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Characterization of novel bacteriophages for effective phage therapy against Vibrio infections in aquaculture
Kira Moon, Sangdon Ryu, Seung Hui Song, Se Won Chun, Nakyeong Lee, Aslan Hwanhwi Lee
J. Microbiol. 2025;63(5):e2502009.   Published online May 27, 2025
DOI: https://doi.org/10.71150/jm.2502009
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AbstractAbstract PDFSupplementary Material

The widespread use of antibiotics in aquaculture has led to the emergence of multidrug-resistant pathogens and environmental concerns, highlighting the need for sustainable, eco-friendly alternatives. In this study, we isolated and characterized three novel bacteriophages from aquaculture effluents in Korean shrimp farms that target the key Vibrio pathogens, Vibrio harveyi, and Vibrio parahaemolyticus. Bacteriophages were isolated through environmental enrichment and serial purification using double-layer agar assays. Transmission electron microscopy revealed that the phages infecting V. harveyi, designated as vB_VhaS-MS01 and vB_VhaS-MS03, exhibited typical Siphoviridae morphology with long contractile tails and icosahedral heads, whereas the phage isolated from V. parahaemolyticus (vB_VpaP-MS02) displayed Podoviridae characteristics with an icosahedral head and short tail.

Whole-genome sequencing produced complete, circularized genomes of 81,710 bp for vB_VhaS-MS01, 81,874 bp for vB_VhaS-MS03, and 76,865 bp for vB_VpaP-MS02, each showing a modular genome organization typical of Caudoviricetes. Genomic and phylogenetic analyses based on the terminase large subunit gene revealed that although vB_VhaS-MS01 and vB_VhaS-MS03 were closely related, vB_VpaP-MS02 exhibited a distinct genomic architecture that reflects its unique morphology and host specificity. Collectively, these comparative analyses demonstrated that all three phages possess genetic sequences markedly different from those of previously reported bacteriophages, thereby establishing their novelty. One-step growth and multiplicity of infection (MOI) experiments demonstrated significant differences in replication kinetics, such as burst size and lytic efficiency, among the phages, with vB_VhaS-MS03 maintaining the most effective bacterial control, even at an MOI of 0.01. Additionally, host range assays showed that vB_VhaS-MS03 possessed a broader spectrum of activity, supporting its potential use as a stand-alone agent or key component of phage cocktails. These findings highlight the potential of region-specific phage therapy as a targeted and sustainable alternative to antibiotics for controlling Vibrio infections in aquaculture.

Journal Article
Thalassotalea aquiviva sp. nov., and Thalassotalea maritima sp. nov., Isolated from Seawater of the Coast in South Korea
Jina Lee, Seung-Hui Song, Kira Moon, Nakyeong Lee, Sangdon Ryu, Hye Seon Song, Sung Moon Lee, Yun Ji Kim, Se Won Chun, Kyung-Min Choi, Aslan Hwanhwi Lee
J. Microbiol. 2024;62(12):1099-1111.   Published online December 10, 2024
DOI: https://doi.org/10.1007/s12275-024-00191-4
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AbstractAbstract
Two novel bacterial strains, 273M-4T and Sam97T, were isolated from seawater in the Yellow Sea, Muan-gun, South Korea, and identified as members of the genus Thalassotalea. Both strains were Gram-stain-negative, aerobic, rod-shaped, non-motile, non-flagellated, and oxidase- and catalase-positive. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains 273M-4T and Sam97T were most closely related to Thalassotalea ponticola KCTC 42155T, with sequence similarities of 97.5% and 98.3%, respectively. Optimal growth for strain 273M-4T occurred at 25-30 °C, pH 7.0, and 2% NaCl, while strain Sam97T grew optimally at 30 °C, pH 8.0, and 2% NaCl. Genome sizes of strains 273M-4T and Sam97T were 3.37 and 3.31 Mb, with DNA G + C contents of 41.0 mol% and 42.9 mol%, respectively. The orthologous average nucleotide identity (OrthoANI) and digital DNA-DNA hybridization (dDDH) values between the two strains were 71.6% and 24.4%, respectively, indicating that they are distinct species. Further genomic analyses of these two strains revealed OrthoANI values of < 73.5% and dDDH values of < 26.7% within the genus Thalassotalea, suggesting their distinctiveness from other Thalassotalea species. The predominate fatty acids of strains 273M-4T and Sam97T were summed feature 3 (consisting of C16:1 ω7c/C16:1 ω6c) and C16:0. All strains contained phosphatidylethanolamine and phosphatidylglycerol as the major polar lipids and ubiquinone-8 (Q-8) as the primary respiratory quinone. Based on phenotypic, phylogenetic, genotypic, and chemotaxonomic data, strains 273M-4T (= KCTC 8644T = LMG 33695T) and Sam97T (= KCTC 8645T = LMG 33694T) represent novel species of the genus Thalassotalea, named Thalassotalea aquiviva sp. nov. and Thalassotalea maritima sp. nov..

Citations

Citations to this article as recorded by  
  • Validation List no. 223. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2025;[Epub]     CrossRef
Review
Metaviromics coupled with phage-host identification to open the viral ‘black box’
Kira Moon , Jang-Cheon Cho
J. Microbiol. 2021;59(3):311-323.   Published online February 23, 2021
DOI: https://doi.org/10.1007/s12275-021-1016-9
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  • 9 Web of Science
  • 9 Crossref
AbstractAbstract
Viruses are found in almost all biomes on Earth, with bacteriophages (phages) accounting for the majority of viral particles in most ecosystems. Phages have been isolated from natural environments using the plaque assay and liquid medium- based dilution culturing. However, phage cultivation is restricted by the current limitations in the number of culturable bacterial strains. Unlike prokaryotes, which possess universally conserved 16S rRNA genes, phages lack universal marker genes for viral taxonomy, thus restricting cultureindependent analyses of viral diversity. To circumvent these limitations, shotgun viral metagenome sequencing (i.e., metaviromics) has been developed to enable the extensive sequencing of a variety of viral particles present in the environment and is now widely used. Using metaviromics, numerous studies on viral communities have been conducted in oceans, lakes, rivers, and soils, resulting in many novel phage sequences. Furthermore, auxiliary metabolic genes such as ammonic monooxygenase C and β-lactamase have been discovered in viral contigs assembled from viral metagenomes. Current attempts to identify putative bacterial hosts of viral metagenome sequences based on sequence homology have been limited due to viral sequence variations. Therefore, culture- independent approaches have been developed to predict bacterial hosts using single-cell genomics and fluorescentlabeling. This review focuses on recent viral metagenome studies conducted in natural environments, especially in aquatic ecosystems, and their contributions to phage ecology. Here, we concluded that although metaviromics is a key tool for the study of viral ecology, this approach must be supplemented with phage-host identification, which in turn requires the cultivation of phage-bacteria systems.

Citations

Citations to this article as recorded by  
  • Long-read powered viral metagenomics in the oligotrophic Sargasso Sea
    Joanna Warwick-Dugdale, Funing Tian, Michelle L. Michelsen, Dylan R. Cronin, Karen Moore, Audrey Farbos, Lauren Chittick, Ashley Bell, Ahmed A. Zayed, Holger H. Buchholz, Luis M. Bolanos, Rachel J. Parsons, Michael J. Allen, Matthew B. Sullivan, Ben Tempe
    Nature Communications.2024;[Epub]     CrossRef
  • Tools and methodology to in silico phage discovery in freshwater environments
    Carlos Willian Dias Dantas, David Tavares Martins, Wylerson Guimarães Nogueira, Oscar Victor Cardenas Alegria, Rommel Thiago Jucá Ramos
    Frontiers in Microbiology.2024;[Epub]     CrossRef
  • A Pseudomonas Lysogenic Bacteriophage Crossing the Antarctic and Arctic, Representing a New Genus of Autographiviridae
    Zhenyu Liu, Wenhui Jiang, Cholsong Kim, Xiaoya Peng, Cong Fan, Yingliang Wu, Zhixiong Xie, Fang Peng
    International Journal of Molecular Sciences.2023; 24(8): 7662.     CrossRef
  • Evaluation of the ABL NGS assay for HIV-1 drug resistance testing
    Thomas Lhossein, Karine Sylvain, Véronique Descamps, Virginie Morel, Baptiste Demey, Etienne Brochot
    Heliyon.2023; 9(11): e22210.     CrossRef
  • Human virome: Implications in cancer
    Rafael Tamayo-Trujillo, Patricia Guevara-Ramírez, Santiago Cadena-Ullauri, Elius Paz-Cruz, Viviana A. Ruiz-Pozo, Ana Karina Zambrano
    Heliyon.2023; 9(3): e14086.     CrossRef
  • Metaviromics analysis of marine biofilm reveals a glycoside hydrolase endolysin with high specificity towards Acinetobacter baumannii
    Georgios E. Premetis, Nikolaos D. Georgakis, Angeliki Stathi, Nikolaos E. Labrou
    Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics.2023; 1871(4): 140918.     CrossRef
  • Comparative genomic analysis of five freshwater cyanophages and reference-guided metagenomic data mining
    Kang Du, Feng Yang, Jun-Tao Zhang, Rong-Cheng Yu, Ziqing Deng, Wei-Fang Li, Yuxing Chen, Qiong Li, Cong-Zhao Zhou
    Microbiome.2022;[Epub]     CrossRef
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    Harilanto Felana Andrianjakarivony, Yvan Bettarel, Fabrice Armougom, Christelle Desnues
    Viruses.2022; 15(1): 76.     CrossRef
  • Omics-based microbiome analysis in microbial ecology: from sequences to information
    Jang-Cheon Cho
    Journal of Microbiology.2021; 59(3): 229.     CrossRef

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