Picorna-like virus associated with mortality and a spongious encephalopathy in grouper Epinephelus malabaricus

Boonyaratpalin, S; Supamattaya, K.; Kasornchandra, Jiraporn; Rw, Hoffmann

doi:10.3354/dao026075

Cited by 42 publications

(34 citation statements)

References 10 publications

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“…Collected fishes exhibited abnormal swimming, abdominal swollen, belly up at rest as well as lethargy, bilateral exophthalmia and haemorrhages in the skin. L. aurata and L. saliens displayed abnormal swimming behaviour which were characteristic of the VNN disease as reported by several workers (Yoshikoshi and Inoue, 1990;Glazebrook et al, 1990;Grotmol et al, 1995;Boonyaratpalin et al, 1996).…”

Section: Discussionsupporting

confidence: 51%

RETRACTED: Isolation and confirmation of viral nervous necrosis (VNN) disease in golden grey mullet (Liza aurata) and leaping mullet (Liza saliens) in the Iranian waters of the Caspian Sea

Zorriehzahra

Ghasemi

Ghiasi

et al. 2016

Veterinary Microbiology

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A B S T R A C TThe present study was conducted on 428 moribund mullet fish samples to isolate and identify the causative agent of a mysterious acute mortality which recently occurred in wild mullets in Iranian waters of Caspian Sea, suspected to be due to viral nervous necrosis (VNN) disease. Disease investigation was carried out employing various diagnostic procedures such as virology, bacteriology, parasitology, haematology, histopathology, IFAT, IHC and nested RT-PCR. Brain and eye samples of affected fishes were collected in sterile conditions and then kept at À80 C for cell culture isolation and nested RT-PCR detection of the causative agent. Other tissue samples were also collected and fixed for histopathology, IHC and EM examinations. CPE was observed in cell cultures at 6 days after inoculation. Nine samples were found positive with virological assay. Nested RT-PCR, performed on suspected tissues and CPE positive samples, showed that about 21 tissue samples and all the CPE positive samples were positive for VNN virus (VNNV). IFAT was selected as a confirmatory method for detecting the presence of Betanodavirus antigen, cell culture isolation results and nested RT-PCR findings. Moreover, VNNV particles with 25-30 nm in diameter were also visualized in the infected brain and retina. In pathogenicity studies, guppy fishes bathed in VNNV-infected tissue culture (10 À4 TCID 50 ) showed clinical signs similar to naturally infected mullet after 15 days post infection (dpi), with mortality rates reaching up to 100% at 30 dpi. Affected organ samples as examined by cell culture isolation, IFAT, IHC and histopathology, revealed the presence of VNNV in the guppy fishes. In conclusion, it was confirmed that VNNV was the main causative agent for the disease outbreak in mullet fish in the Caspian Sea, and this is such first official report of VNN disease from Iran.

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Section: Discussionsupporting

confidence: 51%

RETRACTED: Isolation and confirmation of viral nervous necrosis (VNN) disease in golden grey mullet (Liza aurata) and leaping mullet (Liza saliens) in the Iranian waters of the Caspian Sea

Zorriehzahra

Ghasemi

Ghiasi

et al. 2016

Veterinary Microbiology

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“…Uncoordinated swimming, i.e. spiral swimming and darting, although flatfish tend to have a looping swimming pattern and to rest belly up were reported by [3,[15][16][17][18][19][20][21]. Similar clinical signs were reported during serious mortalities affecting Grey mullet in Iranian coastal water of the Caspian Sea [1,22].…”

Section: Discussionmentioning

confidence: 80%

Vacuolating encephalopathy and retinopathy associated with a nodavirus-like agent: a probable cause of mass mortality of wild Golden grey mullet (Liza aurata) and Sharpnose grey mullet (Liza saliens) in Iranian waters of the Caspian Sea

Zorriehzahra

Nazari

Ghasemi³

et al. 2014

VirusDis.

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“…This is particularly important for virulence assays of piscine nodavirus in fish. Every pathogenicity experiments reported so far (Mori et al 1991, Arimoto et al 1993, Boonyaratpalin et al 1996, Peducasse et al 1999) lacked the quantitative measurement of the inoculated dose. This lack of measurement caused difficulties in comparisons of virulence among virus strains or virus susceptibility among host fish species.…”

Section: Discussionmentioning

confidence: 99%

Cloning of the fish cell line SSN-1 for piscine nodaviruses

Iwamoto¹,

Nakai

Mori³

et al. 2000

Dis. Aquat. Org.

230

194

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Six cell clones were derived from the SSN-1 cell line, which is composed of a mixed cell population and persistently infected with a C-type retrovirus (SnRV). These clones were susceptible to 4 piscine nodavirus strains belonging to different genotypes (SJNNV, RGNNV, TPNNV and BFNNV [striped jack, redspotted grouper, tiger puffer and barfin flounder nervous necrosis viruses]). Three clones, designated A-6, E-9, and E-11, were highly permissive to nodavirus infection and production. The virus-induced cytopathic effects appeared as cytoplasmic vacuoles and intensive disintegration at 3 to 5 d post-incubation. These observations were highly reproducible and formed the basis for a successful virus titration system. Quantitative analysis using the cloned E-11 cell line clearly revealed differences in the optimal growth temperatures among the 4 genotypic variants: 25 to 30°C for strain SGWak97 (RGNNV), 20 to 25°C for strain SJNag93 (SJNNV), 20°C for strain TPKag93 (TPNNV), and 15 to 20°C for strain JFIwa98 (BFNNV). Electron microscopy demonstrated SnRV retrovirus particles only in A-6 and E-9 cells, but PCR amplification for the pol gene and LTR region of the proviral DNA indicated the presence of the retrovirus in the other clones, including E-11. The cell clones obtained in the present study will be more useful for qualitative and quantitative analyses of piscine nodaviruses than the SSN-1 cell line. KEY WORDS: Nodavirus · Viral nervous necrosis, VNN · SSN-1 cell line · Cell cloning · C-type retrovirus · Snakehead retrovirus Resale or republication not permitted without written consent of the publisherDis Aquat Org 43: [81][82][83][84][85][86][87][88][89] 2000 that a new cell line (GF-1) derived from grouper Epinephelus coioides was useful for the isolation and proliferation of a piscine nodavirus (GNNV, grouper nervous necrosis virus).Piscine nodaviruses can be divided into 4 genotypic groups based on partial sequences of the coat protein gene (Nishizawa et al. 1997): SJNNV (striped jack nervous necrosis virus), RGNNV (redspotted grouper nervous necrosis virus), TPNNV (tiger puffer nervous necrosis virus), and BFNNV (barfin flounder nervous necrosis virus). In a previous study, we demonstrated that the SSN-1 cell line was useful for propagating and differentiating 17 isolates of piscine nodavirus collected from 13 host fish species in 5 countries (Iwamoto et al. 1999). However, one problem with the practical use of the SSN-1 cell line was that this cell line was composed of a mixed population of cells, causing inconsistencies in the cytopathic effects (CPE) observed during virus infection. For this reason, FAT was used to titrate the virus instead of CPE as described in our previous study (Iwamoto et al. 1999). FAT was laborious and costly due to the requirement of special test chambers, and it has proved to be inadequate for the quantitative analysis of a large number of samples. In addition, the fact that the SSN-1 cell line is spontaneously infected by a C-type retrovirus designated as SnRV (Frerich...

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Picorna-like virus associated with mortality and a spongious encephalopathy in grouper Epinephelus malabaricus

Cited by 42 publications

References 10 publications

RETRACTED: Isolation and confirmation of viral nervous necrosis (VNN) disease in golden grey mullet (Liza aurata) and leaping mullet (Liza saliens) in the Iranian waters of the Caspian Sea

RETRACTED: Isolation and confirmation of viral nervous necrosis (VNN) disease in golden grey mullet (Liza aurata) and leaping mullet (Liza saliens) in the Iranian waters of the Caspian Sea

Vacuolating encephalopathy and retinopathy associated with a nodavirus-like agent: a probable cause of mass mortality of wild Golden grey mullet (Liza aurata) and Sharpnose grey mullet (Liza saliens) in Iranian waters of the Caspian Sea

Cloning of the fish cell line SSN-1 for piscine nodaviruses

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