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.

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).

This study investigated unrecorded freshwater bacterial species in Korea. Water and sediment samples were collected from the Nakdong River basin from 2020-2022. Bacterial isolates obtained through the conventional culture method with commercial media were subjected to 16S rRNA gene sequencing to identify unrecorded bacterial species. Results of 16S rRNA gene sequencing of the bacterial isolates revealed that a total of 44 bacterial isolates shared 16S rRNA gene sequence similarities of more than 98.65%, with validly published bacterial species not reported in Korea yet. These isolates were phylogenetically assigned to 4 phyla, 7 classes, 21 orders, 33 families, and 42 genera. A total of 2, 6, 12, and 24 species belonged to phyla Bacillota, Bacteroidota, Actinomycetota, and Pseudomonadota, respectively. Here, we provide details of these 44 unrecorded bacterial species, including Gram staining, colony and cellular morphologies, biochemical properties, and phylogenetic position.

In 2022, research for native prokaryotic species in Korea reported 10 unrecorded bacterial strains affiliated to phyla Actinomycetota, Bacillota, and Pseudomonadota. The strains formed monophyletic clades with the most closely related species (with ≥98.7% sequence similarity) in the 16S rRNA gene sequencing. Among them, four species of the phylum Actinomycetota, two species of the phylum Bacillota, and four species of the phylum Pseudomonadota have not been reported in Korea, suggesting unrecorded species in Korea. Information on strains such as Gram staining reaction, colony and cell morphology, biochemical characteristics, and isolation sources were provided in the species description.

Suncheon Bay Ecological Park, possessing abundant fisheries and biological diversity, was registered as a Ramsar wetland in Korea. Approximately 300 bacterial strains were isolated from the Suncheon Bay in a comprehensive study of indigenous prokaryotic species conducted during 2019-2020 in South Korea. A total of 12 bacterial strains were identified using 16S rRNA gene sequencing, demonstrating >98.7% sequence similarity with validly published species. These species were determined to be unrecorded bacterial species in Korea. A total of six strains were isolated from brackish water and Phragmites communis Trin (reed) species. These unrecorded species were phylogenetically diverse and belonged to three classes, six orders, and ten genera. Regarding the genus and class levels, the previously unrecorded species belonged to Jiella, Martelella, Rhizobium, Paracoccus, Rhodovulum, and Altererythrobacter of the class Alphaproteobacteria; Mycolicibacterium, Demequina, and Microbacterium of the class Actinobacteria; Confluentibacter of the class Flavobacteria. The twelve species were further characterized by gram staining, colony and cell morphology, biochemical properties, and phylogenetic position.

As part of the research program “2018 Rapid screening and identification of freshwater microorganisms using MALDI-TOF/MS library” freshwater samples were collected from a branch of the Nakdong River. Almost 300 antibiotic-resistant bacterial strains were isolated from freshwater samples and subsequently identified by 16S rRNA gene sequencing. Seventeen strains among the isolates shared high 16S rRNA gene sequence similarity (>99.0%) with known species that were not previously recorded in Korea, and each of the isolates also formed a robust phylogenetic clade with the closest species. These species were phylogenetically diverse, belonging to four phyla, seven classes, 10 orders, and 13 genera. At the genus and class level, the previously unrecorded species belonged to Rhodovarius, Xanthobacter, and Shinella of the class Alphaproteobacteria; Ottowia, Simplicispira, and Zoogloea of Betaproteobacteria; Pseudomonas, Acinetobacter, and Shewanella of Gammaproteobacteria; Arcobacter of Epsilonproteobacteria; Sphingobacterium of Sphingobacteriia; Trichococcus of Bacilli; and Leucobacter of Actinobacteria. The previously unrecorded species were further characterized by examining their gram-staining, colony and cell morphology, biochemical properties, and phylogenetic position.