- Volume 54, Issue 1, 2004

Five hydrogen-oxidizing, thermophilic, strictly chemolithoautotrophic, microaerophilic strains, with similar (99–100 %) 16S rRNA gene sequences were isolated from terrestrial hot springs at Furnas, São Miguel Island, Azores, Portugal. The strain, designated Az-Fu1T, was characterized. The motile, 0·9–2·0 μm rods were Gram-negative and non-sporulating. The temperature growth range was from 50 to 73 °C (optimum at 68 °C). The strains grew fastest in 0·1 % (w/v) NaCl and at pH 6, although growth was observed from pH 5·5 to 7·0. Az-Fu1T can use elemental sulfur, sulfite, thiosulfate, ferrous iron or hydrogen as electron donors, and oxygen (0·2–9·0 %, v/v) as electron acceptor. Az-Fu1T is also able to grow anaerobically, with elemental sulfur, arsenate and ferric iron as electron acceptors. The Az-Fu1T G+C content was 33·6 mol%. Maximum-likelihood analysis of the 16S rRNA phylogeny placed the isolate in a distinct lineage within the Aquificales, closely related to Sulfurihydrogenibium subterraneum (2·0 % distant). The 16S rRNA gene of Az-Fu1T is 7·7 % different from that of Persephonella marina and 6·8 % different from Hydrogenothermus marinus. Based on the phenotypic and phylogenetic characteristics presented here, it is proposed that Az-Fu1T belongs to the recently described genus Sulfurihydrogenibium. It is further proposed that Az-Fu1T represents a new species, Sulfurihydrogenibium azorense.

Two bacterial isolates from the Baltic Sea, BA1 and BA134T, were characterized for their physiological and biochemical features, fatty acid profiles and phylogenetic position based on 16S rRNA gene sequences. The strains were isolated from surface water of the central Baltic Sea during the decay of a plankton bloom. Phylogenetic analysis of their 16S rRNA gene sequences revealed a clear affiliation to the family ‘Flexibacteriaceae’ and showed highest sequence similarity (91 %) to Cyclobacterium marinum. The G+C content of the DNA was 35·4 mol%. The strains were pink-coloured due to carotinoids, Gram-negative, rod-shaped and catalase- and oxidase-positive. Growth was observed at 0–6 % salinity, with good growth at 0–3 %. Temperature for growth was 4–37 °C, with an optimum around 25 °C. The fatty acid profiles were dominated by branched-chain fatty acids (70 %), with a high abundance of iso-C15 : 0 (29–33 %), iso-C17 : 1 ω9c (7–10 %) and C17 : 1 ω6c (5–10 %). According to their morphology, physiology, fatty acid composition, 16S rRNA gene sequences and DNA–DNA similarity, on one hand, the described bacteria are considered to be members of the same novel species; on the other hand, they are suggested as a novel genus of the family ‘Flexibacteriaceae’. To honour the late aquatic microbiologist Russell T. Bell, the name Belliella baltica gen. nov, sp. nov. is suggested for the Baltic Sea isolates, for which the type strain is BA134T (=DSM 15883T=LMG 21964T=CIP 108006T).

Taxonomic studies were performed on 36 strains that were isolated from microbial mats in Antarctic lakes of the Vestfold Hills, the Larsemann Hills and the McMurdo Dry Valleys. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains are related to members of the genus Flavobacterium; sequence similarity values with their nearest phylogenetic neighbours ranged from 96·8 to 98·5 %. Results of DNA–DNA hybridization and comparison of repetitive extragenic palindromic DNA-PCR fingerprinting patterns revealed that these strains are members of three distinct species. Genotypic results, together with phenotypic characteristics, allowed the differentiation of these species from related Flavobacterium species with validly published names. The isolates are Gram-negative, chemoheterotrophic, rod-shaped cells that are psychrophilic and moderately halotolerant; their DNA G+C contents range from 33·1 to 34·5 mol%. Their whole-cell fatty acid profiles are similar and include C15 : 0, anteiso-C15 : 0, iso-C15 : 0, C15 : 1 ω6c, iso-C16 : 0, iso-C16 : 0 3-OH and summed feature 3 (which comprises iso-C15 : 0 2-OH, C16 : 1 ω7c or both) as major fatty acid components. On the basis of these results, three novel species are proposed, namely Flavobacterium degerlachei sp. nov. (consisting of 14 strains, with LMG 21915T=DSM 15718T as the type strain), Flavobacterium micromati sp. nov. (consisting of three strains, with LMG 21919T=CIP 108161T as the type strain) and Flavobacterium frigoris sp. nov. (consisting of 19 strains, with LMG 21922T=DSM 15719T as the type strain).

Two heterotrophic, aerobic, Gram-negative, pigmented and non-motile marine bacteria that were isolated from the green alga Ulva fenestrata were studied by polyphasic taxonomic methods. 16S rDNA sequence analysis indicated that strain KMM 3912T formed a distinct lineage within the family Flavobacteriaceae. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic analyses, the novel bacteria were classified as Ulvibacter litoralis gen. nov., sp. nov. The type strain is KMM 3912T (=KCTC 12104T=CCUG 47093T).

Three marine strains of the Cytophaga–Flavobacterium–Bacteroides group, designated JC2050T, JC2051T and JC2052T, were obtained from a single sediment sample of getbol, the Korean tidal flat. Comparative 16S rDNA sequence studies revealed that the test strains were not closely related to any validly published genera and that these strains were only distantly related to the genus Cyclobacterium (88·7–91·2 %). Phylogenetic analyses demonstrated that the three getbol isolates formed a distinct monophyletic clade within the family Cytophagaceae. Physiological, biochemical and chemotaxonomic data also indicated that these three getbol isolates differed significantly from members of other genera and were sufficiently different from each other to be recognized as separate species. On the basis of polyphasic evidence, a new genus, Hongiella gen. nov., is proposed, with three novel species, Hongiella mannitolivorans sp. nov. (type strain JC2050T=IMSNU 14012T=DSM 15301T), Hongiella halophila sp. nov. (type strain JC2051T=IMSNU 14013T=DSM 15292T) and Hongiella ornithinivorans sp. nov. (type strain JC2052T=IMSNU 14014T=DSM 15282T). Hongiella mannitolivorans is the type species of the genus.

A novel anaerobic, thermophilic, xylanolytic, motile rod-shaped bacterium with a sheath-like outer structure (toga) was isolated from a Mexican oil well in the Gulf of Mexico. Strain MET12T was a Gram-negative bacterium, reducing elemental sulfur, thiosulfate and sulfite to hydrogen sulfide. Its optimum growth conditions were 55 °C, pH 6·6, 3 % NaCl and 0·15 % MgCl2.6H2O. The DNA G+C content was 36·1 mol%. Phylogenetically, strain MET12T was related to members of genus Petrotoga, with similarities to Petrotoga mobilis, Petrotoga sibirica, Petrotoga miotherma and Petrotoga olearia varying from 97·6 to 98·8 %. However DNA–DNA relatedness values between these species and strain MET12T were lower than 70 %. As strain MET12T (=DSM 14811T=CIP 107371T) was genomically and phenotypically different from existing Petrotoga species, it is proposed as the type strain of a novel species, Petrotoga mexicana sp. nov.

A thermophilic, anaerobic, chemolithoautotrophic bacterium was isolated from the walls of an active deep-sea hydrothermal vent chimney on the East Pacific Rise at 9° 50′ N. Cells of the organism were Gram-negative, motile rods that were about 1·0 μm in length and 0·6 μm in width. Growth occurred between 60 and 80 °C (optimum at 75 °C), 0·5 and 4·5 % (w/v) NaCl (optimum at 2 %) and pH 5 and 7 (optimum at 5·5). Generation time under optimal conditions was 1·57 h. Growth occurred under chemolithoautotrophic conditions in the presence of H2 and CO2, with nitrate or sulfur as the electron acceptor and with concomitant formation of ammonium or hydrogen sulfide, respectively. Thiosulfate, sulfite and oxygen were not used as electron acceptors. Acetate, formate, lactate and yeast extract inhibited growth. No chemoorganoheterotrophic growth was observed on peptone, tryptone or Casamino acids. The genomic DNA G+C content was 54·6 mol%. Phylogenetic analyses of the 16S rRNA gene sequence indicated that the organism was a member of the domain Bacteria and formed a deep branch within the phylum Aquificae, with Thermovibrio ruber as its closest relative (94·4 % sequence similarity). On the basis of phylogenetic, physiological and genetic considerations, it is proposed that the organism represents a novel species within the newly described genus Thermovibrio. The type strain is Thermovibrio ammonificans HB-1T (=DSM 15698T=JCM 12110T).

A thermophilic, marine, anaerobic, chemolithoautotrophic, sulfate-reducing bacterium, strain CIR29812T, was isolated from a deep-sea hydrothermal vent site at the Kairei vent field on the Central Indian Ridge. Cells were Gram-negative motile rods that did not form spores. The temperature range for growth was 55–80 °C, with an optimum at 70 °C. The NaCl concentration range for growth was 10–35 g l−1, with an optimum at 25 g l−1. The pH range for growth was 6–6·7, with an optimum at approximately pH 6·25. H2 and CO2 were the only electron donor and carbon source found to support growth of the strain. However, several organic compounds were stimulatory for growth. Sulfate was used as electron acceptor, whereas elemental sulfur, thiosulfate, sulfite, cystine, nitrate and fumarate were not. No fermentative growth was observed with malate, pyruvate or lactate. The phenotypic characteristics of strain CIR29812T were similar to those of Thermodesulfobacterium hydrogeniphilum, a recently described thermophilic, chemolithoautotrophic sulfate-reducer. However, phylogenetic analyses of the 16S rRNA gene sequences showed that the new isolate was distantly related to members of the family Thermodesulfobacteriaceae (similarity values of less than 90 %). The chemotaxonomic data (fatty acids and polar lipids composition) also indicated that strain CIR29812T could be distinguished from Thermodesulfobacterium commune, the type species of the type genus of the family Thermodesulfobacteriaceae. Finally, the G+C content of the genomic DNA of strain CIR29812T (46·0 mol%) was not in the range of values obtained for members of this family. On the basis of phenotypic, chemotaxonomic and genomic features, it is proposed that strain CIR29812T represents a novel species of a new genus, Thermodesulfatator, of which Thermodesulfatator indicus is the type species. The type strain is CIR29812T (=DSM 15286T=JCM 11887T).

A novel thermophilic bacterium, strain EP1-55-1%T, was isolated from an in-situ colonization system deployed in a superheated, deep-sea, hydrothermal vent emission at the Kairei Field on the Central Indian Ridge in the Indian Ocean. The cells were highly motile rods, each possessing a single polar flagellum. Growth was observed between 35 and 65 °C (optimum temperature, 55 °C; 70 min doubling time) and between pH 4·9 and 7·2 (optimum, pH 5·9). The isolate was a microaerobic-to-anaerobic chemolithoautotroph capable of using molecular hydrogen as the sole energy source and carbon dioxide as the sole carbon source. Molecular oxygen, nitrate or elemental sulfur (S0) could serve as electron acceptors to support growth. The G+C content of the genomic DNA was 34·6 mol%. Phylogenetic analysis based on 16S rDNA sequences indicated that strain EP1-55-1%T represents the first strain for which taxonomic properties have been characterized within the previously uncultivated phylogroup classified as belonging to the uncultivated ε-Proteobacteria group A; the name Hydrogenimonas thermophila gen. nov., sp. nov. is proposed, with strain EP1-55-1%T (=JCM 11971T=ATCC BAA-737T) as the type strain.

A novel moderately thermophilic, microaerobic to anaerobic, chemolithoautotrophic bacterium, designated strain CRT, was isolated from a deep-sea hydrothermal vent site at 36°N on the Mid-Atlantic Ridge. Cells were Gram-negative, non-motile rods. The organism grew at 45–65 °C and pH 6·5–7·4, with optimum growth at 55 °C and pH 6·9–7·1. The NaCl range for growth was 5–50 g l−1 (optimum 30 g l−1). Strain CRT was an obligate chemolithoautotroph, growing with H2 as energy source, sulfur, nitrate or oxygen as electron acceptors and CO2 as carbon source. Hydrogen sulfide and ammonium were the respective products of sulfur and nitrate reduction. The G+C content of the genomic DNA was 32·1 mol%. Based on 16S rRNA gene sequence analysis, this organism was most closely related to Caminibacter hydrogeniphilus (94·9 % similarity). On the basis of phenotypic and phylogenetic data, it is proposed that the isolate represents a novel species, Caminibacter profundus sp. nov. The type strain is CRT (=DSM 15016T=JCM 11957T). The phylogenetic data also correlate well with the significant phenotypic differences between the lineage encompassing the genera Nautilia and Caminibacter and other members of the class ‘Epsilonproteobacteria’. The lineage encompassing the genera Nautilia and Caminibacter is therefore proposed as a new order, Nautiliales ord. nov., represented by a single family, Nautiliaceae fam. nov.

A novel marine sulfate-reducing bacterium, strain CV2803T, which is able to oxidize aliphatic hydrocarbons, was isolated from a hydrocarbon-polluted marine sediment (Gulf of Fos, France). The cells were rod-shaped and slightly curved, measuring 0·6×2·2–5·5 μm. Strain CV2803T stained Gram-negative and was non-motile and non-spore-forming. Optimum growth occurred in the presence of 24 g NaCl l−1, at pH 7·5 and at a temperature between 28 and 35 °C. Strain CV2803T oxidized alkanes (from C13 to C18) and alkenes (from C7 to C23). The DNA G+C content was 41·4 mol%. Comparative sequence analyses of the 16S rRNA gene and dissimilatory sulfite reductase (dsrAB) gene and those of other sulfate-reducing bacteria, together with its phenotypic properties, indicated that strain CV2803T was a member of a distinct cluster that contained unnamed species. Therefore, strain CV2803T (=DSM 15576T=ATCC BAA-743T) is proposed as the type strain of a novel species in a new genus, Desulfatibacillum aliphaticivorans gen. nov., sp. nov.

Strain CJ2T, capable of growth on naphthalene as a sole carbon and energy source, was isolated from coal-tar-contaminated freshwater sediment. The Gram reaction of strain CJ2T was negative. The cells were non-spore-forming, non-motile cocci (without flagella). The isolate was found to be an aerobic heterotroph capable of utilizing glucose and other simple sugars. Growth was observed between 4 and 25 °C (optimum, 20 °C) and between pH 6·0 and 9·0 (optimum, pH 7·0–7·5). The G+C content of the genomic DNA was 61·5 mol% and the major quinone was ubiquinone-8. The peptidoglycan of strain CJ2T was determined as belonging to type A1-γ, meso-diaminopimelic acid. The major fatty acids of strain CJ2T were 16 : 1ω7c (67·0 %), 16 : 0 (19·6 %), 18 : 1ω7c (∼7·9 %) and 10 : 0 3-OH (∼2·5 %). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Mycolic acid and glycolipids could not be detected. Comparative 16S rDNA analysis indicated that strain CJ2T is related to the family Comamonadaceae and that the nearest phylogenetic relative was Polaromonas vacuolata 34-PT (97·1 % similarity). On the basis of the physiological and molecular properties, the naphthalene-degrading isolate was designated Polaromonas naphthalenivorans sp. nov. The type strain is CJ2T (=ATCC BAA-779T=DSM 15660T).

Strain K11T was isolated from activated sludge of a municipal wastewater-treatment plant. Phylogenetic analysis of the 16S rRNA gene sequence revealed that it represents a distinct line of descent within the Comamonadaceae. The novel strain was a Gram-negative, catalase- and oxidase-positive, non-motile, straight to slightly curved rod. Polyhydroxyalkanoate granules were stored intracellularly as reserve material. Colonies on agar plates were small, regular and characterized by a water-insoluble yellow pigment. Unbranched fatty acids 16 : 1ω7c, 16 : 0 and 18 : 1ω7c dominated the cellular fatty acid pattern and ubiquinone-8 (Q-8) was the major component of the respiratory lipoquinones, both traits typical of members of the Comamonadaceae. A distinguishing characteristic was the presence of the two hydroxy fatty acids 10 : 0 3-OH and 12 : 0 2-OH, each in significant amounts. The G+C content of the DNA was 59 mol%. Strain K11T was capable of aerobic chemolithoheterotrophic growth using thiosulfate as an additional substrate, but could not grow autotrophically with thiosulfate or hydrogen. Facultative anaerobic growth was possible with nitrate and nitrite as electron acceptors, but not with ferric iron, sulfate or by fermentation. The sole end product of denitrification was N2O; nitrite accumulated only transiently in small amounts. Based upon phylogenetic and phenotypic evidence, it is proposed to establish the novel taxon Ottowia thiooxydans gen. nov., sp. nov., represented by the type strain K11T (=DSM 14619T=JCM 11629T). Aquaspirillum gracile was among the phylogenetically most closely related species to strain K11T. This species has been wrongly classified, and it is proposed to reclassify it as Hylemonella gracilis gen. nov., comb. nov. The type strain is ATCC 19624T (=DSM 9158T).

Bacterial canker of papaya (Carica papaya) emerged during the 1980s in different islands of the Caribbean. Nineteen strains of Gram-negative, rod-shaped, non-spore-forming bacteria isolated from papaya were compared to 38 reference and type strains of phytopathogenic Enterobacteriaceae and related bacteria. Phylogenetic analysis of 16S rRNA gene sequences showed that the papaya strains belonged to the genus Erwinia. The DNA G+C content of strain CFBP 5189T, 52·5 mol%, is in the range of the genus Erwinia. The 19 papaya strains were all pathogenic to papaya and were differentiated clearly from type or reference strains of phytopathogenic enterobacteria and related bacteria by phenotypic tests. The papaya strains constituted a discrete DNA hybridization group, indicating that they belonged to a unique genomic species. Thus, strains pathogenic to papaya belong to a novel species for which the name Erwinia papayae sp. nov. is proposed, with the type strain CFBP 5189T (=NCPPB 4294T).

An aerobic, heterotrophic, Gram-negative, curved bacterial strain, designated MIL-1T, was isolated by extinction dilution from an n-tetradecane enrichment culture that was established from sea water/sediment samples collected in the harbour of Milazzo, Italy. In the primary enrichment, the isolate formed creamy-white, medium-sized colonies on the surface of the agar. The isolate did not grow in the absence of NaCl; growth was optimal at 2·7 % NaCl. Only a narrow spectrum of organic compounds, including aliphatic hydrocarbons (C7–C20), their oxidized derivatives and acetate, were used as growth substrates. The isolate was not able to grow under denitrifying conditions. The DNA G+C content and genome size of strain MIL-1T were estimated to be 53·2 mol% and 2·2 Mbp, respectively. The major cellular and phospholipid fatty acids were palmitoleic, palmitic and oleic acids (33·5, 29·5 and 11·0 % and 18, 32 and 31 %, respectively). 3-Hydroxy lauric acid was the only hydroxy fatty acid detected. Thirteen different compounds that belonged to two types of phospholipid (phosphatidylethylamine and phosphatidylglycerol) were identified. 16S rRNA gene sequence analysis revealed that this isolate represents a distinct phyletic lineage within the γ-Proteobacteria and has about 94·4 % sequence similarity to Oceanobacter kriegii (the closest bacterial species with a validly published name). The deduced protein sequence of the putative alkane hydrolase, AlkB, of strain MIL-1T is related to the corresponding enzymes of Alcanivorax borkumensis and Pseudomonas oleovorans (81 and 80 % similarity, respectively). On the basis of the analyses performed, Thalassolituus oleivorans gen. nov., sp. nov. is described. Strain MIL-1T (=DSM 14913T=LMG 21420T) is the type and only strain of T. oleivorans.

In conformity with the nomenclature of the genus Rhizobium published by Young et al. (Int J Syst Evol Microbiol 51, 89–103, 2001 ), it is proposed that Agrobacterium larrymoorei be named as Rhizobium larrymoorei comb. nov.

A novel species, Methylocella tundrae, is proposed for three methanotrophic strains (T4T, TCh1 and TY1) isolated from acidic Sphagnum tundra peatlands. These strains are aerobic, Gram-negative, non-motile, dinitrogen-fixing rods that possess a soluble methane monooxygenase and utilize the serine pathway for carbon assimilation. Strains T4T, TCh1 and TY1 are moderately acidophilic organisms capable of growth between pH 4·2 and 7·5 (optimum 5·5–6·0) and between 5 and 30 °C (optimum 15 °C). The major phospholipid fatty acid is 18 : 1ω7c. The DNA G+C content of strain T4T is 63·3 mol%. The three strains possess almost identical 16S rRNA gene sequences and are most closely related to two previously identified species of Methylocella, Methylocella palustris (97 % similarity) and Methylocella silvestris (97·5 % similarity). DNA–DNA hybridization values of strain T4T with Methylocella palustris KT and Methylocella silvestris BL2T were respectively 27 and 36 %. Thus, the tundra strains represent a novel species, for which the name Methylocella tundrae sp. nov. is proposed. Strain T4T (=DSM 15673T=NCIMB 13949T) is the type strain.

A novel species that belongs to the genus Marinospirillum is described on the basis of phenotypic characteristics, phylogenetic analysis of 16S rRNA and gyrB gene sequences and DNA–DNA hybridization. Four strains of helical, halophilic, Gram-negative, heterotrophic bacteria were isolated from kusaya gravy, which is fermented brine that is used for the production of traditional dried fish in the Izu Islands of Japan. All of the new isolates were motile by means of bipolar tuft flagella, of small cell size, coccoid-body-forming and aerophilic; it was concluded that they belong to the same bacterial species, based on DNA–DNA hybridization values (>70 % DNA relatedness). DNA G+C contents of the new strains were 42–43 mol% and they had isoprenoid quinone Q-8 as the major component. Phylogenetic analysis of 16S rRNA gene sequences indicated that the new isolates were members of the genus Marinospirillum; sequence similarity of the new isolates to Marinospirillum minutulum, Marinospirillum megaterium and Marinospirillum alkaliphilum was 98·5, 98·2 and 95·2 %, respectively. Phylogenetic analysis based on the gyrB gene indicated that the new isolates had enough phylogenetic distance from M. minutulum and M. megaterium to be regarded as different species, with 84·7 and 78·7 % sequence similarity, respectively. DNA–DNA hybridization showed that the new isolates had <36 % DNA relatedness to M. minutulum and M. megaterium, supporting the phylogenetic conclusion. Thus, a novel species is proposed: Marinospirillum insulare sp. nov. (type strain, KT=LMG 21802T=NBRC 100033T).

Two strains of bacteria isolated from the blood of French domestic cows were found to be similar to Bartonella species on the basis of phenotypic characteristics. Genotypic analysis based on sequence comparison of the 16S rRNA and citrate synthase (gltA) genes and on DNA–DNA hybridization showed that the two isolates represent a distinct and new species of Bartonella. Moreover, the phylogenetic analysis inferred from comparison of 16S rRNA and gltA sequences demonstrated that the new Bartonella species is related to other ruminant-derived Bartonella species. The name Bartonella chomelii is proposed for the new species. The type strain of Bartonella chomelii sp. nov. is A828T (=CIP 107869T=CCUG47497T).

Three Gram-negative, small, motile, rod-shaped bacteria were isolated from Artemia sp. and sea water in Barcelona, Spain, during 1990 and 1991. They were fermentative, oxidase-positive, sensitive to vibriostatic agent O/129, arginine dihydrolase-positive, lysine and ornithine decarboxylase-negative and grew in the absence of NaCl. They differed from phenotypically related species by their ability to grow at 4 °C and utilize l-rhamnose. Cloning of the 16S rRNA gene of the type strain produced two different 16S rRNA gene sequences, which differed by 15 bases (0·99 %); comparison of these sequences with those deposited in GenBank showed close relationships with Vibrio proteolyticus (97·6 % similarity), Vibrio diazotrophicus (97·9 %), Vibrio campbellii (96·8 %) and Vibrio alginolyticus (96·8 %), among others. DNA–DNA hybridization levels with the closest phylogenetically related Vibrio species were <26·4 %. Sufficient evidence is provided to support the identity of the three strains analysed as members of a novel species of the genus Vibrio, for which the name Vibrio hispanicus sp. nov. is proposed, with the type strain LMG 13240T (=CAIM 525T=VIB 213T).

The Montreal Biodome operates a methanol-fed denitrification system that treats the water in its three million litre marine mesocosm. An unknown bacterium, named strain NL21T, was isolated from this system on TSA and R2A agar. The organism is a Gram-negative, rod-shaped (1×3 μm) facultative aerobe. Optimal growth conditions on R2A agar are 30–35 °C, pH 7–7·5 and 1 % (w/w) NaCl. Phylogenetic analysis of the 16S rDNA sequence reveals that strain NL21T forms a novel lineage in the family ‘Phyllobacteriaceae’ within the α2 subgroup of the Proteobacteria. The closest related genera are Aminobacter, Pseudaminobacter, Mesorhizobium and Defluvibacter. Major cellular fatty acids are C18 : 1 ω7c (75 %), C19 : 0 ω8c cyclopropane (9·4 %) and C18 : 0 (4·2 %). The DNA G+C content of strain NL21T (57 mol%) differs from those of all other described members of the ‘Phyllobacteriaceae’ (60–64 mol%). Strain NL21T reduces nitrate to nitrite, but does not reduce nitrite to nitrogen gas. Only a few sugars and amino acids can serve as carbon sources. Strain NL21T is able to grow without salt and tolerates up to 5 % NaCl. Phylogenetic analysis, as well as physiological and biochemical tests, showed that strain NL21T was different from all other members of the ‘Phyllobacteriaceae’ with validly published names. Strain NL21T therefore represents a novel genus, for which the name Nitratireductor aquibiodomus gen. nov., sp. nov. is proposed, with the type strain NL21T (=DSM 15645T=ATCC BAA-762T).

A group of 42 isolates were isolated from the soil of several disused hay fields, in the Drentse A agricultural research area (The Netherlands), that were taken out of production at different times. The group represents hitherto-uncultured Bacillus lineages that have previously been found, by a non-cultural method, to be predominant in soil. The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA–DNA hybridizations, DNA base-ratio determination, fatty acid analysis and morphological and biochemical characterization. By comparing the groupings obtained by (GTG)5-PCR and 16S rDNA sequence analysis, six clusters of similar strains could be recognized. A DNA–DNA relatedness study showed that these clusters represented five novel genospecies. Further analysis supported the proposal of five novel species in the genus Bacillus, namely Bacillus novalis sp. nov. (type strain IDA3307T=R-15439T=LMG 21837T=DSM 15603T), Bacillus vireti sp. nov. (type strain IDA3632T=R-15447T=LMG 21834T=DSM 15602T), Bacillus soli sp. nov. (type strain IDA0086T=R-16300T=LMG 21838T=DSM 15604T), Bacillus bataviensis sp. nov. (type strain IDA1115T=R-16315T=LMG 21833T=DSM 15601T) and Bacillus drentensis sp. nov. (type strain IDA1967T=R-16337T=LMG 21831T=DSM 15600T).

During a search for xylan-degrading micro-organisms, a sporulated bacterium was recovered from recent and old cow dung and rectal samples. The isolates were identified as members of a novel species of the genus Paenibacillus, based on 16S rRNA gene sequences. According to the results of phylogenetic analysis, the most closely related species was Paenibacillus azoreducens. Phenotypic and chemotaxonomic analyses and DNA–DNA hybridization experiments also showed that the isolates belonged to a novel species of the genus Paenibacillus. The novel species is a facultatively anaerobic, motile, Gram-variable, sporulated rod. The spores of this rod-shaped micro-organism occur in slightly swollen sporangia and are honeycomb-shaped. The main fatty acid is anteiso-branched C15 : 0. Growth was observed with many carbohydrates, including xylan, as the only carbon source and gas production was not observed from glucose. The novel species produces a wide variety of hydrolytic enzymes, such as xylanases, cellulases, amylases, gelatinase, urease and β-galactosidase. On the contrary, it does not produce caseinase, phenylalanine deaminase or lysine decarboxylase. According to the data obtained in this work, the strains belong to a novel species, for which the name Paenibacillus favisporus sp. nov. is proposed (type strain, GMP01T=LMG 20987T=CECT 5760T).

A bacterial strain, GW-9T, which was isolated from groundwater in Korea, was subjected to a polyphasic taxonomic study using phenotypic characterization and phylogenetic and genetic methods. Phylogenetic analysis based on 16S rDNA sequences showed that strain GW-9T forms an evolutionary lineage within the radiation enclosing Nocardioides species and, in particular, a coherent cluster with Nocardioides pyridinolyticus. The cell-wall peptidoglycan type of strain GW-9T was based on ll-diaminopimelic acid as the diamino acid, indicating wall chemotype I. The predominant menaquinone was MK-8(H4). Strain GW-9T had a cellular fatty acid profile containing straight-chain, branched, unsaturated and 10-methyl fatty acids. The major fatty acid was iso-C16 : 0. The DNA G+C content of strain GW-9T was 73 mol%. The 16S rDNA of strain GW-9T was 99·2 % similar to that of the type strain of Nocardioides pyridinolyticus and 94·9–96·0 % similar to sequences of the type strains of other Nocardioides species. Differences in phenotypic characteristics and genetic distinctiveness indicate that strain GW-9T is separate from previously described Nocardioides species. Therefore, on the basis of the data presented, a novel species of the genus Nocardioides, Nocardioides aquiterrae sp. nov., is proposed. The type strain is strain GW-9T (=KCCM 41647T=JCM 11813T).

Three novel strains isolated from brewery environments are described. These strains were Gram-positive, facultatively anaerobic, heterofermentative rods that did not exhibit catalase activity. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that these strains belong to the genus Lactobacillus and are most closely related to Lactobacillus collinoides (approximately 99 % similarity). The novel strains could be differentiated from L. collinoides on the basis of DNA–DNA relatedness, differences in beer-spoilage ability and the inability to utilize d-fructose. These isolates represent a novel species, for which the name Lactobacillus paracollinoides sp. nov. is proposed. The type strain is LA2T (=DSM 15502T=JCM 11969T).

Five strains isolated from two patients with nocardiosis in Japan and three clinical samples from Thailand were found to have morphological, biochemical and chemotaxonomic properties consistent with their classification in the genus Nocardia. DNA–DNA hybridization, coupled with sequence analysis of 16S rDNA, indicated that these strains belong to a novel species of the genus Nocardia, named Nocardia asiatica sp. nov. because the isolation sites were in Asian countries; the type strain is IFM 0245T (=NBRC 100129T=JCM 11892T=DSM 44668T).

A bacterial strain, SL-1T, capable of degrading trichloroethene was isolated from a laboratory enrichment in the Department of Civil and Environmental Engineering, University of Washington, USA. The material in the enrichments was derived from a soil sample from Seattle, WA, USA. Strain SL-1T was capable of using phenol as a source of carbon and energy. Chemotaxonomic, morphological, physiological and phylogenetic analyses showed that strain SL-1T is a member of the genus Pseudonocardia. The ability of strain SL-1T to utilize phenol and degrade trichloroethene, as well as other phenotypic properties and the results from a 16S rRNA phylogenetic analysis, led to the proposal of a novel species, Pseudonocardia chloroethenivorans sp. nov. The type strain is SL-1T (=ATCC BAA-742T=DSM 44698T). Trichloroethene and other chloroethenes are major pollutants at many environmental sites, and P. chloroethenivorans has biodegradation properties that should be of interest to environmental microbiologists and engineers.

The taxonomic position of a marine actinomycete, strain SJS0289/JS1T, was determined using a polyphasic taxonomic approach. The organism, which had phenotypic properties consistent with its classification in the genus Williamsia, formed a distinct clade in the 16S rRNA gene tree together with the type strain of Williamsia muralis, but was readily distinguished from this species using DNA–DNA relatedness and phenotypic data. The genotypic and phenotypic data show that the organism merits recognition as a new species of Williamsia. The name proposed for the new species is Williamsia maris; the type strain is SJS0289/JS1T (=DSM 44693T=JCM 12070T=KCTC 9945T=NCIMB 13945T).

A round-spore-forming Bacillus species that produces an exosporium was isolated from the surface of the Mars Odyssey spacecraft. This novel species has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA–DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus and is a Gram-positive, aerobic, rod-shaped, endospore-forming eubacterium. Ultrathin sections of the spores showed the presence of an exosporium, spore coat, cortex and core. 16S rDNA sequence similarities between this strain, Bacillus fusiformis and Bacillus silvestris were ∼96 % and DNA–DNA reassociation values with these two bacilli were 23 and 17 %, respectively. Spores of the novel species were resistant to desiccation, H2O2 and UV and gamma radiation. Of all strains tested, the spores of this strain were the most consistently resistant and survived all of the challenges posed, i.e. exposure to conditions of desiccation (100 % survival), H2O2 (26 % survival), UV radiation (10 % survival at 660 J m−2) and gamma radiation (0·4 % survival). The name proposed for this novel bacterium is Bacillus odysseyi sp. nov.; the type strain is 34hs-1T (=ATCC PTA-4993T=NRRL B-30641T=NBRC 100172T).

Acidophilic actinomycete strains that represent the two major neutrotolerant clusters defined by numerical taxonomy ( Seong, 1992 ) were the subject of a polyphasic taxonomic study. The centrotypes of each cluster, designated as strain JL164 (=KCTC 9924) of cluster 21 and strain CN732T (=KCTC 9926T=IMSNU 50114T=NRRL B-24245T) of cluster 13, were assigned initially to the genus Streptomyces on the basis of morphological and chemotaxonomic characteristics; this assignation was confirmed by 16S rRNA gene sequence data. Strain CN732T formed a distinct phyletic line within the Streptomyces tree, whereas strain JL164 was related closely to the type strain of Streptomyces mirabilis. It is evident from the present and previous studies that neutrotolerant acidophilic actinomycetes comprise taxonomically diverse groups within the variation encompassed by the genus Streptomyces. It is also apparent that strain CN732T and other members of cluster 13 merit recognition as a novel species, for which the name Streptomyces yeochonensis sp. nov. is proposed.

Based on 16S rRNA gene sequence comparison, an isolate that was detected in sterile-filtered vegetable broth was classified as a novel member of the Erysipelothrix line of descent of the Firmicutes. Strain MF-EP02T resembles members of the two species of Erysipelothrix with validly published names, Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum, in morphology, fatty acid composition, lack of menaquinones in aerobically and anaerobically grown cultures, DNA G+C content and peptidoglycan amino acid composition. Distinct differences in physiological characteristics, however, support the allocation of this isolate to a novel species of the genus Erysipelothrix, for which the name Erysipelothrix inopinata sp. nov. (type strain, MF-EP02T=DSM 15511T=CIP 107935T) is proposed. Members of the Erysipelothrix line of descent are included in the family Erysipelotrichaceae fam. nov.

Strain DSM 44594T, which produces the glycopeptide antibiotic decaplanin, is a member of the genus Amycolatopsis based on 16S rRNA gene sequence analysis and chemotaxonomic properties. It is the first member of this genus that is reported to form pseudosporangia, which resemble those of members of the genus Kibdelosporangium. Phylogenetically, the novel taxon is related to Amycolatopsis orientalis, Amycolatopsis lurida, Amycolatopsis azurea, Amycolatopsis japonica and Amycolatopsis keratiniphila. Morphological, cultural and physiological properties, the production of a unique glycolipid and DNA–DNA similarity of <55 % with phylogenetically related strains reveal that strain DSM 44594T represents a novel species of the genus, for which the name Amycolatopsis decaplanina sp. nov. (type strain, FH 1845T=DSM 44594T=NRRL B-24209T) is proposed.

Four Gram-positive-staining, strictly anaerobic, non-spore-forming, rod-shaped organisms were isolated from a pig manure storage pit. Comparative 16S rRNA gene sequence analysis revealed that the isolates belonged to two related but distinct groups. Sequence analysis showed that the two groups of isolates were highly related to each other (approx. 97 % 16S rRNA gene sequence similarity), forming a distinct cluster within the Clostridium coccoides suprageneric rDNA grouping. Biochemical and physiological studies confirmed the division of the isolates into two related, albeit distinct, groups. Based on both phenotypic and phylogenetic evidence, it is proposed that the unidentified rod-shaped isolates from pig manure should be classified in a novel genus, Hespellia gen. nov., as Hespellia stercorisuis sp. nov. and Hespellia porcina sp. nov. The type species of the novel genus is H. stercorisuis (type strain, PC18T=NRRL B-23456T=CCUG 46279T=ATCC BAA-677T) and the type strain of H. porcina is PC80T (=NRRL B-23458T=ATCC BAA-674T).

An alkaliphilic actinomycete strain, designated YIM 80379T, was isolated from a soil sample collected from the eastern desert of Egypt and subjected to polyphasic taxonomy. The strain produced substrate and aerial mycelia on different media, with an optimum pH for growth of 9·5–10 and scarce or no growth at pH 7. Strain YIM 80379T contained meso-diaminopimelic acid, no diagnostic sugars, type PIII phospholipids and MK-10(H6) and MK-10(H8) as the predominant menaquinones. All of these characters assign isolate YIM 80379T consistently to the genus Nocardiopsis. This was confirmed by 16S rDNA analysis. It can be differentiated from all Nocardiopsis species with validly published names by phenotypic characteristics, phylogenetic analysis and DNA–DNA hybridization results. On the basis of polyphasic evidence, a novel species, Nocardiopsis alkaliphila sp. nov., is proposed. The type strain of the species is YIM 80379T (=CCTCC AA001031T=DSM 44657T).

Ten Gram-positive, aerobic bacterial strains with coccoid cells occurring singly, in pairs, tetrads and small aggregates were isolated from the surface of marble and calcareous stones. The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The major menaquinone was MK-9(H4). The cellular fatty acid pattern consisted mainly of iso-branched chain components. According to their phylogenetic position, the organisms are members of the family Geodermatophilaceae, clustering close to Blastococcus aggregatus DSM 4725T. Based on a combination of phenotypic, chemotaxonomic and genotypic characteristics, strain BC444T and seven relatives group apart from strains BC412 (=DSM 44517) and BC521 (=DSM 44518), which themselves form two individual lineages. DNA–DNA reassociation studies revealed that members of the three lineages were only remotely related to B. aggregatus DSM 4725T; as the strain-rich cluster containing BC444T shared low DNA similarity values with strains BC412 and BC521, it will be described as a novel species of Blastococcus, for which the name Blastococcus saxobsidens sp. nov. is proposed. The type strain is BC444T (=DSM 44509T=NRRL 24246T).

Amycolatopsis orientalis subsp. lurida DSM 43134T differs significantly from the type strain of A. orientalis, A. orientalis subsp. orientalis DSM 40040T, and from other species of the genus in genomic and metabolic properties. Its elevation to species status as Amycolatopsis lurida sp. nov., comb. nov. is justified.

Soil, snow and organic material, collected in November 1997 from the Vestvold Hills, Davis Base, Antarctica, were screened for yeasts. Two isolates, which were shown to be indistinguishable by rDNA sequencing and protein analysis by SDS-PAGE, are described in this communication as a novel species, Cryptococcus watticus sp. nov. (type culture, CBS 9496T=NRRL Y-27556T). Sequence analyses of the 26S rDNA D1/D2 region placed C. watticus in the hymenomycetous yeasts in a cluster with Holtermannia corniformis and Cryptococcus nyarrowii. This species has been allocated to the genus Cryptococcus on the basis of physiological and morphological characteristics.

Bacterial blight is an emerging disease that affects primarily onion, but also garlic and Welsh onion. The present study was undertaken to characterize the causative xanthomonad(s) by a polyphasic approach using a worldwide collection of 33 bacterial strains. Analysis of 16S rRNA gene sequence similarities indicated that the causal agent belongs to the campestris core in the genus Xanthomonas, which is in agreement with results of phenotypic characterization (analyses of carbon source utilization and fatty acid methyl esters). However, DNA–DNA hybridization, thermal stability of DNA reassociation and fluorescent amplified fragment length polymorphism analysis allowed the causal agent to be identified as a pathovar of Xanthomonas axonopodis.

The 16S rRNA gene sequences for 34 strains, including 11 isolates, were determined to classify scab-causing Streptomyces spp. and relatives isolated from potato scab lesions collected in Jeju, Korea. The 16S–23S rDNA internally transcribed spacer (ITS) sequences were determined to investigate whether the 16S–23S ITS region is useful for analysing intra- and interspecific relationships in these bacteria. On the basis of phylogenetic analysis of 16S rRNA gene sequences, most of the isolates were classified as Streptomyces scabiei and Streptomyces acidiscabies. Isolate KJO61 was placed in an ambiguous taxonomic position between Streptomyces reticuliscabiei and Streptomyces turgidiscabies. 16S–23S ITS region sequence analysis showed that tRNA genes were not found in this region of Streptomyces spp. The 16S–23S ITS regions of Streptomyces spp. exhibited various lengths and highly variable sequence similarities (35–100 %) within strains as well as intra- and interspecies. It was revealed that Streptomyces europaeiscabiei could be clearly differentiated from Streptomyces scabiei. However, it was clarified that ITS regions are not useful in phylogenetic analysis of Streptomyces spp.

In the period from January 2001, at least 207 new names proposed in articles in the International Journal of Systematic and Evolutionary Microbiology or cited in Validation Lists are not in accordance with Rules 27(3) and 30 of the Bacteriological Code. The purpose of the present Request for an Opinion is to clarify the status of the names listed and to provide a solution whereby they may be considered to be validly published.

Strains now considered to represent the type strain of Arthrobacter ilicis, described as a pathogen of American holly, are not identical. The designated type strain does not represent this pathogen. However, one of the other strains sourced to the type strain of the pathogen does appear to be authentic, but is not a member of A. ilicis. It is proposed that A. ilicis is an unrelated species, not a pathogen of American holly. The nomenclature of A. ilicis can be rectified by emending the authority and by emending the species description to recognize this species as a novel species that is not a plant pathogen. The pathogen of American holly then becomes a novel pathovar, Curtobacterium flaccumfaciens pv. ilicis. The opinion of the Judicial Commission is sought.

Systematics is the process of characterizing and arranging bacterial diversity in an orderly manner, recognizing groups of similar organisms in a hierarchical scheme whose basic entity is the species. To allow the exchange of scientific knowledge, taxa have to be named. Taxa are not static entities since they are subject to evolution, the direction of which can be inferred by using a wide range of techniques targeting specific traits. However, it is not clear how this dynamism should be reflected in taxonomic nomenclature. In the present report, several considerations are presented that deal with the relationship between the evolution of taxa and their nomenclature; an example is given which concerns the nomenclature of the species Lactobacillus delbrueckii.