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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.


Related in: MedlinePlus

Virus isolation on four fish cell lines. Compared with uninfected fish cell lines, the infected EPC cell line showed typical CPE with cell rounding, detachment and dead cells. The infected FHM and FG cell lines were also observed with CPE. No CPE were observed in CAR cell line. Magnification, 30 × 
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Fig2: Virus isolation on four fish cell lines. Compared with uninfected fish cell lines, the infected EPC cell line showed typical CPE with cell rounding, detachment and dead cells. The infected FHM and FG cell lines were also observed with CPE. No CPE were observed in CAR cell line. Magnification, 30 × 

Mentions: The homogenate supernatants of kidney and spleen tissues induced positive CPE in the EPC, FHM, FG cells at 3 days post inoculation at 15 °C. The CPE was characterized by rounded and granular cells, grape-like clusters, cell detached and lysis. No CPE was observed in the CAR cell line (Fig. 2). In all cell lines, no CPE was induced by the brain and gill samples. At 7 days post inoculation, the EPC cells produced the highest titer of virus with a titer of 1 × 108.5 TCID50/mL, while the titer was 1 × 108.1 TCID50/mL for FHM cells, and 1 × 107.3 TCID50/mL for FG cells, respectively. Virus titers in all cell lines stabilized for the remainder of the infection.Fig. 2


Isolation and identification of a new strain of hirame rhabdovirus (HIRRV) from Japanese flounder Paralichthys olivaceus in China
Virus isolation on four fish cell lines. Compared with uninfected fish cell lines, the infected EPC cell line showed typical CPE with cell rounding, detachment and dead cells. The infected FHM and FG cell lines were also observed with CPE. No CPE were observed in CAR cell line. Magnification, 30 × 
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig2: Virus isolation on four fish cell lines. Compared with uninfected fish cell lines, the infected EPC cell line showed typical CPE with cell rounding, detachment and dead cells. The infected FHM and FG cell lines were also observed with CPE. No CPE were observed in CAR cell line. Magnification, 30 × 
Mentions: The homogenate supernatants of kidney and spleen tissues induced positive CPE in the EPC, FHM, FG cells at 3 days post inoculation at 15 °C. The CPE was characterized by rounded and granular cells, grape-like clusters, cell detached and lysis. No CPE was observed in the CAR cell line (Fig. 2). In all cell lines, no CPE was induced by the brain and gill samples. At 7 days post inoculation, the EPC cells produced the highest titer of virus with a titer of 1 × 108.5 TCID50/mL, while the titer was 1 × 108.1 TCID50/mL for FHM cells, and 1 × 107.3 TCID50/mL for FG cells, respectively. Virus titers in all cell lines stabilized for the remainder of the infection.Fig. 2

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.


Related in: MedlinePlus