Journal of Microbiology

Journal Article

CA‑CAS‑01‑A: A Permissive Cell Line for Isolation and Live Attenuated Vaccine Development Against African Swine Fever Virus: Seung-Chul Lee , Yongkwan Kim , Ji-Won Cha , Kiramage Chathuranga , Niranjan Dodantenna , Hyeok-Il Kwon , Min Ho Kim , Weonhwa Jheong , In-Joong Yoon , Joo Young Lee , Sung-Sik Yoo , Jong-Soo Lee; J. Microbiol. 2024;62(2):125-134. Published online March 13, 2024; DOI: https://doi.org/10.1007/s12275-024-00116-1

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African swine fever virus (ASFV) is the causative agent of the highly lethal African swine fever disease that affects domestic pigs and wild boars. In spite of the rapid spread of the virus worldwide, there is no licensed vaccine available. The lack of a suitable cell line for ASFV propagation hinders the development of a safe and effective vaccine. For ASFV propagation, primary swine macrophages and monocytes have been widely studied. However, obtaining these cells can be time-consuming and expensive, making them unsuitable for mass vaccine production. The goal of this study was to validate the suitability of novel CA-CAS-01-A (CAS-01) cells, which was identified as a highly permissive cell clone for ASFV replication in the MA-104 parental cell line for live attenuated vaccine development. Through a screening experiment, maximum ASFV replication was observed in the CAS-01 cell compared to other sub-clones of MA-104 with 14.89 and log10 7.5 ± 0.15 Ct value and TCID50/ ml value respectively. When CAS-01 cells are inoculated with ASFV, replication of ASFV was confirmed by Ct value for ASFV DNA, HAD50/ ml assay, TCID50/ ml assay, and cytopathic effects and hemadsoption were observed similar to those in primary porcine alveolar macrophages after 5th passage. Additionally, we demonstrated stable replication and adaptation of ASFV over the serial passage. These results suggest that CAS-01 cells will be a valuable and promising cell line for ASFV isolation, replication, and development of live attenuated vaccines.

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Development and characterization of high-efficiency cell-adapted live attenuated vaccine candidate against African swine fever
Min Ho Kim, Ashan Subasinghe, Yongkwan Kim, Hyeok-Il Kwon, Yehjin Cho, Kiramage Chathuranga, Ji-Won Cha, Ji-Yoon Moon, Ji-Hyeon Hong, Jin Kim, Seung-Chul Lee, Niranjan Dodantenna, Nuwan Gamage, W. A. Gayan Chathuranga, Yeonji Kim, In-Joong Yoon, Joo Young
Emerging Microbes & Infections.2024;[Epub] CrossRef

Research Support, Non-U.S. Gov't

NOTE] Microbacterium suwonense sp. nov., Isolated from Cow Dung: Rangasamy Anandham , Tomohiko Tamura , Moriyuki Hamada , Hang-Yeon Weon , Soo-Jin Kim , Yi-Seul Kim , Ken-ichiro Suzuki , Soon-Wo Kwon; J. Microbiol. 2011;49(5):852-856. Published online November 9, 2011; DOI: https://doi.org/10.1007/s12275-011-1036-y

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Abstract: An actinomycete strain, designated M1T8B9T, was isolated from cow dung in Suwon, Republic of Korea. The isolate was a Gram-positive, nonmotile, and non-spore-forming bacterium. Phylogenetic evaluation based on 16S rRNA gene sequence similarity showed that this isolate belongs to the genus Microbacterium, with its closest neighbors being Microbacterium soli DCY17T (98.2%) and Microbacterium esteraromaticum DSM 8609T (98.0%). The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, and one unknown glycolipid. Strain M1T8B9T contained the major fatty acids C15:0 anteiso, C16:0 iso, C17:0 anteiso, and C15:0 iso, and the cell-wall peptidoglycan was of type B2β. According to DNA-DNA hybridization studies, strain M1T8B9T showed 42% and 26% relatedness with M. soli DCY17T and M. esteraromaticum DSM 8609T, respectively. On the basis of the data presented, strain M1T8B9T is considered to represent a novel species of the genus Microbacterium, for which the name Microbacterium suwonense sp. nov. is proposed. The type strain is M1T8B9T (=KACC 14058T =NBRC 106310T).

Journal Article

Symbiotic Relationship between Microbacterium sp. SK0812 and Candida tropicalis SK090404: Seung Won Kang , Bo Young Jeon , Tae Sik Hwang , Doo Hyun Park; J. Microbiol. 2009;47(6):721-727. Published online February 4, 2010; DOI: https://doi.org/10.1007/s12275-009-0146-2

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Abstract: A bacterium growing inside yeast cytoplasm was observed by light microscope without staining. The bacterium was separately stained from yeast cell by a fluorescent dye, 4′,6-diamidino-2-phenylindole (DAPI). The bacterium actively moved inside yeast cytoplasm and propagated in company with the yeast growth. The bacterium was separated from the yeast cytoplasm by selective disruption of yeast cells and the yeast without the intracellular bacterium (YWOB) was obtained by selective inactivation of bacterial cells. The yeast and the intracellular bacterium were identified as Candida tropicalis and Microbacterium sp., respectively. The length of Microbacterium sp. and C. tropicalis measured with SEM image was smaller than 0.5 μm and was larger than 5 μm, respectively. The yeast with the intracellular bacterium (YWIB) grew in a starch-based medium but the YWOB was not C. tropicalis has neither extracellular nor intracellular saccharification enzyme. Glucose was produced from starch by the extracellular crude enzyme (culture fluid) of Microbacterium sp. YWIB produced significantly more ethanol from glucose than YWOB but did not from starch. Conclusively, C. tropicalis is thought to catabolize starch dependent upon Microbacterium sp. growing in its cytoplasm and furnish stable habitat for the Microbacterium sp.

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