During an investigation of indigenous prokaryotic species in the Republic of Korea, 17 bacterial strains belonging to the phylum Bacteroidota and 4 strains belonging to the phylum Bacillota were isolated from freshwater environments. These isolated bacterial strains formed a robust phylogenetic clade with type strains of the closest related bacterial species, with 16S rRNA gene sequences sharing similarities of higher than 98.7%. To date, there have been no official report of these 21 isolates in the Republic of Korea. At the genus level, these unreported species were affiliated with Mucilaginibacter and Pedobacter of class Sphingobacteriia, Flavobacterium and Gillisia of class Flavobacteriia, Hymenobacter of class Cytophagia, and Paenibacillus and Planococcus of class Bacilli. These species were further examined by performing Gram staining, analyzing their colonies and cell morphologies, and determining their basic biochemical characteristics and phylogenetic positions. Detailed descriptions of the 21 previously unreported species are provided.

Freshwater environments serve as crucial habitats for diverse microorganisms, playing essential roles alongside plants and animals within the ecosystem. Bacteria in these environments are particularly important for maintaining ecosystem functions. Among them, members of the phylum Pseudomonadota are known to be involved in processes such as nitrogen fixation, ammonia oxidation, and biofilm formation. Pseudomonadota comprises six classes: Acidithiobacillia, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Hydrogenophilia, and Zetaproteobacteria. This study reports 43 previously unrecorded species belonging to the phylum Pseudomonadota. They were isolated from various freshwater habitats in Republic of Korea. These strains were isolated and cultured using nine different standard media, with classification based on Gram staining, cell morphology, and biochemical characteristics. Species identity was determined using 16S rRNA gene sequencing, with a similarity threshold of 98.7% for designation as unreported species. Phylogenetic analysis revealed that these 43 strains were distributed across three classes, 13 orders, 28 families, and 39 genera. This research emphasizes microbial diversity in freshwater ecosystems and provides valuable insights into ecological roles and potential novel functions of unreported bacterial species in Korea.

As part of the 2024 research initiative, “Investigation and Discovery of Prokaryotes in Freshwater Systems,” samples were collected from diverse freshwater habitats, including both water and soil environments. Approximately 2,000 bacterial strains were isolated as single colonies and identified through 16S rRNA gene sequence analysis. Among these, 38 strains shared ≥98.7% 16S rRNA gene sequence similarities with those of known bacterial species not previously reported in Korea. These strains were thus categorized as newly recorded bacterial species in Korea. These 38 bacterial strains displayed significant phylogenetic diversities, spanning 2 phyla, 4 classes, 15 orders, 24 families, and 34 genera. These unrecorded species were classified into the following classes: Actinomycetia (with genera including Microcella, Conyzicola, Curtobacterium, Leucobacter, Microbacterium, Frigoribacterium, Lysinibacter, Streptomyces, Nonomuraea, Actinocorallia, Ruania, and Actinoplanes), Alphaproteobacteria (Paracoccus, Youngimonas, Loktanella, Corticibacterium, Neorhizobium, Onobrychidicola, Ferranicluibacter, Aureimonas, Asticcacaulis, and Novosphingobium), Betaproteobacteria (Rhodoferax, Rugamonas, and Cupriavidus), and Gammaproteobacteria (Rheinheimera, Shewanella, Kosakonia, Leclercia, Hafnia, Yersinia, Pseudomonas, Lysobacter, and Acinetobacter ). Further characterization included assessment of Gram reaction, colony and cell morphology, biochemical properties, and phylogenetic relationships. This report presents detailed phylogenetic and phenotypic characteristics of these bacterial species.

The Yeongsan River is a prominent inland waterway, alongside the Han River, Nakdong River, and Geum River in South Korea. Numerous bacterial strains were isolated from the Yeongsan River basin for a comprehensive investigation into indigenous prokaryotic species conducted between 2020 and 2023. These bacterial strains were identified using 16S rRNA gene sequencing, wherein 45 bacterial strains shared >98.7% sequence similarities with bacterial species not recorded in Korea thus far. Therefore, this study aimed to catalogue aforementioned unrecorded species and characterize them contingent upon their Gram nature, colony and cell morphologies, biochemical properties, and phylogenetic positions. These bacterial species were determined to be phylogenetically diverse. They were categorized into nine classes, 18 orders, and 25 families. These previously unrecorded species were classified into the following genera and classes: Chitinophaga (class Chitinophagia); Flavobacterium (class Flavobacteriia); Rhodopseudomonas, Gemmobacter, Paracoccus, Azospirillum, Sphingomonas, Novosphingobium, Sphingorhabdus, and Erythrobacter (class Alphaproteobacteria); Bordetella, Pararobbsia, Polynucleobacter, Rhodoferax, Aquabacterium, Malikia, Comamonas, Ideonella, Paucibacter, Undibacterium, Cupriavidus, and Thauera (class Betaproteobacteria); Pectobacterium, Arenimonas, Lysobacter, and Luteimonas (class Gammaproteobacteria); Luteolibacter (class Verrucomicrobiia); Mycolicibacterium, Angustibacter, Ornithinibacter, Janibacter, Schumannella, Aurantimicrobium, Luedemannella, Nocardioides, and Propionicimonas (class Actinomycetes); Geothrix (class Holophagae); and Lactococcus (class Bacilli).