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A sensitive non‐destructive method for detecting IPNV carrier Atlantic salmon, Salmo salar L., by culture of virus from plastic adherent blood leucocytes
Author(s) -
Munro E S,
Gahlawat S K,
Ellis A E
Publication year - 2004
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.2004.00520.x
Subject(s) - percoll , infectious pancreatic necrosis virus , biology , salmo , virus , mucus , lysis , microbiology and biotechnology , virology , centrifugation , fish <actinopterygii> , fishery , ecology , biochemistry
In populations of Atlantic salmon in sea water, infectious pancreatic necrosis virus (IPNV) could be detected by standard virological culture methods in sonicated kidney homogenates and in mucus samples (gill, skin and rectum) from 14 and nine of 25 fish, respectively, but all fish were positive by virus culture from lysates of kidney macrophages and adherent blood leucocytes. In fish which tested negative for IPNV by the standard method of detection, the virus could be detected using adherent blood leucocytes isolated on a Percoll gradient from as little as 10 μ L of blood. The blood sample could be stored for at least 3 days in a heparinized tube on ice before preparing the plastic adherent leucocytes. Furthermore, the latter could be prepared without prior fractionation on Percoll simply by incubating whole blood (33 μ L) in cell culture medium (66 μ L) in 96‐well plates overnight and washing away the non‐adherent cells before lysing the adherent cells and inoculation of the lysate onto CHSE‐214 cells. This highly sensitive method for detecting IPNV‐carriers is therefore very suitable for non‐destructive sampling of fish in the field.