surrounded by the flagellar membrane within a plasma membrane lined pocket or vacuole and the naked axoneme without its membrane, free within the cell cytoplasm. The existence of cytoplasmic microtubules prevented identification of any further dissolution stages of the flagellum. A group of microtubules adjacent to the flagellum but within the cytoplasm was observed in flagellates and also in those cells which possesed enveloped axonemes. The flagellum did not dissociate from the kinetosomes before resorbtion.Immunofluorescence studies with the 6-11-B-1 monoclonal antibody indicated that acetylated microtubules exist in myxamoebae after transformation from flagellates for up to 40 min. Acetylated tubulin is not limited to the centrioles in these cells.     

Structure and Life Cycle of Trypanoplasma beckeri sp. n. (Kinetoplastida), a Parasite of the Cabezon,Scorpaenichthys marmoratus,in Oregon Coastal Waters*,†

SYNOPSIS. Structure of Trypanoplasma beckeri sp. n. from the cabezon, Scorpaenichthys marmoratus (Ayres), is described from living specimens and from both Giemsa-, and protargol-stained smears. Flagellates from fish blood were usually long and slender, averaging 109.0 × 6.5 fan. The anterior flagellum averaged 8.5 μm; the recurrent flagellum bordered the body and terminated as a very short free flagellum, 2.5 μm long on the average. No true undulating membrane was observed, but in living individuals the recurrent flagellum undulated rapidly near its point of origin. The oval nucleus, averaging 8.5 × 4.0 μm, was located near the anterior cad of the body. An argentophilic, aciculum-like structure appeared to connect the nucleus to the area at the base of the flagella. The kinetoplast was not observed in fish blood forms. On the basis of laboratory experiments, the leech, Malmiana diminuta Burreson, was ascertained to be the vector for T. beckeri. Upon entry into the leech, flagellates became rounded, and division commenced within a few hours. Division continued for ?48 h and the flagellates became progressively smaller until reaching a length of ?10.0 μm. After 72 h they were found in high numbers in the proboscis sheath and also in the anterior crop of the leech. When infected leeches fed on an uninfected fish, flagellates were first observed in the fishes’ peripheral circulation 8 days later.     

Desulfovibrio Africans sp. n., a new dissimilatory sulfate-reducing bacterium

Campbell, L. Leon (University of Illinois, Urbana), Mary A. Kasprzycki, and John R. Postgate. Desulfovibrio africanus sp. n., a new dissimilatory sulfate-reducing bacterium. J. Bacteriol. 92:1122-1127. 1966.-The strains Benghazi and Walvis Bay can be distinguished from 40 strains of Desulfovibrio and from D. gigas on the basis of morphological and immunological studies. Electron microscopy revealed polar lophotrichous flagellation similar to that of D. gigas but different from the characteristic single polar flagellum of the 40 strains of Desulfovibrio. Immunological evidence shows that the two strains are related to members of the genus Desulfovibrio but possess several common antigenic components not present in the other strains tested. The deoxyribonucleic acid of both strains has a buoyant density of 1.724 g/cc and a guanine plus cytosine content of 60.2%. Cell-free extracts of both organisms show absorption bands of cytochrome c(3) and desulfoviridin, characteristic for Desulfovibrio. The two organisms carry out the sulfate-linked lactate fermentation and neither will grow in the absence of sulfate. Both strains contain the enzymes of the dissimilatory pathway of sulfate reduction. Therefore, these studies have demonstrated that the Benghazi and Walvis Bay strains should be regarded as taxonomically distinct from other species of Desulfovibrio.     

A light and electron microscopical study of a new, polymorphic free-living amoeba, Phreatamoeba balamuthi n. g., n. sp

A new amoeba, isolated from well water in Gambia, West Africa, is described and named Phreatamoeba balamuthi n. g., n. sp. Requiring anaerobic conditions for growth, it is easily cultured monoxenically with Escherichia coli or axenically in complex, undefined organic media. Three phenotypes have been observed in the life cycle: an amoeba, a flagellate, and a cyst. The amoeba moves by monopodia, is predominantly multinucleate, and varies from 11 to 160 microns in length. The flagellate has a single flagellum and is from 6 to 50 microns long. The cyst is surrounded by a resistant wall that lacks pores and ranges from 9 to 18 microns in diameter. The transformation from amoeba to flagellate can be induced nutritionally, the exact inducing factor(s) being unknown. Sexual reproduction has not been observed.     

Parapedinella reticulata gen. et sp. nov. (Chrysophyceae) from Danish waters

Parapedinella reticulata gen. et sp. nov. (Chrysophyceae) is described on the basis of light microscopy of living material, and also by means of electron microscopy of shadowcast whole mounts prepared from water samples collected in 1981 at some Danish brackish water localities. Additional information is provided by material from a marine habitat in southern Australia. The new taxon comprises small, colourless flagellates possessing a single projecting flagellum which carries tripartite hairs. The membrane of the flagellum is expanded into a sheath which is supported along the edge by a paraxial rod. The cells are covered with delicate scales and possess numerous slender protoplasmic axopodia which are retractable.     

Descriptions of Eumonhystera borealis n. sp., Sphaerolaimus occidentalis n. sp., and a Redescription of S. gracilis de Man 1876 (Nemata) from Bothnian Bay,Baltic Sea

The free-living nematodes Eumonhystera borealis n. sp., Sphaerolaimus occidentalis n. sp., and S. gracilis de Man 1876 from Bothnian Bay in the northern Baltic Sea are described and illustrated. Eumonhystera borealis n. sp. differs from other species by its small body size (314-393 μm), narrow body (a = 37-49), and large anterior amphids. In Sphaerolaimus occidentalis n. sp. the amphids are posterior to the buccal cavity, and it differs from other similar species by having two sclerotized rings in the posterior part of the buccal cavity. An intersex is reported for S. gracilis. Sphaerolaimus gracilis is cannibalistic or a predator of other species, with a preference for E. borealis n. sp. Sphaerolaimus occidentalis n. sp. coexists with S. gracilis at depths of 80 m but not at 12 m.     

A Light and Electron Microscopical Study of a New,Polymorphic Free-Living Amoeba,Phreatamoeba balamuthi n. g., n. sp.1

ABSTRACT. A new amoeba, isolated from well water in Gambia, West Africa, is described and named Phreatamoeba balamuthi n. g., n. sp. Requiring anaerobic conditions for growth, it is easily cultured monoxenically with Escherichia coli or axenically in complex, undefined organic media. Three phenotypes have been observed in the life cycle: an amoeba, a flagellate, and a cyst. The amoeba moves by monopodia, is predominantly multinucleate, and varies from 11 to 160 μm in length. The flagellate has a single flagellum and is from 6 to 50 μm long. The cyst is surrounded by a resistant wall that lacks pores and ranges from 9 to 18 μm in diameter. The transformation from amoeba to flagellate can be induced nutritionally, the exact inducing factor(s) being unknown. Sexual reproduction has not been observed.     

Description of two species of early branching dinoflagellates, Psammosa pacifica n. g., n. sp. and P. atlantica n. sp

In alveolate evolution, dinoflagellates have developed many unique features, including the cell that has epicone and hypocone, the undulating transverse flagellum. However, it remains unclear how these features evolved. The early branching dinoflagellates so far investigated such as Hematodinium, Amoebophrya and Oxyrrhis marina differ in many ways from of core dinoflagellates, or dinokaryotes. Except those handful of well studied taxa, the vast majority of early branching dinoflagellates are known only by environmental sequences, and remain enigmatic. In this study we describe two new species of the early branching dinoflagellates, Psammosa pacifica n. g., n. sp. and P. atlantica n. sp. from marine intertidal sandy beach. Molecular phylogeny of the small subunit (SSU) ribosomal RNA and Hsp90 gene places Psammosa spp. as an early branch among the dinoflagellates. Morphologically (1) they lack the typical dinoflagellate epicone-hypocone structure, and (2) undulation in either flagella. Instead they display a mosa?c of dinokaryotes traits, i.e. (3) presence of bi-partite trychocysts; Oxyrrhis marina-like traits, i.e. (4) presence of flagellar hairs, (5) presence of two-dimensional cobweb scales ornamenting both flagella (6) transversal cell division; a trait shared with some syndineansand Parvilucifera spp. i.e. (7) a nucleus with a conspicuous nucleolus and condensed chromatin distributed beneath the nuclear envelope; as well as Perkinsus marinus -like features i.e. (8) separate ventral grooves where flagella emerge and (9) lacking dinoflagellate-type undulating flagellum. Notably Psammosa retains an apical complex structure, which is shared between perkinsids, colpodellids, chromerids and apicomplexans, but is not found in dinokaryotic dinoflagellates.     

Malawimonas jakobiformis n. gen., n. sp. (Malawimonadidae n. fam.): A Jakoba-like Heterotrophic Nanoflagellate with Discoidal Mitochondrial Cristae

Malawimonas jakobiformis n. gen., n. sp., is established for a bacterivorous heterotrophic nanoflagellate isolated from the Malawi shore of Lake Nyasa (eastern Africa). Trophic stages observed were anteriorly biflagellate and naked. The posterior flagellum of a trophic cell resided in a conspicuous groove on the ventral surface, and bore a prominent vane. A Golgi stack and a mitochondrion with discoidal cristae were present anterior to the nucleus. The kinetid consisted of two short, slightly separated basal bodies, four microtubular roots, and associated fibers and bands. The three microtubular roots associated with the posterior basal body were associated with the ventral groove, while the single root associated with the anterior basal body gave rise to secondary cytoskeletal microtubules. Dividing cells became rounded, with persistent flagella. Cysts were uninucleate, and had thin organic walls without clearly differentiated apertures or ornamentation but with conspicuous attachment pads. Kinetid elements were present within cysts. On the basis of microscopical features, especially those of the kinetid, the nearest relatives of M. jakobiformis are the mitochondriate “jakobid” protists (families Histionidae and Jakobidae) and the amitochondriate retortamonads. Malawimonadidae n. fam. is established to accommodate this species.     

Glossomastix chrysoplasta n. gen., n. sp. (Pinguiophyceae), a new coccoidal, colony-forming golden alga from southern Australia

Glossomastix chrysoplasta gen. et sp. nov. is described from cultures isolated from sandstone rubble, Sorrento Back Beach, Mornington Peninsula, Victoria, Australia. The alga forms wall‐less, coccoidal vegetative cells that congregate in mucilaginous colonies and reproduce by successive bipartition. Plastids have girdle lamellae and partially embedded pyrenoids that are traversed by cytoplasmic channels. Zoospores are uniflagellate and swim poorly; a narrow lingulate pseudopod provides their primary form of motion. The single flagellum, which lacks hairs, a flagellar swelling, and autofluorescence, is the equivalent of the posterior flagellum in other golden algae. The anterior flagellum is absent; the basal body with which it would normally be associated is blind. The flagellar apparatus has two basal bodies, three microtubular roots, and a rhizoplast. The posterior (elder) basal body has a transitional helix that is proximal to the basal plate. Glossomastix chrysoplasta, placed in the Pinguiophyceae on the basis of molecular sequence and biochemical data, shares some ultrastructural features with other members of the class, especially Polypodochrysis teissieri, which has similar zoospores, but it also differs from other pinguiophytes in many respects. Glossomastix chrysoplasta is the pinguiophyte with, on average, the largest cells (exclusive of external materials), and it is the only one with a colonial habit.     

ULTRASTRUCTURAL STUDIES ON BIGELOWIELLA NATANS, GEN. ET SP. NOV., A CHLORARACHNIOPHYTE FLAGELLATE

Three isolates from the Provasoli-Guillard National Center for Culture of Marine Phytoplankton at Bigelow Laboratory, previously labeled Pedinomonas sp. and Pedinomonas minutissima from the green algal class Pedinophyceae, have been examined by light microscopy and TEM and shown to belong to the Chlorarachniophyceae, a class of nucleomorph-containing amebae. The three isolates represent the first chlorarachniophycean flagellates to be discovered. The ultrastructure of the cells has been examined in detail, with particular emphasis on the flagellar apparatus, a feature not examined in detail in chlorarachniophytes before. Cells are basically biflagellate, but the second flagellum is represented by a very short basal body only. Flagellar replication has shown this flagellum to be the mature stage, that is, the no. 1 flagellum, whereas the long emergent flagellum is the no. 2 flagellum that shortens into a short basal body during cell division. Mitosis is open with a pair of centrioles at each pole. Emergent flagella are absent during mitosis. Cells may form cysts, and the flagellar basal bodies and part of the flagellar roots are maintained in the cysts. Four microtubular roots emanate from the basal bodies, and the path of one of them is very unusual and very unlike any other known flagellate. No striated roots were observed. Other fine-structural features of the cell include a very unusual type of pyrenoid and a special type of extrusome. Cells are mixotrophic. The three isolates are very similar and are described as Bigelowiella natans , gen. et sp. nov. Ultrastructurally, chlorarachniophytes do not show close relationship to any known group of algae or other protists.     

Fermentation of maleate by a gram-negative strictly anaerobic non-spore-former,Propionivibrio dicarboxylicus gen. nov., sp. nov.

Three strains of maleate-fermenting anaerobic curved rods were isolated in pure culture from anaerobic freshwater mud samples. Among the isolates, strain CreMal1 was studied in detail. It was a mesophilic non-sporing gram-negative strict anaerobe, and grew not only on maleate but also on fumarate and l-malate, producing propionate and acetate stoichiometrically as end products. Succinate was an intermediate in the degradation of maleate. Nitrate, sulfate, and other sulfur compounds were not utilized as electron acceptors. It had 61 mol% guanine-plus-cytosine content, but possessed a single polar flagellum and did not utilize carbohydrates and lactate, unlike the genus Selenomonas. Therefore, strain CreMal1 is described as a member of Propionivibrio dicarboxylicus gen. nov., sp. nov., in the family Bacteroidaceae. Strain CreMal1 was deposited as type strain in the Japan Collection of Microorganisms and in Deutsche Sammlung von Mikroorganismen.Dedicated to Professor Dr. Norbert Pfennig on the occasion of his 65th birthday     

Brachiola vesicularum, N. G., N. Sp., a New Microsporidium Associated with AIDS and Myositis

Brachiola vesicularum, n. g., n. sp., is a new microsporidium associated with AIDS and myositis. Biopsied muscle tissue, examined by light and electron microscopy, revealed the presence of organisms developing in direct contact with muscle cell cytoplasm and fibers. No other tissue types were infected. All parasite stages contain diplokaryotic nuclei and all cell division is by binary fission. Sporogony is disporoblastic, producing 2.9 times 2 μm diplokaryotic spores containing 8-10 coils of the polar filament arranged in one to three rows, usually two. Additionally, this microsporidium produces electron-dense extracellular secretions and vesiculotubular appendages similar to Nosema algerae. However, the production of protoplasmic extensions which may branch and terminate in extensive vesiculotubular structures is unique to this parasite. Additionally, unlike Nosema algerae , its development occurred at warm blooded host temperature (37-38° C) and unlike Nosema connori , which disseminates to all tissue types, B. vesicularum infected only muscle cells. Thus, a new genus and species is proposed. Because of the similarities with the genus Nosema , this new genus is placed in the family Nosematidae. Successful clearing of this infection (both clinically and histologically) resulted from treatment with albendazole and itraconozole.     

Candida silvae sp.n., a yeast isolated from humans and horses

A new yeast,Candida silvae sp.n., isolated from humans and horses is described. The new species has some important properties in common withCandida norvegensis andTorulopsis inconspicua, but its assimilation pattern and maximum temperature for growth are different.This work was supported by the Gulbenkian Foundation, Lisboa, Portugal.