Korean Journal of Environmental Biology (환경생물)

Korean Society of Environmental Biology (한국환경생물학회)
권호 표지
DOI QR코드

DOI QR Code

Quantitative analysis of viral hemorrhagic septicemia virus in tissues of infected olive flounder(Paralichthys olivaceus)

바이러스성 출혈성 패혈증 바이러스(Viral Hemorrhagic Septicemia Virus) 감염 넙치(Paralichthys olivaceus)의 조직별 바이러스 정량분석

  • Jang, Jin Hyeon (Aquatic Disease Control Division, National Fishery Products Quality Management Services) ;
  • Hwang, Seong Don (Aquaculture Industry Division, East Sea Fisheries Research Institute) ;
  • Jung, Ji Min (Aquatic Disease Control Division, National Fishery Products Quality Management Services) ;
  • Kwon, Mun-Gyoung (Aquatic Disease Control Division, National Fishery Products Quality Management Services) ;
  • Hwang, Jee Youn (Aquatic Disease Control Division, National Fishery Products Quality Management Services)
  • 장진현 (국립수산물품질관리원 수산방역과) ;
  • 황성돈 (국립수산과학원 동해수산연구소 양식산업과) ;
  • 정지민 (국립수산물품질관리원 수산방역과) ;
  • 권문경 (국립수산물품질관리원 수산방역과) ;
  • 황지연 (국립수산물품질관리원 수산방역과)
  • Received : 2020.05.26
  • Accepted : 2021.03.09
  • Published : 2021.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

A diagnostic test for viral hemorrhagic septicemia virus (VHSV), which infects more than 80 species of freshwater and marine fish at home and abroad, causing mass mortality, was conducted to provide quantitative data on the amount of virus expression in various tissues of flounder in chronological order. The tissues were collected in chronological order after the intraperitoneal injection of 3.0E+07 tissue culture infective dose50 (TCID50) per 0.1mL per fish of VHSV to randomly selected flounder. As a result of relative quantification through real-time PCR, the highest levels of virus expression were found in the spleen, kidney, gill, and liver on day 5. This study proved that the spleen was an appropriate site for the final diagnosis of VHSV in the early stages of infection and will provide important information for the diagnosis of legal infectious diseases in Korea.

국내외에서 80여 종이 넘는 담수 및 해산어류를 감염시켜 대량폐사를 발생시키는 바이러스성 출혈성 패혈증 바이러스(VHSV) 진단검사를 위해 넙치의 여러 조직의 바이러스 발현량에 대한 정량적 데이터를 시간순서에 따라 분석하였다. 무작위 선별된 넙치에 3.0E+07 TCID50 per 0.1mL per fish의 VHSV를 복강 주사하여 시간순서(0시간, 6시간, 12시간, 1일, 2일, 3일, 5일, 7일)에 따라 조직(아가미, 간, 신장, 비장, 근육)을 채취하였다. Real-time PCR 법을 통해 상대 정량한 결과 5일차 아가미, 간, 신장, 비장에서 바이러스의 발현량이 가장 높게 나타났다. 이번 연구를 통해 감염 초기단계에서 비장이 VHSV 확정진단을 위한 적정조직임을 입증하였으며, 국내 법정전염병 진단에 중요한 정보를 제공할 것이다.

Keywords

Acknowledgement

이 논문은 2021년도 국립수산물품질관리원 수산생물방역기술 개발 및 프로그램 운영(R2021071)의 지원으로 수행된 연구입니다.

References

  1. Ahn SJ, MY Cho, BY Jee and MA Park. 2013. Phylogenetic analysis of Viral Haemorrhagic Septicaemia Virus (VHSV) isolates from Asia. J. Fish Pathol. 26:149-161. https://doi.org/10.7847/JFP.2013.26.3.149
  2. Ariel E and NJ Olesen. 2001. Assessment of a commercial kit collection for diagnosis of the fish viruses: IHNV, IPNV, SVCV and VHSV. Bull. Eur. Assoc. Fish Pathol. 21:6-11.
  3. Bernard J, M Bremont and JR Winton. 1992. Nucleocapsid gene sequence of a North-American isolate of viral hemorrhagic septicemia virus, a fish Rhabdovirus. J. Gen. Virol. 73:1011-1014. https://doi.org/10.1099/0022-1317-73-4-1011
  4. Bland F, M Snow, KA Garver and I Matejusova. 2012. Genotype-specific Taqman assays for the detection and rapid characterization of European strains of viral haemorrhagic septicaemia virus. J. Virol. Methods 187:209-214. https://doi.org/10.1016/j.jviromet.2012.10.005
  5. Duesund H, S Nylund, K Watanabe, KF Ottem and A Nylund. 2010. Characterization of a VHS virus genotype III isolated from rainbow trout(Oncorhychus mykiss) at a marine site on the west coast of Norway. Virol. J. 7:19. https://doi.org/10.1186/1743-422X-7-19
  6. Einer-Jensen K, P Ahrens, R Forsberg and N Lorenzen. 2004. Evolution of the fish rhabdovirus viral haemorrhagic septicaemia virus. J. Gen. Virol. 85:1167-1179. https://doi.org/10.1099/vir.0.79820-0
  7. Elsayed E, M Falsal, M Thomas, G Whelan, W Satts and J Wlnton. 2006. Isolation of viral haemorrhagic septicaemia virus from muskellunge, Esox masquinongy, in lake St. Clari, Michigan, USA reveals a new sublineage of the North American genotype. J. Fish Dis. 29:611-619. https://doi.org/10.1111/j.1365-2761.2006.00755.x
  8. Gadd T, M Jakava-Viljanen, H Tapiovaara, P Koski and L Sihvonen. 2011. Epidemiological aspects of viral haemorrhagic septicaemia virus genotype II isolated from Baltic herring, Clupea harengus membras L. J. Fish Dis. 34:517-529. https://doi.org/10.1111/j.1365-2761.2011.01264.x
  9. Gagne N, AM MacKinnon, L Boston, B Souter, M Cook-Versloot, S Griffiths and G Olivier. 2007. Isolation of viral haemorrhagic septicaemia virus from mummichog, stickleback, striped bass and brown trout in eastern Canada. J. Fish Dis. 30:213-223. https://doi.org/10.1111/j.1365-2761.2007.00802.x
  10. Garver KA, LM Hawley, CA Mcclure, T Schroeder, S Aldous, F Doig, M Snow, S Edes, C Baynes and J Richard. 2011. Development and validation of a reverse transcription quantitative PCR for universal detection of viral hemorrhagic septicemia virus. Dis. Aquat. Org. 95:97-112. https://doi.org/10.3354/dao02344
  11. Guomundsdottir S, N Vendramin, A Cuenca, H Siguroardottir, A Kristmundsson, TM Iburg and NJ Olesen. 2019. Outbreak of viral haemorrhagic septicaemia(VHS) in lumpfish(Cyclopterus lumpus) in Iceland caused by VHS virus genotype IV. J. Fish Dis. 42:47-62. https://doi.org/10.1111/jfd.12910
  12. Hedrick RP, WN Batts, S Yun, GS Traxler, J Kaufman and JR Winton. 2003. Host and geographic range extensions of the North American strain of viral hemorrhagic septicemia virus. Dis. Aquat. Org. 55:211-220. https://doi.org/10.3354/dao055211
  13. Hwang JY, JH Jang, DJ Kim, MG Kwon, JS Seo, SD Hwang and MH Son. 2017. Detection of specific antibodies against viral hemorrhagic septicemia virus in infected olive flounder Paralichthys olivaceus using enzyme-linked immunosorbent assay. Korean J. Fish. Aquat. Sci. 50:547-552. https://doi.org/10.5657/KFAS.2017.0547
  14. Jonstrup SP, S Kahns, HF Skall, TS Boutrup and NJ Olesen. 2013. Development and validation of a novel Taqman-based real-time RT-PCR assay suitable for demonstrating freedom from viral haemorrhagic septicaemia virus. J. Fish Dis. 36:9-23. https://doi.org/10.1111/j.1365-2761.2012.01416.x
  15. Kim HS, KW Baek, EJ Ko, NT Luan, Y Lim, HJ Roh and HJ Cha. 2020. Genome based quantification of VHSV in multiple organs of infected olive flounder (Paralichthys olivaceus) using real-time PCR. Genes Genom. 42:773-777. https://doi.org/10.1007/s13258-020-00951-7
  16. Kim JO, WS Kim, SW Kim, HJ Han, J Kim, M Park and MJ Oh. 2014. Development and application of quantitative detection method for viral hemorrhagic septicemia virus (VHSV) genogroup IVa. Viruses 6:2204-2213. https://doi.org/10.3390/v6052204
  17. Kim SM, JI Lee, MJ Hong, HS Park and SI Park. 2003. Genetic relationship of the VHSV (Viral hemorrhagic septicemia virus) isolated from cultured olive flounder, Paralichthys olivaceus in Korea. J. Fish Pathol. 16:1-12.
  18. Lee WL, HM Yun, SR Kim, SJ Jung and MJ Oh. 2007. Detection of viral hemorrhagic septicemia virus (VHSV) from marine fish in the South Western Coastal Area and East China Sea. J. Fish Pathol. 20:201-209.
  19. Livak KJ and TD Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402-408. https://doi.org/10.1006/meth.2001.1262
  20. Mackay IM. 2004. Real-time PCR in the microbiology laboratory. Clin. Microbiol. Infect. 10:190-212. https://doi.org/10.1111/j.1198-743X.2004.00722.x
  21. OIE. 2017. Viral Haemorrhagic Septicaemia. Manual of Diagnostic Tests for Aquatic Animals. World Organization for Animal Health. Paris, France. Retrieved from http://www.oie.int/international-standard-setting/aquatic-manual/access-online on Nov. 2.
  22. Park HK, LJ Jun, SM Kim, M Park, MY Cho, SD Hwang and JB Jeong. 2016. Monitoring of VHS and RSIVD in cultured Paralichthys olivaceus of Jeju in 2015. Korean J. Fish. Aquat. Sci. 49:176-183. https://doi.org/10.5657/KFAS.2016.0176
  23. Park HK, SM Kim, DW Lee, LJ Jun and JB Jeong. 2015. Monitoring of VHS and RSIVD in cultured Paralichthys olivaceus of Jeju in 2014. J. Fish. Mar. Sci. Edu. 27:879-889. https://doi.org/10.13000/JFMSE.2015.27.3.879
  24. Pierce LR and CA Stepien. 2012. Evolution and biogeography of an emerging quasispecies: Diversity patterns of the fish Viral Hemorrhagic Septicemia virus (VHSV). Mol. Phylogenet. Evol. 63:327-341. https://doi.org/10.1016/j.ympev.2011.12.024
  25. Reed LJ and H Muench. 1938. A simple method of estimating fifty per cent endpoints. Am. J. Epidemiol. 27:493-497. https://doi.org/10.1093/oxfordjournals.aje.a118408
  26. Skall HF, NJ Olesen and S Mellergaard. 2005. Viral haemorrhagic septicaemia virus in marine fish and its implications for fish farming - a review. J. Fish Dis. 28:509-529. https://doi.org/10.1111/j.1365-2761.2005.00654.x
  27. Soliman H and M El-Matbouli. 2006. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of viral hemorrhagic septicaemia virus (VHS). Vet. Microbiol. 114:205-213. https://doi.org/10.1016/j.vetmic.2005.11.063
  28. Thompson TM, WN Batts, M Faisal, P Bowser, JW Casey, K Phillips and G Kurath. 2011. Emergence of viral hemorrhagic septicemia virus in the North American Great Lakes region is associated with low viral genetic diversity. Dis. Aquat. Org. 96:29-43. https://doi.org/10.3354/dao02362
  29. Traxler GS, D Kieser and J Richard. 1999. Mass mortality of pilchard and herring associated with viral hemorrhagic septicemia virus in British Columbia, Canada. FHS Newsl. 27:4-5.
  30. Trdo N, A Benmansour, C Calisher, RG Dietzgen, RX Fang, AO Jackson, G Kurath, S Nadin-Davis, RB Tesh and PJ Walker. 2005. Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses. Academic Press. Cambridge, MA. pp. 623-644.
  31. Winton J, G Kurath and W Batts. 2007. Detection of Viral Hemorrhagic Septicemia Virus. No. 2007-3055. United States Geological Survey. Reston, VA.