Yanghanlia caeni gen. nov., sp. nov., a novel taxon within the family Alcaligenaceae isolated from sludge of a pesticide-manufacturing factory.

A Gram-stain-negative bacterium, designated LG-2T, was isolated from sludge collected at a pesticide-manufacturing factory in Jiangsu Province, PR China. Cells of strain LG-2T were strictly aerobic, non-motile and spherical. Growth was observed at 15-42 °C (optimum, 30 °C), pH 6.0-9.0 (optimum, pH 7.0) and 0-3.0 % (w/v) NaCl (optimum, 1.0 %). LG-2T showed 95.5-96.9 % 16S rRNA sequence similarity to type strains in the genera Pusillimonas, Bordetella, Parapusillimonas, Candidimonas and Paracandidimonas of the family Alcaligenaceae. The phylogenomic tree indicated that strain LG-2T was clustered in the family Alcaligenaceae and formed a clade with Paracandidimonas soli IMT-305T, while the phylogenetic trees based on 16S rRNA gene sequences indicated that strain LG-2T formed a distinct clade within the family Alcaligenaceae. The average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity values between LG-2T and its closely related type strains in the genera Pusillimonas, Bordetella, Parapusillimonas, Candidimonas and Paracandidimonas were 70.8-75.3, 18.9-23.7 and 59.6 %-69.3 %, respectively. The major cellular fatty acids were C16 : 0, C17 : 0 cyclo, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and summed feature 2 (C12 : 0 aldehyde and/or unknown 10.928). The predominant menaquinone was Q-8. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, two aminophospholipids, three aminolipids and nine unknown polar lipids. The genome size of strain LG-2T was 3.2 Mb and the DNA G+C content was 63.4 mol%. On the basis of the phenotypic, phylogenetic and genomic results from this study, strain LG-2T represents a novel species of a new genus in the family Alcaligenaceae, for which the name Yanghanlia caeni gen. nov., sp. nov. is proposed, with strain LG-2T (=KCTC 8084T= CCTCC AB 2023123T) as the type strain.

Pusillimonas maritima sp. nov., isolated from surface seawater.

Two Gram-stain-negative, short rod-shaped and non-flagellated strains, designated 17-4AT and L52-1-41, were isolated from the surface seawater of the Indian Ocean and South China Sea, respectively. The 16S rRNA genes of the two strains shared sequence similarity of 99.45 %. Strain 17-4AT shared the highest 16S rRNA gene similarity of 98.02 % with Pusillimonas caeni EBR-8-1T, followed by Pusillimonas noertemannii BN9T (97.47 %), Pusillimonas soli MJ07T (96.93 %), Parapusillimonas granuli Ch07T (96.68 %), Pusillimonas ginsengisoli DCY25T (96.65 %), Eoetvoesia caeni PB3-7BT (96.63 %), Paracandidimonas caeni 24T (96.34 %), Castellaniella defragrans 54PinT (96.28 %) and Pusillimonas harenae B201T (96.05 %). L52-1-41 shared the highest 16S rRNA gene similarity of 97.74 % with Pusillimonas caeni EBR-8-1T, followed by Pusillimonas noertemannii BN9T (97.47 %), Pusillimonas soli MJ07T (96.65 %), Parapusillimonas granuli Ch07T (96.41 %), Pusillimonas ginsengisoli DCY25T (96.37 %), Eoetvoesia caeni PB3-7BT (96.35 %), Pusillimonas harenae B201T (96.28 %), and Paracandidimonas caeni 24T (96.06 %). The results of phylogenetic analyses indicated that 17-4AT and L52-1-41 formed a stable, distinct and highly supported lineage affiliated to the genus Pusillimonas. The results of the digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) analyses indicated that they represented a single species. They featured similar genomic DNA G+C contents of 53.2-53.4 mol%. Activities of catalase and oxidase were negative for both strains. The fatty acids patterns of 17-4AT and L52-1-41 were most similar, mostly comprised of C16 : 0, C17 : 0cyclo, C18 : 0, C18 : 1ω9c and summed feature 8 (C18 : 1ω7c and/or C18 : 1 ω6c). The major polar lipids of the two strains were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and unidentified aminolipids. The respiratory quinone of the two strains was Q-8. Hence, on the basis of the phenotypic, chemotaxonomic and genotypic data presented in this study, we proposed the classification of both strains as representatives of a novel species named Pusillimonas maritima sp. nov., with the type strain 17-4AT (=MCCC 1A12670T=KCTC 62121T=NBRC 113794T), and another strain L52-1-41 (=MCCC 1A05046=KCTC 52313).

Corticimicrobacter populi gen. nov., sp. nov., a member of the family Alcaligenaceae, isolated from symptomatic bark of Populus × euramericana canker.

One Gram-negative aerobic bacterial strain was isolated from the bark tissue of Populus × euramericana and investigated using a polyphasic approach including 16S rRNA gene sequencing and both phenotypic and chemotaxonomic assays. The 16S rRNA gene and housekeeping gene phylogenies suggest that the novel isolate is different from the other genera of the family Alcaligenaceae. The G+C content, major fatty acids, physiological and biochemical data supported the distinctiveness of the novel strain from reference species. The major fatty acids detected in the novel isolate were C16 : 1ω7c and/or C16 : 1ω6c, C16 : 0, C14 : 0 3OH and/or C16 : 1isoI and C18 : 1ω7c. The phospholipid profile of strain d3-2-2T contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminolipid, aminophospholipid and an unidentified lipid. The quinone of the novel isolate was Q-8. Therefore, based on the data, the strain constitutes a novel species of a novel genus within the family Alcaligenaceae, for which the name Corticimicrobacter populi gen. nov., sp. nov. is proposed. The type strain is 3d-2-2T (=CFCC 11891T=KCTC 52807T).

Parvibium lacunae gen. nov., sp. nov., a new member of the family Alcaligenaceae isolated from a freshwater pond.

A bacterial strain designated KMB9T was isolated from a freshwater pond in Taiwan and characterized using a polyphasic taxonomy approach. Cells of strain KMB9T were Gram-stain-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, motile by means of a monopolar flagellum, non-spore-forming and rods surrounded by a thick capsule and forming white-coloured colonies. Growth occurred at 20-40 °C (optimum, 25-37 °C), at pH 6.5-7.5 (optimum, pH 7.0) and with 0-0.5 % NaCl (optimum, 0 %). Phylogenetic analyses based on 16S rRNA gene and four housekeeping gene sequences (recA, rpoA, rpoB and atpD) showed that strain KMB9T forms a distinct phyletic line within the family Alcaligenaceae, and the levels of 16S rRNA gene sequence similarity to its closest relatives with validly published names were less than 93.3 %. The predominant fatty acids were summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C18 : 1ω7c. The major isoprenoid quinone was Q-8. The major polyamine was putrescine. The polar lipid profile revealed the presence of phosphatidylethanolamine, phosphatidylglycerol and several uncharacterized aminophospholipids, aminolipids, phospholipids and lipids. The genomic DNA G+C content of strain KMB9T was 54.5 mol%. On the basis of the genotypic and phenotypic data, strain KMB9T represents a novel species of a new genus in the family Alcaligenaceae, for which the name Parvibium lacunae gen. nov., sp. nov. is proposed. The type strain is KMB9T (=BCRC 81053T=LMG 30055T=KCTC 52814T).

Analysis of bacterial FAMEs using gas chromatography - vacuum ultraviolet spectroscopy for the identification and discrimination of bacteria.

The identification of microorganisms is very important in different fields and alternative methods are necessary for a rapid and simple identification. The use of fatty acids for bacterial identification is gaining attention as phenotypic characteristics are reflective of the genotype and are more easily analyzed. In this work, gas chromatography-vacuum ultraviolet spectroscopy (GC-VUV) was used to determine bacteria fatty acid methyl esters (FAMEs), to identify and discriminate different environmental bacteria based on their fatty acid profile. Microorganisms were grown in agar and their fatty acids extracted, saponified, and esterified before analysis. Unique FAME profiles were obtained for each microorganism mainly composed of branched, cyclopropane, hydroxy, saturated, and unsaturated fatty acid methyl esters. S. maltophilia showed a higher diversity of fatty acids while Bacillus species showed higher complexity in terms of branched-chain FAMEs, with several iso and anteiso forms. 12 different bacteria genera and 15 species were successfully differentiated based on their fatty acid profiles after performing PCA and cluster analysis. Some difficult to differentiate species, such as Bacillus sp., which are genetically very similar, were differentiated with the developed method.

Castellaniella fermenti sp. nov., isolated from a fermented meal.

A polyphasic taxonomic approach was used to characterize a presumably novel bacterium, designated strain CC-YTH191T, isolated from a fermented meal in Taiwan. Cells of strain CC-YTH191T were Gram-stain-negative aerobic rods, which grew at 15-40 °C (optimal 25-30 °C), pH 6.0-9.0 (optimal 7.0) and 1-2 % (w/v) NaCl (optimal 1 %). On the basis of 16S rRNA gene sequence analysis, strain CC-YTH191T appeared to belong to the genus Castellaniella, and was closely related to Castellaniella hirudinis (96.7 % similarity), Castellaniella ginsengisoli (96.7 %) and Castellaniella caeni (96.0 %), while with other related species it shared <96.0 % similarity. The major cellular fatty acids of the isolate were C16 : 0, C17 : 0cyclo, C14 : 0 3OH/C16 : 1iso I and C18 : 1ω7c/C18 : 1ω6c. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, three unidentified phospholipids, an unidentified aminolipid and an unidentified aminophospholpid. Putrescine was the predominant polyamine followed by spermidine. The DNA G+C content was 62.2 mol% and the predominant quinone system was ubiquinone 8 (Q-8). All these features confirmed the placement of the strain CC-YTH191T as a novel species within the genus Castellaniella, for which the name Castellaniella fermenti sp. nov. is proposed. The type strain is CC-YTH191T (=BCRC 81023T=JCM 31755T).

Pusillimonas caeni sp. nov., isolated from a sludge sample of a biofilm reactor.

A polyphasic taxonomic study was carried out on strain EBR-8-1T isolated from a biofilm reactor in Korea. The cells of the strain were Gram-stain negative, non-spore-forming, non-motile, and short rod-shaped. Comparative 16S rRNA gene sequence studies showed a clear affiliation of this strain with Betaproteobacteria, which showed high pairwise sequence similarities with Pusillimonas noertemannii BN9T (99.1 %), Pusillimonas soli MJ07T (97.3 %), Pusillimonas ginsengisoli DCY25T (97.2 %), and Pusillimonas harenae B201T (96.8 %). The phylogenetic analysis based on 16S rRNA gene sequences showed that the strain formed a clear phylogenetic lineage within the genus Pusillimonas. The major fatty acids were identified as C16:0, C17:0 cyclo and C19:0 cyclo ω8c. The major cellular polar lipids were identified as phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unidentified aminolipid. The respiratory quinone was identified as Q-8 and the genomic DNA G+C content was determined to be 63.3 mol%. On the basis of polyphasic evidence, it is proposed that strain EBR-8-1T should be placed in a new species, Pusillimonas caeni sp. nov. The type stain is EBR-8-1T (=KCTC 42353T = JCM 30463T).

Eoetvoesia caeni gen. nov., sp. nov., isolated from an activated sludge system treating coke plant effluent.

A novel bacterium, PB3-7B(T), was isolated on phenol-supplemented inorganic growth medium from a laboratory-scale wastewater purification system that treated coke plant effluent. 16S rRNA gene sequence analysis revealed that strain PB3-7B(T) belonged to the family Alcaligenaceae and showed the highest pairwise sequence similarity to Parapusillimonas granuli Ch07(T) (97.5%), Candidimonas bauzanensis BZ59(T) (97.3%) and Pusillimonas noertemannii BN9(T) (97.2%). Strain PB3-7B(T) was rod-shaped, motile and oxidase- and catalase-positive. The predominant fatty acids were C(16 : 0), C(17 : 0) cyclo, C(19 : 0) cyclo ω8c and C(14 : 0) 3-OH, and the major respiratory quinone was Q-8. The G+C content of the genomic DNA of strain PB3-7B(T) was 59.7 mol%. The novel bacterium can be distinguished from closely related type strains based on its urease activity and the capacity for assimilation of glycerol and amygdalin. On the basis of the phenotypic, chemotaxonomic and molecular data, strain PB3-7B(T) is considered to represent a new genus and species, for which the name Eoetvoesia caeni gen. nov., sp. nov. is proposed. The type strain of Eoetvoesia caeni is PB3-7B(T) ( = DSM 25520(T) = NCAIM B 02512(T)).

Description of Pelistega indica sp. nov., isolated from human gut.

A Gram-stain-negative, motile, non-spore-forming, coccoid bacterium was isolated from a stool sample of a healthy human subject and formed cream colour colonies on tryptic soy agar. Almost full-length (1500 bp) small subunit rRNA (16S rRNA) gene sequences were generated and a similarity search was conducted by blast. The results of the similarity search indicated that the bacterium belongs to the class Betaproteobacteria, family Alcaligenaceae. It showed maximum sequence similarity (96.5 %) with Pelistega europaea CCUG 39967(T) followed by Advenella mimigardefordensis DSM 17166(T) (96.1 %) and Taylorella asinigenitalis LMG 19572(T) (95.3 %). The DNA G+C content of strain HM-7(T) was 42 mol%. Strain HM-7(T) contained C14 : 0, C16 : 0, C16 : 0 3-OH and C18 : 0 as the dominant fatty acids. Morphological, physiological and biochemical data also indicated that strain HM-7(T) represents a member of the genus Pelistega, but at the same time distinguished it from Pelistega europaea CCUG 39967(T), the only species of the genus with a validly published name. Based on polyphasic characterization we conclude that the bacterium represents a novel species of the genus Pelistega and propose the name Pelistega indica sp. nov., with strain HM-7(T) ( = MCC 2185(T) = DSM 27484(T)) as the type strain of the species.

Characterization of oral bacterial diversity of irradiated patients by high-throughput sequencing.

The objective of this study was to investigate the compositional profiles and microbial shifts of oral microbiota during head-and-neck radiotherapy. Bioinformatic analysis based on 16S rRNA gene pyrosequencing was performed to assess the diversity and variation of oral microbiota of irradiated patients. Eight patients with head and neck cancers were involved in this study. For each patient, supragingival plaque samples were collected at seven time points before and during radiotherapy. A total of 147,232 qualified sequences were obtained through pyrosequencing and bioinformatic analysis, representing 3,460 species level operational taxonomic units (OTUs) and 140 genus level taxa. Temporal variations were observed across different time points and supported by cluster analysis based on weighted UniFrac metrics. Moreover, the low evenness of oral microbial communities in relative abundance was revealed by Lorenz curves. This study contributed to a better understanding of the detailed characterization of oral bacterial diversity of irradiated patients.

Isolation of Kerstersia gyiorum from a patient with cholesteatomatous chronic otitis media.

We describe the first case of a Kerstersia gyiorum strain isolated from a patient with cholesteatomatous chronic otitis media. We emphasize the isolation of members of the family Alcaligenaceae in serious infections and unusual sites and the importance of polyphasic identification addressing the definitive identification.

Candidimonas nitroreducens gen. nov., sp. nov. and Candidimonas humi sp. nov., isolated from sewage sludge compost.

Two bacterial strains (SC-089(T) and SC-092(T)) 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-089(T) and SC-092(T), 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-089(T) = LMG 24812(T) = CCUG 55806(T)) and Candidimonas humi sp. nov. (type strain, SC-092(T) = LMG 24813(T) = CCUG 55807(T)).

Pusillimonas soli sp. nov., isolated from farm soil.

A Gram-negative, motile, non-spore-forming bacterial strain, designated MJ07(T), was isolated from a farm soil and was characterized to determine its taxonomic position by using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain MJ07(T) belongs to the family Alcaligenaceae, class Betaproteobacteria, and is related most closely to Pusillimonas ginsengisoli KCTC 22046(T) (98.6 % sequence similarity) and Pusillimonas noertemannii BN9(T) (96.9 %). The levels of 16S rRNA gene sequence similarity between strain MJ07(T) and members of all other recognized species of the family Alcaligenaceae were below 95.2 %. The G+C content of the genomic DNA of strain MJ07(T) was 59.4 mol%. The detection of a quinone system with ubiquinone Q-8 as the major respiratory lipoquinone, putrescine as the predominant polyamine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and two unknown aminolipids as major polar lipids and a fatty acid profile with C16:0 (32.0 %), C17:0cyclo (24.7 %) and C19:0cyclo ω8c (11.5 %) as the major components supported the affiliation of strain MJ07(T) to the genus Pusillimonas. Strain MJ07(T) exhibited relatively low levels of DNA-DNA relatedness with respect to P. ginsengisoli KCTC 22046(T) (50 ± 8 %) and P. noertemannii KACC 13183(T) (18 ± 7 %). On the basis of its phenotypic and genotypic properties together with its phylogenetic distinctiveness, strain MJ07(T) (=KCTC 22455(T) =JCM 16386(T)) should be classified in the genus Pusillimonas as the type strain of a novel species, for which the name Pusillimonas soli sp. nov. is proposed.

Parapusillimonas granuli gen. nov., sp. nov., isolated from granules from a wastewater-treatment bioreactor.

A novel betaproteobacterium, designated strain Ch07(T), was isolated from granules from the wastewater-treatment bioreactor of an alcohol fermentation factory in South Korea. In order to determine its taxonomic position, the novel strain was characterized using a polyphasic approach. The new strain was Gram-negative, facultatively anaerobic, non-spore-forming, motile and short rod-shaped. 16S rRNA gene sequence analysis revealed that strain Ch07(T) belonged to the class Betaproteobacteria, being related to Pusillimonas noertemannii BN9(T) (gene sequence similarity 97.30 %), Achromobacter xylosoxidans subsp. xylosoxidans DSM 10346(T) (97.09 %), Bordetella pertussis DSM 5571(T) (97.01 %), Pigmentiphaga kullae DSM 13608(T) (96.68 %) and Castellaniella defragrans DSM 1214(T) (96.47 %). The results of DNA-DNA hybridization tests showed that reassociation values were less than 62 % with respect to these closely related type strains. Chemotaxonomic data showed that strain Ch07(T) possessed ubiquinone Q-8. The G+C content of the genomic DNA was 67.9+/-0.1 mol%. The major polyamine of strain Ch07(T) was putrescine. The major polar lipids of strain Ch07(T) were phosphatidylethanolamine, followed by diphosphatidylglycerol and phosphatidylglycerol. When strain Ch07(T) was incubated on tryptic soy agar, the major cellular fatty acids were C(16 : 0), C(17 : 0) cyclo, summed feature 3 (C(16 : 1)omega7c/iso C(15 : 0) 2-OH) and summed feature 5 (C(18 : 1)omega7c/omega9t/omega12t). The results of DNA-DNA hybridizations, in combination with the chemotaxonomic and physiological data, demonstrated that strain Ch07(T) represents a novel species of a new genus, for which the name Parapusillimonas granuli gen. nov., sp. nov. is proposed. The type strain of the type species is Ch07(T) (=KCTC 12668(T)=LMG 24012(T)).

Castellaniella daejeonensis sp. nov., isolated from soil.

A Gram-negative, non-spore-forming, motile, facultatively anaerobic bacterium, designated strain MJ06(T), was isolated from oil-contaminated soil and was characterized taxonomically by using a polyphasic approach. Strain MJ06(T) contained ubiquinone Q-8 as the major respiratory lipoquinone, putrescine as the predominant polyamine and phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. The G+C content of the genomic DNA of strain MJ06(T) was 66.2 mol%. The major fatty acids were summed feature 4 (C(16 : 1)ω 7c and/or iso-C(15 : 0) 2-OH; 32.5 %), C(16 : 0) (22.8 %) and summed feature 7 (one or more of C(18 : 1)ω 7c, C(18 : 1)ω 9t and C(18 : 1)ω 12t; 14.9 %). Comparative 16S rRNA gene sequence analysis showed that strain MJ06(T) belonged to the family Alcaligenaceae, class Betaproteobacteria, and joined the evolutionary radiation enclosed by the genus Castellaniella. Levels of 16S rRNA gene sequence similarity between strain MJ06(T) and its phylogenetically closest relatives, Castellaniella denitrificans NKNTAU(T), Castellaniella defragrans 54Pin(T), Castellaniella ginsengisoli DCY36(T) and Castellaniella caeni Ho-11(T), were 98.6, 98.3, 97.8 and 97.3 %, respectively. Levels of similarity between strain MJ06(T) and the type strains of all other recognized species in the family Alcaligenaceae were below 95.0 %. Strain MJ06(T) exhibited relatively low levels of DNA-DNA relatedness with respect to C. defragrans DSM 12141(T) (52 %), C. denitrificans DSM 11046(T) (31 %), C. ginsengisoli KCTC 22398(T) (18 %) and C. caeni KCTC 12197(T) (15 %). On the basis of its phenotypic and genotypic properties together with phylogenetic distinctiveness, strain MJ06(T) is considered to represent a novel species of the genus Castellaniella, for which the name Castellaniella daejeonensis sp. nov. is proposed. The type strain is MJ06(T) (=KCTC 22454(T) =JCM 16240(T)).

Castellaniella ginsengisoli sp. nov., a beta-glucosidase-producing bacterium.

A Gram-negative, motile bacterium, designated DCY36T, was isolated from soil of a ginseng field in South Korea and was characterized using a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence analysis showed that strain DCY36T belongs to genus Castellaniella in the family Alcaligenaceae of the class Betaproteobacteria. The 16S rRNA gene sequence similarities between strain DCY36T and the three recognized representatives of the genus, Castellaniella caeni Ho-11T, Castellaniella defragrans 54PinT and Castellaniella denitrificans NKNTAUT, were 98.4, 97.5 and 98.1%, respectively. Strain DCY36T exhibited relatively low levels of DNA-DNA relatedness with respect to these three species. The G+C content of the genomic DNA was 63.7 mol%. Strain DCY36T contained ubiquinone Q-8. The major fatty acids were C16:0 (27.4%), C18:1omega7c (16.9%) and summed feature 4 (C16:1omega7c and C15:0 iso 2-OH, 32.5%). On the basis of phenotypic and genotypic properties and phylogenetic distinctiveness, strain DCY36T (=KCTC 22398T=JCM 15515T) should be classified in the genus Castellaniella as the type strain of a novel species, for which the name Castellaniella ginsengisoli sp. nov. is proposed.

Castellaniella caeni sp. nov., a denitrifying bacterium isolated from sludge of a leachate treatment plant.

A Gram-negative, non-motile, facultatively anaerobic, denitrifying bacterial strain, designated Ho-11(T), was isolated from sludge of a leachate treatment plant and was characterized taxonomically by using a polyphasic approach. The G+C content of the genomic DNA was 63.5 mol%. Strain Ho-11(T) contained ubiquinone Q-8 as the major respiratory lipoquinone and putrescine as the predominant polyamine. The major fatty acids were summed feature 4 (C(16:1)omega7c and/or iso-C(15:0) 2-OH; 29.3%), C(16:0) (28.0%) and summed feature 7 (C(18:1)omega7c, C(18:1)omega9t and/or C(18:1)omega12t; 19.8%). Comparative 16S rRNA gene sequence analysis showed that strain Ho-11(T) belonged to the family Alcaligenaceae, class Betaproteobacteria, and joined the evolutionary radiation enclosed by the genus Castellaniella. 16S rRNA gene sequence similarities between strain Ho-11(T) and the type strains of the two recognized species of the genus, Castellaniella denitrificans DSM 11046(T) and Castellaniella defragrans DSM 12141(T), were 97.8 and 97.4%, respectively. Levels of similarity between strain Ho-11(T) and all other recognized species of the family Alcaligenaceae were below 95.6%. Strain Ho-11(T) exhibited relatively low levels of DNA-DNA relatedness with respect to C. denitrificans DSM 11046(T) (33%) and C. defragrans DSM 12141(T) (28%). On the basis of its phenotypic and genotypic properties together with phylogenetic distinctiveness, strain Ho-11(T) (=KCTC 12197(T)=LMG 23411(T)) should be classified in the genus Castellaniella as the type strain of a novel species, for which the name Castellaniella caeni sp. nov. is proposed.

Pigmentiphaga daeguensis sp. nov., isolated from wastewater of a dye works, and emended description of the genus Pigmentiphaga.

A Gram-negative, non-spore-forming, rod-shaped Pigmentiphaga-like bacterial strain, K110(T), was isolated from wastewater collected from a dye works in Korea and was subjected to a polyphasic taxonomic analysis. Strain K110(T) grew optimally at pH 7.0--8.0 and 37 degrees C in the presence of 0.5 % (w/v) NaCl. It contained Q-8 as the predominant ubiquinone and C(16 : 0), cyclo C(17 : 0) and cyclo C(19 : 0)omega8c as the major fatty acids. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine and two unidentified aminolipids. The DNA G+C content was 67.4 mol%. In a neighbour-joining phylogenetic tree constructed on the basis of 16S rRNA gene sequences, strain K110(T) joined Pigmentiphaga kullae, the sole species of the genus, at a bootstrap confidence level of 100 %. Strain K110(T) exhibited a 16S rRNA gene sequence similarity of 99.4 % with respect to the type strain of P. kullae. Although strain K110(T) was found to be similar to P. kullae in terms of phenotypic properties, it differed in terms of motility, polar lipids, DNA-DNA relatedness and repetitive extragenic palindromic PCR genomic fingerprinting patterns. On the basis of phenotypic, phylogenetic and genetic data, strain K110(T) represents a novel species of the genus Pigmentiphaga, for which the name Pigmentiphaga daeguensis sp. nov. is proposed. The type strain is K110(T) (=KCTC 12838(T)=JCM 14330(T)).

Advenella incenata gen. nov., sp. nov., a novel member of the Alcaligenaceae, isolated from various clinical samples.

A polyphasic taxonomic study of 14 isolates recovered from various human and veterinary clinical samples was performed. Phenotypically these isolates shared several characteristics with members of the Alcaligenaceae and related genera. Random amplified polymorphic DNA fingerprinting and whole-cell protein analysis suggested the presence of multiple genomic groups, which was confirmed by DNA-DNA hybridization experiments. 16S rRNA gene sequence analysis indicated that these isolates were related to the genera Pelistega, Taylorella, Oligella, Pigmentiphaga, Alcaligenes, Kerstersia, Achromobacter and Bordetella and belonged to the family Alcaligenaceae. Based on the results of the present study the organisms were classified in a novel genus, Advenella gen. nov. This genus comprises one named species, Advenella incenata sp. nov. (type strain LMG 22250T=CCUG 45225T) and five currently unnamed genomic species. The DNA G+C content of members of the novel genus Advenella is between 54.0 and 57.7 mol%.