Cell Culture Isolation of Piscine Nodavirus (Betanodavirus) in Fish-Rearing Seawater

Nishi, Seiki; Yamashita, Hirofumi; Kawato, Yasuhiko; Nakai, Toshiko

doi:10.1128/aem.03834-15

Cited by 28 publications

(29 citation statements)

References 48 publications

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“…The infection in the BGA cell line showed severe CPE and vacuolation in the cytoplasm of the cells. The CPE in the cell line is similar to other cell lines as reported earlier in CSK, GB and ICF (Abdul Majeed, Nambi, Taju, & Sahul Hameed, ; Chi, Hu, & Lo, ; Nishi et al, ). The significant CPEs in the infected cells confirmed that betanodavirus could infect and replicate in BGA cells.…”

Section: Discussionsupporting

confidence: 87%

“…Till date, more than 40 marine and freshwater fishes have been found to be susceptible to betanodavirus infection (Nishi, Yamashita, Kawato, & Nakai, ). Betanodavirus infection in freshwater species such as Guppies, European eel, Chinese catfish and Mosquitofish has also been reported (Chi, Shieh, & Lin, ; Hedge, Teh, Lam, & Sin, ; Hegde, Lam, & Sin, ; Praveenraj, Ezhil Praveena, Bhuvaneswari, Navaneeth Krishnan, & Jithendran, ).…”

Section: Introductionmentioning

confidence: 99%

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Susceptibility of betanodavirus in a newly established vertebra‐derived cell line from Mosquitofish (Gambusia affinis)

Jyotsna

Vijayakumar

Mani

et al. 2019

Journal of Fish Diseases

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In the present study, a new cell line from the vertebra of mosquitofish Gambusia affinis was successfully established and characterized. The cell line is named as bone Gambusia affinis (BGA) and subcultured for more than 55 passages in Leibovitz's/L15 medium supplemented with 15% FBS at 28°C. The cell line has a modal chromosome number of 48. Molecular characterization of the partial sequence of the coi gene confirmed the origin of the BGA cell line from mosquitofish. These cells exhibited epithelial morphology confirmed by the cytokeratin marker. The BGA cells showed mineralization of their extracellular matrix when stained with alizarin red and von Kossa stain. BGA cells were found to be susceptible to RGNNV and SJNNV strains of betanodavirus (NNV) showing cytopathic effect with multiple vacuolations in the cells. The RT-PCR confirmed the betanodavirus infections in BGA cells. The SEM micrograph showed the morphological changes observed in the cell during virus infection. The in vivo challenge experiment also showed the viral replicating efficiency in the Gambusia affinis with increasing viral titre. Thus, our present results show that the BGA cell line is a useful tool for isolating betanodavirus and could be used to investigate bone cell differentiation and extracellular matrix mineralization. K E Y W O R D S betanodavirus, cell line, Gambusia affinis, mineralization, vertebra S U PP O RTI N G I N FO R M ATI O N Additional supporting information may be found online in the Supporting Information section. How to cite this article: Jyotsna, Vijayakumar P, Mani R, Raja S, Dhas TS. Susceptibility of betanodavirus in a newly established vertebra-derived cell line from Mosquitofish (Gambusia affinis).

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Section: Discussionsupporting

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Susceptibility of betanodavirus in a newly established vertebra‐derived cell line from Mosquitofish (Gambusia affinis)

Jyotsna

Vijayakumar

Mani

et al. 2019

Journal of Fish Diseases

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“…The drainage challenge method used in Exp. 2 is less stressful to fish and mimics natural infection, compared with the injection, bath, or oral gavage challenge methods, although it inevitably resulted in relatively lower mortality as in the previous study (Nishi et al, 2016). However, the use of one drainage as an infection source is superior in ensuring the uniformity of infection pressure among tanks as compared to a similar challenge method by cohabitation with diseased fish in the same tank.…”

Section: Resultsmentioning

confidence: 89%

“…2). In the drainage method (Nishi et al, 2016), fish (recipient) were exposed to drainage from an aquarium of fish (donor, n = 20) infected with the virus by IM injection (10 5.5 TCID 50 /fish), and dead fish were removed from the tank on the day. During immunization and challenge periods, fish were kept in 45-L plastic tanks with a flow-through system at a density of 20-25 individuals/tank.…”

Section: Immunization and Virus Challengementioning

confidence: 99%

An Oral Vaccination Method with the Aid of Capsaicin against Viral Nervous Necrosis (VNN)

et al. 2018

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“…However, much remains to be determined about the host-specificity of SJNNVs and virus transmission mechanisms in natural environments. In this regard, a recently reported cell culture isolation method from seawater (Nishi et al, 2016) will be useful for future epidemiological studies on VNN or betanodaviruses.…”

Section: Discussionmentioning

confidence: 99%

Pathogenicity of Striped Jack Nervous Necrosis Virus (SJNNV) Isolated from Asymptomatic Wild Japanese Jack Mackerel <i>Trachurus japonicus</i>

et al. 2016

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Cell Culture Isolation of Piscine Nodavirus (Betanodavirus) in Fish-Rearing Seawater

Cited by 28 publications

References 48 publications

Susceptibility of betanodavirus in a newly established vertebra‐derived cell line from Mosquitofish (Gambusia affinis)

Susceptibility of betanodavirus in a newly established vertebra‐derived cell line from Mosquitofish (Gambusia affinis)

An Oral Vaccination Method with the Aid of Capsaicin against Viral Nervous Necrosis (VNN)

Pathogenicity of Striped Jack Nervous Necrosis Virus (SJNNV) Isolated from Asymptomatic Wild Japanese Jack Mackerel <i>Trachurus japonicus</i>

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