Cryptococcus lacticolor sp. nov. and Rhodotorula oligophaga sp. nov., novel yeasts isolated from the nasal smear microbiota of Queensland koalas kept in Japanese zoological parks

A total of 515 yeast strains were isolated from the nasal smears of Queensland koalas and their breeding environments in Japanese zoological parks between 2005 and 2012. The most frequent species in the basidiomycetous yeast biota isolated from koala nasal passages was Cryptococcus neoformans, followed by Rhodotorula minuta. R. minuta was the most frequent species in the breeding environments, while C. neoformans was rare. Seven strains representing two novel yeast species were identified. Analyses of the 26S rDNA (LSU) D1/D2 domain and nuclear ribosomal DNA internal transcribed spacer region sequences indicated that these strains represent new species with close phylogenetic relationships to Cryptococcus and Rhodotorula. A sexual state was not found for either of these two novel yeasts. Key phenotypic characters confirmed that these strains could be placed in Cryptococcus and Rhodotorula. The names Cryptococcus lacticolor sp. nov. (type strain TIMM 10013^T = JCM 15449^T = CBS 10915^T = DSM 21093^T, DDBJ/EMBL/Genbank Accession No.; AB375774 (ITS) and AB375775 (26S rDNA D1/D2 region), MycoBank ID; MB 802688, Fungal Barcoding Database ID; 3174), and Rhodotorula oligophaga sp. nov. (type strain TIMM 10017^T = JCM 18398^T = CBS 12623^T = DSM 25814^T, DDBJ/EMBL/Genbank Accession No.; AB702967 (ITS) and AB702967 (26S rDNA D1/D2 region), MycoBank ID; MB 802689, Fungal Barcoding Database ID; 3175) are proposed for these new species.     

On the description of Tisochrysis lutea gen. nov. sp. nov. and Isochrysis nuda sp. nov. in the Isochrysidales, and the transfer of Dicrateria to the Prymnesiales (Haptophyta)

The Isochrysidaceae is a family of non-calcifying organisms within the haptophyte order Isochrysidales. Isochrysis galbana, a species widely used as a food source in aquaculture, is the best-known representative of this family that contains three genera but only six described species. We sequenced partial nuclear small subunit (SSU) and large subunit rDNA and mitochondrial cytochrome oxidase 1 genes of 34 isochrysidacean culture strains (including authentic strains when available) and compared molecular phylogenetic inferences with cytological and ultrastructural observations. The isochrysidaceaen culture strain Isochrysis affinis galbana (Tahiti isolate), widely used in aquaculture and commonly known as T-Iso, is clearly genetically distinct from Isochrysis galbana, despite seemingly being morphologically identical. A strain with a similar ultrastructure to that of Isochrysis galbana except for the lack of body scales had sequences that were more similar to but still distinct from those of Isochrysis galbana. Dicrateria inornata, a species that lacks body scales, is classified within the Isochrysidaceae, but the SSU rDNA sequence of the authentic strain of this species matches that of Imantonia rotunda within another haptophye order, the Prymnesiales. D. inornata and Imantonia rotunda have similar ultrastructure except for the respective absence/presence of scales. These results lead us to propose the erection of one new genus (Tisochrysis gen. nov.) and two new species (Tisochrysis lutea sp. nov. and Isochrysis nuda sp. nov.). D. inornata is reclassified within the Prymnesiales, and Imantonia rotunda is transferred to this genus (Dicrateria rotunda comb. nov.).     

Two new species of the genus Candida in the Zygoascus clade, Candida lundiana sp. nov. and Candida suthepensis sp. nov., isolated from raw honey in Thailand

During a survey of yeasts associated with raw honey collected in Thailand, two strains of the Zygoascus clade were isolated from the Asian cavity-nesting honeybee Apis cerana and the stingless bee Homotrigona fimbriata. Phylogeny based on 26S rDNA D1/D2 sequences placed these yeasts as members of a clade including Candida bituminiphila, Candida patagonica and Candida polysorbophila. The strains of the two novel species, CBS 12271^T and CBS 12270^T, respectively, could be unquestionably distinguished from their relatives by rDNA sequences and other taxonomic characteristics. Therefore, the novel anamorphic species, Candida lundiana sp. nov. (type strain CBS 12271^T = JCM 16823^T) and Candida suthepensis sp. nov. (type strain CBS 12270^T = JCM 16822^T) are described.     

Gilliamella intestini sp. nov., Gilliamella bombicola sp. nov., Gilliamella bombi sp. nov. and Gilliamella mensalis sp. nov.: Four novel Gilliamella species isolated from the bumblebee gut

Spectra of five isolates (LMG 28358^T, LMG 29879^T, LMG 29880^T, LMG 28359^T and R-53705) obtained from gut samples of wild bumblebees of Bombus pascuorum, Bombus lapidarius and Bombus terrestris were grouped into four MALDI-TOF MS clusters. RAPD analysis revealed an identical DNA fingerprint for LMG 28359^T and R-53705 which also grouped in the same MALDI-TOF MS cluster, while different DNA fingerprints were obtained for the other isolates.Comparative 16S rRNA gene sequence analysis of the four different strains identified Gilliamella apicola NCIMB 14804^T as nearest neighbour species. Average nucleotide identity values of draft genome sequences of the four isolates and of G. apicola NCIMB 14804^T were below the 96% threshold value for species delineation and all four strains and G. apicola NCIMB 14804^T were phenotypically distinct. Together, the draft genome sequences and phylogenetic and phenotypic data indicate that the four strains represent four novel Gilliamella species for which we propose the names Gilliamella intestini sp. nov., with LMG 28358^T as the type strain, Gilliamella bombicola sp. nov., with LMG 28359^T as the type strain, Gilliamella bombi sp. nov., with LMG 29879^T as the type strain and Gilliamella mensalis sp. nov., with LMG 29880^T as the type strain.     

Three new beetle-associated yeast species in the Pichia guilliermondii clade

New yeasts in the Pichia guilliermondii clade were isolated from the digestive tract of basidiocarp-feeding members of seven families of Coleoptera. A molecular phylogeny and unique traits placed eight isolates in Candida fermentati and three undescribed taxa in the genus Candida. The new species and type strains are C. smithsonii (type strain NRRL Y-27642^T), C. athensensis (type strain NRRL Y-27644^T), and C. elateridarum (type strain NRRL Y-27647^T). Based on comparison of small-and large-subunit rDNA sequences, C. smithsonii and C. athensensis form a statistically well-supported subclade with P. guilliermondii, C. xestobii, and C. fermentati; C. elateridarum is basal to this subclade.     

Sabulilitoribacter multivorans gen. nov., sp. nov., a polysaccharide-degrading bacterium of the family Flavobacteriaceae isolated from seashore sand

A Gram-negative, aerobic, non-flagellated, non-gliding and rod-shaped bacterial strain, designated M-M16^T, was isolated from seashore sand around a seaweed farm on the South Sea, South Korea, and its taxonomic position was investigated by using a polyphasic study. Strain M-M16^T grew optimally at 30 °C, at pH 7.0–8.0 and in the presence of 2 % (w/v) NaCl. Strain M-M16^T exhibited the highest 16S rRNA gene sequence similarity values to the type strains of Gaetbulibacter lutimaris (96.5 %) and Flaviramulus basaltis (95.8 %). Neighbour-joining and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences revealed that strain M-M16^T clustered with the type strains of Gaetbulibacter species and F. basaltis. Strain M-M16^T contained MK-6 as the predominant menaquinone and iso-C_15:1 G, iso-C_15:0 and iso-C_17:0 3-OH as the major fatty acids. The major polar lipids detected in strain M-M16^T were phosphatidylethanolamine and one unidentified lipid. The DNA G+C content of strain M-M16^T was 37.4 mol%. The phylogenetic and chemotaxonomic data and other phenotypic properties revealed that strain M-M16^T represents a novel genus and species within the family Flavobacteriaceae, for which the name Sabulilitoribacter multivorans gen. nov., sp. nov. is proposed. The type strain of S. multivorans is M-M16^T (= KCTC 32326^T = CCUG 63831^T).     

Leifsonia gen. nov., a genus for 2,4-diaminobutyric acid-containing actinomycetes to accommodate "Corynebacterium aquaticum" Leifson 1962 and Clavibacter xyli subsp. cynodontis Davis et al. 1984

"Corynebacterium aquaticum" was first proposed by Leifson in 1962 but not included in the approved lists of bacterial names in 1980. This species has been left from reclassification of the genus Corynebacterium because of the unusual chemotaxonomic characteristics such as 2,4-diaminobutyric acid (DAB) in the peptidoglycan and menaquinones of MK-10 and MK-11. A close relationship of "C. aquaticum" to the genera Agromyces and Rathayibacter has been pointed out from the viewpoint of chemotaxonomic profiles and phylogeny based on the 16S rDNA sequences. An analysis of DAB isomers of the peptidoglycan distinguished "C. aquaticum" clearly from these genera by possessing both L-DAB and D-DAB. We also found that the type strain of Clavibacter xyli subsp. cynodontis and two strains of amine-decomposing bacteria showed the similar chemotaxonomic features and formed a cluster with "C. aquaticum" in the phylogenetic tree based on 16S rDNA sequences in the family Microbacteriaceae. Considering these results, we propose a new genus Leifsonia to accommodate the four strains. The four species, Leifsonia aquatica sp. nov., nom. rev., comb. nov. (type species, type strain=JCM 1368), Leifsonia shinshuensis sp. nov. (type strain=DB102=JCM 10591), Leifsonia naganoensis sp. nov. (type strain=DB103=JCM 10592), and Leifsonia cynodontis comb. nov. (type strain=JCM 9733=ICMP 8790), were proposed here for the strains.     

Aliarcobacter vitoriensis sp. nov., isolated from carrot and urban wastewater

Two isolates, one recovered from a carrot and another one from urban wastewater, were characterized using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that both isolates clustered together, and were most closely related to Aliarcobacter lanthieri. Multilocus phylogenetic analysis (MLPA) using the concatenated sequences of five housekeeping genes (atpA, gyrA, gyrB, hsp60 and rpoB) suggested that these isolates formed a distinct phylogenetic lineage among the genera derived from the former genus Arcobacter. Whole-genome sequence, in silico DNA-DNA hybridization (isDDH) and the average nucleotide identity (ANI) value between the genome of strain F199^T and those of related species confirmed that these isolates represent a novel species. These strains can be differentiated from its phylogenetically closest species A. lanthieri by its inability to growth on 1% glycine and by their enzyme activity of esterase lipase (C8) and acid phosphatase. Our results, by the application of a polyphasic analysis, confirmed that these two isolates represent a novel species of the genus Aliarcobacter, for which the name Aliarcobacter vitoriensis sp. nov. is proposed. The type strain is F199^T (=CECT 9230^T=LMG 30050^T).     

New species and new combinations in the yeast genera Kregervanrija gen. nov., Saturnispora and Candida

Analysis of nucleotide sequences from the domains 1 and 2 of the large-subunit rDNA demonstrated species of the Pichia fluxuum clade to be phylogenetically isolated, and domains 1 and 2 and internal transcribed spacer rDNA sequence analyses of strains phenotypically identified as P. fluxuum resulted in the discovery of three new species. From this work, the following new genus, new species, and new combinations are proposed: Kregervanrija gen. nov.; type species Kregervanrija fluxuum comb. nov. (type strain NRRL YB-4273, CBS 2287); Kregervanrija delftensis comb. nov. (type strain NRRL Y-7119, CBS 2614); Kregervanrija pseudodelftensis sp. nov. (type strain NRRL Y-5494, CBS 10105); Saturnispora besseyi comb. nov. (NRRL YB-4711, CBS 6343); Saturnispora mendoncae sp. nov. (type strain NRRL Y-11515, CBS 5620); and Candida abiesophila sp. nov. (type strain NRRL Y-11514, CBS 5366).     

Rhizobium croatiense sp. nov. and Rhizobium redzepovicii sp. nov., two new species isolated from nodules of Phaseolus vulgaris in Croatia

Phaseolus vulgaris is a legume indigenous to America which is nodulated by strains of genus Rhizobium in Croatia. Four of these strains, 13T^T, 9T, 18T^T and 8Z are phylogenetically close to the species from the Rhizobium leguminosarum phylogenetic complex in the 16S rRNA gene analysis. The results of both the analyses of the concatenated recA and atpD genes and whole genomes revealed that the strains 13T^T and 9T clustered with Rhizobium sophoriradicis CCBAU 03470^T and the strains 18T^T and 8Z with Rhizobium ecuadorense CNPSO 671^T. Whole genome average nucleotide identity blast (ANIb) and dDDH values between the strains 13T^T and the type strain of R. sophoriradicis and between the strains 18T^T and the type strain of R. ecuadorense were lower than 95% and 70%, respectively, which are the threshold values recommended for bacterial species differentiation. These results combined with those of chemotaxonomic and phenotypic analyses support the affiliation of these strains to two novel species within the genus Rhizobium for which we propose the names Rhizobium croatiense sp. nov. 13T^T (=LMG 32397^T, = HAMBI 3740^T) as type strain and Rhizobium redzepovicii sp. nov. 18T^T (=LMG 32398^T, = HAMBI 3741^T) as type strain.     

Agrobacterium vaccinii sp. nov. isolated from galls on blueberry plants (Vaccinium corymbosum)

Four non-pathogenic strains isolated from the galls on blueberry plants (Vaccinium corymbosum) were characterized by using polyphasic taxonomic methods. Based on 16S rRNA gene phylogeny, strains were clustered within the genus Agrobacterium. Furthermore, multilocus sequence analysis (MLSA) based on the partial sequences of atpD, recA and rpoB housekeeping genes and whole-genome-based phylogeny indicated that the strains studied form a novel Agrobacterium species. Analyses showed that the strains belong to “rubi” sub-clade of Agrobacterium genus and their closest relatives are Agrobacterium rubi and “Agrobacterium bohemicum”. Average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) comparisons between genome sequences of representative strains B7.6^T and B19.1.4, and their closest relatives, confirmed the distinct phylogenetic position of studied strains, because obtained values were considerably below the proposed thresholds for the species delineation. The four strains studied were phenotypically distinguishable from other species of the genus Agrobacterium. Overall, polyphasic characterization showed that the strains studied represent a novel species of the genus Agrobacterium, for which the name Agrobacterium vaccinii sp. nov. is proposed. The type strain of A. vaccinii is B7.6^T (=CFBP 8740^T = LMG 31849^T).     

Tahibacter aquaticus gen. nov., sp. nov., a new gammaproteobacterium isolated from the drinking water supply system of Budapest (Hungary)

Three Gram-stain negative, aerobic, non-motile, non-spore-forming, rod-shaped bacterial strains, PYM5-11^T, RaM5-2 and PYM5-8, were isolated from the drinking water supply system of Budapest (Hungary) and their taxonomic positions were investigated by a polyphasic approach. All three strains grew optimally at 20-28 °C and pH 5-7 without NaCl. The G+C content of the DNA of the type strain was 65.4 mol%. On the basis of 16S rRNA gene sequence analysis, the isolates showed 94.5-94.9% sequence similarity to the type strain of Dokdonella koreensis and a similarity of 93.0-94.1% to the species of the genera Aquimonas and Arenimonas. The major isoprenoid quinone of the strains was ubiquinone Q-8. The predominant fatty acids were iso-C_15:0, iso-C_17:1ω9c, C_16:1ω7c, and C_16:0. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylmonomethylethanolamine, as well as several unidentified aminolipids and phospholipids were present. The 16S rRNA gene sequence analysis, the predominant fatty acids, the polar lipid composition, RiboPrint patterns, physiological and biochemical characteristics showed that the three strains were related but distinct from the type strains of the four recognized species of the genus Dokdonella, and indicated that the strains represented a new genus within the Gammaproteobacteria. The strain PYM5-11 (=DSM 21667^T=NCAIM B 02337^T) is proposed as the type strain of a new genus and species, designated as Tahibacter aquaticus gen. nov., sp. nov.     

Gluconacetobacter maltaceti sp. nov., a novel vinegar producing acetic acid bacterium

Comparison of HaeIII- and HpaII-restriction profiles of PCR-amplified 16S-23S rDNA ITS regions of Gluconacetobacter sp. LMG 1529^T and SKU 1109 with restriction profiles of reference strains of acetic acid bacteria described by Tr?ek and Teuber [34] revealed the same but unique restriction profiles for LMG 1529^T and SKU 1109. Further analyses of nearly complete 16S rRNA gene sequences, nearly complete 16S-23S rDNA ITS sequences, as well as concatenated partial sequences of the housekeeping genes dnaK, groEL and rpoB, allocated both strains to a single phylogenetic cluster well separated from the other species of the genus Gluconacetobacter. DNA–DNA hybridizations confirmed their novel species identity by 73% DNA–DNA relatedness between both strains, and values below the species level (<70%) between SKU 1109 and the type strains of the closest phylogenetic neighbors. The classification of strains LMG 1529^T and SKU 1109 into a single novel species was confirmed also by AFLP and (GTG)₅-PCR DNA fingerprinting data, as well as by phenotypic data. Strains LMG 1529^T and SKU 1109 can be differentiated from their closely related Gluconacetobacter species, Gluconacetobacter entanii and Gluconacetobacter hansenii, by their ability to form 2-keto-d-gluconic acid from d-glucose, their ability to use d-mannitol, d-gluconate and glycerol as carbon source and form acid from d-fructose, and their ability to grow without acetic acid. The major fatty acid of LMG 1529^T and SKU 1109 is C_18:1ω7c (60.2–64.8%). The DNA G + C content of LMG 1529^T and SKU 1109 is 62.5 and 63.3 mol% respectively. The name Gluconacetobacter maltaceti sp. nov. is proposed. The type strain is LMG 1529^T (= NBRC 14815^T = NCIMB 8752^T).     

Saccharomyces bacillaris is not a synonym of Candida stellata: reinstatement as Starmerella bacillaris comb. nov.

Torulopsis bacillaris (Kroemer and Krumbholz) Lodder (basionym Saccharomyces bacillaris Kroemer and Krumbholz) was frequently detected in oenological works on yeast ecology conducted in the mid-1950s in different wine regions of the world, before its unification with Torulopsis stellata (Kroemer and Krumbholz) Lodder. Most of the phenotypic characteristics pointed out for T. bacillaris are currently attributed to Candida zemplinina Sipiczki. In the present work isoenzyme profiles and rDNA restriction profiles of the neotype of S. bacillaris from two yeast culture collections (CBS 843 and PYCC 3044) and of the type strain of C. zemplinina (CBS 9494) were determined and similar profiles were detected. Moreover, the sequences of the D1/D2 region of the 26S rRNA gene of the three strains were 100 % identical. Different profiles were observed for the type strain of C. stellata (CBS 157) both for isoenzyme and rDNA restriction analysis and only 91 % similarity was found between the D1/D2 sequence of this strain and that of the neotype of S. bacillaris. In view of the newly obtained data and the fact that all above-mentioned species belong to the Starmerella clade, only distantly related to Candida tropicalis (the type species of the genus), S. bacillaris is hereby reinstated as Starmerella bacillaris comb. nov., with C. zemplinina as an obligate synonym.     

Isoptericola salitolerans sp. nov., a halotolerant filamentous actinobacterium isolated from a salt lake, China

A novel bacterial strain, TRM F109^T, was isolated from hypersaline habitat in Sichuan Province, China. It was a Gram-positive, aerobic, non-motile, halotolerant, filamentous bacterium. Phylogenetic analysis based on 16S rRNA gene sequences revealed levels of similarity of 97.0–98.4 % to the type strains of recognized species of the genus Isoptericola. Chemotaxonomic data also supported the placement of strain TRM F109^T within the genus Isoptericola. The low levels of DNA–DNA relatedness between the novel strain and the type strains of recognized species of the genus Isoptericola, in combination with differential phenotypic data, demonstrate that strain TRM F109^T represents a novel species of the genus Isoptericola, for which the name Isoptericola salitolerans sp. nov. is proposed. The type strain is TRM F109^T (=JCM 15901^T=KCTC 19617^T).     

Systematics of Crepidostomum species from the Russian Far East and northern Japan,with description of a new species and validation of the genus Stephanophiala

Current article touched upon the issue of the complicated taxonomic status of some species from the genus Crepidostomum collected from the freshwater fish in the rivers of Primorsky region, Sakhalin, and Hokkaido Islands. Primary morphological analyses showed affiliation of the worms to the species C. farionis (Müller, 1784) Lühe, 1909; C. metoecus Braun, 1900b; C. chaenogobii Yamaguti and Matsumura, 1942; C. nemachilus Krotov, 1959. We described the new species Crepidostomum achmerovi sp. nov. that is a sibling species of C. nemachilus. Molecular-genetic investigation have shown that C. nemachilus and C. achmerovi sp. nov. are closely related to C. metoecus in both 28S rDNA and cox1 mtDNA markers. Crepidostomum nemachilus forms a separate branch within the C. metoecus clade on the 28S BI tree with strong statistical support and separate clade in relation to C. metoecus clade on the cox1 BI tree. Values of p-distances between Crepidostomum species were at intergeneric level. Crepidostomum metoecus species complex including five species (C. metoecus, C. nemachilus, C. oschmarini, C. brinkmanni, and C. achmerovi sp. nov.) was reconsidered as independent genus Crepidostomum sensu stricto. Minimum Spanning Network showed that C. nemachilus, C. metoecus and C. achmerovi sp. nov. were separated by large number of mutational events and represent independent phyletic lines. An amended diagnosis is provided for the subfamily Crepidostomatinae, the genera Crepidostomum s. str. and Stephanophiala Nicoll, 1909, along with keys to species of both genera.     

Sequence Diversity of the Bacillus thuringiensis and B. cereus Sensu Lato Flagellin (H Antigen) Protein: Comparison with H Serotype Diversity

We set out to analyze the sequence diversity of the Bacillus thuringiensis flagellin (H antigen [Hag]) protein and compare it with H serotype diversity. Some other Bacillus cereus sensu lato species and strains were added for comparison. The internal sequences of the flagellin (hag) alleles from 80 Bacillus thuringiensis strains and 16 strains from the B. cereus sensu lato group were amplified and cloned, and their nucleotide sequences were determined and translated into amino acids. The flagellin allele nucleotide sequences for 10 additional strains were retrieved from GenBank for a total of 106 Bacillus species and strains used in this study. These included 82 B. thuringiensis strains from 67 H serotypes, 5 B. cereus strains, 3 Bacillus anthracis strains, 3 Bacillus mycoides strains, 11 Bacillus weihenstephanensis strains, 1 Bacillus halodurans strain, and 1 Bacillus subtilis strain. The first 111 and the last 66 amino acids were conserved. They were referred to as the C1 and C2 regions, respectively. The central region, however, was highly variable and is referred to as the V region. Two bootstrapped neighbor-joining trees were generated: a first one from the alignment of the translated amino acid sequences of the amplified internal sequences of the hag alleles and a second one from the alignment of the V region amino acid sequences, respectively. Of the eight clusters revealed in the tree inferred from the entire C1-V-C2 region amino acid sequences, seven were present in corresponding clusters in the tree inferred from the V region amino acid sequences. With regard to B. thuringiensis, in most cases, different serovars had different flagellin amino acid sequences, as might have been expected. Surprisingly, however, some different B. thuringiensis serovars shared identical flagellin amino acid sequences. Likewise, serovars from the same H serotypes were most often found clustered together, with exceptions. Indeed, some serovars from the same H serotype carried flagellins with sufficiently different amino acid sequences as to be located on distant clusters. Species-wise, B. halodurans, B. subtilis, and B. anthracis formed specific branches, whereas the other four species, all in the B. cereus sensu lato group, B. mycoides, B. weihenstephanensis, B. cereus, and B. thuringiensis, did not form four specific clusters as might have been expected. Rather, strains from any of these four species were placed side by side with strains from the other species. In the B. cereus sensu lato group, B. anthracis excepted, the distribution of strains was not species specific.