TY - JOUR
T1 - Propagation and isolation of ranaviruses in cell culture
AU - Ariel, Ellen
AU - Nicolajsen, Nicole
AU - Christophersen, Maj-Britt
AU - Holopainen, Riikka
AU - Tapiovaara, Hannele
AU - Jensen, Ann Britt Bang
PY - 2009
Y1 - 2009
N2 - 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.
AB - 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.
U2 - 10.1016/j.aquaculture.2009.05.019
DO - 10.1016/j.aquaculture.2009.05.019
M3 - Journal article
SN - 0044-8486
VL - 294
SP - 159
EP - 164
JO - Aquaculture
JF - Aquaculture
ER -