Investigation of laboratory methods for characterization of aquatic viruses in fish infected experimentally with infectious salmon anemia virus.

Autor:	Eckstrand CD; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Torrevillas BK; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Wolking RM; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Bradway DS; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Warg JV; National Veterinary Services Laboratories, U.S. Department of Agriculture, Ames, IA, USA., Clayton RD; National Veterinary Services Laboratories, U.S. Department of Agriculture, Ames, IA, USA., Williams LB; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Pessier AP; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Reno JL; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., McMenamin-Snekvik KM; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Thompson J; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Baszler T; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA., Snekvik KR; Washington Animal Disease Diagnostic Laboratory, Washington State University, Pullman, WA, USA.
Jazyk:	angličtina
Zdroj:	Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc [J Vet Diagn Invest] 2024 May; Vol. 36 (3), pp. 319-328. Date of Electronic Publication: 2023 May 19.
DOI:	10.1177/10406387231173332
Abstrakt:	Rapid growth in aquaculture has resulted in high-density production systems in ecologically and geographically novel conditions in which the emergence of diseases is inevitable. Well-characterized methods for detection and surveillance of infectious diseases are vital for rapid identification, response, and recovery to protect economic and food security. We implemented a proof-of-concept approach for virus detection using a known high-consequence fish pathogen, infectious salmon anemia virus (ISAV), as the archetypal pathogen. In fish infected with ISAV, we integrated histopathology, virus isolation, whole-genome sequencing (WGS), electron microscopy (EM), in situ hybridization (ISH), and reverse transcription real-time PCR (RT-rtPCR). Fresh-frozen and formalin-fixed tissues were collected from virus-infected, control, and sham-infected Atlantic salmon ( Salmo salar ). Microscopic differences were not evident between uninfected and infected fish. Viral cytopathic effect was observed in cell cultures inoculated with fresh-frozen tissue homogenates from 3 of 3 ISAV-infected and 0 of 4 uninfected or sham-infected fish. The ISAV genome was detected by shotgun metagenomics in RNA extracted from the medium from 3 of 3 inoculated cell cultures, 3 of 3 infected fish, and 0 of 4 uninfected or sham-infected fish, yielding sufficient coverage for de novo assembly. An ISH probe against ISAV revealed ISAV genome in multiple organs, with abundance in renal hematopoietic tissue. Virus was detected by RT-rtPCR in gill, heart, kidney, liver, and spleen. EM and metagenomic WGS from tissues were challenging and unsuccessful. Our proof-of-concept methodology has promise for detection and characterization of unknown aquatic pathogens and also highlights some associated methodology challenges that require additional investigation. Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to research, authorship, and/or publication of this article.
Databáze:	MEDLINE
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