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Isolation and identification of a new strain of hirame rhabdovirus (HIRRV) from Japanese flounder Paralichthys olivaceus in China

View Article: PubMed Central - PubMed

ABSTRACT

Background: Hirame rhabdovirus virus (HIRRV) is a rhabdovirus that causes acute hemorrhage disease in fish culture, resulting in a great economic loss in parts of Asia and Europe.

Methods: In this study, we isolated a virus strain named as CNPo2015 from cultured Japanese flounder in Shandong province, China. Cell isolation, electron microscopic observation, RT-PCR detection and phylogenetic analysis were used for virus identification. Further, artificial infection experiment was conducted for virulence testing.

Results: The gross signs included abdominal distension, fin reddening and yellow ascitic fluid in the abdominal cavity. Histopathological examination revealed marked cell degeneration and necrosis in the kidney. The tissue homogenates induced obvious cytopathic effects in EPC, FHM and FG cell lines. Electron microscopic observation showed the virus had a bullet-like shape with a capsule membrane. RT-PCR and sequencing analysis revealed that CNPo2015 belonged to the HIRRV with high sequence identity to HIRRV isolates. Infection experiment confirmed that the HIRRV CNPo2015 strain was virulent to flounder juveniles with a LD50 value of 1.0 × 105.9 TCID50/fish.

Conclusion: In conclusion, we described the first isolation and characterization of a HIRRV from Japanese flounder in China. This will provide a candidate material for further research on the infection mechanism and preventive strategies of HIRRV.

No MeSH data available.


RT-PCR analysis. a RT-PCR amplification from infected EPC cells using specific primers of HIRRV, VHSV, IHNV, VNNV and LCDV. b The flounder β-actin was used as an internal control gene
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Fig4: RT-PCR analysis. a RT-PCR amplification from infected EPC cells using specific primers of HIRRV, VHSV, IHNV, VNNV and LCDV. b The flounder β-actin was used as an internal control gene

Mentions: As shown in Fig. 4, a specific band was amplified from virus culture supernatants using HIRRV specific primers, while no product was detected in the normal EPC cell samples (data not shown). No product was detected using primers of VHSV, IHNV, VNNV or LCDV. The G gene fragment of CNPo2015 revealed 100% sequence identity with the known HIRRV isolates.Fig. 4


Isolation and identification of a new strain of hirame rhabdovirus (HIRRV) from Japanese flounder Paralichthys olivaceus in China
RT-PCR analysis. a RT-PCR amplification from infected EPC cells using specific primers of HIRRV, VHSV, IHNV, VNNV and LCDV. b The flounder β-actin was used as an internal control gene
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5384145&req=5

Fig4: RT-PCR analysis. a RT-PCR amplification from infected EPC cells using specific primers of HIRRV, VHSV, IHNV, VNNV and LCDV. b The flounder β-actin was used as an internal control gene
Mentions: As shown in Fig. 4, a specific band was amplified from virus culture supernatants using HIRRV specific primers, while no product was detected in the normal EPC cell samples (data not shown). No product was detected using primers of VHSV, IHNV, VNNV or LCDV. The G gene fragment of CNPo2015 revealed 100% sequence identity with the known HIRRV isolates.Fig. 4

View Article: PubMed Central - PubMed

ABSTRACT

Background: Hirame rhabdovirus virus (HIRRV) is a rhabdovirus that causes acute hemorrhage disease in fish culture, resulting in a great economic loss in parts of Asia and Europe.

Methods: In this study, we isolated a virus strain named as CNPo2015 from cultured Japanese flounder in Shandong province, China. Cell isolation, electron microscopic observation, RT-PCR detection and phylogenetic analysis were used for virus identification. Further, artificial infection experiment was conducted for virulence testing.

Results: The gross signs included abdominal distension, fin reddening and yellow ascitic fluid in the abdominal cavity. Histopathological examination revealed marked cell degeneration and necrosis in the kidney. The tissue homogenates induced obvious cytopathic effects in EPC, FHM and FG cell lines. Electron microscopic observation showed the virus had a bullet-like shape with a capsule membrane. RT-PCR and sequencing analysis revealed that CNPo2015 belonged to the HIRRV with high sequence identity to HIRRV isolates. Infection experiment confirmed that the HIRRV CNPo2015 strain was virulent to flounder juveniles with a LD50 value of 1.0 × 105.9 TCID50/fish.

Conclusion: In conclusion, we described the first isolation and characterization of a HIRRV from Japanese flounder in China. This will provide a candidate material for further research on the infection mechanism and preventive strategies of HIRRV.

No MeSH data available.