Platyamoeba stenopodia. n. g., n. sp., a Freshwater Amoeba*

SYNOPSIS. Platyamoeba stenopodia n. g., n. sp., isolated from a lake in Alabama, is an amoeba with flattened form, usually more than 2.5 × as long as broad, the anterior half hyaline and the posterior half granular, with a single vesiculate nucleus at the posterior edge of the hyaline region. It forms a transitory floating stage with slender, hyaline pseudopods. Its cyst is spherical, uninucleate, with a smooth inner wall and a thinner, closely applied outer wall, which is often slightly wrinkled. Length of locomotive amoeba 15–36 μ diameter of cyst 7–11.5 μ Reproduction by binary fission with mesomitotic nuclear division, the nuclear membrane disappearing in prophase. The genus Platyamoeba is erected for amoebae including this organism; the type species is P. placida.     

Thiomicrospira pelophila,gen. n., sp. n., a new obligately chemolithotrophic colourless sulfur bacterium

From marine mud flats a very thin, comma- or spiral-shaped bacterium was isolated. The new organism was an obligately chemolithotrophic sulfur bacterium. Its physiology was found to be essentially similar to that ofThiobacillus thioparus. Because of the spirillum-like appearance it was proposed to classify this bacterium into a new genusThiomicrospira, with the species nameTms. pelophila. Tms. pelophila and a marineT. thioparus, which was isolated from the same mud, occupy different niches in this habitat.Tms. pelophila has a remarkable sulfide-tolerance as compared withT. thioparus. This property could be used for the specific enrichment ofTms. pelophila. The organism was also readily isolated in pure culture by filtering mud suspensions through a 0.22 Μm membrane filter.     

Ovipleistophora gen. n., a new genus for Pleistophora mirandellae-like microsporidia

Based on ultrastructural study and molecular analysis, a new genus, Ovipleistophora, is established for Pleistophora mirandellae-like microsporidia from roach and ruff oocytes. Unlike Pleistophora, Ovipleistophora has a thick additional envelope around the meront. This envelope breaks open to release the cells into the host cell cytoplasm. The cells, becoming multinuclear sporogonic plasmodia, already have a surface coat that transforms into the sporont wall and eventually into the sporophorous vesicle wall. The surface coat and its transformation differ from those of Pleistophora, but bear some resemblance to those of Trachipleistophora. In Trachipleistophora the sporonts, however, do not form plasmodia, as they do in Ovipleistophora and Pleistophora. Small subunit ribosomal DNA analysis supports the establishment of the new genus and assignment of P. mirandellae from 2 different fish hosts to the same species. The same small subunit ribosomal DNA analysis lends support for transferring P. ovariae into the genus Ovipleistophora.     

Chamaesiphonosira cymbellicola n.gen., n.sp., ein knospenbildendes Bakterium,und sein spezialisierter Epiphytismus

The organism shows long-conical, colourless cells of bacterial size, affixed with the broader basis and cutting off buds from the free apex in numbers as high as 30, which form a chain and later separate. Multiplication is entirely by budding. Some resemblance to Cyanophycean cells, in particular to the exosporangia of Chamaesiphon, seems to be cytologically not verified. The organism grows exclusively on living cells of the diatom Cymbella cesati, and is fixed to them principally on the edges of the valvae. In spite of the presence of many other similar diatom species, the organism represents a remarkable case of extremely specialized epiphytism. There are no signs of damage of the “host” cell, in cell division the epiphyte is taken over by the epithecae of the daughter cells. Systematically the species because of practical reasons can be included into the Caulobacterales sensu Henrici and Johnson (1935). A second species, obviously belonging in here, is represented by Chamaesiphon hyalinus Scherffel (1907).     

HALOSTYLODINIUM ARENARIUM, GEN ET SP. NOV. (DINOPHYCEAE), A COCCOID SAND-DWELLING DINOFLAGELLATE FROM SUBTROPICAL JAPAN

A new genus and species of marine coccoid dinoflagellate from subtropical Japan, Halostylodinium arenarium Horiguchi et Yoshizawa-Ebata, gen. et sp. nov., is described. The dominant stage of the dinoflagellate is a nonmotile ovoidal to spheroidal cell with a distinct stalk. The stalk consists of an upper thick tubule, a lower thin tubule, and a discoidal holdfast. The dinoflagellate possesses a yellowish-brown chloroplast with multiple lobes radiating from a central pyrenoid. It reproduces by the formation of two motile cells, which swim for a short period and then transform directly into the stalked nonmotile cell. The stalk is produced during transformation from the apical stalk complex present in the apex of the motile cell. The apical stalk complex consists of a double-folded apical pore plate and doughnut-shaped holdfast-building material. The ultrastructure of the apical stalk complex is compared with those of Bysmatrum arenicola and Stylodinium littorale. Halostylodinium arenarium possesses delicate thecal plates, and the thecal plate formula is Po, 5', 2a, 7", 7c, 6s, 5"', 1p, 2"". A phylogenetic study based on the 18S ribosomal RNA gene did not show any clear affinities between this organism and any species included in the analysis.     

Subdoligranulum variabile gen. nov., sp. nov. from human feces

During studies on the microflora of human feces we have isolated a strictly anaerobic, non-spore-forming, Gram-negative staining organism which exhibits a somewhat variable coccus-shaped morphology. Comparative 16S ribosomal RNA gene sequencing studies show the unidentified organism is phylogenetically a member of the Clostridium leptum supra-generic rRNA cluster and displays a close affinity to some rDNA clones derived from human and pig feces. The nearest named relatives of the unidentified isolate corresponded to Faecalibacterium prausnitzii (formerly Fusobacterium prausnitzii) displaying a 16S rRNA sequence divergence of approximately 9%, with Anaerofilum agile and A. pentosovorans the next closest relatives of the unidentified bacterium (sequence divergence approximately 10%). Based on phenotypic and phylogenetic considerations, it is proposed that the unusual coccoid-shaped organism be classified as a new genus and species, Subdoligranulum variabile. The type strain of S. variabile is BI 114(T) (=CCUG 47106(T)=DSM 15176(T)).     

A new species, Aphanomyces salsuginosus sp. nov., isolated from ice fish Salangichthys microdon

A species of Aphanomyces was isolated from the ice fish Salangichithys microdon living in brackish water in Japan. White cotton-like growth was found on the heads and fins of the fish. Hyphae penetrated into the dermal layers, subcutaneous tissues, muscular layers, and cartilaginous tissue of the mandible and maxilla; these hyphae were associated with cellular debris and lesions in host tissue. White fluffy colonies from subcultures of these growths were isolated on glucose–yeast agar plates with 0.5% sodium chloride (NaCl). These isolates consisted of delicate, slightly wavy, and moderately branched hyphae. Zoosporangia were isodiametric with the vegetative hyphae. Oogonia were abundant and approximately 21–33 μm in diameter, with irregular short papillae. Generally they were spherical or subspherical and only rarely pyriform. Individual oogonia usually contained a single oospore, which was spherical and 19–27 μm in diameter, with a large shiny vesicle. Antheridial branches, when present, were usually androgynous; however, they were sometimes monoclinous or diclinous. The optimal growth temperature of the isolates was 20°C, and cultures grew well at low salinity (0–0.5% NaCl). Phylogenic analysis based on the internal transcribed space 1-5.8S-ITS 2 of the ribosomal RNA gene indicates that these isolates will be an as-yet unidentified species of Aphanomyces.     

Monopylocystis visvesvarai n. gen., n. sp. and Sawyeria marylandensis n. gen., n. sp.: two new amitochondrial heterolobosean amoebae from anoxic environments

Two new species of heterolobosean amoebae from anoxic environments, Monopylocystis visvesvarai and Sawyeria marylandensis, are described on the basis of light microscopy, electron microscopy, and their phylogenetic affiliation based on analyses of nuclear small-subunit ribosomal RNA gene sequences. Both species lack mitochondria but have organelles provisionally interpreted as hydrogenosomes, and neither can tolerate aerobic conditions. As their conditions of culture do not exclude all oxygen, they may be microaerophiles rather than strict anaerobes. Both species have unusual nucleolar morphologies. Monopylocystis visvesvarai, from a marine sediment, has nucleolar material distributed around the nuclear periphery. It is the first non-aerobic heterolobosean protist for which a cyst is known; the cyst is unmineralized and unornamented except for a single, raised, plugged pore. Sawyeria marylandensis, from an iron-rich freshwater stream, has nucleolar material distributed in one or two parietal masses, which persist during mitosis. In phylogenetic analyses of small-subunit rRNA gene sequences, Monopylocystis visvesvarai, Sawyeria marylandensis and Psalteriomonas lanterna converge to form a single clade of non-aerobic (anaerobic/microaerophilic) heteroloboseans.     

Genetics and Morphology Characterize the Dinoflagellate Symbiodinium voratum,n. sp., (Dinophyceae) as the Sole Representative of Symbiodinium Clade E

Dinoflagellates in the genus Symbiodinium are ubiquitous in shallow marine habitats where they commonly exist in symbiosis with cnidarians. Attempts to culture them often retrieve isolates that may not be symbiotic, but instead exist as free‐living species. In particular, cultures of Symbiodinium clade E obtained from temperate environments were recently shown to feed phagotrophically on bacteria and microalgae. Genetic, behavioral, and morphological evidence indicate that strains of clade E obtained from the northwestern, southwestern, and northeastern temperate Pacific Ocean as well as the Mediterranean Sea constitute a single species: Symbiodinium voratum n. sp. Chloroplast ribosomal 23S and mitochondrial cytochrome b nucleotide sequences were the same for all isolates. The D1/D2 domains of nuclear ribosomal DNA were identical among Western Pacific strains, but single nucleotide substitutions differentiated isolates from California (USA) and Spain. Phylogenetic analyses demonstrated that S. voratum is well‐separated evolutionarily from other Symbiodinium spp. The motile, or mastigote, cells from different cultures were morphologically similar when observed using light, scanning, and transmission electron microscopy; and the first complete Kofoidian plate formula for a Symbiodinium sp. was characterized. As the largest of known Symbiodinium spp., the average coccoid cell diameters measured among cultured isolates ranged between 12.2 (± 0.2 SE) and 13.3 (± 0.2 SE) μm. Unique among species in the genus, a high proportion (approximately 10–20%) of cells remain motile in culture during the dark cycle. Although S. voratum occurs on surfaces of various substrates and is potentially common in the plankton of coastal areas, it may be incapable of forming stable mutualistic symbioses.     

Characterization of three endophytic, indole-3-acetic acid-producing yeasts occurring in Populus trees

Three endophytic yeast, one isolated from stems of wild cottonwood (Populus trichocarpa), two from stems of hybrid poplar (P. trichocarpa × Populus deltoides), were characterized by analyzing three ribosomal genes, the small subunit (18S), internal transcribed spacer (ITS), and D1/D2 region of the large subunit (26S). Phenotypic characteristics of the yeast isolates were also obtained using a commercial yeast identification kit and used for assisting the species identification. The isolate from wild cottonwood was identified to be closest to species Rhodotorula graminis. The two isolates from hybrid poplar were identified to be species Rhodotorula mucilaginosa. In addition, the three yeast isolates were observed to be able to produce indole-3-acetic acid (IAA), a phytohormone which can promote plant growth, when incubated with l-tryptophan. To our knowledge, the yeast strains presented in this study were the first endophytic yeast strains isolated from species of Populus.     

Rhodopseudomonas globiformis,sp. n., a new species of the Rhodospirillaceae

Enrichments of pH: 5.1, inoculated from a warm, acidic sulfur spring of the Yellowstone Park, yielded purple red cultures of a mobile, spherical organism belonging to the Rhodospirillaceae, described herein as a new species Rhodopseudomonas globiformis. Under favourable conditions, the cells are 1.6–1.8 μm in diameter, smaller and larger cells may occur. The photopigments consist of bacterio-chlorophyll a_P and new aliphatic methoxylated ketocarotenoids. The organism grows either under anaerobic conditions in the light or under microaerophilic conditions in the dark. Biotin, p-aminobenzoic acid and a source of reduced sulfur are required as growth factors. Gluconate, mannitol, fructose and ethanol are the best carbon sources at a pH of 4.9. Growth is inhibited by low concentrations of sulfide.     

Prototheca tumulicola sp. nov., a novel achlorophyllous,yeast-like microalga isolated from the stone chamber interior of the Takamatsuzuka Tumulus

Two achlorophyllous microalgal strains were isolated from the soil and white moldy colony collected inside the stone chamber of the Takamatsuzuka Tumulus in Japan. Phylogenetic analyses of the small subunit ribosomal RNA (SSU rRNA) and Dl/D2 large subunit ribosomal RNA (LSU rRNA) gene sequences, and concatenated gene sequences of the SSU and D1/D2 LSU rRNA genes indicated that our two isolates were the members of the non-photosynthetic, yeast-like microalgal Chlorellaceous genus Prototheca (Chlorellales, Trebouxiophyceae, Chlorophyta) but well distinguished from known species. Based on phenotypic and genotypic characteristics, isolates T6713-13-10^T and T61213-7-11 are proposed to represent a novel species in Prototheca, P. tumulicola, with the type strain JCM 31123^T (isolate T6713-13-10^T).     

Characteristics of Mycoplasma meleagridis sp. n., Isolated from Turkeys.

Yamamoto, R. (University of California, Davis), C. H. Bigland, and H. B. Ortmayer. Characteristics of Mycoplasma meleagridis sp. n., isolated from turkeys. J. Bacteriol. 90:47-49. 1965.-A designation is proposed for a pathogenic Mycoplasma species isolated from turkeys. The organism originally was recovered from the air-sac lesion of a turkey poult in 1957, and was designated the "N" strain. Mycoplasma species with identical characteristics have since been recovered from the sinus, trachea, oviduct, vagina, semen, and bursa of Fabricius of turkeys. The organism has been recovered from many turkey flocks throughout the country. Many investigators have confirmed the original finding that this organism is antigenically distinct from other known serotypes of Mycoplasma found in poultry. The species proposed is Mycoplasma meleagridis sp. n.     

Pseudomonas creosotenesis sp. n., a Creosote-tolerant Marine Bacterium

In a study of the marine biological environment in which creosoted pilings are located, a previously unreported species of bacteria was isolated. This species was detected on creosoted piling from 11 widely differing locations and was the predominant species of bacteria found on these piling. The new organism was identified as a gram-negative rod belonging to the genus Pseudomonas and has been named Pseudomonas creosotensis. It has been completely described by the standard morphological and biochemical tests.     

Diversity of culturable actinobacteria isolated from marine sponge <Emphasis Type="Italic">Haliclona</Emphasis> sp.

This study describes actinobacteria isolated from the marine sponge Haliclona sp. collected in shallow water of the South China Sea. A total of 54 actinobacteria were isolated using media selective for actinobacteria. Species diversity and natural product diversity of isolates from marine sponge Haliclona sp. were analysed. Twenty-four isolates were selected on the basis of their morphology on different media and assigned to the phylum Actinobacteria by a combination of 16S rRNA gene based restriction enzymes digestion and 16S rRNA gene sequence analysis. The 16S rRNA genes of 24 isolates were digested by restriction enzymes TaqI and MspI and assigned to different groups according to their restriction enzyme pattern. The phylogenetic analysis based on 16S rRNA gene sequencing showed that the isolates belonged to the genera Streptomyces, Nocardiopsis, Micromonospora and Verrucosispora; one other isolate was recovered that does not belong to known genera based on its unique 16S rRNA gene sequence. To our knowledge, this is the first report of a bacterium classified as Verrucosispora sp. that has been isolated from a marine sponge. The majority of the strains tested belong to the genus Streptomyces and three isolates may be new species. All of the 24 isolates were screened for genes encoding polyketide synthases (PKS) and nonribosomal peptide synthetases (NRPS). PKS and NRPS sequences were detected in more than half of the isolates and the different "PKS-I-PKS-II-NRPS" combinations in different isolates belonging to the same species are indicators of their potential natural product diversity and divergent genetic evolution.     

Mesorhizobial strains nodulating Anagyris latifolia and Lotus berthelotii in Tamadaya ravine (Tenerife, Canary Islands) are two symbiovars of the same species, Mesorhizobium tamadayense sp. nov

Barranco de Tamadaya is a deep ravine located in southern Tenerife, which is included within a protected area where several endemic plants grow. Among them, two legumes are catalogued as critically endangered, Anagyris latifolia and Lotus berthelotii. Rhizobial strains isolated from their root nodules grown in soil samples from this ravine harboured symbiotic genes belonging to two distant symbiovars, but they shared identical 16S rRNA gene sequences (rrs). The phylogeny based on the rrs sequences placed these isolates in a separate subbranch that did not include any of the currently recognised Mesorhizobium species, but the resolution of the ribosomal tree did not permit further taxonomic conclusions. Nevertheless, multilocus sequence analysis (MLSA) of four housekeeping genes (atpD, recA, glnII and dnaK) and the rrs gene generated a highly supported Bayesian phylogeny, identifying these isolates as a new Mesorhizobium lineage. DNA-DNA hybridisation homology percentages were lower than 30% compared to type strains of the closest related species, and supported the phylogenetic data. Phenotypic characterisation also distinguished this lineage from the other closest Mesorhizobium species. The polyphasic approach thus confirmed that the isolates represented a novel species for which we propose the name Mesorhizobium tamadayense sp. nov. The type strain is Ala-3(T) (CECT 8040(T), LMG 26736(T)).     

Thraustochytrium gaertnerium sp. nov.: a new thraustochytrid stramenopilan protist from mangroves of Goa, India

Thraustochytrids are ubiquitous, chemo-organotrophic, marine stramenipilan protists belonging to the class Labyrinthulomycetes. Their taxonomy is largely based on life cycle development stages. We describe here a new species of thraustochytrid isolated from mangroves of Goa, India. The organism is characterized by large zoosporangia with two distinct development cycles. In one, typical thalli with ectoplasmic net elements mature into zoosporangia that divide to form heterokont biflagellate zoospores, leaving behind a proliferation body. In the second type, the thalli develop into amoeboid cells, reminiscent of the genus Ulkenia Gaertner. Unlike Ulkenia, however, the 'amoebae' do not immediately produce zoospores, but round up prior to division into zoospores. The two types of development occur simultaneously in single cell-derived in- vitro cultures. Molecular characterization of the new isolate involving 18S rRNA gene typing and comparative phylogenetic analysis further establish it to be a new and distinct thraustochytrid species with Schizochytrium aggregatum Goldstein and Belsky and Thraustochytrium kinnei Gaertner as the closest forms. We have named this new species as Thraustochytrium gaertnerium, deriving its species name in honour of Dr Alwin Gaertner, a pioneer in the studies of taxonomy and ecology of thraustochytrids.     

Tsukubamonas globosa n. gen., n. sp., a novel excavate flagellate possibly holding a key for the early evolution in "Discoba"

We report the ultrastructure and phylogenetic position of a free-living heterotrophic flagellate, Tsukubamonas globosa n. gen., n. sp. This flagellate was isolated from a pond in the University of Tsukuba, Japan. Under light microscopy, the spherical vegetative cells were naked and highly vacuolated, and always swam with rotating motion. Electron microscopic observations revealed that T. globosa possessed a ventral feeding groove, which is one of the hallmark characteristics of the supergroup Excavata. The position of T. globosa was unresolved in the small subunit ribosomal RNA phylogeny. On the other hand, a multigene phylogeny using α-tubulin, β-tubulin, actin, heat shock protein 90, and translation elongation factor 2 robustly united T. globosa with members of the "Discoba" clade of Excavata, composed of jakobids, euglenozoans, and heteroloboseans, although the precise position of T. globosa in this clade remained unresolved. Our detailed morphological comparisons elucidated that T. globosa possessed a novel set of morphological features, and could not be classified into any taxa in the Discoba clade. Instead we classified T. globosa into Tsukubamonadidae n. fam. under Tsukubamonadida n. ord.