Paenalcaligenes faecalis sp. nov., a novel species of the family Alcaligenaceae isolated from chicken faeces.

A Gram-negative, motile, rod-shaped aerobic and alkalogenic bacterium, designated as strain YLCF04T, was isolated from chicken faeces. Its growth was optimal at 28 °C (range, 10-40 °C), pH 8 (range, pH 6-9) and in 1 % (w/v) NaCl (range, 0-10 %). It was classified to the genus Paenalcaligenes and was most closely related to Paenalcaligenes hominis CCUG 53761AT (97.5 % similarity) based on 16S rRNA gene sequence analysis. Average nucleotide identity and digital DNA-DNA hybridization values between YLCF04T and P. hominis CCUG 53761AT were 76.3 and 18.2 %, respectively. Strain YLCF04T has a genome size of 2.7 Mb with DNA G+C content of 46.3 mol%. Based on its phylogenetic, genomic, phenotypic and biochemical characteristics, strain YLCF04T represents a novel species of the genus Paenalcaligenes, for which the name Paenalcaligenes faecalis sp. nov. is proposed. The type strain is YLCF04T (=CCTCC AB 2022359T= KCTC 92789T).

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.

The phylogeny of the genus Azohydromonas supports its transfer to the family Comamonadaceae.

The genus Azohydromonas encompasses five validly described species belonging to the betaproteobacterial class. Recognized for their potential biotechnological uses, they were first described as belonging to the genus Alcaligenes. The phylogeny of the 16S rRNA gene of the original strains as well as newly described species led to a description of these strains within a new bacterial genus, Azohydromonas. However, the phylogenetic position of this genus remains described as part of the family Alcaligenaceae, even those some authors have placed it within the order Burkholderiales. To unravel the precise position of the genus Azohydromonas, a wide phylogenomic analysis was performed. The results of 16S rRNA gene phylogeny, as well as those obtained by the multilocus analysis of homologous proteins and overall genome relatedness indices, support the reclassification of Azohydromonas in the Rubrivivax-Ideonella lineage of the family Comamonadaceae, so the transfer of this genus is proposed.

Orrella amnicola sp. nov., isolated from a freshwater river, reclassification of Algicoccus marinus as Orrella marina comb. nov., and emended description of the genus Orrella.

A novel Gram-negative, aerobic, non-motile, ovoid to rod-shaped bacterium, designated NBD-18T, was isolated from a freshwater river in Taiwan. Optimal growth occurred at 30 °C, at pH 6 and in the absence of NaCl. The predominant fatty acids of strain NBD-18T were C16 : 0, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C17 : 0 cyclo and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidyldimethylethanolamine. The major polyamine was putrescine. The major isoprenoid quinone was Q-8. The genomic DNA G+C content of strain NBD-18T was 50.9 %. Strain NBD-18T was most closely related to Orrella dioscoreae LMG 29303T and Algicoccus marinus HZ20T at a 16S rRNA gene sequence similarity of 97.7 %. 16S rRNA gene sequence similarity between O. dioscoreae LMG 29303T and A. marinus HZ20T was 97.7 %. Phylogenetic analyses based on 16S rRNA gene sequences and an up-to-date bacterial core gene set indicated that strain NBD-18T, O. dioscoreae LMG 29303T and A. marinus HZ20T are affiliated with the same genus. Digital DNA-DNA hybridization, average nucleotide identity and average amino acid identity values among these three strains supported that they belong to the same genus and that strain NBD-18T represents a novel species. Thus, A. marinus HZ20T should be reclassified as Orrella marina comb. nov. based on the rules for priority of publication and validation. On the basis of the genotypic, chemotaxonomic and phenotypic data, strain NBD-18T represents a novel species in the genus Orrella, for which the name Orrella amnicola sp. nov. is proposed. The type strain is NBD-18T (=BCRC 81197T=LMG 31338T).

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).

The applicability of gene sequencing and MALDI-TOF to identify less common gram-negative rods (Advenella, Castellaniella, Kaistia, Pusillimonas and Sphingobacterium) from environmental isolates.

Our aim was to identify less common non-fermenting gram-negative rods during the bioremediation process. Five genera were found: Advenella, Castellaniella, Kaistia, Pusillimonas and Sphingobacterium, for a total of 15 isolates. Therefore, we evaluated the applicability of four methods currently available for bacteria identification: (1) conventional biochemical methods, (2) the VITEK®-2 system, (3) MALDI-TOF mass spectrometry and (4) 16S rRNA gene sequencing. The biochemical methods and the VITEK®-2 system were reliable only for the Sphingobacterium isolate and solely at the genus level. Both MALDI-TOF mass spectrometry platforms (Bruker and VITEK® MS) did not achieve reliable identification results for any of these genera. 16S rRNA gene sequencing identified eight isolates to the species level but not to the subspecies level, when applicable. The remaining seven isolates were reliably identified through 16S rRNA gene sequencing to the genus level only. Our findings suggest that the detection and identification of less common genera (and species) that appeared at certain moments during the bioremediation process can be a challenge to microbiologists considering the most used techniques. In addition, more studies are required to confirm our results.

Purification, characterization, and biological cytotoxic activity of the extracellular cholesterol oxidase produced by Castellaniella sp. COX.

A new bacterial strain producing extracellular cholesterol oxidase (ChOx) was isolated and identified as Castellaniella sp. COX. The ChOx was purified by salting-out and ion-exchange chromatography up to 10.4-fold, with a specific activity of 15 U/mg with a molecular mass of 59 kDa. The purified ChOx exhibited pH 8.0 and temperature 40°C for its optimum activity. The enzyme showed stability over a wide pH range and was most stable at pH value 7.0, and at pH 8.0, it retained almost 86% of its initial activity after 3 h of incubation at 37°C. The enzyme possessed a half-life of 8 h at 37°C, 7 h at 40°C, and 3 h at 50°C. A Lineweaver-Burk plot was calibrated to determine its Km (0.16 mM) and Vmax (18.7 µmol·mg-1 ·min-1 ). The ChOx activity was enhanced with Ca2+ , Mg2+ , and Mn2+ while it was inhibited by Hg2+ , Ba2+ , Fe2+ , Cu2+ , and Zn2+ ions. Organic solvents like acetone, n-butanol, toluene, dimethyl sulfoxide, chloroform, benzene, and methanol were well tolerated by the enzyme while iso-propanol and ethanol were found to enhance the activity of purified ChOx. ChOx induced cytotoxicity with an IC50 value of 1.78 and 1.88 U/ml against human RD and U87MG established cell lines, respectively, while broadly sparing the normal human cells.

Algicoccus marinus gen. nov. sp. nov., a marine bacterium isolated from the surface of brown seaweed Laminaria japonica.

A Gram-staining-negative, strictly aerobic, non-motile, ovoid- to rod-shaped bacterium, designated as HZ20T, was isolated from the surface of a brown seaweed (Laminaria japonica) sample collected from the East China Sea. Colonies are 1.0-2.0 mm in diameter, smooth, circular, convex and yellow after grown on MA at 28 °C for 72 h. The strain was found to grow at 4-50 °C (optimum, 37 °C), pH 5.0-9.5 (optimum, pH 7.0-7.5) and with 0-10% (w/v) NaCl (optimum, 1.0-1.5%). Chemotaxonomic analysis showed ubiquinone-8 as the only quinone, C17:0 cyclo, C16:0, summed feature 8 (C18:1ω7c and/or C18:1ω6c) and summed feature 2 (C12:0 aldehyde/unknown 10.9525/C16:1 iso I/C14:0 3OH) as the major fatty acids (> 5%), and diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, one unidentified amino phospholipid, two unidentified phospholipids, five unidentified glycolipid and two unidentified lipids as the polar lipids. The DNA G + C content was 55.5 mol %. 16S rRNA gene sequences of the isolate showed highest similarities to Bordetella flabilis AU10664T (97.1%), Parapusillimonas granuli Ch07T (97.1%), Paracandidimonas soli IMT-305T (97.1%), Kerstersia gyiorum LMG5906T (97.0%) and Bordetella sputigena LMG 28641T (97.0%). The phylogenetic trees using 16S rRNA gene and genome sequences both showed that the strain HZ20T formed a deep branch separated from other related genera, indicating that it represents a novel species of a novel genus. The calculated average nucleotide identity (ANI) and percent of conserved proteins (POCP) values using genome sequences of strain HZ20T and related strains also support this conclusion. Based on the phenotypic properties and phylogenetic distinctiveness, we propose strain HZ20T (= MCCC 1K03465T = KCTC 62330T) to represent a novel species of a novel genus with the name Algicoccus marinus gen. nov. sp. nov.

Pusillimonas thiosulfatoxidans sp. nov., a thiosulfate oxidizer isolated from activated sludge.

A Gram-stain-negative, motile bacterium, designated strain YE3T, was isolated from activated sludge obtained from a municipal wastewater treatment plant in Daejeon Metropolitan City, Republic of Korea. The cells were oxidase- and catalase-positive, and grew under aerobic conditions at 10-40 °C (optimum, 30 °C), with 1.0-8.0 % (w/v) NaCl (1.0 %) and at pH 5.5-9.0 (pH 7.0). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YE3T was most closely related to Pusillimonasharenae KACC 14927T (98.2 % sequence similarity) and Pusillimonasginsengisoli KCTC 22046T (98.0 %). DNA-DNA relatedness values for strain YE3T and P. harenae KACC 14927T, P. ginsengisoli KCTC 22046T and P. soli KCTC 22455T were 28.7±2.27 %, 21.3±1.16 %, and 14.0±0.67 %, respectively. The genomic G+C content of the type strain YE3T was 59.3 mol%, as determined by whole-genome sequencing. The dominant fatty acids were C16 : 0 (39.2 %) and C17 : 0cyclo (37.5 %). The major polar lipids of strain YE3T were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Two aminophospholipids and four unidentified lipids were also detected. Furthermore, strain YE3T was able to oxidize thiosulfate under heterotrophic conditions. Based on the phenotypic, genotypic, chemotaxonomic and phylogenetic analyses, strain YE3T represents a novel species of the genus Pusillimonas, for which the name Pusillimonas thiosulfatoxidans sp. nov. is proposed. The type strain is YE3T (=KCTC 62737T=NBRC 113113T).

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).

Paracandidimonas caeni sp. nov., isolated from sludge.

A beige-pigmented, Gram-stain-negative, aerobic, non-motile, rod-shaped bacterium, named 24T, was isolated from sludge of a pesticide manufacturing factory in Nantong, Jiangsu Province, China. 16S rRNA gene sequence analysis revealed that strain 24T shared highest similarity with Parapusillimonas granuli Ch07T (98.20 %), followed by Candidimonas nitroreducens SC-089T (98.07 %) and Paracandidimonas soli IMT-305T (98.03 %). Phylogenetic trees showed that strain 24T formed a distinct clade with Paracandidimonas soli IMT-305T. The results of DNA-DNA hybridization tests showed that reassociation values were less than 45 % with respect to these closely related type strains. Strain 24T contained Q-8 and putrescine as the major respiratory quinone and polyamine, respectively. The main cellular fatty acids were summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), summed feature 2 (iso-C16 : 1 I/C14 0 3-OH/C12 : 0 aldehyde), summed feature 8 (C18 : 1ω7c/C18 : 1ω6c) and C12 : 0. The polar lipid profile included phosphatidylmethylethanolamin, phosphatidylethanolamine, phosphatidylglycerol, one unidentified phospholipid and one unidentified aminolipid. The G+C content was 56.83 mol%. Combined data from phenotypic, phylogenetic and DNA-DNA relatedness studies demonstrated that strain 24T represents a novel species of the genus Paracandidimonas, for which the name Paracandidimonas caeni sp. nov. is proposed. The type strain is 24T (=CCTCC AB 2018057T=KACC 19692T).

Pigmentiphaga humi sp. nov., isolated from soil amended with humic acid.

A slightly beige-pigmented, Gram-stain-negative, rod-shaped bacterium, strain IMT-318T, was isolated from soil in a field located in Malvern, Alabama, USA. Phylogenetic analysis based on the 16S rRNA gene placed the strain within the genus Pigmentiphaga with highest 16S rRNA gene sequence similarity of 98.74 % and 98.67 % to the type strains of Pigmentiphaga kullae and Pigmentiphaga daeguensis, respectively. Sequence similarities to all other species of the genus were below 98.0 %. Results of the chemotaxonomic analysis, however, showed clear similarities to the genus Pigmentiphaga. The main cellular fatty acids of the strain were C16 : 0, C18 : 1 ω7c, C17 : 0 cyclo and C19 : 0 cyclo ω8c. The major quinone was ubiquinone Q-8. The polar lipid profile was composed of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified aminophospholipid. In the polyamine pattern, putrescine and 2-hydroxyputrescine were predominant. The diamino acid of the peptidoglycan was meso-diaminopimelic acid. Based on phylogenetic, chemotaxonomic and phenotypic analyses, we propose a new species of the genus Pigmentiphaga, with the name Pigmentiphaga humi sp. nov. and strain IMT-318T (=LMG 30658T=CIP 111626T=CCM 8859T) as the type strain.

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).

Mesosutterella multiformis gen. nov., sp. nov., a member of the family Sutterellaceae and Sutterella megalosphaeroides sp. nov., isolated from human faeces.

Two novel, obligately anaerobic, Gram-stain-negative, rod or coccoid-shaped bacteria, designated strains 4NBBH2T and 6FBBBH3T, were isolated from faecal samples of a healthy Japanese woman and man. The 16S rRNA gene sequence analysis showed that these strains represent a distinct lineage within the family Sutterellaceae. Strain 4NBBH2T formed a monophyletic branch between the genera Parasutterella and Sutterella, with sequence similarity to Sutterella stercoricanis CCUG 47620T (92.6 %), followed by Sutterella wadsworthensis WAL 7877 (92.4 %), Sutterella parvirubra YIT 11816T (92.1 %) and Parasutterella secunda YIT 12071T (91.8 %). Strain 6FBBBH3T was affiliated to the genus Sutterella, with highest similarity to S. stercoricanis CCUG 47620T (97.1 %), followed by S. parvirubra YIT 11816T (96.6 %) and S. wadsworthensis WAL 7877 (95.2 %). Strains 4NBBH2T and 6FBBBH3T were asaccharolytic. Analysis of fatty acids revealed that strain 4NBBH2T could be differentiated from Sutterella species (including strain 6FBBBH3T) by the presence of a low concentration of C16 : 1ω7c. The major respiratory quinones of strain 4NBBH2T were menaquinone (MK)-6 and methylmenaquinone (MMK)-6, whereas those of strain 6FBBBH3T were MK-5 and MMK-5. The G+C content of the genomic DNA of strains 4NBBH2T and 6FBBBH3T were 56.9 and 62.8 mol%, respectively. On the basis of the collected data, strain 4NBBH2T represents a novel species in a novel genus of the family Sutterellaceae, for which the name Mesosutterella multiformis gen. nov., sp. nov. is proposed. The type strain is 4NBBH2T (=JCM 32464T=DSM 106860T). We also propose a novel Sutterella species, Sutterellamegalosphaeroides sp. nov., for strain 6FBBBH3T (=JCM 32470T=DSM 106861T).

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).

The first case report of emphysematous pyelonephritis and bacteremia due to Oligella urethralis.

Oligella urethralis (O. urethralis) is an organism that rarely causes infections in humans. We report the case of a 90-year-old bedridden woman with progressive dementia who was placed in a long-term-care facility. She was admitted to our hospital due to fever and unconsciousness with pyuria. The abdominal computed tomography showed left pneumatosis and urinary stone. Fluoroquinolones-resistant O. urethralis, which was identified by the Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) and the 16S rRNA gene sequencing, was isolated form the blood and urine cultures at admission. To the best of our knowledge, this is the first case of emphysematous pyelonephritis caused by O. urethralis.

Caenimicrobium hargitense gen. nov., sp. nov., a new member of the family Alcaligenaceae (Betaproteobacteria) isolated from activated sludge.

A new betaproteobacterium, CGII-59m2T, was isolated from an activated sludge bioreactor which treated landfill leachate. The 16S rRNA gene sequence analysis revealed that strain CGII-59m2T belonged to the family Alcaligenaceae and shared the highest pairwise similarity values with Parapusillimonas granuli LMG 24012T (97.7 %), various species of the genus Bordetella (97.3-97.0 %) and Candidimonas nitroreducens LMG 24812T (97.0 %). Cells of strain CGII-59m2T were rod-shaped, non-motile, and oxidase- and catalase-positive. The predominant fatty acids were C16 : 1ω7c, C16 : 0, cyclo C17 : 0 and C18 : 1ω7c, the major respiratory quinone was Q-8, and the main polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown phospholipid. The G+C content of the genomic DNA of strain CGII-59m2T was 62.3 mol%. The new bacterium can be distinguished from the closely related type strains based on its non-motile cells and its high C16 : 1ω7c fatty acid content. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGII-59m2T is considered to represent a novel species of a new genus, for which the name Caenimicrobium hargitense gen. nov., sp. nov. is proposed. The type strain is CGII-59m2T (=DSM 29806T=NCAIM B.02615T).

Quisquiliibacterium transsilvanicum gen. nov., sp. nov., a novel betaproteobacterium isolated from a waste-treating bioreactor.

A new betaproteobacterium, CGI-09T, was isolated from an activated sludge bioreactor which treated landfill leachate. Based on 16S rRNA gene sequence analysis, the new strain shared the highest pairwise similarity values with members of the order Burkholderiales: Derxia gummosa IAM 13946T (family Alcaligenaceae), 93.7 % and Lautropia mirabilis DSM 11362T (family Burkholderiaceae), 93.6 %. Cells of strain CGI-09T were rod-shaped and non-motile. The new strain was oxidase and catalase positive and capable of reducing nitrate to nitrite. The predominant fatty acids were C16 : 1 ω7c, C16 : 0, cycloC17 : 0 and C18 : 1 ω7c, the major respiratory quinone was Q-8, and the detected polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unknown phospholipid. The G+C content of the genomic DNA of strain CGI-09T was 70.2 mol%. The new bacterium can be distinguished from the members of genera Derxia and Lautropia based on its non-motile cells, arginine dihydrolase activity, its high cyclo C17 : 0 fatty acid content and the lack of hydroxy fatty acids. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGI-09T is considered to represent a new genus and species within the family Burkholderiaceae, for which the name Quisquiliibacterium transsilvanicum gen. nov., sp. nov. is proposed. The type strain is CGI-09T (=DSM 29781T=JCM 31785T).

Saccharedens versatilis gen. nov., sp. nov., a sugar-degrading member of the Burkholderiales isolated from Cephalotes rohweri ant guts.

Cephalotes 'turtle' ants host a core group of gut-associated symbionts, but their potential contributions to ant nutrition and disease resistance remain uncharacterized in vitro. To gain a better understanding of the metabolic capability of core symbionts belonging to the Burkholderiales, we cultivated and characterized strain CAG32T from the guts of Cephalotes rohweri ants. Strain CAG32T was rod-shaped, Gram-stain-negative, motile and formed pale-white colonies on trypticase soy agar. Optimum growth occurred under an atmosphere of 20 % O2 supplemented with 1 % CO2. Strain CAG32T grew under NaCl concentrations of 0-2.0 %, temperatures of 23-47 °C and pH values of 4.0-8.0, and was capable of producing n-butyric acid and degrading carbohydrates for growth. The G+C content of the genomic DNA was 59.2±0.6 mol% and the major fatty acids were C16 : 0, C16 : 1ω7c/C16 : 1ω6c, C17 : 0 cylcopropane, C12 : 0 and C14 : 0 3-OH/C16 : 1 iso I. The only respiratory quinone detected was ubiquinone-8 (Q-8) and the major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Based on phylogenetic analysis of the 16S rRNA gene sequence, strain CAG32T shared 96.9 % nucleotide similarity with its closest cultivated neighbours Bordetella petrii Se-1111RT and Bordetella bronchiseptica ATCC 19395T. This, combined with differences in the phenotypic and biochemical profile from neighbouring strains, warrants the classification of strain CAG32T as representing a novel species of a new genus within the Burkholderiales family Alcaligenaceae. The name Saccharedens versatilis gen. nov., sp. nov. is proposed. The type strain of Saccharedens versatilis is CAG32T (=NCIMB 15010T=DSM 100909T).

Pigmentiphaga aceris sp. nov., isolated from tree sap.

Two Gram-stain-negative bacterial strains, SAP-32T and SAP-36, were isolated from sap drawn from the Acer pictum from Mount Halla in Jeju, Republic of Korea. The organisms were strictly aerobic, non-sporulating, motile rods and showed growth at 10-30 °C, pH 7-8 and with 0-2 % NaCl. The major isoprenoid quinone was Q-8. The predominant fatty acids were C16 : 0, cyclo-C17 : 0, summed feature 3 and C18 : 0. The polar lipids contained phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, an unknown aminophosphoglycolipid, an unknown glycolipid, an unknown phospholipid and two unknown lipids. The DNA G+C content was 64.4 mol%. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that SAP-32T and SAP-36 formed a distinct cluster with members of the genus Pigmentiphaga within the family Alcaligenaceae. Both strains showed 16S rRNA gene sequence similarity of 100 % to each other. The closest relatives of the isolates were Pigmentiphaga daeguensis (97.08 % sequence similarity), Pigmentiphaga kullae (97.01 %) and Pigmentiphaga litoralis (96.73 %). On the basis of data from phenotypic, chemotaxonomic and phylogenetic analyses, SAP-32T (=KCTC 52619T=DSM 104039T) and SAP-36 (=KCTC 52620=DSM 104072) represent members of a novel species of the genus Pigmentiphaga, for which the name Pigmentiphaga aceris sp. nov. is proposed.