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Two novel, non-motile, Gram-stain-negative, rod-shaped bacterial strains, designated AS60T and WG47T, were isolated from freshwater in South Korea. To clarify their taxonomic positions, both strains were characterized based on genomic information, including 16S rRNA gene and draft genome sequences. Phylogenetic analyses revealed that both isolates belong to the genus Flavobacterium. Based on 16S rRNA gene sequencing, AS60T clustered with Flavobacterium silvisoli KACC 21178T (96.09%), Flavobacterium cheonhonense KACC 14967T (96.07%) and Flavobacterium sangjuense KACC 17473T (95.96%). Strain WG47T showed the highest similarity to Flavobacterium dankookense KACC 23179T (97.14%), F. cheonhonense KACC 14967T (97.07%) and Flavobacterium chungnamense KACC 14971T (96.71%). The draft genomes of AS60T and WG47T were 3.07 and 3.30 Mb, with G+C contents of 35.6 and 38.9 mol%, respectively. Average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values were calculated. For AS60T, ANI values compared to F. silvisoli KACC 21178T, F. cheonhonense KACC 14967T and WG47T were 76.67, 79.38 and 76.01%, respectively, with dDDH values of 20, 19.6 and 22.1%. For WG47T, ANI values compared to F. dankookense KACC 23179T and F. cheonhonense KACC 14967T were 75.12 and 74.75%, with dDDH values of 19.9 and 19.2%. Both strains exhibited MK-6 as the predominant respiratory quinone, and their fatty acid profiles included iso-C15:1, iso-C15:0 3OH, iso-C15:1 G, iso-C16:0, iso-C16:0 3OH and iso-C17:0 3OH. Phosphatidylethanolamine was the major polar lipid. Strain WG47T was found to produce flexirubin-type pigments, while AS60T did not. These results, combined with phenotypic and biochemical data, suggest that strains AS60T and WG47T represent two novel species of Flavobacterium, for which the names Flavobacterium anseongense sp. nov. (AS60T=KACC 22413T=LMG 32491T) and Flavobacterium wongokense sp. nov. (WG47T=KACC 22407T=LMG 32500T) are proposed.
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