Fluviispira sanaruensis sp., nov., Isolated from a Brackish Lake in Hamamatsu, Japan.

Strain RF1110005T, which was isolated from brackish lake water sampled at Lake Sanaru in Japan as a "filterable" bacterial strain, was characterized as a novel species in the genus Fluviispira, family Silvanigrellaceae, order Silvanigrellales, the class Oligoflexia and the phylum Bdellovibrionota. Cells of RF1110005T were aerobic, Gram stain negative, and show a pleomorphic morphology of spiral, filamentous and rod shapes. Catalase reaction was positive. Strain RF1110005T grew optimally at 30 °C, pH 7.0-8.0 and 0.5% NaCl (w/v). The major polar lipids in RF1110005T were phosphatidylethanolamine and phosphatidylglycerol. The predominant cellular fatty acids were iso-C15:0 and anteiso-C15:0. Phylogenetic analysis based on 16S rRNA gene sequences and concatenates of core gene sequence showed that the nearest neighbor of strain RF1110005T was Fluviispira multicolorata strain 33A1-SZDPT with 98.4% 16S rRNA gene sequence similarity. The genome size of strain RF1110005T was 3.5 Mbp with two plasmids (80 kb and 69 kb), and the G + C content was 33.7 mol%. Comparisons with genome-wide analyses and chemotaxonomic characters clearly showed that strain RF1110005T differed from F. multicolorata. Therefore, a novel species in Fluviispira sanaruensis, sp. nov., is proposed for strain RF1110005T (= JCM 31447 T = LMG 30360 T).

Chryseotalea sanaruensis gen. nov., sp., nov., a Member of the Family Cytophagaceae, Isolated from a Brackish Lake in Hamamatsu Japan.

A strain-designated YsT was isolated as a filterable bacterial strain from Lake Sanaru, a brackish water lake in Hamamatsu Japan. YsT is aerobic, Gram-negative, and slender rod shaped. YsT grew optimally at 30 °C, pH 7.0-8.0 and without the addition of NaCl. MK-7 was the sole isoprenoid quinone. The main cellular polar lipids were phosphatidylethanolamine and unidentified amino- and polar-lipids. The predominant cellular fatty acids were C18:0, iso-C14:0 and iso-C15:0. Phylogenetic analysis of 16S rRNA gene sequence revealed the nearest neighbours of strain YsT to be members of the Ohtaekwangia and Chryseolinea genera with 91.2-92.1% sequence similarity. The percentages of conserved proteins (POCP) between the genomes of YsT and related strains were less than 50%. Phenotypic analyses suggested that YsT could not metabolize glucose and related sugars, which was discriminative from its phylogenetic relatives. We, therefore, propose a novel species in a new genus, Chryseotalea sanaruensis gen. nov., sp. nov. in the family Cytophagaceae (= JCM 30318T = LMG 30359T), based on cell size, the predominant cellular fatty acid composition, and the DNA GC content (38.9 mol%).

Algoriphagus sanaruensis sp. nov., a member of the family Cyclobacteriaceae, isolated from a brackish lake in Hamamatsu, Japan.

Strain M8-2T, which was isolated from brackish lake water (Lake Sanaru) in Japan, was characterized for representation of a novel species in the genus Algoriphagus. Cells of strain M8-2T were aerobic, Gram-stain-negative and curved-rod-shaped (0.2-0.5 µm wide and 0.7-1.9 µm long). Strain M8-2T grew optimally at 30 °C, pH 6.5-7.5 and in the presence of 0.5-1.0 % (w/v) NaCl. MK-7 was the sole isoprenoid quinone. The major polar lipids were phosphatidylethanolamine, an unidentified phospholipid and an unidentified polar lipid. The predominant cellular fatty acids were iso-C15 : 0 and anteiso-C15 : 0. Phylogenetic analysis based on its 16S rRNA gene sequence showed that strain M8-2T belonged to the genus Algoriphagus and was closely related to Algoriphagus aquatilis A8-7T, Algoriphagus boseongensis BS-R1T, Algoriphagus aquaeductus T4T, Algoriphagus olei CC-Hsuan-617T, Algoriphagusshivajiensis NIO-S3T and Algoriphagus mannitolivorans DSM 15301T with sequence similarities of 96.6-97.4 %. Results of average nucleotide identity (<75 %) and digital DNA-DNA hybridization (<19 %) studies showed that M8-2T was distinct from its phylogenetic relatives. Based on the results of tests for acid production, the predominant cellular fatty acid composition, the DNA G+C content and phylogenetic position, a novel species in the genus Algoriphagus, with the name Algoriphagussanaruensis sp. nov., is proposed for strain M8-2T (=JCM 31446T=LMG 29969T).

Novel Self-Transmissible and Broad-Host-Range Plasmids Exogenously Captured From Anaerobic Granules or Cow Manure.

Novel self-transmissible plasmids were exogenously captured from environmental samples by triparental matings with pBBR1MCS-2 as a mobilizable plasmid and Pseudomonas resinovorans as a recipient. A total of 272 recipients were successfully obtained as plasmid host candidates from granules of an anaerobic methane fermentation plant and from cow manure. The whole nucleotide sequences of six plasmids were determined, including one IncP-1 plasmid (pSN1104-59), four PromA-like plasmids (pSN1104-11, pSN1104-34, pSN0729-62, and pSN0729-70), and one novel plasmid (pSN1216-29), whose incompatibility group has not been previously identified. No previously known antibiotic resistance genes were found in these plasmids. In-depth phylogenetic analyses showed that the PromA-like plasmids belong to subgroups of PromA (designated as PromAγ and PromAδ) different from previously proposed subgroups PromAα and PromAß. Twenty-four genes were identified as backbone genes by comparisons with other PromA plasmids. The nucleotide sequences of pSN1216-29 share high identity with those found in clinical isolates. A minireplicon of pSN1216-29 was successfully constructed from repA encoding a replication initiation protein and oriV. All the captured plasmids were found to have a broad host range and could be transferred to and replicated in different classes of Proteobacteria. Notably, repA and oriV of pSN1216-29 showed high similarity with one of two replication systems of pSRC119-A/C, known as a plasmid with multidrug resistance genes found in Salmonella enterica serovar Senftenberg. Our findings suggest that these "cryptic" but broad-host-range plasmids may be important for spreading several genes as "vehicles" in a wider range of bacteria in natural environments.

Proteobacteria and Bacteroidetes are major phyla of filterable bacteria passing through 0.22 µm pore size membrane filter, in Lake Sanaru, Hamamatsu, Japan.

141 filterable bacteria that passed through a 0.22 µm pore size filter were isolated from Lake Sanaru in Hamamatsu, Japan. These belonged to Proteobacteria, Bacteroidetes, Firmicutes, or Actinobacteria among which the first two phyla comprised the majority of the isolates. 48 isolates (12 taxa) are candidates assignable to new bacterial species or genera of Proteobacteria or Bacteroidetes.