Propagation and isolation of ranaviruses in cell culture

Ariel, Ellen, Nicolajsen, Nicole, Christophersen, Maj-Britt, Holopainen, Riikka, Tapiovaara, Hannele, and Jensen, Britt Bang (2009) Propagation and isolation of ranaviruses in cell culture. Aquaculture, 294 (3). pp. 159-164.

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Abstract

The optimal in vitro propagation procedure for a panel of ranavirus isolates and the best method for isolation of Epizootic haematopoietic necrosis virus (EHNV) from organ material in cell-culture were investigated. The panel of ranavirus isolates included: Frog virus 3 (FV3), Bohle iridovirus (BIV), Pike-perch iridovirus (PPIV), European catfish virus (ECV), European sheatfish virus (ESV), EHNV, Doctor fish virus (DFV), Guppy virus 6 (GF6), short-finned eel virus (SERV) and Rana esculenta virus Italy 282/102 (REV 282/102). Each isolate was titrated in five cell lines: bluegill fry (BF-2), epithelioma papulosum cyprini (EPC), chinook salmon embryo (CHSE-214) rainbow trout gonad (RTG-2) and fathead minnow (FHM), and incubated at 10, 15, 20, 24 and 28 °C for two weeks.

BF-2, EPC and CHSE-214 cells performed well and titers obtained in the three cell lines were similar, whereas FHM and RTG-2 cells consistently produced lower titers than the other cell lines at all temperatures. The optimal temperature for propagating the isolates collectively to high titers in vivo was 24 °C.

Additionally, three established methods for re-isolation of virus from EHNV-infected organ material were compared. Challenged fish were sampled twice weekly and 7 organs were processed separately according to the three methods. Samples incubated on BF-2 cells at 22 °C for 2 weeks + 1 week sub-cultivation (method 1) provided more positive results than the other 2 methods and when using the EPC cell line. Virus was most frequently isolated from the kidney, followed by brain, muscle, heart, liver, gills and lastly spleen.

Item ID: 10634
Item Type: Article (Refereed Research - C1)
Keywords: Ranavirus; EHNV; propagation; isolation; cell culture; fish virus
ISSN: 1873-5622
Date Deposited: 16 Apr 2010 05:54
FoR Codes: 07 AGRICULTURAL AND VETERINARY SCIENCES > 0707 Veterinary Sciences > 070712 Veterinary Virology @ 100%
SEO Codes: 83 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 8301 Fisheries - Aquaculture > 830102 Aquaculture Fin Fish (excl. Tuna) @ 50%
92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920109 Infectious Diseases @ 50%
Citation Count from Web of Science Web of Science 16
Downloads: Total: 3
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