Viral replication in excised fin tissues (VREFT) corresponds with prior exposure of Pacific herring, Clupea pallasii (Valenciennes), to viral haemorrhagic septicaemia virus (VHSV).
Procedures for a viral replication in excised fin tissue (VREFT) assay were adapted to Pacific herring, Clupea pallasii, and optimized both to reduce processing time and to provide the greatest resolution between naïve herring and those previously exposed to viral haemorrhagic septicaemia virus (VHSV), Genogroup IVa. The optimized procedures included removal of the left pectoral fin from a euthanized fish, inoculation of the fin with >10(5) plaque-forming units (PFU) mL(-1) VHSV for 1 h, rinsing the fin in fresh medium six times to remove unadsorbed virions, incubation of the fin in fresh medium for 4 days and enumeration of the viral titre in a sample of the incubation medium by plaque assay. The optimized VREFT assay was effective at identifying the prior exposure history of laboratory-reared Pacific herring to VHSV. The geometric mean VREFT value was significantly greater (P < 0.01) among naïve herring (1.2 × 10(3) PFU mL(-1) ) than among groups that survived exposure to VHSV (1.0-2.9 × 10(2) PFU mL(-1) ); additionally, the proportion of cultures with no detectable virus was significantly greater (P = 0.0002) among fish that survived exposure to VHSV (39-47%) than among naïve fish (3.3%). The optimized VREFT assay demonstrates promise for identifying VHSV exposure history and forecasting disease potential in populations of wild Pacific herring.
U.S. Geological Survey-Western Fisheries Research Center, Marrowstone Marine Field Station, Nordland, WA 98358, USA.
C A
CA
Grady
J L
JL
Gregg
R M
RM
Wade
J R
JR
Winton
P K
PK
Hershberger
D058500:Animal Fins / Q000821:virology*; D000818:Animals; D046508:Culture Techniques / Q000662:veterinary*; D004198:Disease Susceptibility / Q000662:veterinary; D005393:Fish Diseases / Q000276:immunology / Q000821:virology*; D005399:Fishes; D031941:Hemorrhagic Septicemia, Viral; D029141:Novirhabdovirus / Q000276:immunology / Q000502:physiology*; D018353:Rhabdoviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D010948:Viral Plaque Assay / Q000379:methods; D014779:Virus Replication