Heart and skeletal muscle inflammation (HSMI) is a significant and often fatal disease of cultured Atlantic salmon in Norway. The consistent presence of Piscine orthoreovirus (PRV) in HSMI diseased fish along with the correlation of viral load and antigen with development of lesions has supported the supposition that PRV is the etiologic agent of this condition; yet the absence of an in vitro culture system to demonstrate disease causation and the widespread prevalence of this virus in the absence of disease continues to obfuscate the etiological role of PRV with regard to HSMI. In this study, we explore the infectivity and disease causing potential of PRV from western North America—a region now considered endemic for PRV but without manifestation of HSMI—in challenge experiments modeled upon previous reports associating PRV with HSMI. We identified that western North American PRV is highly infective by intraperitoneal injection in Atlantic salmon as well as through cohabitation of both Atlantic and Sockeye salmon. High prevalence of viral RNA in peripheral blood of infected fish persisted for as long as 59 weeks post-challenge. Nevertheless, no microscopic lesions, disease, or mortality could be attributed to the presence of PRV, and only a minor transcriptional induction of the antiviral Mx gene occurred in blood and kidney samples during log-linear replication of viral RNA. Comparative analysis of the S1 segment of PRV identified high similarity between this North American sequence and previous sequences associated with HSMI, suggesting that factors such as viral co-infection, alternate PRV strains, host condition, or specific environmental circumstances may be required to cause this disease.
Understanding how pathogenic organisms spread in the environment is crucial for the management of disease, yet knowledge of propagule dispersal and transmission in aquatic environments is limited. We conducted empirical studies using the aquatic virus, infectious hematopoietic necrosis virus (IHNV), to quantify infectious dose, shedding capacity, and virus destruction rates in order to better understand the transmission of IHN virus among Atlantic salmon marine net-pen aquaculture. Transmission of virus and subsequent mortality in Atlantic salmon post-smolts was initiated with as low as 10 plaque forming units (pfu) ml−1. Virus shedding from IHNV infected Atlantic salmon was detected before the onset of visible signs of disease with peak shed rates averaging 3.2×107 pfu fish−1 hour−1 one to two days prior to mortality. Once shed into the marine environment, the abundance of free IHNV is modulated by sunlight (UV A and B) and the growth of natural biota present in the seawater. Virus decayed very slowly in sterilized seawater while rates as high as k = 4.37 d−1 were observed in natural seawater. Decay rates were further accelerated when exposed to sunlight with virus infectivity reduced by six orders of magnitude within 3 hours of full sunlight exposure. Coupling the IHNV transmission parameter estimates determined here with physical water circulation models, will increase the understanding of IHNV dispersal and provide accurate geospatial predictions of risk for IHNV transmission from marine salmon sites.
A survey of wild fishes captured around marine net-pen salmon farms and from open waters for certain salmonid pathogens was conducted in the coastal waters of British Columbia. Viral hemorrhagic septicemia virus was detected in Pacific herring Clupea pallasi, shiner perch Cymatogaster aggregata, and threespine sticklebacks Gasterosteus aculeatus. Infectious hematopoietic necrosis (IHN) virus was detected in one Pacific herring (collected well away from the farms) and in tube-snouts Aulorhynchus flavidus and shiner perch collected from a farm experiencing an IHN outbreak. Renibacterium salmoninarum was observed in moribund Pacific hakes Merluccius productus collected from within a net-pen and was also detected in several ocean-caught salmon. Aeromonas salmonicida subsp. salmonicida (typical strain) was isolated from a juvenile chinook salmon Oncorhynchus tshawytscha, whereas the atypical strain of this organism was isolated from a lingcod Ophiodon elongatus. Loma salmonae (Microsporea) was observed in chinook salmon, chum salmon Oncorhynchus keta, coho salmon O. kisutch, sockeye salmon O. nerka, and pink salmon O. gorbuscha, all of which were captured well away from net-pens. Loma spp. (Microsporea) were observed in the gills of shiner perch, lingcod, Pacific tomcod Microgadus proximus, Pacific cod Gadus macrocephalus, walleye pollock Theragra chalcogramma, and sablefish Anoplopoma fimbria; all but the first species represent new hosts for Loma. Epitheliocystis, caused by a chlamydia-like organism, was detected in the gills of chinook salmon, chum salmon, coho salmon, pink salmon, lingcod, Pacific cod, Pacific hakes, Pacific tomcod, walleye pollock, sablefish, shiner perch, Dover soles Microstomus pacificus, Pacific sanddabs Citharichthys sordidus, and various species of rockfish Sebastes spp., most of which represent new host records for this infection. * Corresponding author: kent@dfo-mpo.gc.ca rhagic septicemia (VHS) virus, Renibacterium salmoninarum, Aeromonas salmonicida, Loma spp. (Microsporea), and the epitheliocystis organism.
Viral hemorrhagic septicemia virus (VHSV) infects over 70 fish species inhabiting marine, brackish or freshwater environments throughout the Northern Hemisphere. Over its geographic range, 4 VHSV genotypes and multiple subtypes exist. Here, we describe the development and validation of a rapid, sensitive and specific real-time reverse transcription quantitative PCR assay (RT-qPCR) that amplifies sequence from representative isolates of all VHSV genotypes (I, II, III and IV). The pan-specific VHSV RT-qPCR assay reliably detects 100 copies of VHSV nucleoprotein RNA without cross-reacting with infectious hematopoietic necrosis virus, spring viremia of carp virus or aquatic birnavirus. Test performance characteristics evaluated on experimentally infected Atlantic salmon Salmo salar L. revealed a diagnostic sensitivity (DSe) ≥93% and specificity (DSp) = 100%. The repeatability and reproducibility of the procedure was exceptionally high, with 93% agreement among test results within and between 2 laboratories. Furthermore, proficiency testing demonstrated the VHSV RT-qPCR assay to be easily transferred to and performed by a total of 9 technicians representing 4 laboratories in 2 countries. The assay performed equivalent to the traditional detection method of virus isolation via cell culture with the advantage of faster turnaround times and high throughput capacity, further suggesting the suitability of the use of this VHSV RT-qPCR in a diagnostic setting.
Epizootics of Ichthyophthirius multifiliis occurred in adult prespawning and spawning sockeye salmon Oncorhynchus nerka during the 1994 and 1995 spawning seasons in the Skeena River watershed in northern British Columbia, Canada. Exceedingly high mortalities occurred at Fulton River and in three spawning channels adjacent to Babine Lake. During these 2 years, the infection and subsequent death of prespawning adults resulted in an estimated 153.6 million fewer sockeye salmon fry produced from Fulton River and the three spawning channels adjacent to Babine Lake than the historical average. Resident fish were the most likely source of the parasite in the watershed because several species were found with light infections of I. multifiliis. Transmission of the parasite to anadromous sockeye salmon was enhanced by the high density of fish held below the spawning grounds for days or weeks prior to moving into the spawning channel. This is the first report of an epizootic of ichthyophthiriasis in wild spawning salmon.
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