- Volume 55, Issue 3, 2005

A moderately halophilic, Gram-positive, rod-shaped bacterium (BH030004T) was isolated from a solar saltern in Korea. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BH030004T belonged to the genus Pontibacillus. Chemotaxonomic data (DNA G+C content, 42 mol%; major isoprenoid quinone, MK-7; cell-wall type, A1γ-type meso-diaminopimelic acid; major fatty acids, iso-C15 : 0 and anteiso-C15 : 0) also supported the affiliation of the isolate to the genus Pontibacillus. Although the 16S rRNA gene sequence similarity between strain BH030004T and Pontibacillus chungwhensis DSM 16287T was relatively high (99·1 %), physiological properties and DNA–DNA hybridization (about 7 % DNA–DNA relatedness) allowed genotypic and phenotypic differentiation of strain BH030004T from the type strain of P. chungwhensis. Therefore, strain BH030004T represents a novel species of the genus Pontibacillus, for which the name Pontibacillus marinus sp. nov. is proposed. The type strain is BH030004T (=KCTC 3917T=DSM 16465T).

Thirteen strains of endospore-forming bacteria were isolated from geothermal soils at Cryptogam Ridge, the north-west slope of Mt Melbourne, and at the vents and summit of Mt Rittmann in northern Victoria Land, Antarctica. 16S rRNA gene sequencing, SDS-PAGE and routine phenotypic characterization tests indicated that the seven isolates from the north-west slope of Mt Melbourne represent a novel species of Brevibacillus and that the six isolates from Cryptogam Ridge and the vents and summit of Mt Rittmann represent a novel species of Aneurinibacillus. Brevibacillus strains were not isolated from the sites at Mt Rittmann or Cryptogam Ridge and Aneurinibacillus strains were not isolated from the north-west slope of Mt Melbourne. Preliminary metabolic studies revealed that l-glutamic acid, although not essential for growth, was utilized by both species. The Brevibacillus species possessed an uptake system specific for l-glutamic acid, whereas the Aneurinibacillus species possessed a more general uptake system capable of transporting other related amino acids. Both species utilized a K+ antiport system and similar energy systems for the uptake of l-glutamic acid. The rate of uptake by the Brevibacillus species type strain was 20-fold greater than that shown by the Aneurinibacillus species type strain. The names Brevibacillus levickii sp. nov. and Aneurinibacillus terranovensis sp. nov. are proposed for the novel taxa; the type strains are Logan B-1657T (=LMG 22481T=CIP 108307T) and Logan B-1599T (LMG 22483T=CIP 108308T), respectively.

Eight mycobacterial strains isolated during an 11 year period from the sputum of independent patients with various pulmonary disorders and, in one case, from a lymph node of a young girl, were found to present identical features. Phenotypic and genotypic characteristics revealed that the most closely related species to these test isolates were Mycobacterium triplex and Mycobacterium lentiflavum. However, the lipids of the cell wall of the test isolates differed from those of the latter species by TLC and presented unique profiles by both GC and HPLC. Genotypic analysis showed that they had unique 16S rRNA gene and internal transcribed spacer (ITS) sequences, and could be differentiated from all other mycobacterial strains by PCR restriction analysis of hsp65. The strains presented high resistance to antimycobacterial drugs. The name Mycobacterium florentinum sp. nov. is proposed for this taxon, with strain FI-93171T (=DSM 44852T=CIP 108409T) as the type strain.

Strain Con a/3T is a Gram-positive, motile, endospore-forming, rod-shaped and arsenic-resistant bacterium, which was isolated from a concretion of arsenic ore obtained from a bore-hole. The bacterium grew in the presence of 20 mM arsenate and 0·5 mM arsenite. Diaminopimelic acid was present in the cell wall peptidoglycan, MK-7 was the major menaquinone, and iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0 and C16 : 1(δ7cis) were the major fatty acids. Based on its phenotypic, chemotaxonomic and phylogenetic characteristics, strain Con a/3T was identified as a member of the genus Bacillus. It exhibited maximum similarity (97 %) at the 16S rRNA gene level with Bacillus barbaricus (DSM 14730T); however, the DNA–DNA relatedness value with B. barbaricus was 60 %. Strain Con a/3T also exhibited a number of phenotypic differences from B. barbaricus (DSM 14730T). Strain Con a/3T was therefore identified as representing a novel species of the genus Bacillus, for which the name Bacillus arsenicus sp. nov. is proposed. The type strain is Con a/3T (=MTCC 4380T=DSM 15822T=JCM 12167T).

Strains of a Gram-positive bacterium were isolated from the Irish smear-ripened cheese Gubbeen, and assigned to a new species, Corynebacterium mooreparkense, in 2001. During a further study on the same cheese, no additional isolates from this species could be found. Instead, multiple isolates of its nearest phylogenetic neighbour, Corynebacterium variabile, were found. A first screening with rep-PCR and SDS-PAGE pointed to a similarity between C. mooreparkense and C. variabile. Following this peculiar result, attempts were made to collect all type strains deposited at different culture collections and all strains described by Brennan et al. [Int J Syst Evol Microbiol (2001) 51, 843–852] . Subsequently, 16S rRNA gene sequencing and DNA–DNA hybridizations were performed. All C. mooreparkense strains had a 16S rRNA gene sequence similarity of at least 99·5 % with C. variabile and the DNA–DNA relatedness was 95 %. On the basis of these results, it is concluded that C. mooreparkense is a later heterotypic synonym of C. variabile.

A Gram-positive, aerobic, non-motile, mesophilic strain, designated YIM 70056T, was isolated from a forest soil sample in Yunnan Province, China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that this isolate had less than 97·0 % similarity to any Rhodococcus species with validly published names, with the exception of Rhodococcus fascians (DSM 20669T), which was found to be its closest neighbour (98·9 % similarity). Chemotaxonomic data, including peptidoglycan type, diagnostic sugar compositions, fatty acid profiles, menaquinones, polar lipids and mycolic acids, were determined for this isolate; the results supported the affiliation of strain YIM 70056T to the genus Rhodococcus. The DNA G+C content was 63·5 mol%. The results of DNA–DNA hybridization with R. fascians DSM 20669T, in combination with chemotaxonomic and physiological data, demonstrated that isolate YIM 70056T represents a novel Rhodococcus species, for which the name Rhodococcus yunnanensis sp. nov. is proposed, with YIM 70056T (=CCTCC AA 204007T=KCTC 19021T=DSM 44837T) as the type strain.

A group of slowly growing photochromogenic mycobacteria was isolated from Chesapeake Bay striped bass (Morone saxatilis) during an epizootic of mycobacteriosis. Growth characteristics, acid-fastness and 16S rRNA gene sequencing results were consistent with those of the genus Mycobacterium. Biochemical reactions, growth characteristics and mycolic acid profiles (HPLC) resembled those of Mycobacterium shottsii, a non-pigmented mycobacterium also isolated during the same epizootic. Sequencing of the 16S rRNA genes, the gene encoding the exported repeated protein (erp) and the gene encoding the 65 kDa heat-shock protein (hsp65) and restriction enzyme analysis of the hsp65 gene demonstrated that this group of isolates is unique. Insertion sequences associated with Mycobacterium ulcerans, IS2404 and IS2606, were detected by PCR. These isolates could be differentiated from other slowly growing pigmented mycobacteria by their inability to grow at 37 °C, production of niacin and urease, absence of nitrate reductase, negative Tween 80 hydrolysis and resistance to isoniazid (1 μg ml−1), p-nitrobenzoic acid, thiacetazone and thiophene-2-carboxylic hydrazide. On the basis of this polyphasic study, it is proposed that these isolates represent a novel species, Mycobacterium pseudoshottsii sp. nov. The type strain, L15T, has been deposited in the American Type Culture Collection as ATCC BAA-883T and the National Collection of Type Cultures (UK) as NCTC 13318T.

Clostridium sardiniense Prévot 1938 and Clostridium absonum Nakamura et al. 1973 have long been considered similar in terms of their biological and biochemical properties, but their taxonomic positions have not been clarified by DNA–DNA hybridization studies or rigorous analysis of 16S rRNA genes. In the present study, DNA–DNA hybridization analysis revealed that C. absonum strains DSM 599T, DSM 600 and KZ 1544 shared 83·0–86·3 % DNA relatedness with C. sardiniense DSM 2632T. 16S rRNA gene sequence analysis showed that the C. absonum strains also shared high identity with C. sardiniense DSM 2632T (99·7, 99·3 and 99·8 % for DSM 599T, DSM 600 and KZ 1544, respectively), implying that C. absonum and C. sardiniense are synonyms. In addition, alignment of the inferred amino acid sequences for phospholipase C (PLC) indicated 96·5 % identity between PLCs from C. sardiniense and C. absonum, but relatively low identity with other clostridial species. These results strongly suggest that the species C. sardiniense and C. absonum should be united, with the name C. sardiniense having priority.

Two thermophilic, spore-forming strains, TM1T and TM5, were isolated from deep-sea sediment (4000 m below sea level) of the Ayu Trough in the western Pacific Ocean. Cells of the two strains were Gram-positive, motile and rod-shaped. Their spores were ellipsoidal, subterminal to terminal and occurred in swollen sporangia. The two strains grew at temperatures up to 65 °C and in the pH range 6·5–9·0. The NaCl concentration for optimal growth was 3·0 % (w/v) and growth was inhibited by 5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains TM1T and TM5 belonged to the genus Bacillus, and that strain TM1T was most closely related to Bacillus aeolius DSM 15084T (96·7 %), Bacillus smithii DSM 4216T (96·1 %), Bacillus methanolicus NCIMB 13113T (95·8 %) and Bacillus pallidus DSM 3670T (95·7 %). Between the 16S rRNA gene sequences of strains TM1T and TM5 there were only three nucleotide differences, implying that the two strains were of the same species. The cellular fatty acid profiles of the two strains were also very similar, with iso-C15 : 0, iso-C16 : 0, C16 : 0, iso-C17 : 0 and anteiso-C17 : 0 as the major components. The G+C content of strain TM1T was 38·7 %. On the basis of phenotypic and molecular data, strains TM1T and TM5 represent a novel species of the genus Bacillus, for which the name Bacillus alveayuensis sp. nov. is proposed. The type strain is TM1T (=KCTC 10634T=JCM 12523T).

Morphological, biochemical and molecular genetic studies were carried out on an unknown non-spore-forming, Gram-positive, rod-shaped bacterium that was isolated from dog faeces. The bacterium grew under strictly anaerobic conditions, was asaccharolytic, and possessed a relatively high G+C content of 61 mol%. Phylogenetic analysis based on comparative 16S rRNA gene sequencing showed that the unidentified bacterium was a member of the family Coriobacteriaceae and represents a hitherto unknown subline within the genus Slackia. Based on the presented findings, a novel species, Slackia faecicanis sp. nov., is described. The type strain of Slackia faecicanis is 5WC12T (=CCUG 48399T=CIP 108281T).

Three lactic acid bacterial (LAB) strains obtained from a Malaysian acid-fermented condiment, tempoyak (made from pulp of the durian fruit), showed analogous but distinct patterns after screening by SDS-PAGE of whole-cell proteins and comparison with profiles of all recognized LAB species. 16S rRNA gene sequencing of one representative strain showed that the taxon belongs phylogenetically to the genus Leuconostoc, with its nearest neighbour being Leuconostoc fructosum (98 % sequence similarity). Biochemical characteristics and DNA–DNA hybridization experiments demonstrated that the strains differ from Leuconostoc fructosum and represent a single, novel Leuconostoc species for which the name Leuconostoc durionis sp. nov. is proposed. The type strain is LMG 22556T (=LAB 1679T=D-24T=CCUG 49949T).

Two Gram-positive, rod-shaped, endospore-forming bacteria (strains CR-119T and CR-95T) were isolated from brackish sediments in the mouth of the river Vélez in Málaga, southern Spain, and subjected to a polyphasic taxonomic study. Phenotypic tests showed that these strains were related to other Bacillus species at a similarity level of less than 87·6 %. Both strains are halotolerant, aerobic, chemoheterotrophic, motile with peritrichous flagella and biosurfactant producers. Their endospores are oval, subterminal and non-deforming structures. The predominant menaquinone in both strains is MK-7. The fatty-acid profiles of both strains contain large quantities of branched and saturated fatty acids. The major fatty acids (%) are 15 : 0 anteiso (32·4), 15 : 0 iso (16·8), 17 : 0 iso (13·4), 16 : 0 (11·5) and 17 : 0 anteiso (10·2) in strain CR-119T and 15 : 0 anteiso (37·5), 17 : 0 iso (16·0) and 17 : 0 anteiso (15·8) in strain CR-95T. The G+C contents of strains CR-119T and CR-95T are 41·0 and 42·5 mol%, respectively. RAPD analysis confirmed the low degree of similarity between the two strains and also amongst other Bacillus species. 16S rRNA gene analysis of strain CR-119T showed the highest sequence similarity to be 97·4 %, with Bacillus mojavensis and Bacillus subtilis subsp. spizizenii. In the case of strain CR-95T, the maximum similarity value was 99·5 %, with B. mojavensis. DNA–DNA hybridization of strains CR-119T and CR-95T with the above species produced values lower than 46·9 %. Therefore, on the basis of phenotypic characteristics, phylogenetic data and genomic distinctiveness, we conclude that these Bacillus strains merit classification as novel species, for which we propose the names Bacillus axarquiensis sp. nov. (type strain CR-119T=CECT 5688T=LMG 22476T) and Bacillus malacitensis sp. nov. (type strain CR-95T=CECT 5687T=LMG 22477T).

Two sporulating bacterial strains designated CECAP06T and CECAP16 were isolated from the rhizosphere of the legume Cicer arietinum in Argentina. Almost-complete 16S rRNA gene sequences identified the isolates as a Paenibacillus species. It was most closely related to Paenibacillus cineris LMG 18439T (99·6 % sequence similarity), Paenibacillus favisporus LMG 20987T (99·4 % sequence similarity) and Paenibacillus azoreducens DSM 13822T (97·7 % sequence similarity). The cells of this novel species were motile, sporulating, rod-shaped, Gram-positive and strictly aerobic. The predominant fatty acids were anteiso-C15 : 0, C16 : 0 and iso-C16 : 0. The DNA G+C content of strains CECAP06T and CECAP16 was 51·3 and 50·9 mol%, respectively. Growth was observed from many carbohydrates, but gas production was not observed from glucose. Catalase and oxidase activities were present. The isolates produced β-galactosidase and hydrolysed aesculin. Gelatinase, caseinase and urease were not produced. The results of DNA–DNA hybridization showed that the strains from this study constitute a novel species of the genus Paenibacillus, for which the name Paenibacillus rhizosphaerae sp. nov. is proposed. The type strain is CECAP06T (=LMG 21955T=CECT 5831T).

The taxonomic status of Pediococcus urinaeequi is described, and the transfer of the species to the genus Aerococcus with the name Aerococcus urinaeequi comb. nov. is proposed, on the basis of the analysis of 16S rRNA gene sequence and DNA–DNA hybridization data.

An alkaliphilic actinomycete, strain YIM 80305T, which was isolated from a muddy sample in Chaka salt lake, Qinghai Province of China, was characterized using a polyphasic approach. The isolate produced light-yellow substrate and yellow–white aerial mycelia on most tested media. Optimum pH for growth was 9·0–10·0 with scant growth at pH 7·0. Results showed that strain YIM 80305T was obligately Na+-dependent, and showed sensitivity to K+. The DNA G+C content was 70·5 mol%. 16S rRNA gene sequence analysis together with these characteristics consistently assigned strain YIM 80305T to the genus Streptomyces. It formed a distinct clade based on analyses of the almost-complete and 120-nucleotide variable γ region of the 16S rRNA gene. It could be differentiated by phenotypic and genotypic analysis from all the Streptomyces species whose names have been validly published. On the basis of polyphasic evidence, Streptomyces sodiiphilus sp. nov. is proposed. The type strain is YIM 80305T (=CCTCC AA 203015T=CIP 107975T).

A Gram-positive, spore-forming and moderately halophilic bacterium, strain BH139T, was isolated from saline sediment of Xinjiang Province, China. Cells of strain BH139T were motile, flagellated rods. The organism grew optimally at 30–35 °C in the presence 12–14 % (w/v) NaCl. The major fatty acids were branched saturated fatty acids such as iso-C16 : 0, anteiso-C15 : 0, iso-C15 : 0, iso-C14 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was about 43 mol% and the predominant isoprenoid quinone was MK-7. Comparative 16S rRNA gene sequence analyses showed that strain BH139T was most closely related to Lentibacillus salicampi KCCM 41560T (96·9 % 16S rRNA gene sequence similarity) and formed a distinct phyletic line from that species. On the basis of physiological and molecular properties, the isolate represents a novel species within the genus Lentibacillus, for which the name Lentibacillus salarius sp. nov. is proposed. The type strain is BH139T (=KCTC 3911T=DSM 16459T).

The type strain of Nocardia corynebacterioides was the subject of a polyphasic taxonomic study. The 16S rRNA gene sequence was aligned with the sequences of representatives of the genera Corynebacterium, Dietzia, Gordonia, Mycobacterium, Nocardia, Rhodococcus, Skermania, Tsukamurella and Williamsia, and phylogenetic trees were constructed by using maximum-parsimony, maximum-likelihood and neighbour-joining methods. It was evident from the phylogenetic analysis that N. corynebacterioides represents a distinct phyletic line within the genus Rhodococcus. Menaquinone analysis showed that the organism contained dihydrogenated menaquinone with eight isoprene units, MK-8(H2), as the major isoprenologue. The genealogical evidence, together with chemotaxonomic and phenotypic data from this and previous studies, indicates that N. corynebacterioides DSM 20151T (=CIP 104510T) should be reclassified in the genus Rhodococcus as Rhodococcus corynebacterioides comb. nov.

Physiological characters, mating compatibility, PCR-RAPD fingerprints, mol% G+C content, DNA–DNA relatedness, and large-subunit and internal transcribed spacer rRNA gene sequences of strains assigned to the genus Zygoascus were re-examined. On the basis of those data, and after phylogenetic analyses, an emendation of Zygoascus hellenicus (type material is a cross of CBS 6736T×CBS 5839T) is proposed, comprising two novel anamorphic varieties, Candida steatolytica var. steatolytica (CBS 6736T) and C. steatolytica var. inositophila (CBS 5839T). A novel teleomorphic species, Zygoascus meyerae sp. nov. (type material is a cross of CBS 4099T×CBS 7521T) is described, together with two novel anamorphic varieties corresponding to it, Candida hellenica var. hellenica (CBS 4099T) and C. hellenica var. acidophila (CBS 7115T).

Cryptococcus taeanensis, a new anamorphic yeast species originating from a salt farm on the Taean peninsula in Korea, is described. Strain 3-12T grew by budding, contained ubiquinone Q-10 and xylose in cell hydrolysates, utilized d-glucuronate and did not ferment d-glucose. A molecular phylogenetic analysis based on the large-subunit rRNA D1/D2 domain and ITS region sequences placed C. taeanensis near Auriculibuller fuscus and Bullera japonica, recently proposed taxa of the Tremellales. However, these species were distinguishable based on standard physiological tests used for yeast identification, with characteristics including the assimilation of l-sorbose, absence of ballistoconidia, no arbutin hydrolysis and no growth in the presence of 0·01 % cycloheximide. The isolate exhibited the typical physiology of the genus Cryptococcus Vuillemin, but its large-subunit rRNA D1/D2 domain sequence was clearly distinct from previously described species in the genus. Therefore, on the basis of these results, Cryptococcus taeanensis sp. nov. is proposed; the type strain is 3-12T (=KCTC 17149T=CBS 9742T).

Three heterothallic, haplontic yeast species, Metschnikowia hamakuensis, Metschnikowia kamakouana and Metschnikowia mauinuiana, are described from isolates associated with endemic nitidulid beetles living on various endemic plants on three Hawaiian islands. As morphospecies, they are similar to Metschnikowia hawaiiensis, but based on mating compatibility and ascospore formation, they can be assigned clearly to distinct biological species. Analysis of ITS/5·8S and D1/D2 large subunit rDNA sequences shows that, with M. hawaiiensis and two other isolates, these species form a distinct subclade within the large-spored Metschnikowia species, indicating that they are Hawaiian endemics. Type cultures are: M. hamakuensis, UWOPS 04-207.1T=CBS 10056T=NRRL Y-27834T (type, h+) and UWOPS 04-204.1=CBS 10055=NRRL Y-27833 (allotype, h−); M. kamakouana, UWOPS 04-112.5T=CBS 10058T=NRRL Y-27836T (type, h+) and UWOPS 04-109.1=CBS 10057=NRRL Y-27835 (allotype, h−); and M. mauinuiana, UWOPS 04-190.1T=CBS 10060T=NRRL Y-27838T (type, h+) and UWOPS 04-110.4=CBS 10059=NRRL Y-27837 (allotype, h−).