Life Cycle, Morphology, Ontogenesis, and Phylogeny of Bromeliothrix metopoides nov. gen., nov. spec., a Peculiar Ciliate (Protista, Colpodea) from Tank Bromeliads (Bromeliaceae)

Bromeliothrix metopoides was discovered in tank bromeliads from Central and South America. Pure cultures could be established in various media stimulating growth of its food, i.e. bacteria and heterotrophic flagellates of the genus Polytomella. The new ciliate was investigated in the light- and scanning electron microscope, with various silver impregnation techniques, and with molecular methods, using the small-subunit rDNA. The morphology and its changes during the life cycle are documented by 167 figures and a detailed morphometry. Bromeliothrix metopoides is about 27-55 × 22-36 μm in size and has a complex life cycle with Metopus-shaped, bacteriophagous theronts and trophonts (microstomes) and obovate, flagellate-feeding macrostomes having a large, triangular oral apparatus. The thin-walled resting cysts of the theronts and trophonts are uniquely ellipsoidal, while the thick-walled cyst of the macrostome morph is globular. Reproduction occurs in freely motile condition either by binary fission or polytomy, producing a unique, motile "division chain" composed of four globular offspring, of which the central ones are connected by a curious, plug-like holdfast. Division is associated with a complete reorganization of the parental oral and somatic infraciliature. Stomatogenesis is merotelokinetal as in other members of the order Colpodida. The right polykinetid is generated by the rightmost postoral kinety, while the left polykinetid is produced by the two left postoral kineties and five left side kineties. The division in freely motile condition resembles the Exocolpodidae Foissner et al., 2002, to which Bromeliothrix is tentatively assigned, differing from Exocolpoda mainly by the formation of a macrostome morph and a division chain. Bromeliothrix has a ciliary and silverline pattern typical for members of the family Colpodidae. This matches the molecular classification which, however, hardly reflects the outstanding division and life cycle, suggesting some decoupling of morphological and molecular evolution. The specific morphological and ontogenetic traits of Bromeliothrix are interpreted as adaptations to the highly competitive habitat, favouring r-selected life strategies. Bromeliothrix metopoides is widespread in various tank bromeliads and can be easily cultivated in a wide variety of limnetic and terrestrial media. Thus, it remains obscure why this ciliate is restricted to tank bromeliads, i.e. did not occur in about 2,000 soil and freshwater samples investigated globally, including some 100 samples from Central and South America.     

A 'living fossil' eel (Anguilliformes: Protanguillidae, fam. nov.) from an undersea cave in Palau

We report the discovery of an enigmatic, small eel-like fish from a 35 m-deep fringing-reef cave in the western Pacific Ocean Republic of Palau that exhibits an unusual suite of morphological characters. Many of these uniquely characterize the Recent members of the 19 families comprising the elopomorph order Anguilliformes, the true eels. Others are found among anguilliforms only in the Cretaceous fossils, and still others are primitive with respect to both Recent and fossil eels. Thus, morphological evidence explicitly places it as the most basal lineage (i.e. the sister group of extant anguilliforms). Phylogenetic analysis and divergence time estimation based on whole mitogenome sequences from various actinopterygians, including representatives of all eel families, demonstrate that this fish represents one of the most basal, independent lineages of the true eels, with a long evolutionary history comparable to that of the entire Anguilliformes (approx. 200 Myr). Such a long, independent evolutionary history dating back to the early Mesozoic and a retention of primitive morphological features (e.g. the presence of a premaxilla, metapterygoid, free symplectic, gill rakers, pseudobranch and distinct caudal fin rays) warrant recognition of this species as a 'living fossil' of the true eels, herein described as Protanguilla palau genus et species nov. in the new family Protanguillidae.     

Ultrastructural description of a new chytrid genus of caecum anaerobe, Caecomyces equi gen. nov., sp. nov., assigned to the Neocallimasticaceae

Vegetative and reproductive stages of Caecomyces equi gen. nov., sp. nov. isolated from the horse caecum were examined by light and electron microscopy. This organism, which is similar to isolates known as Sphaeromonas communis, produces uniflagellate, uninucleate zoospores whose perikinetosomal structures, i.e. circumflagellar ring, spur, struts and scoop, are similar in many respects to those described in species of Neocallimastix. Microtubular roots extend basally from the spur and associate with hydrogenosomes and the nucleus. Another group of microtubules radiates laterally in a fan-shaped array close to the plasmalemma. Zoospores encyst, shedding their flagella with basal bodies, and germinate to diglobular thalli. Either coralloid or bulbous rhizoids form in plant material, but only the latter in axenic culture. Incipient zoospores are produced from a multinucleate eucarpic thallus and devlop within cleavage vacuoles containing flagella. An isolate from the cow rumen was found to be similar to C. equi in morphology and zoospore ultrastructure. On the basis of zoospore ultrastructure, we assign the new genus to the Neocallimasticaceae of the order Spizellomycetales. Organisms previously described as Sphaeromonas communis and Piromonas communis are renamed Caecomyces communis and Piromyces communis and assigned to the same family.     

Proposal of Monorhizochytrium globosum gen. nov., comb. nov. (Stramenopiles,Labyrinthulomycetes) for former Thraustochytrium globosum based on morphological features and phylogenetic relationships

Thraustochytrium is the type genus of the family Thraustochytriaceae in the class Labyrinthulomycetes. This genus is characterized by zoospore formation, namely, shape of the cell wall of sporangia and presence or absence of a proliferous body. However, there are several issues associated with the taxonomy of this genus, and these include polyphyletic taxa and overlapping of taxonomic features among species. In particular, the first and second species, T. proliferum and T. globosum, were described based on observations of the morphological features of natural samples in the absence of culture conditions. Before addressing the taxonomic issues associated with this genus, it is important to consider the taxonomic features of each species, i.e., the life history under culture conditions and the phylogenetic position. The objective of the present study was to isolate T. globosum, the second described species in the genus Thraustochytrium, from the type locality. We successfully isolated strain NBRC 112723, which exhibited characteristic features of T. globosum. Under culture conditions, strain NBRC 112723 exhibited taxonomic features observed in other thraustochytrid species. Our molecular phylogeny indicated that this strain isolated from the type locality was located in an unidentified thraustochytrid group; moreover, some strains located in this group exhibited characteristic features of strain NBRC 112723. We clearly distinguished T. globosum based on the taxonomic criteria used to classify the T. proliferum type species. Therefore, we propose the establishment of a new genus, Monorhizochytrium, for the species T. globosum in the family Thraustochytriaceae.     

Corynoplastis japonica gen. et sp. nov. and Dixoniellales ord. nov. (Rhodellophyceae,Rhodophyta) based on morphological and molecular evidence

A new unicellular red alga, Corynoplastis japonica gen. et sp. nov., is described from Tobishima, Japan. Cells are spherical, 18–33 µm in diameter, pale purple to brownish red and surrounded by a mucilaginous sheath. A single chloroplast with many lobes extends from the cell periphery to the cell center. A peripheral thylakoid is present. A pyrenoid occurs at each innermost chloroplast lobe end and one or two thylakoids are present in the pyrenoid matrix. The nucleus is eccentric to peripheral and Golgi bodies are scattered throughout the cell and associated with endoplasmic reticulum. Cells have a slow random gliding motility. The low molecular weight carbohydrate mannitol is present in the cells. Molecular phylogenetic analysis indicates that this alga is closely related to members of the genus Rhodella. A new order, Dixoniellales, is established for Dixoniella, Neorhodella and Glaucosphaera based on molecular and ultrastructural evidence (Golgi bodies associated only with the nucleus). The redefined order Rhodellales in which Rhodella and Corynoplastis are placed is characterized ultrastructurally by Golgi bodies scattered throughout the cytoplasm and associated with endoplasmic reticulum.     

The hyphomycete Teberdinia hygrophila gen. nov., sp. nov. and related anamorphs of Pseudeurotium species

A hyphomycetous fungus isolated from montane fen soil in the Caucasus Mountains, Russia, had obscurely sympodial conidiogenous cells that suggested a link to the heterogeneous genus Leptodontidium. Sequence analysis of the nuclear ribosomal small subunit and internal transcribed spacer region, however, disclosed that the fungus was an anamorphic member of a clade containing the cleistothecial ascomycetous genus Pseudeurotium. Teberdinia, gen. nov., is proposed for the blastic, generally sympodially proliferating anamorphs in this group, and Teberdinia hygrophila, sp. nov., is proposed for the species from upland fens. Binomials are not proposed for the remaining Teberdinia anamorphs of Pseudeurotium species. Purely anamorphic isolates in this clade are difficult to recognize using current morphological keys and might be more widely distributed and ecologically significant than is currently evident.     

Nocardia testaceus sp. nov. and Nocardia senatus sp. nov., isolated from patients in Japan

Two actinomycete strains isolated from sputum between 1999 and 2001 in Japan were provisionally assigned to the genus Nocardia based on morphological criteria. These isolates were further studied in order to determine their specific taxonomic status. Detailed chemotaxonomic characterization and 16S rDNA gene sequence analysis of these isolates also confirmed that they belong to the genus Nocardia. The 16S rDNA sequence data of the two strains showed that they are most similar to that of Nocardia carnea and Nocardia flavorosea. However, DNA-DNA relatedness data showed that the two strains could be distinguished from N. carnea and N. flavorosea and therefore represented two new species within the genus Nocardia. The designation of the two isolated strains are Nocardia testaceus for IFM 0937(T) (=JCM 12235(T), DSM 44765(T)) and Nocardia senatus for IFM 10088(T) (=JCM 12236(T), DSM 44766(T)).     

Further studies on Nephroselmis and its allies (Prasinophyceae). II. Mamiella gen. nov., Mamiellaceae fam. nov., Mamiellales ord. nov.

The marine prasinophycean flagellate presently known as Nephroselmis gilva has been examined, using both the type culture and material from temperate (Denmark, New Zealand) and tropical waters (Thailand). All cell surfaces are covered with unmineralized scales, two types on the body, two on the flagella including flagellar hairs. The detailed structure of the scales is described, using high power electron microscopy of detached positive–stained scales. Previously overlooked organelles within the cell include large numbers of extrusive bodies, a rare type of organelle in chlorophyll–a– and –b–containing organisms, and an eyespot. N. gilva differs profoundly from the type species of Nephroselmis , and is transferred to the new genus Mamiella. Mamiella gilva is closely related to Mantoniella squamata. Together with the genus Dolichomastix they form the new family, Mamiellaceae, a small group of marine flagellates of worldwide distribution.
The members of the new family probably represent the most primitive extant prasinophytes. When compared with other members of the class, its species stand out, particularly by the lack of small square or diamond–shaped scales on the flagella and cell body. It is suggested that the Mamiellaceae should be referred to a separate order, Mamiellales.     

Ultrastructure and identification of the predatory flagellateColpodella pugnax Cienkowski (Apicomplexa) with a description ofColpodella turpis n. sp. and a review of the genus

A flagellated predator of the chlorophyte algaDunaliella spp. was examined by light and electron microscopy. Although this predator had previously been identified as a species of the kinetoplastid genusBodo, the present study revealed the flagellate to be indistinguishable fromColpodella pugnax, the type-species for its genus. The flagellate lacks a kinetoplast, a microtubule supported cytopharynx and paraxial rods in the flagella — characters universally found in kinetoplastid flagellates. The cell has mitochondria with vesicular cristae. Multiple membranes surround the cell and are underlain by longitudinal microtubules not originating from the flagellar region. Most notably, the flagellate has micropores and an apical complex including a conoid, sacculate rhoptries and, apparently, a polar ring. This study hs confirmed thatColpodella is the genus with free-living species most closely related to the apicomplexan parasites (i.e. the “Euapicomplexa” andPerkinsus). No unambiguous synapomorphy supports an “apicomplexan parasites” clade: Inclusion ofColpodella is necessary to secure the Apicomplexa as a monophyletic (=holophyletic) taxon. A new family, the Colpodellidae, is erected for this genus. Colpodella turpis, a previously undescribed species that also consumesDunaliella spp., was isolated from the same samples asC. pugnax. A diagnosis for this species is presented together with a brief review of the genus, in which we recognise seven species. The generic namesAlphamonas Aléxéieff,Nephromonas Droop andDingensia Patterson & Zölffel are rendered into synonomy withColpodella.     

Molecular Characterization of “Candidatus Similichlamydia latridicola” gen. nov., sp. nov. (Chlamydiales: “Candidatus Parilichlamydiaceae”), a Novel Chlamydia-Like Epitheliocystis Agent in the Striped Trumpeter,Latris lineata (Forster)

Histological analysis of gill samples taken from individuals of Latris lineata reared in aquaculture in Tasmania, Australia, and those sampled from the wild revealed the presence of epitheliocystis-like basophilic inclusions. Subsequent morphological, in situ hybridization, and molecular analyses were performed to confirm the presence of this disease and discovered a Chlamydia-like organism associated with this condition, and the criteria set by Fredericks and Relman''s postulates were used to establish disease causation. Three distinct 16S rRNA genotypes were sequenced from 16 fish, and phylogenetic analyses of the nearly full-length 16S rRNA sequences generated for this bacterial agent indicated that they were nearly identical novel members of the order Chlamydiales. This new taxon formed a well-supported clade with “Candidatus Parilichlamydia carangidicola” from the yellowtail kingfish (Seriola lalandi). On the basis of sequence divergence over the 16S rRNA region relative to all other members of the order Chlamydiales, a new genus and species are proposed here for the Chlamydia-like bacterium from L. lineata, i.e., “Candidatus Similichlamydia latridicola” gen. nov., sp. nov.     

Identification of a new‐to‐science cyanobacterium,Toxifilum mysidocida gen. nov. & sp. nov. (Cyanobacteria,Cyanophyceae)

Cyanobacteria occupy many niches within terrestrial, planktonic, and benthic habitats. The diversity of habitats colonized, similarity of morphology, and phenotypic plasticity all contribute to the difficulty of cyanobacterial identification. An unknown marine filamentous cyanobacterium was isolated from an aquatic animal rearing facility having mysid mortality events. The cyanobacterium originated from Corpus Christi Bay, TX. Filaments are rarely solitary, benthic mat forming, unbranched, and narrowing at the ends. Cells are 2.1 × 3.1 μm (width × length). Thylakoids are peripherally arranged on the outer third of the cell; cyanophycin granules and polyphosphate bodies are present. Molecular phylogenetic analysis in addition to morphology (transmission electron microscopy and scanning electron microscopy) and chemical composition all confirm it as a new genus and species we name Toxifilum mysidocida. At least one identified Leptolyngbya appears (based on genetic evidence and TEM) to belong to this new genus.     

Phylogeny and Establishment of a New Ciliate Family,Wilbertomorphidae fam. nov. (Ciliophora,Karyorelictea), a Highly Specialized Taxon Represented by Wilbertomorpha colpoda gen. nov., spec. nov.

The morphology and molecular phylogeny of a new marine interstitial karyorelictean ciliate, Wilbertomorpha colpoda gen. nov., spec. nov., isolated from a sandy beach at Qingdao, China, are investigated. The ciliature is mainly on the right surface with the left surface barren except for a single marginal kinety, which is typical of the order Loxodida. Nevertheless, other morphological features, such as the highly reduced buccal structure, the absence of a buccal cavity and the presence of extremely specialized cortical granules, prevent its assignation to any known family. Furthermore, the secondary structure of the variable region 2 of the SSU rRNA gene is unique among loxodids. Thus, we establish a new family, Wilbertomorphidae fam. nov. for the genus Wilbertomorpha. Phylogenetic analyses based on SSU rRNA gene sequence data suggest that this taxon is a sister group to the family Geleiidae which belongs to the order Protoheterotrichida. However, morphologically Wilbertomorpha bears a much stronger resemblance to Loxodidae which belongs to the order Loxodida. Based on these findings, we tentatively place Wilbertomorphidae nov. fam. incertae sedis within Karyorelictea.     

Heterobasidion occidentale sp. nov. and Heterobasidion irregulare nom. nov.: a disposition of North American Heterobasidion biological species

The genus Heterobasidion includes some of the most important pathogens of conifers in the world, and as such it is one of the most intensely studied genera of fungi. Because of the remarkable paucity of distinguishing morphological traits, the taxonomy of species within this genus has always been problematic. A partial resolution of the taxonomic issues regarding this genus was achieved by defining the most important and first described species within it, Heterobasidion annosum, as a species complex containing at least two partially intersterile biological species defined as intersterility groups (ISGs). With time, the number of ISGs has increased to include at least two distinct North American and three distinct Eurasian ISGs. Two additional, yet unnamed, taxonomic groups within Heterobasidion have been recently described in Japan. ISGs are distinguishable either by minor morphological differences, by partial intersterility, by ecological traits including host preference, and/or by their geographic range. Several studies employing a variety of molecular tools and analyses have confirmed the distinct genetic divergence among ISGs, identifying each of them as a monophyletic group. Using genetic markers, genotypes can always be unambiguously assigned to one ISG, and very few inter-ISG hybrids have been identified. In this paper, we summarize the available information, both genetic and ecological, that differentiates the two North American ISGs from each other and from other taxonomic units within the genus. We demonstrate that morphometric characteristics such as pore density and pore shape differentiate the two ISGs. Based on the cumulative genetic, ecological, and morphological evidence, we propose a disposition of ISGs of the North American H. annosum by replacing the P ISG with Heterobasidion irregulare, and the S ISG with Heterobasidion occidentale.     

Agolohymena aspidocauda nov. gen., nov. spec., a histophagous freshwater tetrahymenid ciliate in the family Deltopylidae (Ciliophora,Hymenostomatia), from Idaho (northwest U.S.A.): Morphology,ontogenesis and molecular phylogeny

Morphology, ontogeny and the molecular phylogeny of Agolohymena aspidocauda nov. gen., nov. spec., a new freshwater tetrahymenid ciliate from Idaho, U.S.A, are described. The ontogeny and histophagous mode of nutrition are similar to those of Deltopylum rhabdoides Fauré-Fremiet and Mugard, 1946. The new genus is placed with Deltopylum in the resurrected family Deltopylidae Song and Wilbert, 1989. We emend the diagnostic features of the family to include division by polytomy, right and left somatic kineties extending into the preoral suture, crook-shaped or sigmoid adoral membranelles 1 and 2, markedly reduced adoral membranelle 3 and a tetrahymenid silverline pattern. The main diagnostic features of the new genus are a disc-shaped caudal ciliary array and formation of two types of resting cysts, one smooth and the other bearing tangled tubular or cylindrical lepidosomes. Nuclear small subunit ribosomal RNA gene and mitochondrial cytochrome oxidase subunit 1 gene sequences place the new genus basal within the order Tetrahymenida, well separated from members of the family Tetrahymenidae (Lambornella and Tetrahymena) and also from other tetrahymenids (Colpidium, Dexiostoma, Glaucoma). The genetic divergences between this species and other genera in Tetrahymenida are large enough to suggest placement of the new genus in a separate family. This corroborates the morphological data, since the elaborate caudal ciliary array and the lepidosome-covered resting cyst of this species are not found in other Tetrahymenidae.     

A mid-Cretaceous trichomycete,Priscadvena corymbosa gen. et sp. nov., in Burmese amber

Priscadvena corymbosa gen. et sp. nov., is described from thalli and sporangia emerging from the oral cavity of a click beetle (Coleoptera: Elateridae) in mid-Cretaceous Burmese amber. The fossil contains several features unknown in extant Trichomycetes including a click beetle (Coleoptera: Elateridae) host, spiny, aerial thalli with the entire thallus bearing numerous small uninucleate globular spores and stalks attached to the oral cavity of its host. Based on these features, P. corymbosa gen. et sp. nov. is placed in a new family, Priscadvenaceae fam. nov., and new order, Priscadvenales ord. nov. The new morphological and behavioral features of the fossil add to the diversity of the trichomycetes as currently defined.     

Actinomadura mexicana sp. nov. and Actinomadura meyerii sp. nov., two novel soil sporoactinomycetes

The taxonomic position of two soil isolates, strains A288(T) and A290(T) [provisionally assigned to the genus Actinomadura] was clarified in a polyphasic study. The organisms showed a combination of chemotaxonomic and morphological properties typical of actinomadurae. They also formed distinct phyletic lines in the 16S rRNA Actinomadura gene tree; strain A288(T) was associated with A. nitritigenes whereas strain A290(T) was closely related to a group that consisted of A. citrea, A. coerulea, A. glauciflava, A. luteofluorescens and A. verrucosospora. Strains A288(T) and A290(T) showed key phenotypic features which readily distinguish them from one another and from representatives of related validly described species of Actinomadura. It is proposed that the two organisms be classified as new species of the genus Actinomadura. The names proposed for the new taxa are Actinomadura mexicana (A290(T) = DSM 44485(T) = NRRL B-24203(T)), and Actinomadura meyerii (A288(T) = DSM 44485(T) = NRRL B-24203(T)).