- Volume 61, Issue 9, 2011

Three chitin-degrading strains representing two novel species were isolated from mangrove forests in Okinawa, Japan. The isolates, ABABA23T, ABABA211 and ABABA212T, were Gram-negative, non-spore-forming, strictly aerobic chemo-organotrophs. The novel strains produced Q-8 as the major isoprenoid quinone component. The predominant fatty acids were iso-C15 : 0 and C16 : 0. On the basis of 16S rRNA gene sequence analysis, the isolates were closely affiliated with members of the genus Microbulbifer. The DNA G+C contents of strains ABABA23T and ABABA212T were 57.8 and 60.2 mol%, respectively. DNA–DNA relatedness values between these two strains and Microbulbifer reference strains were significantly lower than 70 %, the generally accepted threshold level below which strains are considered to belong to separate species. Based on differences in taxonomic characteristics, the three isolates represent two novel species of the genus Microbulbifer, for which the names Microbulbifer chitinilyticus sp. nov. (type strain, ABABA212T = JCM 16148T = NCIMB 14577T) and Microbulbifer okinawensis sp. nov. (type strain, ABABA23T = JCM 16147T = NCIMB 14576T; reference strain, ABABA211) are proposed.

A bacterial strain, designated KTW-16T, was isolated from the reef-building coral Stylophora pistillata, collected from southern Taiwan. Strain KTW-16T was a Gram-negative, facultatively anaerobic, pale-yellow, non-motile short rod. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KTW-16T belonged to the genus Paracoccus in the Alphaproteobacteria and exhibited 93.7–96.9 % 16S rRNA gene sequence similarity with type strains of species of the genus Paracoccus (96.9 % with Paracoccus alcaliphilus JCM 7364T). Strain KTW-16T grew at 15–40 °C (optimum 35 °C), at pH 6.0–10.0 (optimum pH 8.0) and with 0–9 % NaCl (optimum 5 %). The predominant cellular fatty acids were C18 : 1ω7c, C19 : 0 cyclo ω8c and C18 : 0. The major respiratory quinone was Q-10 and the DNA G+C content was 69.1 mol%. The polar lipid profile consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and several unknown polar lipids. The physiological and biochemical tests allowed clear phenotypic differentiation of the isolate from the type strains of already described Paracoccus species. It is evident from the genotypic, phenotypic and chemotaxonomic analysis that strain KTW-16T should be classified in a novel species of the genus Paracoccus, for which the name Paracoccus stylophorae sp. nov. is proposed. The type strain is KTW-16T ( = LMG 25392T  = BCRC 80106T).

A Gram-reaction-negative, strictly aerobic, motile, rod-shaped bacterium, designated strain BZ78T, was isolated from soil from an industrial site. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BZ78T belonged to the family Rhodospirillaceae and formed a coherent cluster with the type strain of Tistrella mobilis (98.3 % pairwise similarity). The predominant cellular fatty acids of strain BZ78T were C18 : 1ω7c (58.3 %), C19 : 0ω8c cyclo (11.5 %), C18 : 1 2-OH (10.9 %) and C14 : 0 3-OH (6.4 %). The predominant ubiquinone was Q-10. The genomic DNA G+C content of strain BZ78T was 65.8 mol%. On the basis of phenotypic characteristics, phylogenetic analysis and DNA–DNA relatedness data, strain BZ78T is considered to represent a novel species of the genus Tistrella, for which the name Tistrella bauzanensis sp. nov. is proposed. The type strain is BZ78T ( = DSM 22817T  = CGMCC 1.10188T  = LMG 26047T).

A polyphasic taxonomic approach was applied to the study of a Gram-negative bacterium (B2T) isolated from soil by selective enrichment with pentachlorophenol. 16S rRNA gene sequence analysis of strain B2T showed that the strain belongs to the genus Achromobacter within the Betaproteobacteria. The 16S rRNA gene sequence displayed more than 99 % similarity to the sequences of the type strains of all species of Achromobacter, with the highest sequence similarity to those of Achromobacter spanius CCM 7183T and A. piechaudii CCM 2986T (99.8 %). On the basis of phylogenetic analysis, genomic DNA–DNA relatedness and phenotypic characteristics, including chemotaxonomic (cellular fatty acid profile) analysis, a novel species is proposed, Achromobacter marplatensis sp. nov., with the type strain B2T ( = CCM 7608T  = CCUG 56371T  = CECT 7342T).

Two bacterial strains (SC-089T and SC-092T) isolated from sewage sludge compost were characterized by using a polyphasic approach. The isolates were Gram-negative short rods, catalase- and oxidase-positive, and showed good growth at 30 °C, at pH 7 and with 1 % (w/v) NaCl. Ubiquinone 8 was the major respiratory quinone, and phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol were amongst the major polar lipids. On the basis of 16S rRNA gene sequence analysis, the strains were observed to be members of the family Alcaligenaceae, but could not be identified as members of any validly described genus. The low levels of 16S rRNA gene sequence similarity to other recognized taxa, together with comparative analysis of phenotypic traits and chemotaxonomic markers, supported the proposal of a new genus within the family Alcaligenaceae, for which the name Candidimonas gen. nov. is proposed. Strains SC-089T and SC-092T, which shared 99.1 % 16S rRNA gene sequence similarity, could be differentiated at the phenotypic level, and DNA–DNA hybridization results supported their identification as representing distinct species. The names proposed for these novel species are Candidimonas nitroreducens sp. nov. (type strain, SC-089T = LMG 24812T = CCUG 55806T) and Candidimonas humi sp. nov. (type strain, SC-092T = LMG 24813T = CCUG 55807T).

Five isolates, designated TA2, TA4, TA25T, KOxT and NS15T were isolated in previous studies by enrichment in mineral medium with potassium oxalate as the sole carbon source and were characterized using a polyphasic approach. The isolates were Gram-reaction-negative, aerobic, non-spore-forming rods. Phylogenetic analyses based on 16S rRNA and DNA gyrase B subunit (gyrB) gene sequences confirmed that the isolates belonged to the genus Pandoraea and were most closely related to Pandoraea sputorum and Pandoraea pnomenusa (97.2–99.7 % 16S rRNA gene sequence similarity). The isolates could be differentiated from their closest relatives on the basis of several phenotypic characteristics. The major cellular fatty acid profiles of the isolates comprised C16 : 0, C18 : 1ω7c, C17 : 0 cyclo and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). On the basis of DNA–DNA hybridization studies and phylogenetic analyses, the isolates represent three novel species within the genus Pandoraea, for which the names Pandoraea oxalativorans sp. nov. (TA25T  = NBRC 106091T  = CCM 7677T  = DSM 23570T), Pandoraea faecigallinarum sp. nov. (KOxT  = NBRC 106092T  = CCM 2766T  = DSM 23572T) and Pandoraea vervacti sp. nov. (NS15T  = NBRC 106088T  = CCM 7667T  = DSM 23571T) are proposed.

Two Gram-reaction-negative, rod-shaped, gliding, yellow-pigmented bacterial strains, designated ZLD-17T and ZLD-29T, were isolated from arid soil samples collected from Xinjiang Province, north-west China, and subjected to analysis using a polyphasic taxonomic approach. Both novel strains required 1.0–2.0 % (w/v) sea salts for optimal growth. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these two strains belong to the genus Lysobacter within the class Gammaproteobacteria. Strain ZLD-17T showed highest 16S rRNA gene sequence similarities to Lysobacter capsici KCTC 22007T (96.9 %), Lysobacter spongiicola DSM 21749T (96.8 %) and Lysobacter koreensis KCTC 12204T (96.8 %), whereas strain ZLD-29T showed highest sequence similarities to Lysobacter niastensis DSM 18481T (96.0 %) and Lysobacter enzymogenes DSM 2043T (95.9 %). 16S rRNA gene sequence similarity between ZLD-17T and ZLD-29T was 96.1 %. The DNA G+C contents of strains ZLD-17T and ZLD-29T were 67.9 and 68.2 mol%, respectively. The major cellular fatty acids of both strains were summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1ω7c), iso-C17 : 1ω9c, iso-C16 : 0, C16 : 0 and iso-C11 : 0 3-OH; their predominant isoprenoid quinone was Q-8 and their major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Based on their phenotypic characteristics, phylogenetic position as determined by 16S rRNA gene sequence analysis and chemotaxonomic data, strains ZLD-17T ( = CCTCC AB 207174T  = KCTC 23076T) and ZLD-29T ( = CCTCC AB 207175T = KCTC 23077T) represent two novel species of the genus Lysobacter, for which the names Lysobacter korlensis sp. nov. and Lysobacter bugurensis sp. nov. are proposed, respectively.

Two Gram-negative, rod-shaped, non-spore-forming strains, designated 08RB2639T and 08RB2781-1, were isolated from a sheep (Ovis aries) and a domestic boar (Sus scrofa domestica), respectively. By 16S rRNA gene sequencing, the isolates revealed identical sequences and were shown to belong to the Alphaproteobacteria. They exhibited 97.8 % 16S rRNA gene sequence similarity with Ochrobactrum rhizosphaerae PR17T, O. pituitosum CCUG 50899T, O. tritici SCII24T and O. haematophilum CCUG 38531T and 97.4 % sequence similarity with O. cytisi ESC1T, O. anthropi LMG 3331T and O. lupini LUP21T. The recA gene sequences of the two isolates showed only minor differences (99.5 % recA sequence similarity), and strain 08RB2639T exhibited the highest recA sequence similarity with Ochrobactrum intermedium CCUG 24694T (91.3 %). The quinone system was ubiquinone Q-10, with minor amounts of Q-9 and Q-11, the major polyamines were spermidine, putrescine and sym-homospermidine and the major lipids were phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine, with moderate amounts of the Ochrobactrum-specific unidentified aminolipid AL2. The major fatty acids (>20 %) were C18 : 1ω7c and C19 : 0 cyclo ω8c. These traits were in excellent agreement with the assignment of the isolates to the genus Ochrobactrum. DNA–DNA relatedness and physiological and biochemical tests allowed genotypic and phenotypic differentiation from other members of the genus Ochrobactrum. Hence, it is concluded that the isolates represent a novel species, for which the name Ochrobactrum pecoris sp. nov. is proposed (type strain 08RB2639T  = DSM 23868T  = CCUG 60088T  = CCM 7822T).

A bacterial strain designated NAA16T was isolated from a freshwater spring in Taiwan and was characterized using a polyphasic taxonomic approach. Strain NAA16T was aerobic, Gram-staining-negative, rod-shaped, non-spore-forming and motile by means of a single polar flagellum. Growth occurred at 20–40 °C (optimum, 25 °C), at pH 7.0–8.0 (optimum, pH 7.5) and with up to 1 % NaCl (optimum, 0.5 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that the closest relatives of strain NAA16T were Singularimonas variicoloris MN28T, Sinobacter flavus CW-KD 4T and Solimonas soli DCY12T, with respective sequence similarities of 96.7, 96.6 and 96.2 %. Phylogenetic trees reconstructed from 16S rRNA gene or rpoB sequences (encoding the β-subunit of the RNA polymerase) revealed that the novel strain NAA16T and these three closest relatives formed an independent phylogenetic clade within the Gammaproteobacteria. Strain NAA16T contained C16 : 0, C18 : 1ω7c and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as predominant fatty acids and possessed phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an uncharacterized aminophospholipid as dominant polar lipids. The major isoprenoid quinone was Q-8. The DNA G+C content of strain NAA16T was 66.2 mol%. The taxonomic relationship of strain NAA16T, Singularimonas variicoloris DSM 15731T, Sinobacter flavus DSM 18980T and Solimonas soli LMG 24014T was clarified by means of a direct experimental comparison. Based on phenotypic, chemotaxonomic and phylogenetic data, the descriptions of the genus Solimonas and its type species Solimonas soli are emended. Members of the genus are Gram-negative, oxidase- and catalase-positive, aerobic or facultatively anaerobic and chemo-organotrophic. Chemotaxonomically, members of the genus Solimonas possess Q-8 as the major respiratory quinone, C16 : 0 and C18 : 1ω7c as predominant fatty acids and phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an uncharacterized aminophospholipid as dominant polar lipids; the DNA G+C content is 64.9–68.4 mol%. Phylogenetic evidence, supported by chemotaxonomic and phenotypic data, allowed us to assign strain NAA16T to the genus Solimonas within the novel species Solimonas aquatica sp. nov. (type strain NAA16T  = BCRC 17835T  = LMG 24500T). The reclassification of Sinobacter flavus as Solimonas flava comb. nov. (type strain CW-KD 4T  = DSM 18980T  = KCTC 12881T  = CCTCC AB 206145T) and Singularimonas variicoloris as Solimonas variicoloris comb. nov. (type strain MN28T  = DSM 15731T  = LMG 22844T) is also proposed.

Here, we describe a novel bacterium, strain TUD-YJ37T, which can accumulate polyhydroxybutyrate (PHB) to more than 85 % (w/w) dry cell weight. The bacterium was isolated from a mixed-culture bioreactor by using a feast–famine regime and its properties were characterized. Phylogenetic analysis based on full 16S rRNA gene sequences revealed that the isolate is a member of the Gammaproteobacteria, forming an independent, deep phylogenetic lineage. It is most closely related to members of the genera Methylocaldum, Methylococcus and Natronocella, with sequence similarities below 91 %. Strain TUD-YJ37T was an obligately aerobic, ovoid, Gram-negative bacterium, motile by means of a polar flagellum. It utilized C2–C10 fatty acids as carbon and energy sources. The temperature range for growth was 20–35 °C, with an optimum of 30 °C; the pH range was 6.0–8.0, without a clear optimum. The major respiratory quinone was Q-8. Polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, three unidentified phospholipids, an unidentified aminolipid and another unidentified lipid. The predominant fatty acids in the membrane polar lipids were C16 : 1ω7c, C16 : 0 and C18 : 1ω7c. The G+C content of the genomic DNA was 67.4 mol%. On the basis of phenotypic, chemotaxonomic and molecular data, the isolate is proposed to represent a novel genus and species, for which the name Plasticicumulans acidivorans gen. nov., sp. nov. is proposed. The type strain of Plasticicumulans acidivorans is TUD-YJ37T ( = DSM 23606T  = CBS 122990T).

Recent rDNA sequencing of 25 isolates from a previous study, during which limonene-utilizing yeasts were isolated from monoterpene-rich environments by using 1,4-disubstituted cyclohexanes as sole carbon sources, led to the identification of four hitherto unknown Rhodotorula species. Analyses of the 26S rDNA D1/D2 region as well as the internal transcribed spacer (ITS) domain indicated that two isolates (CBS 8499T and CBS 10736) were identical and were closely related to Rhodotorula cycloclastica, a previously described limonene-utilizing yeast. These novel isolates differed from known yeast species and could be distinguished from R. cycloclastica by standard physiological tests. The other three isolates represent three novel Rhodotorula species, closely related to Sporobolomyces magnisporus. These three species could also be distinguished from other Rhodotorula species by standard physiological tests. Based on these results, we suggest that the new isolates represent novel species, for which the names Rhodotorula eucalyptica sp. nov. (type strain CBS 8499T  = NRRL Y-48408T), Rhodotorula pini sp. nov. (type strain CBS 10735T  = NRRL Y-48410T), Rhodotorula bloemfonteinensis sp. nov. (type strain CBS 8598T  = NRRL Y-48407T) and Rhodotorula orientis sp. nov. (type strain CBS 8594T  = NRRL Y-48719T) are proposed. R. eucalyptica and R. pini can also utilize limonene.