<Emphasis Type="Italic">Streptomyces sudanensis</Emphasis> sp. nov., a new pathogen isolated from patients with actinomycetoma

Nine strains isolated from mycetoma patients and received as Streptomyces somaliensis were the subject of a polyphasic taxonomic study. The organisms shared chemical markers consistent with their classification in the genus Streptomyces and formed two distinct monophyletic subclades in the Streptomyces 16S rRNA gene tree. The first subclade contained four organisms, including the type strain of S. somaliensis, and the second clade the remaining five strains which had almost identical 16S rRNA sequences. Members of the two subclades were sharply separated using DNA:DNA relatedness and phenotypic data which also showed that the subclade 1 strains formed an heterogeneous group. In contrast, the subclade 2 strains were assigned to a single genomic species and had identical phenotypic profiles. It is evident from these data that the subclade 2 strains should be recognised as a new species of Streptomyces. The name proposed for this new species is Streptomyces sudanensis sp. nov. The type strain is SD 504^T (DSM = 41923^T = NRRL B-24575^T). Erika T. Quintana and Katarzyna Wierzbicka contributed equally to this work. The GenBank accession numbers for the 16S rRNA gene sequences of Streptomyces somaliensis DSM 40738^T and Streptomyces sudanensis DSM 41607, DSM 41608, DSM 41609, SD 504^T and SD 509 are EF540897, EF540898, EF540999, EF515876 and EF540900.     

Streptomyces poriferorum sp. nov., a novel marine sponge-derived Actinobacteria species expressing anti-MRSA activity

Marine sponges represent a rich source of uncharacterized microbial diversity, and many are host to microorganisms that produce biologically active specialized metabolites. Here, a polyphasic approach was used to characterize two Actinobacteria strains, P01-B04^T and P01-F02, that were isolated from the marine sponges Geodia barretti (Bowerbank, 1858) and Antho dichotoma (Esper, 1794), respectively. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains P01-B04^T and P01-F02 are closely related to Streptomyces beijiangensis DSM 41794^T, Streptomyces laculatispora NRRL B-24909^T, and Streptomyces brevispora NRRL B-24910^T. The two strains showed nearly identical 16S rRNA gene sequences (99.93%), and the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) relatedness values were 99.96% and 99.6%, respectively, suggesting that these strains are affiliated with the same species. Chemotaxonomic and culture characteristics of both strains were also consistent with the genus Streptomyces, while phenotypic properties, genome-based comparisons, and phylogenomic analyses distinguished strains P01-B04^T and P01-F02 from their closest phylogenetic relatives. In silico analysis predicted that the 8.9 Mb genome of P01-B04^T contains at least 41 biosynthetic gene clusters (BGCs) encoding secondary metabolites, indicating that this strain could express diverse bioactive metabolites; in support of this prediction, this strain expressed antibacterial activity against Gram-positive bacteria including a clinical isolate of methicillin-resistant Staphylococcus aureus (MRSA) EAMC30. Based on these results, the marine sponge-associated isolates represent a novel species of the genus Streptomyces, for which the name Streptomyces poriferorum sp. nov. is proposed, with P01-B04^T (=DSM 111306^T = CCM 9048^T) as the type strain.     

Metalloid Reducing Bacteria Isolated from Deep Ocean Hydrothermal Vents of the Juan de Fuca Ridge, Pseudoalteromonas telluritireducens sp. nov. and Pseudoalteromonas spiralis sp. nov

Five strains of Gram-negative, rod, curved rod and spiral-shaped bacteria were isolated from the vicinity of deep ocean hydrothermal vents along the Main Endeavour Segment of the Juan de Fuca Ridge in the Pacific Ocean. All strains showed remarkable resistance to high levels of toxic metalloid oxyanions, and were capable of reducing the oxyanions tellurite and selenite to their less toxic elemental forms. Phylogenetic analysis of four strains identified these isolates as close relatives of the genus Pseudoalteromonas within the class Gammaproteobacteria. Pseudoalteromonas agarivorans was the closest relative of strains Te-1-1 and Se-1-2-red^T, with, respectively, 99.5 and 99.8% 16S rDNA sequence similarity. Strain Te-2-2^T was most closely related to Pseudoalteromonas paragorgicola, with 99.8% 16S rDNA sequence similarity. The DNA G+C base composition was 39.6 to 41.8 mol%, in agreement with other members of the genus Pseudoalteromonas. However, the isolates showed important morphological and physiological differences from previously described species of this genus, with one group forming rod-shaped bacteria typical of Pseudoalteromonas and the other forming vibrioid- to spiral-shaped cells. Based on these differences, and on phylogenetic data, we propose the creation of the new species Pseudoalteromonas telluritireducens sp. nov., with strain Se-1-2-red^T (DSMZ=16098^T=VKM B-2382^T) as the type strain, and Pseudoalteromonas spiralis sp. nov., with strain Te-2-2^T (DSMZ=16099^T=VKM B-2383^T) as the type strain. The EMBL accession numbers for the 16S rDNA sequences are: Te-1-1, AJ314843; Te-2-2^T, AJ314842; Se-1-2-or, AJ314844; Se-1-2-red^T, AJ314845.     

Anionic carbohydrate-containing cell wall polymers of <Emphasis Type="Italic">Streptomyces melanosporofaciens</Emphasis> and related species

The structures of cell wall anionic carbohydrate-containing polymers in Streptomyces melanosporofaciens VKM Ac-1864^T and phylogenetically close organisms—S. hygroscopicus subsp. hygroscopicus VKM Ac-831^T, S. violaceusniger VKM Ac-583^T, S. endus VKM Ac-1331^T, S. endus VKM Ac-129, and S. rutgersensis subsp. castelarensis VKM Ac-832^T—have been comparatively studied by chemical and NMR spectroscopic methods. The natural polymer of a new, previously unknown structure, Kdn (3-deoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid) with β-galactose residues at C-9, has been found in the cell walls of all the strains under study. The cell walls of all the studied organisms contain three teichoic acids (TA): a predominant TA (1,3-poly(glycerol phosphate) with N-acetylated α-glucosaminyl substitutes by C-2 of glycerol, and minor TAs, 1,3-and 2,3-poly(glycerol phosphate) polymers without substitution. Their chains have O-acetyl and O-lysyl groups. Microorganisms of the above-mentioned species differ in the number of α-glucosaminyl substitutes and in the degree of their acetylation in the predominant teichoic acid.     

Molecular and culture dependent characterization of endolithic bacteria in two beach sand samples and description of Rhizobium endolithicum sp. nov.

The taxonomic position of a streptomycete isolated from a potato tubercle was determined by using a polyphasic approach. The organism had chemotaxonomic and morphological properties consistent with its classification in the genus Streptomyces and formed a distinct phyletic line in the Streptomyces 16S rRNA gene tree. It was found to be closely related to Streptomyces celluloflavus NRRL B-2493^T (99.4 % 16S rRNA gene similarity) and shared a 99.0 % 16S rRNA gene similarity value with Streptomyces albolongus NRRL B-3604^T and Streptomyces cavourensis subsp. cavourensis NBRC 13026^T; low levels of DNA–DNA relatedness with these organisms showed that the isolate belonged to a distinct genomic species. The isolate was distinguished readily from the type strains of these species using a combination of morphological and other phenotypic properties. On the basis of these results, it is proposed that isolate ASBV-1^T (= CBMAI 1465^T = CCMA 894^T = NRRL B-24922^T) be classified as the type strain of Streptomyces araujoniae sp. nov.     

Selective isolation and characterisation of members of the Streptomyces violaceusniger clade associated with the roots of Paraserianthes falcataria

Large numbers of putatively novel streptomycetes were isolated from environmental samples collected from in and around the root system of the tropical angiosperm, Paraserianthes falcataria. Representative isolates were assigned to 37 multi-membered and 107 single membered colour groups based on their ability to form pigments on oatmeal and peptone yeast extract iron agars. The largest taxon, colour group 3, encompassed 94 isolates which had morphological properties typical of members of the Streptomyces violaceusniger clade. Twelve representatives of this taxon chosen on the basis of Curie-point pyrolysis mass spectrometric data were compared with representatives of the validly described species which constitute the Streptomyces violaceusniger clade. Six out of the twelve representative strains were readily distinguished from one another and from the marker strains using a combination of genotypic and phenotypic properties. These organisms were consequently considered to merit species status as Streptomyces asiaticus sp. nov., Streptomyces cangkringensis sp. nov., Streptomyces indonesiensis sp. nov., Streptomyces javensis sp. nov., Streptomyces rhizosphaerius sp. nov. and Streptomyces yogyakartensis sp. nov.     

Streptomyces deserti sp. nov., isolated from hyper-arid Atacama Desert soil

The taxonomic position of a Streptomyces strain isolated from a hyper-arid desert soil was established using a polyphasic approach. The organism had chemical and morphological properties typical of the genus Streptomyces and formed a phyletic line at the periphery of the Streptomyces coeruleorubidus subcluster in the 16S rRNA gene tree. DNA:DNA relatedness values between the isolate and its nearest phylogenetic neighbours, Streptomyces lomondensis NRRL 3252^T and Streptomyces lusitanus NRRL B-12501^T were 42.5 (±0.48)% and 25.0 (±1.78)%, respectively. The isolate was readily distinguished from these organisms using a combination of morphological and phenotypic properties. On the basis of these results, it is proposed that isolate C63^T (CGMCC 4.6997^T, = KACC 15425^T) be classified as the type strain of Streptomyces deserti sp. nov.     

Classification of Streptomyces phylogroup pratensis (Doroghazi and Buckley, 2010) based on genetic and phenotypic evidence,and proposal of Streptomyces pratensis sp. nov.

The Streptomyces phylogroup pratensis (Doroghazi and Buckley, 2010) contains isolates obtained from grassy fields, as well as Streptomyces flavogriseus ATCC 33331 and strain CGMCC 4.1868. This latter strain was received as Streptomyces griseoplanus but was subsequently found to be mislabeled, and S. flavogriseus ATCC 33331 (=IAF-45-CD) was shown to be clearly distinct from the type strain S. flavogriseus ATCC 25452^T (=CGMCC 4.1884^T). In order to evaluate the taxonomic position of phylogroup pratensis further, sequences of the 16S rRNA gene and five protein-coding housekeeping genes (atpD, gyrB, recA, rpoB and trpB) were determined for six strains of the phylogroup and type strains of 19 related species, which were selected by a BLAST search based on the sequences of the phylogroup. The 16S rRNA gene sequences for the phylogroup were identical to those of eight species belonging to cluster I of the S. griseus clade. However, in all the individual protein-coding gene and MLSA phylogenies, the phylogroup strains without exception formed an obviously distinct cluster that could be equated with a new species status. The phylogenetic evidence for the new species assignment was also supported by corresponding DNA–DNA hybridization values and by phenotypic characteristics. It is therefore proposed that the phylogroup should be classified as Streptomyces pratensis sp. nov., and the type strain is ch24^T (=CGMCC 4.6829^T = NRRL B-24916^T).     

Erratum to: Two novel species Enterococcus lemanii sp. nov. and Enterococcus eurekensis sp. nov., isolated from a swine-manure storage pit

A polyphasic taxonomic study using morphological, biochemical, chemotaxonomic and molecular genetic methods was performed on six strains of an unknown Gram-positive, nonspore-forming, facultative anaerobic coccus-shaped bacterium isolated from a swine-manure storage pit. On the basis of 16S rRNA, RNA polymerase-subunit (rpoA), and the 60-kilodalton chaperonin (cpn60) gene sequence analyses, it was shown that all the isolates were enterococci but formed two separate lines of descent. Pairwise 16S rRNA sequence comparisons demonstrated that the two novel organisms were most closely related to each other (97.9 %) and to Enterococcus aquimarinus (97.8 %). Both organisms contained major amounts of C_16:0, C_16:1 ω7c, and C_18:1 ω7c/12t/9t as the major cellular fatty acids. Based on biochemical, chemotaxonomic, and phylogenetic evidence, the names Enterococcus lemanii sp. nov. (type strain PC32^T = CCUG 61260^T = NRRL B-59661^T) and Enterococcus eurekensis sp. nov. (type strain PC4B^T = CCUG 61259^T = NRRL B-59662^T) are proposed for the hitherto undescribed species.     

Evaluation of the antibiotic biosynthetic potential of the genus Amycolatopsis and description of Amycolatopsis circi sp. nov., Amycolatopsis equina sp. nov. and Amycolatopsis hippodromi sp. nov

Aims: To describe three new Amycolatopsis strains and assess the antibiotic biosynthetic potential of the genus. Methods and Results: Three strains, designated S1·3^T, S3·6^T and SE(8)3^T, belonging to the genus Amycolatopsis were isolated and found to cluster together by 16S rRNA and gyrB gene‐based phylogenetic analysis. Genetic distance values, based on the gyrB gene, were calculated between the strains and their closest relatives and were all above the threshold value of 0·02 that has been proposed to distinguish Amycolatopsis type strains. DNA–DNA hybridization experiments against related type strains confirmed that strain S3·6^T represents a unique genomic species. Strain S3·6^T was also found to be distinct from strains S1·3^T and SE(8)3^T, the latter two of which were also shown to be distinct from each other. Antibiotic biosynthetic genes were identified from multiple Amycolatopsis strains, and their presence was found to be phylogenetically associated. Conclusions: The data presented in this study indicate that strains S1·3^T, SE(8)3^T and S3·6^T belong to three novel species, for which the names Amycolatopsis circi sp. nov. (= DSM 45561^T = NRRL B‐24841^T), Amycolatopsis equina sp. nov. (= DSM 45563^T = NRRL B‐24842^T) and Amycolatopsis hippodromi sp. nov. (= DSM 45562^T = NRRL B‐24843^T) are proposed. Significance and Impact of the Study: Three new species of Amycolatopsis are described, and the knowledge of the antibiotic biosynthetic potential of the genus has been extended.     

<Emphasis Type="Italic">Streptomyces shenzhenensis</Emphasis> sp. nov., a novel actinomycete isolated from mangrove sediment

An actinomycete strain, which was designated 172115^T, was isolated from mangrove soil in Shenzhen, China. Strain 172115^T fell within the genus Streptomyces in the 16S rRNA gene tree and could be grouped into this genus based on its chemotaxonomic and morphological data. The strain shared the highest 16S rRNA gene sequence similarities with Streptomyces lanatus NBRC 12787^T (AB184845) (98.29%) and Streptomyces lucensis NBRC 13056^T (AB184280) (98.26%). The DNA–DNA hybridization values between strain 172115^T and the two most closely related type strains were low enough to justify the assignment of the strain to a novel species. On the basis of these phenotypic, phylogenetic and chemotaxonomic characteristics, 172115^T represents a novel species of the genus Streptomyces, for which, the name Streptomyces shenzhenensis sp. nov. is proposed for strain 172115^T (=CCTCC AA 2011001^T=DSM 42034^T).     

Streptomyces polyrhachii sp. nov., a novel actinomycete isolated from an edible Chinese black ant (Polyrhachis vicina Roger)

A novel actinomycete, designated strain NEAU-ycm1^T, was isolated from an edible Chinese black ant (Polyrhachis vicina Roger) and characterized with a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of streptomycetes. Phylogenetic analysis based on the almost complete 16S rRNA gene sequence show that the novel isolate belongs to the genus Streptomyces and forms a separate subclade. The closest phylogenetic relatives were identified as the type strains of Streptomyces intermedius NBRC 13049^T (97.74 %), Streptomyces aureoverticillatus NRRL B-3326^T (97.69 %), Streptomyces rutgersensis NBRC 12819^T (97.68 %), Streptomyces gougerotii NBRC 3198^T (97.68 %) and Streptomyces diastaticus subsp. diastaticus NBRC 3714^T (97.68 %). Similarities to other type strains of the genus Streptomyces were lower than 97.55 %. A comparison between strain NEAU-ycm1^T and the closest related Streptomyces type strains revealed that it is different from them in morphological, physiological and biochemical characteristics. Therefore, it is proposed that NEAU-ycm1^T (=CGMCC 4.7094^T = DSM 42102^T) represents a novel species of the genus of Streptomyces, for which the name Streptomyces polyrhachii sp. nov. is proposed.     

Two novel species Enterococcus lemanii sp. nov. and Enterococcus eurekensis sp. nov., isolated from a swine-manure storage pit

A polyphasic taxonomic study using morphological, biochemical, chemotaxonomic and molecular genetic methods was performed on six strains of unknown Gram-positive, nonspore-forming, facultative anaerobic coccus-shaped bacteria isolated from a swine-manure storage pit. On the basis of the 16S rRNA, RNA polymerase α-subunit (rpoA) and 60 kDa chaperonin (cpn60) gene sequence analyses, it was shown that all the isolates were enterococci but formed two separate lines of descent. Pairwise 16S rRNA gene sequence comparisons demonstrated that the two novel organisms were most closely related to each other (97.9 %) and to Enterococcus aquimarinus (97.8 %). Both organisms contained major amounts of C_16:0, C_16:1 ω7c, C_16:1 ω7c, and C_18:1 ω7c/12t/9t as the major cellular fatty acids. Based on biochemical, chemotaxonomic and phylogenetic evidence, the names Enterococcus lemanii sp. nov. (type strain PC32^T = CCUG 61260^T = NRRL B-59661^T) and Enterococcus eurekensis sp. nov. (type strain PC4B^T = CCUG 61259^T = NRRL B-59662^T) are proposed for these hitherto undescribed species.     

Amycolatopsis bartoniae sp. nov. and Amycolatopsis bullii sp. nov., mesophilic actinomycetes isolated from arid Australian soils

The status of two mesophilic filamentous actinomycetes isolated from an arid Australian soil sample was determined using a polyphasic taxonomic approach. The isolates had chemical and morphological properties consistent with their classification in the genus Amycolatopsis, assignments that were supported by analysis of 16S rRNA gene sequence data. Isolate SF26^T formed a distinct phyletic line and hence was sharply separated from its nearest phylogenetic neighbour, Amycolatopsis sacchari DSM 44468^T. In contrast, isolate SF27^T formed a subclade in the Amycolatopsis tree with Amycolatopsis vancoresmycina DSM 44592^T but was separated readily from the latter by DNA:DNA pairing data. The two isolates were distinguished from one another and from their respective nearest phylogenetic neighbours using a range of phenotypic properties. These data indicate that the two isolates should be recognized as new species in the genus Amycolatopsis. The names proposed for these new taxa are Amycolatopsis bartoniae sp. nov. and Amycolatopsis bullii sp. nov. with isolates SF26^T (=NCIMB 14706^T = NRRL B-2846^T) and SF27^T (=NCIMB 14707^T = NRRL B-24847^T) as the respective type strains.     

Streptomyces leeuwenhoekii sp. nov., the producer of chaxalactins and chaxamycins,forms a distinct branch in Streptomyces gene trees

A polyphasic study was carried out to establish the taxonomic status of an Atacama Desert isolate, Streptomyces strain C34^T, which synthesises novel antibiotics, the chaxalactins and chaxamycins. The organism was shown to have chemotaxonomic, cultural and morphological properties consistent with its classification in the genus Streptomyces. Analysis of 16S rRNA gene sequences showed that strain C34^T formed a distinct phyletic line in the Streptomyces gene tree that was very loosely associated with the type strains of several Streptomyces species. Multilocus sequence analysis based on five house-keeping gene alleles underpinned the separation of strain C34^T from all of its nearest phylogenetic neighbours, apart from Streptomyces chiangmaiensis TA-1^T and Streptomyces hyderabadensis OU-40^T which are not currently in the MLSA database. Strain C34^T was distinguished readily from the S. chiangmaiensis and S. hyderabadensis strains by using a combination of cultural and phenotypic data. Consequently, strain C34^T is considered to represent a new species of the genus Streptomyces for which the name Streptomyces leeuwenhoekii sp. nov. is proposed. The type strain is C34^T (= DSM 42122^T = NRRL B-24963^T). Analysis of the whole-genome sequence of S. leeuwenhoekii, with 6,780 predicted open reading frames and a total genome size of around 7.86 Mb, revealed a high potential for natural product biosynthesis.     

Saccharothrix yanglingensis sp. nov., an antagonistic endophytic actinomycete isolated from cucumber plant

An endophytic actinomycete strain, designated Hhs.015^T, was isolated from roots of cucumber seedlings. The endophytic isolate was identified by means of a polyphasic taxonomic approach. On the basis of 16S rRNA gene sequence similarities, strain Hhs.015^T was closely related to members of the genus Saccharothrix. DNA–DNA hybridization with the four closest relatives, Saccharothrix longispora NRRL B-16116^T, Saccharothrix xinjiangensis NRRL B-24321^T, Saccharothrix autraliensis CGMCC 4.1355^T and Saccharothrix espanaensis CGMCC 4.1714^T, gave similarity values of 33.8, 28.2, 44.1 and 29.5%, respectively, which indicated that strain Hhs.015^T represents a novel species of the genus Saccharothrix. This is consistent with the morphological, physiological and chemotaxonomic data. As a whole, these results suggest that strain Hhs.015^T represents a novel Saccharothrix species. The name Saccharothrix yanglingensis sp. nov. is proposed, with the type strain Hhs.015^T (=CGMCC 4.5627^T = KCTC 19722^T).     

<Emphasis Type="Italic">Paenibacillus seodonensis</Emphasis> sp. nov., isolated from a plant of the genus <Emphasis Type="Italic">Campanula</Emphasis>

Strain DCT-19^T, representing a Gram-stain-positive, rodshaped, aerobic bacterium, was isolated from a native plant belonging to the genus Campanula on Dokdo, the Republic of Korea. Comparative analysis of the 16S rRNA gene sequence showed that this strain was closely related to Paenibacillus amylolyticus NRRL NRS-290^T (98.6%, 16S rRNA gene sequence similarity), Paenibacillus tundrae A10b^T (98.1%), and Paenibacillus xylanexedens NRRL B-51090^T (97.6%). DNADNA hybridization indicated that this strain had relatively low levels of DNA-DNA relatedness with P. amylolyticus NRRL NRS-290^T (30.0%), P. xylanexedens NRRL B-51090^T (29.0%), and P. tundrae A10b^T (24.5%). Additionally, the genomic DNA G + C content of DCT-19^T was 44.8%. The isolated strain grew at pH 6.0–8.0 (optimum, pH 7.0), 0–4% (w/v) NaCl (optimum, 0%), and a temperature of 15–45°C (optimum 25–30°C). The sole respiratory quinone in the strain was menaquinone-7, and the predominant fatty acids were C_15:0 anteiso, C_16:0 iso, and C_16:0. In addition, the major polar lipids were diphosphatidylglycerol and phosphatidylethanolamine. Based on its phenotypic properties, genotypic distinctiveness, and chemotaxonomic features, strain DCT-19^T is proposed as a novel species in the genus Paenibacillus, for which the name Paenibacillus seodonensis sp. nov. is proposed (=KCTC 43009^T =LMG 30888^T). The type strain of Paenibacillus seodonensis is DCT-19^T.     

<Emphasis Type="Italic">Kribbella podocarpi</Emphasis> sp. nov., isolated from the leaves of a yellowwood tree (<Emphasis Type="Italic">Podocarpus latifolius</Emphasis>)

An endophytic actinobacterial strain was isolated from a yellowwood tree growing on the slope of Devil’s Peak, Cape Town, South Africa. Analysis of the 16S rRNA gene showed that the strain belongs to the genus Kribbella. Phylogenetic analyses using the 16S rRNA gene and multilocus sequence analysis using the concatenated gene sequences of the gyrB, rpoB, relA, recA and atpD genes showed that strain YPL1^T is closely related to the type strains of Kribbella karoonensis and Kribbella shirazensis. DDH experiments showed that strain YPL1^T is a distinct genomic species from its close phylogenetic relative, K. karoonensis Q41^T. Physiological comparisons further showed that strain YPL1^T is phenotypically distinct from the type strains of Kribbella jejuensis, Kribbella aluminosa, K. karoonensis, K. shirazensis and Kribbella swartbergensis. Strain YPL1^T is thus presented as the type strain of a novel species, for which the name Kribbella podocarpi sp. nov. (= DSM 29424^T = NRRL B-65063^T), is proposed.     

Halophilic and halotolerant aerobic methylobacteria from the technogenic Solikamsk biotopes

Seven strains of moderately halophilic and halotolerant aerobic methylobacteria from the technogenic Solikamsk biotopes (Perm krai, Russia) were isolated in pure cultures and characterized. The isolates were represented by gram-negative and gram-positive (strain 2395B) cells. All the cells were shown to multiply by binary fission without formation of spores or prosthecae. All isolates except strain 2395B were able to oxidize methanol by a classical methanol dehydrogenase. The ribulose monophosphate (RMP) (strain LS), serine (strains S12, S3, 2395A), or ribulose bisphosphate (strains SK15 and S3270) pathways of C₁-assimilation were used. In strain 2395B, the key enzymes of the RMP and serine metabolic pathways were determined. Using polyphasic taxonomy, three strains were identified as representatives of the known species: Arthrobacter protophormiae 2395B, Methylophaga thalassica LS, and Ancylobacter rudongensis S3270. Three more strains were identified as members of new species: Methylopila oligotropha sp. nov. (strain 2395AT; VKM B-2788^T = CCUG 63805^T), Ancylobacter defluvii sp. nov. (strain SK15^T; VKM B-2789^T = CCUG 63806^T), and Paracoccus communis sp. nov. (strain S3^T; VKM B-2787^T = CCUG 63804^T). According to the results of 16S rRNA gene sequencing, the obligately methylotrophic strain S12 had less than 94% similarity with the known genera of the Proteobacteria and was probably a representative of a novel genus.