Two Gram-negative, aerobic, pale pink/rose-coloured, rod-shaped, non-motile bacterial strains, designated T and RG-4, were isolated from a freshwater fish culture pond and a freshwater spring sample, respectively. Based on characterization by using a polyphasic approach, the two strains showed highly similar phenotypic, physiological and genetic characteristics. They shared 99.9 % 16S rRNA gene sequence similarity and 89-94 % DNA-DNA relatedness, suggesting that they represent a single genomic species. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains GFA-11 T and RG-4 formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Flectobacillus within the family Flexibacteraceae. Their closest neighbours were Flectobacillus major DSM 103 T (97.7 % 16S rRNA gene sequence similarity) and Flectobacillus lacus CL-GP79 T (95.9 %). Levels of DNA-DNA relatedness between the two novel strains and the type strains of F. major and F. lacus were less than 70 %. The results of physiological and biochemical tests allowed clear phenotypic differentiation of strains GFA-11 T and RG-4 from recognized members of the genus Flectobacillus. The predominant fatty acid constituents of strain GFA-11 T were C 16 : 1 v5c (40.2 % of the total) and iso-C 15 : 0 (15.0 %). The DNA G+C content of strain GFA-11 T was 39.7 mol%. On the basis of 16S rRNA gene sequence analysis and chemotaxonomic and physiological data, strains GFA-11 T and RG-4 are considered to represent a novel species of the genus Flectobacillus, for which the name Flectobacillus roseus sp. nov. is proposed. The type strain is GFA-11 T (5BCRC 17834 T 5LMG 24501 T ).
A bacterial strain designated GFC-1 T was isolated from a fish-culture pond in Taiwan and was characterized by using the polyphasic taxonomic approach. Strain GFC-1 T was Gram-negative, aerobic, rod-shaped, motile and non-spore-forming. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain belonged to the genus Andreprevotia of the family Neisseriaceae and its closest neighbour was Andreprevotia chitinilytica JS11-7 T (97.0 % sequence similarity). The results of physiological and biochemical tests allowed clear phenotypic differentiation of isolate GFC-1 T from A. chitinilytica JS11-7 T . The major fatty acids were summed feature 3 (C 16 : 1 v7c and/or iso-C 15 : 0 2-OH), C 16 : 0 and C 18 : 1 v7c. The DNA G+C content was 63.0 mol%. On the basis of 16S rRNA gene sequence analysis and chemotaxonomic and physiological data, strain GFC-1 T should be classified as representing a novel species and a second member of the genus Andreprevotia, for which the name Andreprevotia lacus sp. nov. is proposed. The type strain is GFC-1 T (5BCRC 17832 T 5LMG 24502 T ).The genus Andreprevotia, a member of the family Neisseriaceae, currently encompasses only one species, namely Andreprevotia chitinilytica (type strain JS11-7 T ), isolated from Halla Mountain forest soil, Jeju Island, Korea (Weon et al., 2007). During our investigations of the biodiversity of bacteria associated with a fish-culture pond located in Taoyuan County, Taiwan, several ivory-coloured bacterial colonies were isolated on R2A agar (BD Difco).Strains showing similar colony morphology were selected and a representative strain, GFC-1 T , was selected for detailed analyses. Strain GFC-1 T was maintained on R2A agar at 25 u C. Subcultivation was performed on R2A agar at 25 u C for between 48 and 72 h. On this medium, strain GFC-1 T was able to grow at 15-37 u C, but not at 10 or 40 u C. The strain was preserved at 280 u C in nutrient broth (NB; Difco) with 20 % (v/v) glycerol or by lyophilization.Cell morphology was observed by phase-contrast microscopy (DM 2000; Leica) at lag, exponential and stationary phases of growth. Motility was tested by the hanging-drop method. Spot Test Flagella Stain (BD Difco) was used for flagellum staining. A Gram Stain Set S kit (BD Difco) and the Ryu non-staining KOH method (Powers, 1995) were adopted for testing the Gram-staining reaction. Poly-bhydroxybutyrate granule accumulation was observed under light microscopy after staining the cells with Sudan black. Details of cell morphology are provided in the species description. The pH range for growth was determined by measuring the OD 600 of the culture grown in NB that had been adjusted prior to sterilization to various pH values (pH 4.0-10.0 at intervals of 0.5 pH units), using appropriate biological buffers (Chung et al., 1995). Verification of the pH values after autoclaving revealed only minor changes. Tolerance to NaCl was tested in NB prepared according to the formula of the BD Difco medium, but adjusted to different NaCl concentrations [0, 0.5 and 1....
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