- Volume 62, Issue Pt_6, 2012

A re-evaluation of the taxonomic position of five strains, one assigned to Cronobacter sakazakii (strain 1330T, isolated from spiced meat purchased in Slovakia), two previously assigned to Cronobacter genomospecies 1 (strains NCTC 9529T and 731, isolated from water and a leg infection, respectively) and two previously assigned to Cronobacter turicensis (strains 96 and 1435, isolated from onion powder and rye flour, respectively) was carried out. The analysis included phenotypic characterization, 16S rRNA gene sequencing and multilocus sequence analysis (MLSA) of seven housekeeping genes (atpD, fusA, glnS, gltB, gyrB, infB, ppsA; 3036 bp). 16S rRNA gene sequence analysis and MLSA showed that strain 1330T formed an independent phylogenetic lineage in the MLSA, with Cronobacter dublinensis LMG 23823T as the closest neighbour. DNA–DNA reassociation and phenotypic analysis revealed that strain 1330T represented a novel species, for which the name Cronobacter condimenti sp. nov. is proposed (type strain 1330T = CECT 7863T = LMG 26250T). Strains NCTC 9529T, 731, 96 and 1435 clustered together within an independent phylogenetic lineage, with C. turicensis LMG 23827T as the closest neighbour in the MLSA. DNA–DNA reassociation and phenotypic analysis confirmed that these strains represent a novel species, for which the name Cronobacter universalis sp. nov. is proposed (type strain NCTC 9529T = CECT 7864T = LMG 26249T).

Three Gram-negative, motile and slightly curved rod-shaped bacteria, strains SUEMI03T, SUEMI08T and SUEMI10T, were isolated from an old volcanic mountain soil on Tenerife (Canary Islands). The three strains were related phylogenetically to Herbaspirillum seropedicae . 16S rRNA gene sequence similarity was 99.2–99.6 % among strains SUEMI03T, SUEMI08T and SUEMI10T, which presented 97.5, 97.8 and 97.7 % identity, respectively, with respect to H. seropedicae DSM 6445T. The three strains grew optimally in TSB at 28 °C and contained summed features 3 (C16 : 1ω6c and/or C16 : 1ω7c) and 8 (C18 : 1ω6c and/or C18 : 1ω7c) and C16 : 0 as major cellular fatty acids. The DNA G+C contents of strains SUEMI03T, SUEMI08T and SUEMI10T were 61.6, 60.4 and 61.9 mol%, respectively. Strains SUEMI03T, SUEMI08T and SUEMI10T presented less than 60 % interstrain DNA relatedness and less than 30 % relatedness with respect to H. seropedicae DSM 6445T. In spite of their common geographical origin, the three strains isolated in this study presented several phenotypic differences, presenting phenotypic profiles highly divergent from that of H. seropedicae . Therefore, we propose that the strains isolated in this study represent three novel species of the genus Herbaspirillum , named Herbaspirillum canariense sp. nov. (type strain SUEMI03T  = LMG 26151T  = CECT 7838T), Herbaspirillum aurantiacum sp. nov. (type strain SUEMI08T  = LMG 26150T  = CECT 7839T) and Herbaspirillum soli sp. nov. (type strain SUEMI10T  = LMG 26149T  = CECT 7840T).

A yellowish pigmented, Gram-negative, rod-shaped, non-spore-forming bacterium (strain CC-TBT-3T), was isolated on marine agar 2216 from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan. 16S rRNA gene sequence analysis of strain CC-TBT-3T showed a relatively low similarity (<95.5 %) to representatives of the genera Novosphingobium , Sphingosinicella and Sphingomonas of the Sphingomonadaceae , with the most related strain being the type strain of Novosphingobium soli . In addition to the relatively low 16S rRNA gene sequence similarity to members of established species, the isolate also showed some unique chemotaxonomic features, including the presence of some glycolipids with unusual chromatographic behaviour. The major components of the polar lipid profile were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid and three unidentified glycolipids. The major respiratory quinone was ubiquinone Q-10. The polyamine pattern was characterized by the triamine sym-homospermidine as a major component. Although the predominant fatty acids were C18 : 1ω7c and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH), the isolate did not show the typical hydroxyl fatty acids, such as C14 : 0 2-OH, C15 : 0 2-OH and C16 : 0 2-OH, found in members of the genera Novosphingobium , Sphingomonas and Sphingosinicella , but showed instead high amounts of C18 : 1 2-OH (12.0 %). The DNA G+C content of strain CC-TBT-3T was 63.4 mol%. 16S rRNA gene sequence, chemotaxonomic and physiological analyses revealed that strain CC-TBT-3T represents a novel species in a new genus in the family Sphingomonadaceae for which the name Sphingomicrobium lutaoense gen. nov., sp. nov. is proposed; the type strain is of the type species S. lutoaense, CC-TBT-3T ( = DSM 24194T = CCM 7794T).

The taxonomic status of a methyl-parathion-degrading strain, OP-1T, isolated from a wastewater-treatment system in China, was determined using a polyphasic approach. The rod-shaped cells were Gram-staining-negative, non-spore-forming and non-motile. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the novel strain belonged to the genus Burkholderia , as it appeared closely related to Burkholderia glathei ATCC 29195T (97.4 % sequence similarity), Burkholderia sordidicola KCTC 12081T (96.5 %) and Burkholderia bryophila LMG 23644T (96.3 %). The major cellular fatty acids, C16 : 0, C17 : 0 cyclo and C18 : 1ω7c, were also similar to those found in established members of the genus Burkholderia . The genomic DNA G+C content of strain OP-1T was 59.4 mol%. The level of DNA–DNA relatedness between the novel strain and the closest recognized species, Burkholderia glathei ATCC 29195T, was only 30 %. Based on the phenotypic, genotypic and phylogenetic evidence, strain OP-1T represents a novel species of the genus Burkholderia , for which the name Burkholderia zhejiangensis sp. nov. is proposed. The type strain is OP-1T ( = CCTCC AB 2010354T = KCTC 23300T).

A novel Gram-negative, aerobic, catalase- and oxidase-positive, non-sporulating, non-motile, rod-shaped bacterium, designated strain UST081027-248T, was isolated from seawater of the Red Sea. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain UST081027-248T fell within the genus Erythrobacter . Levels of 16S rRNA gene sequence similarity between the novel strain and the type strains of Erythrobacter species ranged from 95.3 % (with Erythrobacter gangjinensis ) to 98.2 % (with Erythrobacter citreus ). However, levels of DNA–DNA relatedness between strain UST081027-248T and the type strains of closely related species were below 70 %. Optimal growth of the isolate occurred in the presence of 2.0 % NaCl, at pH 8.0–9.0 and at 28–36 °C. The isolate did not produce bacteriochlorophyll a. The predominant cellular fatty acids were C17 : 1ω6c, summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) and C15 : 0 2-OH. The genomic DNA G+C content of strain UST081027-248T was 60.4 mol%. Phenotypic properties and phylogenetic distinctiveness clearly indicated that strain UST081027-248T represents a novel species of the genus Erythrobacter , for which the name Erythrobacter pelagi sp. nov. is proposed. The type strain is UST081027-248T ( = JCM 17468T = NRRL 59511T).

A Gram-stain-negative, non-motile, ovoid or rod-shaped bacterial strain, designated L-6T, was isolated from seawater of Baekdo harbour of the East Sea in Korea and its taxonomic position was investigated by using a polyphasic study. Strain L-6T grew optimally at 30 °C, at pH 7.5–8.0 and in the presence of 2 % (w/v) NaCl. In the neighbour-joining phylogenetic tree based on 16S rRNA gene sequences, strain L-6T formed a cluster with the type strain of Celeribacter neptunius at a bootstrap resampling value of 100 %. Strain L-6T exhibited 16S rRNA gene sequence similarity values of 97.7 % to C. neptunius H 14T and of less than 96.2 % to the type strains of other species used in the phylogenetic analysis. The G+C content of the chromosomal DNA of strain L-6T was 60.9 mol%. The predominant ubiquinone found in strain L-6T and C. neptunius CIP 109922T was ubiquinone-10 (Q-10). The predominant fatty acid of strain L-6T and C. neptunius CIP 109922T was C18 : 1ω7c. The major polar lipids of strain L-6T were phosphatidylglycerol, one unidentified aminolipid and one unidentified lipid. The mean level of DNA–DNA relatedness between strain L-6T and C. neptunius CIP 109922T was 17 %. Differential phenotypic properties, together with phylogenetic and genetic distinctiveness, demonstrated that strain L-6T is distinguishable from C. neptunius . On the basis of the data presented, strain L-6T is considered to represent a novel species of the genus Celeribacter , for which the name Celeribacter baekdonensis sp. nov. is proposed. The type strain is L-6T ( = KCTC 23497T  = CCUG 60799T).

Pasteurella species B has so far only been reported from the oral cavity of dogs, cats and a ferret. In the present study, information from 15 recent isolates from different sources, including African hedgehogs (Atelerix albiventris), banded mongoose (Mungos mungo), Moholi bushbabies (Galago moholi) and pneumonia of a cat, were compared to five strains investigated previously from bite wounds in humans inflicted by a cat and dog and from gingiva of a cat. rpoB gene sequence comparison showed that 17 isolates, including the reference strain (CCUG 19794T), had identical sequences, whereas two were closely related and demonstrated 97.9 and 99.6 % similarity to strain CCUG 19794T, respectively; the type strain of Pasteurella stomatis was the most closely related strain, with 92.3 % similarity. This is within the mean range (76–100 %) of rpoB gene sequence similarity between species of the same genus within the family Pasteurellaceae . 16S rRNA gene sequencing of four strains selected based on rpoB sequence comparison showed at least 99.7 % similarity between strains of Pasteurella species B, with 96.2 % similarity to the type strain of the closest related species ( Pasteurella canis ), indicating that Pasteurella species B should have separate species status. Separate species status was also documented when recN sequence comparisons were converted to a genome similarity of 93.7 % within Pasteurella species B and 59.0 % to the type strain of the closest related species ( P. canis ). Based on analysis of the phylogenetic and phenotypic data, and since most isolates originate from the oral cavities of a diverse group of animals, it is suggested that these bacteria be classified as Pasteurella oralis sp. nov.; the type strain is P683T ( = CCUG 19794T = CCM 7950T = strain 23193T = MCCM 00102T), obtained from a cat. Previous reports of the type strain have shown ubiquinone-8, demethylmenaquinone-8 and menaquinone-8 as the major quinones. Polyamines in the type strain were reported as diaminopropane, putrescine, cadaverine, sym-norspermidine, spermidine and spermine in a previous investigation, and the major fatty acids of the type strain were reported to be C16 : 0, C16 : 1ω7c and C14 : 0, with minor amounts of C18 : 0 and C18 : 1ω9c. The DNA G+C content of the type strain has been reported to be 40.0 mol%.

Strains of the recently proposed species Bacteroides chinchillae share more than 99.4 % 16S rRNA gene sequence similarity with the type strain of Bacteroides sartorii although these two species do not appear to be similar from their published descriptions. The aim of this study was to perform phenotypic and genetic analyses of both species to clarify their taxonomic position. B. chinchillae JCM 16497T exhibited high hsp60 gene sequence similarity with B. sartorii JCM 17136T (100 %) as well as B. chinchillae JCM 16498 (100 %). The hsp60 gene sequence analysis and levels of DNA–DNA relatedness observed demonstrated B. sartorii JCM 17136T, B. chinchillae JCM 16497T, and B. chinchillae JCM 16498 are members of a single species. Based on these data, we propose Bacteroides chinchillae as a later heterotypic synonym of Bacteroides sartorii . An emended description of B. sartorii is provided.

A Gram-stain-negative, aerobic, non-flagellated, non-spore-forming, motile (by gliding) bacterial strain, designated M-M6T, was isolated from marine sand of Geoje island, Korea. Strain M-M6T grew optimally at 25 °C, at pH 7.0–8.0 and in the presence of 2 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain M-M6T fell within the clade comprising Cellulophaga species, forming a coherent cluster with Cellulophaga lytica ATCC 23178T and Cellulophaga fucicola NN015860T, with which it shared 16S rRNA gene sequence similarities of 98.1 and 98.2 %, respectively. Sequence similarities between strain M-M6T and the type strains of other recognized Cellulophaga species were in the range 92.4–93.8 %. Strain M-M6T contained MK-6 as the predominant menaquinone and iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH, and C16 : 1ω7c and/or iso-C15 : 0 2-OH as the major fatty acids. The major polar lipids detected in strain M-M6T and the type strains of C. lytica and C. fucicola were two unidentified lipids, one unidentified aminolipid and one unidentified aminophospholipid. The DNA G+C content of strain M-M6T was 35.4 mol%. Levels of DNA–DNA relatedness between strain M-M6T and C. lytica JCM 8516T and C. fucicola JCM 21778T were 33 and 35 %, respectively. Differential phenotypic properties and phylogenetic and genetic distinctiveness distinguished strain M-M6T from all recognized Cellulophaga species. On the basis of the data presented, strain M-M6T is considered to represent a novel species of the genus Cellulophaga , for which the name Cellulophaga geojensis sp. nov. is proposed. The type strain is M-M6T ( = KCTC 23498T = CCUG 60801T).

Three Gram-staining-negative non-endospore-forming strains were isolated from farmed fish in Chile: one (LM-09-FpT) from a rainbow trout (Oncorhynchus mykiss) and the others (LM-19-FpT and LM-20-Fp) from two Atlantic salmon (Salmo salar). Phylogenetic analyses based on 16S rRNA gene sequences indicated that all three isolates belonged to the genus Flavobacterium . In these analyses, strain LM-09-FpT appeared most closely related to the type strains of Flavobacterium chungangense (98.5 % sequence similarity), Flavobacterium glaciei (98.2 %), Flavobacterium aquidurense (97.6 %), Flavobacterium saccharophilum (97.6 %) and Flavobacterium hercynium (97.6 %). The 16S rRNA gene sequences of strains LM-19-FpT and LM-20-Fp were found to be identical and most similar to the corresponding sequences of the type strains of Flavobacterium aquidurense (98.6 %), Flavobacterium frigidimaris (98.5 %), Flavobacterium hercynium (97.9 %), Flavobacterium saccharophilum (97.7 %) and Flavobacterium pectinovorum (97.7 %). For each of the three novel strains, menaquinone (MK-6) was the predominant respiratory quinone and the major compounds in the polar lipid profile were phosphatidylethanolamine, an unidentified aminolipid, phosphatidylserine and two or three unknown lipids. The fatty acid profile of each strain, which comprised major amounts of iso-C15 : 0, C15 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) as well as smaller amounts of various hydroxylated fatty acids (e.g. iso-C16 : 0 3-OH, iso-C17 : 0 3-OH, C16 : 0 3-OH and C15 : 0 3-OH), indicated that each belonged to the genus Flavobacterium . Based on their physiological and biochemical characteristics and the results of DNA–DNA hybridizations, which showed relatively low levels of relatedness between the novel strains and the most closely related Flavobacterium species, strain LM-09-FpT ( = LMG 26360T = CCM 7940T) represents a novel species within the genus Flavobacterium , for which the name Flavobacterium chilense sp. nov. is proposed, and strains LM-19-FpT ( = LMG 26359T = CCM 7939T) and LM-20-Fp ( = LMG 26331) represent a second novel species within the same genus, for which the name Flavobacterium araucananum sp. nov. is proposed.

A Gram-stain-positive, strictly aerobic, spherical, non-motile red-pigmented bacterial strain, designated MJ27T, was isolated from a sludge sample of the Daejeon sewage disposal plant in South Korea. A polyphasic approach was used to study the taxonomic position of strain MJ27T. Strain MJ27T shared highest 16S rRNA gene sequence similarity with Deinococcus grandis DSM 3963T (98.8 %), Deinococcus caeni Ho-08T (97.5 %) and Deinococcus aquaticus PB314T (96.6 %.); levels of sequence similarity with the type strains of other Deinococcus species were less than 96.0 %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain MJ27T belonged to the clade formed by members of the genus Deinococcus in the family Deinococcaceae . The G+C content of the genomic DNA of strain MJ27T was 67.6 mol%. The chemotaxonomic characteristics of strain MJ27T were typical of members of the genus Deinococcus , with MK-8 as the predominant respiratory quinone, C16 : 1ω7c, C15 : 1ω6c, C16 : 0 and C15 : 0 as major fatty acids (>12 %), ornithine as the diamino acid in the cell-wall peptidoglycan and resistance to gamma radiation [D10 (dose required to reduce the bacterial population by tenfold) >9 kGy]. The low levels of DNA–DNA relatedness reported here (5.3±1.5–29.2±2.3 %) indicate that strain MJ27T represents a species that is separate from its closest relatives in the genus Deinococcus . On the basis of phylogenetic inference, fatty acid profile and other phenotypic properties, strain MJ27T is considered to represent a novel species of the genus Deinococcus , for which the name Deinococcus daejeonensis sp. nov. is proposed. The type strain is MJ27T ( = KCTC 13751T = JCM 16918T).

Six strains with the typical characteristics of mycoplasmas were isolated from the tracheae of six Canarian Egyptian vultures (Neophron percnopterus majorensis). The results of biochemical, serological and molecular genetic studies showed that the isolates were nearly identical and that they could be considered as representing a novel species of the genus Mycoplasma . Colonies possessed the typical fried-egg appearance and electron micrographs revealed a pleomorphic cellular morphology with the lack of a cell wall. The isolates hydrolysed arginine and required sterol for growth but did not ferment glucose or hydrolyse urea. We propose that the isolates be assigned to a novel species,Mycoplasma neophronis sp. nov. The type strain is G.A.T ( = DSM 24097T = ATCC BAA-2157T). The antiserum of strain G.A.T has been deposited in the Mollicutes collection at Purdue University (Indiana, USA).

Strain SulfLac1T, a thermophilic, anaerobic and slightly halophilic, rod-shaped bacterium with a sheath-like outer structure (toga), was isolated from a whey digester in Tunisia. The strain’s non-motile cells measured 3–30×1 µm and appeared singly, in pairs or as long chains. The novel strain reduced thiosulfate and elemental sulfur, but not sulfate or sulfite, into sulfide. It grew at 37–65 °C (optimum 55 °C), at pH 6.5–7.9 (optimum pH 6.9) and with 0.2–3 % (w/v) NaCl (optimum 0.5 %). The G+C content of the strain’s genomic DNA was 33.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SulfLac1T was most closely related to Petrotoga mobilis (91.4 % sequence similarity). Based on phenotypic, phylogenetic and chemotaxonomic evidence, strain SulfLac1T represents a novel species of a new genus within the order Thermotogales , for which the name Defluviitoga tunisiensis gen. nov., sp. nov. is proposed. The type strain of the type species is SulfLac1T ( = DSM 23805T  = JCM 17210T).

The majority of environmental micro-organisms identified with the rRNA approach have never been visualized. Thus, their reliable classification and taxonomic assignment is often difficult or even impossible. In our preliminary 18S rRNA gene sequencing work from the world’s largest anoxic marine environment, the Cariaco Basin (Caribbean Sea, Venezuela), we detected a ciliate clade, designated previously as CAR_H [Stoeck, S., Taylor, G. T. & Epstein, S. S. (2003). Appl Environ Microbiol 63, 5656–5663]. Here, we combine the traditional rRNA detection method of fluorescent in situ hybridization (FISH) with scanning electron microscopy (SEM) and confirm the phylogenetic separation of the CAR_H sequences from all other ciliate classes by showing an outstanding morphological feature of this group: a unique, archway-shaped kinety surrounding the oral apparatus and extending to the posterior body end in CAR_H cells. Based on this specific feature and the molecular phylogenies, we propose a novel ciliate class, Cariacotrichea nov. cl.

Three apiculate yeast strains, EJ7M09T, GJ5M15 and GJ15M04, isolated from mushrooms in Taiwan were found to represent a novel species of the genus Kloeckera. The phylogenetically closest relative of this novel species is Hanseniaspora occidentalis, but the type strain of H. occidentalis differed by 4.6 % divergence (25 substitutions; 5 gaps) in the sequence of the D1/D2 domain of the large subunit rRNA gene. This difference clearly suggests that the three strains represent a distinct species. As none of the strains that were examined in this study produced ascospores or exhibited conjugation on common sporulation medium either alone or in a pairwise mixture, this species could be considered as an anamorphic member of the genus Hanseniaspora, and a novel species, Kloeckera taiwanica sp. nov., is proposed, with EJ7M09T ( = BCRC 23182T = CBS 11434T) as the type strain.

Five strains of a novel yeast species were isolated from rotting wood samples collected in an Amazonian forest site in the state of Roraima, northern Brazil. The sequences of the D1/D2 domains of the large subunit of the rRNA gene showed that this species belongs to the Scheffersomyces clade and is related to Candida coipomoensis, Candida lignicola and Candida queiroziae. The novel species Candida amazonensis sp. nov. is proposed to accommodate these isolates. The type strain of C. amazonensis sp. nov. is UFMG-HMD-26.3T ( = CBS 12363T = NRRL Y-48762T).