- Volume 56, Issue 5, 2006

A gas vacuolate bacterium, designated strain 37T, was isolated from a sea ice core collected from Point Barrow, Alaska, USA. Cells of strain 37T were large (6–14 μm in length), rod-shaped, contained gas vacuoles of two distinct morphologies, and grew well at NaCl concentrations of 1–10 % and at temperatures of −12 to 10 °C. The DNA G+C content was 40 mol%. Whole-cell fatty acid analysis showed that 16 : 1ω7c comprised 67 % of the total fatty acid content. Phylogenetic analysis of 16S rRNA gene sequences indicated that this bacterium was closely related to members of the genus Psychromonas, with highest sequence similarity (96.8 %) to Psychromonas antarctica. Phenotypic analysis differentiated strain 37T from P. antarctica on the basis of several characteristics, including cell morphology, growth temperature range and the ability to hydrolyse polymers. DNA–DNA hybridization experiments revealed a level of relatedness of 37 % between strain 37T and P. antarctica, providing further support that it represents a distinct species. The name Psychromonas ingrahamii sp. nov. is proposed for this novel species. The type strain is 37T (=CCUG 51855T=CIP 108865T).

A novel alkalitolerant, sulphate-reducing bacterium (strain RT2T) was isolated from alkaline district heating water. Strain RT2T was a motile vibrio (0.5–0.8 μm wide and 1.4–1.9 μm long) and grew at pH 6.9–9.9 (optimum at pH 9.0–9.4) and at 16–47 °C (optimum at 43 °C). The genomic DNA G+C content was 64.7 mol%. A limited number of compounds were used as electron donors with sulphate as electron acceptor, including lactate, pyruvate, formate and hydrogen/acetate. Sulphite and thiosulphate also served as electron acceptors. Based on physiological and genotypic properties, the isolate was considered to represent a novel species of the genus Desulfovibrio, for which the name Desulfovibrio alkalitolerans sp. nov. is proposed. The type strain is RT2T (=DSM 16529T=JCM 12612T). The strain is the first alkali-tolerant member of the genus Desulfovibrio to be described.

A polyphasic taxonomic study was carried out to determine the taxonomic position of a newly isolated denitrifying bacterium, designated Slu-05T, which had been isolated from sludge from the main aerobic treatment tanks of a municipal sewage treatment plant. Phylogenetic analysis based on comparative 16S rRNA gene sequencing indicated that strain Slu-05T was closely related to Azonexus fungiphilus LMG 19178T (96.4 % sequence similarity), the sole species in the genus Azonexus. Strain Slu-05T comprised Gram-negative, motile, non-spore-forming and slightly curved rods. The predominant respiratory lipoquinone was Q-8. The major fatty acids were C16 : 1 ω7c, C16 : 0, C18 : 1 isomers and C10 : 0 3-OH. The G+C content of the genomic DNA was 65.6 mol%. The results of DNA–DNA hybridization (15.6 %) together with phenotypic determination showed that strain Slu-5T could be distinguished from A. fungiphilus. Moreover, some phenotypic properties concerning enzyme activity, the substrates utilized as carbon sources and growth conditions distinguish strain Slu-5T from A. fungiphilus. On the basis of the results obtained in this study, Slu-05T (=DSM 17719T=KCTC 12530T=CCBAU 10199T) is the type strain of a novel species of Azonexus, for which the name Azonexus caeni sp. nov. is proposed.

A halotolerant bacterium was isolated from a saline lake located in Mallorca, Spain. Cells of the strain, designated MACL01T, were Gram-negative, rod-shaped and motile by means of polar flagella. Colonies of strain MACL01T were white to cream in TSA medium, turning brown after 7 days of incubation; they were blue in thiosulphate/citrate/bile salts/sucrose agar medium. A neighbour-joining phylogenetic analysis based on 16S rRNA gene sequences showed that strain MACL01T belongs to the genus Photobacterium, in which it forms a distinct lineage together with Photobacterium rosenbergii and Photobacterium ganghwense (showing 96.9 and 96.2 % similarity, respectively). The most closely related taxon according to phylogenetic analysis of the rpoA gene is also P. rosenbergii (90 % similarity). The recA gene also showed low similarity (83.7, 83.4 and 82.4 %, respectively) with respect to those of Vibrio proteolyticus LMG 3772T, Photobacterium leiognathii LMG 4228T and P. rosenbergii LMG 22223T. Neighbour-joining phylogenetic analysis of the rpoA and recA genes confirms that strain MACL01T belongs to the genus Photobacterium, forming a branch together with P. rosenbergii. Strain MACL01T was able to grow in 0–8 % NaCl. Growth occurred between 4 and 37 °C (optimum, 28 °C) and at pH 5–8.5. Luminescence was negative on marine agar. Strain MACL01T was found to be sensitive to the vibriostatic agent O/129. It reduced nitrate to nitrite, produced β-galactosidase and hydrolysed gelatin, but did not produce arginine dihydrolase, indole or acetoin. Strain MACL01T used several carbohydrates and fermented glucose, l-arabinose and sucrose. The most abundant fatty acids were summed feature 3 (32.6 %; comprising C16 : 1 ω7c and/or C15 : 0 iso 2-OH), C16 : 0 (21.2 %) and C18 : 1 ω7c (19.9 %). The G+C content of the genomic DNA was 49.8 mol%. On the basis of genotypic, phenotypic, chemotaxonomic and phylogenetic results, strain MACL01T (=LMG 22194T=CECT 5860T) should be classified as the type strain of a novel species of the genus Photobacterium, for which the name Photobacterium halotolerans sp. nov. is proposed.

A Gram-negative, non-motile, non-spore-forming, slightly halophilic bacterial strain, DSW-74T, was isolated from sea water off the island of Dokdo, Korea, and its taxonomic position was investigated by a polyphasic study. Strain DSW-74T grew optimally at 37 °C and in the presence of 2 % (w/v) NaCl. It contained Q-10 as the predominant ubiquinone and C17 : 1 ω6c and C18 : 1 ω7c as the major fatty acids. Its DNA G+C content was 65.8 mol%. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain DSW-74T was most closely related to Porphyrobacter species. Similarity values between the 16S rRNA gene sequence of strain DSW-74T and those of the type strains of recognized Porphyrobacter species and of Erythromicrobium ramosum were in the range 97.4–98.7 %. Strain DSW-74T exhibited 16S rRNA gene sequence similarity values of <97.5 % to recognized Erythrobacter species and the other species used in the phylogenetic analysis. DNA–DNA relatedness levels and differential phenotypic properties made it possible to categorize strain DSW-74T as representing a novel Porphyrobacter species. On the basis of the taxonomic data presented, it is proposed that DSW-74T (=KCTC 12395T=DSM 17193T) should be classified in the genus Porphyrobacter as the type strain of a novel species, Porphyrobacter dokdonensis sp. nov.

An aerobic, mesophilic bacterium, strain JA40T, was isolated from soil contaminated with polycyclic aromatic hydrocarbons and polychlorinated biphenyls collected from Johnston Atoll in the North Pacific Ocean. The strain formed yellow-pigmented colonies on heterotrophic media. The cells were Gram-negative, non-motile, non-sporulating rods. The strain reduced nitrite to nitrous oxide, the DNA G+C content was 64 mol% and the dominant fatty acids were 15 : 0 iso, 17 : 1 iso cis7 and 11 : 0 iso 3-OH. DNA sequencing of 1457 nt of the 16S rRNA gene established that JA40T belongs in the genus Pseudoxanthomonas within the Xanthomonadaceae branch of the Gammaproteobacteria. Strain JA40T can be differentiated from other mesophilic species in the genus on the basis of its physiological and biochemical characteristics and distinctive fatty acid profile. Thus strain JA40T (=ATCC BAA-1031T=CIP 108476T) is the type strain of a novel species of the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas kalamensis sp. nov. is proposed.

Two psychrophilic, Gram-negative, rod-shaped, motile bacteria (strains 112T and 102T) that conserved energy from dissimilatory Fe(III) reduction concomitant with acetate oxidation were isolated from permanently cold Arctic marine sediments. Both strains grew at temperatures down to −2 °C, with respective temperature optima of 14 °C and 14–17 °C for strains 112T and 102T. The isolated strains reduced Fe(III) using common fermentation products such as acetate, lactate, propionate, formate or hydrogen as electron donors, and they also grew with fumarate as the sole substrate. As alternatives to Fe(III), they reduced fumarate, S0 and Mn(IV). Based on 16S rRNA gene sequence similarity, strain 112T was most closely related to Desulfuromonas acetoxidans (97.0 %) and Desulfuromonas thiophila NZ27T (95.5 %), and strain 102T to Malonomonas rubra Gra Mal 1T (96.3 %) and Desulfuromusa succinoxidans GylacT (95.9 %) within the Deltaproteobacteria. Strains 112T and 102T therefore represent novel species, for which the names Desulfuromonas svalbardensis sp. nov. (type strain 112T=DSM 16958T=JCM 12927T) and Desulfuromusa ferrireducens sp. nov. (type strain 102T=DSM 16956T=JCM 12926T) are proposed.

The novel species Metschnikowia aberdeeniae is described to accommodate five isolates recovered from insects of morning glory flowers in the Serengeti National Park, Tanzania. On the basis of rDNA ITS and D1/D2 large-subunit sequences, these yeasts form, together with six other isolates, a novel subclade of large-spored Metschnikowia species. The exact position of the subclade within the Metschnikowiaceae cannot be determined with any confidence from these sequences or from small-subunit rDNA sequences, as the variable sites of the sequences are excessively divergent. However, in morphological and physiological terms, the novel isolates are typical of the genus Metschnikowia in general and of the large-spored group in particular. The type strain of Metschnikowia aberdeeniae sp. nov. is strain SUB 05-213.1T (=CBS 10289T=NRRL Y-27921T) (h−) and the allotype is strain SUB 05-213.2 (=CBS 10290=NRRL Y-27922) (h+).

Strains belonging to three novel yeast species, Candida heliconiae (four isolates), Candida picinguabensis (three isolates) and Candida saopaulonensis (two isolates), were recovered in the year 2000 from water of flower bracts of Heliconia velloziana L. Emigd. (Heliconiaceae) found in a forest ecosystem site in an Atlantic rainforest of south-eastern Brazil. C. picinguabensis and C. saopaulonensis were nearly identical in morphology and physiology, but sequence divergence in the D1/D2 domain of the large-subunit rDNA indicated that they should be regarded as different species. They belong to the Metschnikowiaceae clade. C. heliconiae had affinities to Pichia mexicana and related species, but was genetically isolated from all currently accepted species in that group. The type strains are C. heliconiae UNESP 00-91C1T (=CBS 10000T=NRRL Y-27813T), C. picinguabensis UNESP 00-89T (=CBS 9999T=NRRL Y-27814T) and C. saopaulonensis UNESP 00-99T (=CBS 10001T=NRRL Y-27815T).

Three anamorphic, ascomycetous yeast strains isolated from plant samples collected in Linzhi District, Tibet, China, were revealed as representing two novel species by 26S rRNA gene D1/D2 domain sequence and physiological property comparisons. The names Candida tibetensis sp. nov. and Candida linzhiensis sp. nov. are proposed for these novel species, with XZ 41-6T (=AS 2.3072T=CBS 10298T) and XZ 92-1T (=AS 2.3073T=CBS 10299T) as the respective type strains. D1/D2 sequence analysis showed that C. tibetensis and C. linzhiensis are closely related to Candida caryicola and Candida sequanensis, respectively.

Two protein-coding genes, actin and translation elongation factor-1α (EF-1α), as well as two ribosomal gene regions, D1/D2 domains of the large subunit and both internal transcribed spacers including the 5.8S gene region, were evaluated regarding their usefulness for reconstruction of phylogenetic relationships in the Hanseniaspora–Kloeckera species group. This included analyses of sequence divergence values, heterogeneity of evolutionary rates and the reliability of the inferred trees. Both protein-coding genes showed greater capacities to resolve at the strain level and between the closely related species of Hanseniaspora–Kloeckera, compared with the ribosomal gene regions. However, to obtain a fully resolved and reliable phylogenetic tree that reflected the biological relationships it was necessary to combine three congruent sequence datasets. The novel species Hanseniaspora pseudoguilliermondii sp. nov. (type strain CBS 8772T) is described as a result of the application of various molecular approaches to delimit species. Furthermore, incongruent gene genealogies of genetically divergent strains of Hanseniaspora occidentalis, as determined by amplified fragment length polymorphism analysis and DNA–DNA reassociation measurements, indicated the presence of two novel varieties, H. occidentalis var. occidentalis (type strain CBS 2592T) and H. occidentalis var. citrica var. nov. (type strain CBS 6783T), which could be distinguished by habitat preference.

A thermotolerant, Gram-positive, aerobic, endospore-forming, acidophilic bacterium (strain Kr1T) was isolated from the pulp of a gold-containing sulfide concentrate processed at 40 °C in a gold-recovery plant (Siberia). Cells of strain Kr1T were straight to slightly curved rods, 0.8–1.2 μm in diameter and 1.5–4.5 μm in length. Strain Kr1T formed spherical and oval, refractile, subterminally located endospores. The temperature range for growth was 20–60 °C, with an optimum at 40 °C. The pH range for growth on medium containing ferrous iron was 1.2–2.4, with an optimum at pH 2.0; the pH range for growth on medium containing S0 was 2.0–5.0, with an optimum at pH 2.5. Strain Kr1T was mixotrophic, oxidizing ferrous iron, S0, tetrathionate or sulfide minerals as energy sources in the presence of 0.02 % yeast extract or other organic substrates. The G+C content of the DNA of strain Kr1T was 48.2±0.5 mol%. Strain Kr1T showed a low level of DNA–DNA reassociation with the known Sulfobacillus species (11–44 %). 16S rRNA gene sequence analysis revealed that Kr1T formed a separate phylogenetic group with a high degree of similarity between the nucleotide sequences (98.3–99.6 %) and 100 % bootstrap support within the phylogenetic Sulfobacillus cluster. On the basis of its physiological properties and the results of phylogenetic analyses, strain Kr1T can be affiliated to a novel species of the genus Sulfobacillus, for which the name Sulfobacillus thermotolerans sp. nov. is proposed. The type strain is Kr1T (=VKM B-2339T=DSM 17362T).

A novel anaerobic, moderately thermophilic, spore-forming bacterium, designated strain BRT, was isolated from deep-sea hydrothermal core samples collected at the Rainbow vent field on the Mid-Atlantic Ridge (36° 14′ N 33° 54′ W). The cells were found to be rod-shaped, non-motile, Gram-positive and spore-forming. The organism grew in the temperature range 37–60 °C, with an optimum at 55 °C, and at pH values in the range 6–8.5, with an optimum around pH 7. NaCl concentrations for growth were in the range 10–40 g l−1, with an optimum at 20–30 g l−1. Strain BRT grew chemo-organoheterotrophically with carbohydrates, proteinaceous substrates and organic acids with nitrate as electron acceptor. The novel isolate was not able to ferment. The G+C content of the genomic DNA was 34.5 mol%. Phylogenetic analysis of the 16S rRNA gene sequence placed strain BRT in the Bacillaceae within the class ‘Bacilli’. On the basis of the phenotypic and phylogenetic data, this isolate should be described as a member of a novel genus, for which the name Vulcanibacillus gen. nov. is proposed. The type species is Vulcanibacillus modesticaldus sp. nov., with the type strain BRT (=DSM 14931T=JCM 12998T).

A strictly aerobic, rod-shaped bacterium (0.6–0.8×2–3 μm), designated strain Kh10-101T, was isolated from a saltpan (22° 15′ N, 69° 1′ E) in the vicinity of Port Okha, India. The creamish pigmented colonies of strain Kh10-101T were round, flat and translucent with irregular margins and a smooth surface. The strain possessed up to three subpolar flagella, and was motile by a corkscrew motion. The strain grew optimally at 37 °C (temperature growth range 25–40 °C) in a complex glucose-containing medium with 5 % NaCl (NaCl growth range 0–10 %) at pH 9 (pH growth range pH 7–10), indicating that it was a mesophilic halotolerant alkaliphile. The strain was sensitive to lincomycin, meticillin, cefuroxime and cephalexin, but resistant to gentamicin, tetracycline and cotrimazine. Spores were not detected and cells were heat sensitive. The isolate metabolized a range of carbohydrates and hydrolysed casein, gelatin and starch. Growth was not observed on aromatic compounds, Tween 40 or Tween 80. Nitrate was not reduced and catalase was produced. Electron microscopic examination of thin sections revealed a single thick Gram-positive cell wall. The DNA G+C content was 41±1 mol%. Phylogenetic analyses of the 16S rRNA gene sequence revealed that strain Kh10-101T was a member of the sixth rRNA group of the genus Bacillus, which includes alkalitolerant, alkaliphilic and halotolerant species. The halotolerant obligate alkaliphile Bacillus krulwichiae is the closest relative of strain Kh10-101T (96 % similarity) but a number of phenotypic differences suggest that strain Kh10-101T (=JCM 13040T=ATCC BAA-1137T) should be designated the type strain of a new species, for which the name Bacillus okhensis sp. nov. is proposed.

An unidentified strain of the viridans group of streptococci was isolated from a human blood sample. It was distinguished from all other recognized species of the Streptococcus sanguinis group by several biochemical characteristics. Phylogenetic analysis based on 16S rRNA gene sequence comparisons clustered this strain with Streptococcus ferus (mutans group) but phylogenetic analysis based on rpoB and sodA gene sequence comparisons included it in the S. sanguinis group. The isolate showed 95.4 and 95.2 % 16S rRNA gene sequence similarity to S. ferus and S. sanguinis, respectively, confirming it as belonging to a novel taxon, for which the name Streptococcus massiliensis sp. nov. is proposed. The type strain is 4401825T (=CIP 108498T=CCUG 49690T).

Rhizobial bacteria almost exclusively nodulate members of the families Fabaceae, Mimosaceae and Caesalpiniaceae, but are found on a single non-legume taxon, Parasponia (Ulmaceae). Based on their host-range, their nitrogen-fixing ability and strain competition experiments, bacterial strains isolated from Parasponia were thought to constitute a separate lineage that would account for their exceptional host affinity. This hypothesis was investigated by focusing on four isolates that are representative of the morphological and cultural types of Parasponia-nodulating bradyrhizobia. Their evolutionary relationships with other rhizobia were analysed using 16S rRNA gene sequences and their nodulation properties were explored using the nodA gene as a proxy for host-range specificity. Phylogenetic analyses of the 16S rRNA and nodA gene sequences revealed that bacterial isolates from Parasponia species are embedded among other bradyrhizobia. They did not cluster together in topologies based on the 16S rRNA or nodA gene sequences, but were scattered among other bradyrhizobia belonging to either the Bradyrhizobium japonicum or the Bradyrhizobium elkanii lineages. These data suggest that the ability of some bradyrhizobia to nodulate species of the genus Parasponia does not represent a historical relationship that predates the relationship between rhizobia and legumes, but is probably a more recent host switch for some rhizobia.

In order to develop a detection method for the rice pathogens Burkholderia plantarii, Burkholderia glumae and Burkholderia gladioli, the phylogeny of six plant-pathogenic Burkholderia species was analysed using the combined nucleotide sequences of gyrB and rpoD. B. plantarii, B. glumae and B. gladioli formed tight monophyletic branches supported by high bootstrap probabilities. The high sequence similarity revealed a close phylogenetic relationship between B. glumae and B. plantarii. B. plantarii strains were divided into three subclusters comprising rice strains, whereas the single Vanda strain occupied a unique position in the phylogenetic tree. The gyrB and rpoD sequences of all B. glumae strains examined were highly conserved. In contrast, B. gladioli strains demonstrated a far greater sequence diversity, but this diversity did not correlate with pathovar, host plant or geographical origin of the strains. A multiplex-PCR protocol using specific primers from the gyrB sequences was designed that allowed the specific detection and identification of B. plantarii, B. glumae and B. gladioli in rice seeds infected with these pathogenic species.