Freshwater environments are rich ecosystems that support diverse microbial communities, including members of the phylum Actinomycetota critical for nutrient cycling, organic matter decomposition, and water quality maintenance. Actinomycetota known to produce numerous bioactive secondary metabolites are valuable in biotechnology, medicine, and agriculture. Despite their significance, the diversity and distribution of Actinomycetota in freshwater habitats, especially in the Republic of Korea, are underexplored. This study aimed to report the isolation and characterization of 22 previously unrecorded bacterial species of Actinomycetota from various freshwater environments in Korea. Using standard dilution plating techniques on six different culture media, 22 bacterial strains were isolated, incubated, and characterized based on colony and cellular morphologies, Gram staining, and biochemical properties. Genomic DNA was extracted and the 16S rRNA gene was sequenced to determine species identity using the EzBioCloud service with a cutoff of 98.7% sequence similarity for classification as unreported species. These strains were phylogenetically diverse, belonging to two classes, ten orders, and eighteen genera. This study enhances our understanding of bacterial diversity in freshwater ecosystems and underscores the importance of exploring microbial diversity in underexplored habitats, potentially leading to discovery of novel bioactive compounds. Findings of this study contribute valuable insights into ecological roles and biotechnological potential of Actinomycetota in freshwater environments.

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).

As part of the research program “Freshwater Prokaryotic Organisms Research and Discovery,” freshwater samples were collected from the Nakdonggang River. After plating the samples on several culture media and incubating aerobically, approximately 900 bacterial strains were isolated and identified using 16S rRNA gene sequences. Among the bacterial isolates showing higher than 98.7% 16S rRNA gene sequence similarity with those of already confirmed bacterial species previously unreported in Korea, 29 strains were selected. These strains were phylogenetically diverse and belonged to 3 phyla, 6 classes, 13 orders, and 21 genera. At the genus level, these previously unreported species were found to be affiliated with Novosphingobium, Sphingomonas, Polymorphobacter, Croceibacterium, Devosia, Endobacterium, Agaricicola, Bradyrhizobium, Paracoccus, and Pseudotabrizicola of the class Alphaproteobacteria; Undibacterium, Azonexus, and Dechloromonas of the class Betaproteobacteria; Acinetobacter and Budvicia of the class Gammaproteobacteria; Streptomyces, Nocardioides, Mycobacterium, and Cellulomonas of the phylum Actinomycetota; Flavobacterium and Pedobacter of the phylum Bacteroidota. These species were further characterized by examining their Gram reaction, colony and cell morphologies, biochemical properties, and phylogenetic positions. Detailed descriptions of these 29 previously unreported species are provided.

Fungal contaminant in the indoor air is one of risk factors that could damage valuable paper-based records preserved in libraries. In the process of monitoring airborne fungi at the Nara Repository, the National Archives, Seoul, Korea, three fungal strains, DUCC 16098, DUCC 17764, and DUCC17767 were isolated from the archive’s air samples. Fungal identification was performed based on the morphological characteristics and phylogenetic analysis of the internal transcribed spacer (ITS), the 28S LSU rDNA, and β-tubulin gene (BenA), and TEF1-α gene. These DUCC 16098, DUCC 17764 and DUCC17767 strains were identified as Clonostachys farinosa, Penicillium cosmopolitanum, and Cephalotrichum purpureofuscum. These species have not been recorded before in Korea. Information on these fungi will help the monitoring and management of airborne fungi in the archival facilities.

Air conditioner filters purify the air of indoor environments by removing air pollutants and supporting the efficiency of the unit’s cooling function. However, an air conditioner filter can become a microenvironment in which some fungi can grow as dust continues to accumulate and favorable humidity conditions are formed. Fungal growth in air conditioner filters could lead to fungal allergies or fungal diseases, in addition to emitting a foul odor. In an effort to understand what species causes this malodorous problem, we investigated the diversity of fungi found in air conditioners. Fungi were sampled from the collected air conditioner filters and grown on DG18 agar media. After purification for pure isolates, species identification was undertaken. Colony morphology was observed on PDA, MEA, CYA, and OA media. Microstructures of fruiting body, mycelia, and spores were examined using a light microscope. Molecular identification was performed by PCR and sequencing of PCR amplicons, and molecular phylogenetic analysis of sequenced DNA markers, including the Internal Transcribed Spacer (ITS), the 28S large subunit of the nuclear ribosomal RNA (LSU rDNA), the β-tubulin (BenA) gene, the Calmodulin (CaM) gene, and the DNA-directed RNA polymerase II subunit 2 (RPB2) gene. Through this identification process, we found two fungal species, Aspergillus miraensis and Dichotomopilus ramosissimus, which are unrecorded species in Korea. We will now report their morphological and molecular features.

To understand microorganism effects on wild mushroom fruiting bodies, we investigated the fungi in hyphosphere soil supporting wild mushroom species Cortinarius violaceus, Amanita hemibapha, Laccaria vinacelavellanea, and Amanita verna found in the Gotjawal area of Jeju Island. Fungal species identification based on morphological traits and molecular analysis of ITS, LSU rDNA, and -tubulin gene sequences resulted in isolation and identification of eleven fungal species previously unrecorded in Korea. These newly-recorded species are: Arthrinium kogelbergensis, Kalmusia longisporum, Keithomyces carneum, Neopyrenochaeta cercidis, Penicillium ranomafanaense, Phomatodes nebulosa, Pyrenochaeta nobilis, Tolypocladium album, Talaromyces kendrickii, Talaromyces qii, and Umbelopsis gibberispora, and their morphological characteristics and phylogenetic positions are described.

In the genus Acrobasis Zeller, 1839, there are 14 species known in Korea. In this study, we present Acrobasis rubrizonella (Ragonot, 1893) and Acrobasis subceltifoliella Yamanaka, 2006 from Korea for the first time. We hear provide diagnoses, descriptions, and images of the adults and genitalia of the two species.

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.

A total of 31 bacterial strains were isolated from the Geum River basin in the Republic of Korea during our investigation of indigenous prokaryotic species. The isolated bacterial strains had high 16S rRNA gene sequence similarity (>98.7%) with those of validly published bacterial species, which have not been reported in Republic of Korea. The 31 bacterial strains were phylogenetically diverse and assigned to 4 phyla, 8 classes, 18 orders, 21 families, and 27 genera. At the genus level, the unreported species were affiliated with Kineococcus, Pedococcus, Rhodoluna, Salinibacterium, Rhodoluna, Arthrobacter, Williamsia, Nakamurella, Nocardioides of the class Actinobacteria, Patulibacter of the class Thermoleophilia, Pontibacter, Hymenobacter of the class Cytophagia, Flavobacterium of the class Flavobacteriia, Geomicrobium of the class Bacilli, Brevundimonas, Gellertiella, Rhizobium, Paracoccus, Taonella, Sphingomonas of the class Alphaproteobacteria, Burkholderia, Polaromonas, Hydrogenophaga, Chitinilyticum, Azospira, Zoogloea of the class Betaproteobacteria, and Pseudomonas of the class Gammaproteobacteria. The unreported bacterial species were further characterized by examining their morphological, cultural, physiological, and biochemical properties. The detailed descriptions of the 31 bacterial strains were provided.

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.

Free-living nematodes feed on bacteria, algae, fungi, dead organisms, and living tissues. They release nutrients for plant use and improve soil structure and water holding capacity. They are usually the most abundant type of nematodes in soil and marine environments. This study has been conducted to survey the distribution and identification of soil nematodes from rhizosphere soil of plant for investigation of indigenous species according to Convention on Biological Diversity. Three species of free-living nematodes belonging to the order of Dorylaimida were found from paddy soil and soybean fields, Miryang, Gyeongsangnam-do and one species was found from soil, Dokdo island, Gyeongsangbuk-do, Korea. Aporcelaimellus obtusicaudatus, Eudorylaimus arcus and Mesodorylaimus spengelii are newly collected from Korea. A. obtusicaudatus can be recognized by its large body, short odontostyle being much thicker than cuticle at the same level, well developed gonads, shape and length of the tail. E. arcus is distinctive because of the wide lateral field, tail form and supplement arrangement. M. spengelii is smaller species with short odontostylet, simple guiding ring. Female tail more or less uniformly attenuated, tip is not swollen, nor curved dorsally.

A single juvenile malacanthid specimen (Hoplolatilus chlupatyi) was collected from the South Sea of Korea and identified by DNA barcoding. This species is readily distinguished from other malacanthid species in having well developed rostral spine and elongated spines in the posttemporal and preopercle. A molecular analysis based on mitochondrial DNA COI sequences showed that this species is matched to adult H. chlupatyi (genetic distance = 0.005). Therefore, this is the first record of H. chlupatyi in Korea. We propose new Korean names, “Mu-ji-gae-og-dom-sog” for the genus Hoplolatilus and “Mu-ji-gae-og-dom” for the species H. chlupatyi.

Nine fish and one clam species were collected from freshwater environments in Korea, including four lakes, two streams, and the Nakdong River, to investigate the host-associated bacteria. Hundreds of bacterial strains were isolated from the samples using a cell sorter and a dilution plating method. After identification of the bacterial strains using 16S rRNA gene sequences, 42 strains with greater than 98.7% sequence similarity with validly published species were determined to be unrecorded bacterial species in Korea. These strains were phylogenetically diverse and assigned to four phyla, six classes, 17 orders, 27 families, and 32 genera. At the genus level, the unrecorded species were classified as Corynebacterium, Mycobacterium, Mycolicibacterium, Gordonia, Williamsia, Modestobacter, Brachybacterium, Sanquibacter, Arthrobacter, and Mycolicibacterium of the class Actinobacteria; Empedobacter, and Flavobacterium of the class Flavobacteriia; Fictibacillus, Psychrobacillus, Cohnella, Paenibacillus, Rummeliibacillus, Enterococcus, and Vagococcus of the class Bacilli; Aquamicrobium, Paracoccus, and Sphingomonas of the class Alphaproteobacteria; Achromobacter, Delftia, and Deefgea of the class Betaproteobacteria; and Aeromonas, Providencia, Yersinia, Marinomonas, Acinetobacter, and Pseudomonas of the class Gammaproteobacteria. The 42 unrecorded species were subjected to further taxonomic characterization using gram staining, cellular and colony morphological determination, biochemical analyses, and phylogenetic analyses. This paper provides detailed descriptions of the 42 previously unrecorded bacterial species.

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.