Piscine reovirus, but not Jaundice Syndrome, was transmissible to Chinook Salmon, Oncorhynchus tshawytscha (Walbaum), Sockeye Salmon, Oncorhynchus nerka (Walbaum), and Atlantic Salmon, Salmo salar L.

A Jaundice Syndrome occurs sporadically among sea-pen-farmed Chinook Salmon in British Columbia, the westernmost province of Canada. Affected salmon are easily identified by a distinctive yellow discolouration of the abdominal and periorbital regions. Through traditional diagnostics, no bacterial or viral agents were cultured from tissues of jaundiced Chinook Salmon; however, piscine reovirus (PRV) was identified via RT-rPCR in all 10 affected fish sampled. By histopathology, Jaundice Syndrome is an acute to peracute systemic disease, and the time from first clinical signs to death is likely <48 h; renal tubular epithelial cell necrosis is the most consistent lesion. In an infectivity trial, Chinook Salmon, Sockeye Salmon and Atlantic Salmon, intraperitoneally inoculated with a PRV-positive organ homogenate from jaundiced Chinook Salmon, developed no gross or microscopic evidence of jaundice despite persistence of PRV for the 5-month holding period. The results from this study demonstrate that the Jaundice Syndrome was not transmissible by injection of material from infected fish and that PRV was not the sole aetiological factor for the condition. Additionally, these findings showed the Pacific coast strain of PRV, while transmissible, was of low pathogenicity for Atlantic Salmon, Chinook Salmon and Sockeye Salmon.

Development of a multiplex assay to measure the effects of shipping and storage conditions on the quality of RNA used in molecular assays for detection of viral haemorrhagic septicemia virus.

Abstract In routine diagnostics, real-time reverse transcriptase quantitative PCR (RT-qPCR) has become a powerful method for fish health screening. Collection, transportation, and storage conditions of specimens could dramatically affect their integrity and could consequently affect RT-qPCR test results. In this study, to assess the expression profile of elongation factor 1 alpha (ELF-1α) gene, head kidney (HK) tissues from Atlantic Salmon Salmo salar were exposed at room temperature, 4°C, -20°C, and -80°C as well as in 70% ethanol for 6, 12, 24, 48, and 72 h. Data showed a significant increase of RT-qPCR cycle threshold (Ct) values for ELF-1α ranging from 14.7 to 26.5 cycles for tissues exposed to room temperature. In order to mimic the sample transportation conditions, different temperatures of storage were used and tissue quality was evaluated using ELF-1α gene expression. Data showed that Ct values for ELF-1α increased significantly when the tissues were transported on ice for 2 h, stored at -20°C, thawed on ice for 6 h, and stored again at -80°C. The HK tissues collected from Atlantic Salmon challenged with viral hemorrhagic septicemia virus (VHSV) through intraperitoneal injection were exposed at room temperature for 0, 6, 12, 24, 48, 72, and 96 h. Data showed a good correlation of values for ELF-1α and VHSV Ct although the ELF-1α mRNA of the host degraded faster than the RNA of VHSV. Based on these data, HK tissues could be transported on ice or ice packs without the quality of the tissue being affected when stored at -80°C upon arrival at the laboratory. In addition, 70% ethanol could be used as a preservative for long-distance transportation. For an efficient diagnostic test, a duplex VHSV-ELF-1α was developed and optimized. Data showed that the sensitivity of the duplex assay for VHSV was similar to the singleplex. Received November 25, 2013; accepted February 14, 2014.

Parasites and hepatic lesions among pink salmon, Oncorhynchus gorbuscha (Walbaum), during early seawater residence.

Juvenile pink salmon, Oncorhynchus gorbuscha (Walbaum), in the Broughton Archipelago region of western Canada were surveyed over 2 years for sea lice (Lepeophtheirus salmonis and Caligus clemensi), gross and microscopic lesions and evidence of infections with viruses and bacteria. The 1071 fish examined had an approximate ocean residence time no longer than 3 months. A high prevalence of degenerative liver lesions, renal myxosporean parasites and a low prevalence of skin lesions and sea lice were observed. No indications of viral or bacterial diseases were detected in either year. The monthly prevalence of sea lice in 2007 (18-51%) was higher than in 2008 (1-26%), and the infestation density exceeded the lethal threshold in only two fish. Degenerative hepatic lesions and renal myxosporean parasites occurred in approximately 40% of the pink salmon examined in June of both years, and the peak monthly prevalence of hepatocellular hydropic degeneration was greater in 2007 (32%, in May) than in 2008 (12%, in June). Logistic regression analysis found skin lesions and hepatocellular hydropic degeneration significantly associated with sea lice. Most parasites and lesions occurred during both years, but the prevalence was often higher in 2007. Fish weight was 35% less in June 2007 than in June 2008, but condition factor was not different. Further research is required to monitor inter-annual variations and aetiology of the liver lesions and to assess their potential role on pink salmon survival.

Differences in Lepeophtheirus salmonis abundance levels on Atlantic salmon farms in the Broughton Archipelago, British Columbia, Canada.

Sea lice data collected from Atlantic salmon farms in the Broughton Archipelago between 2003 and 2005 were examined for inter-regional differences in mobile Lepeophtheirus salmonis (Krøyer) abundance using the generalized linear model procedure. Factors such as age of the salmon populations, location of farms and time of year had a significant effect on the abundance of the mobile stages of L. salmonis whereas water temperature and salinity did not. Separate evaluation of SLICE treatment data found no significant difference in treatment frequency among the areas but did show that there were significantly lower numbers of farm treatments during the summer months when compared with other seasons. The role of environment and wild fish in influencing sea lice abundance on the farmed salmon is discussed. The findings suggest that effective management programmes for sea lice should not only be based on geographical location but should take into account other factors which could influence lice abundance levels.

Infectious haematopoietic necrosis epidemic (2001 to 2003) in farmed Atlantic salmon Salmo salar in British Columbia.

I investigated a recent infectious haematopoietic necrosis disease (IHN) epidemic in farmed Atlantic salmon Salmo salar in British Columbia (BC), Canada. All companies with infected farms (n = 36) participated in the study. Over 12 million Atlantic salmon on infected farms died or were culled during the epidemic with cumulative mortality on the farms averaging 58%. The first reported case of IHN occurred in August 2001 and the last outbreak in June 2003. Outbreaks on the farms lasted between 20 and 22 wk. Genetic sequencing by other researchers, revealed that 2 different IHN isolates contributed to this epidemic, one linked to all cases in 4 areas, the other associated with all cases in a fifth area. Spatial and temporal patterns of the farm outbreaks were examined to determine possible methods of spread between the farms. Evidence presented herein appears to show that farming practices themselves contributed significantly to the spread between farms both within and between areas. Natural waterborne transmission may have played a role in the spread of the virus between farms located in close proximity to each other. The data collected from this epidemic are compared with reports which examined the first reported epidemic in Atlantic salmon in BC (1992 to 1996). Evidence is presented for the hypothesis that wild fish species may have been the source of introduction of the virus into the farmed Atlantic salmon population.

Questionnaire-based risk assessment for amoebic gill disease (AGD) and evaluation of freshwater bathing efficacy of reared Atlantic salmon Salmo salar.

Factors causing amoebic gill disease (AGD), the main disease affecting the salmonid industry in Tasmania, are largely unknown. Managers of 57 sites, from 4 countries, in both the Southern and Northern Hemisphere were questioned regarding the 2000 smolt year-class Atlantic salmon. Questions included site characteristics, fish population characteristics, husbandry and management programs, through to freshwater bathing characteristics. Possible risk factors for AGD outbreaks that were identified in this study using farm managers' opinions were mixed sex stocks (Tasmania), downstream positioning from other salmonid farming sites (pooled), tidal sites (Tasmania), depth of site (pooled), rate of salinity change in halocline and depth of halocline (pooled, Tasmania), phytoplankton blooms (pooled, Tasmania), antifouling paints (Tasmania, overseas) and automated feeding (pooled, overseas). The information from farm managers suggested that freshwater bathing is less effective with smaller bathing cages and lower levels of saturated oxygen in the bathing water. Risk factors could not always be determined due to the complex interactions between the biological and chemical environment, host and pathogenic factors, coupled with low sample numbers.

A field evaluation of an indirect immunofluorescent antibody test developed to diagnose plasmacytoid leukemia in chinook salmon (Oncorhynchus tshawytscha).

An immunofluorescent antibody test (IFAT) developed for the diagnosis for plasmacytoid leukemia was evaluated against histology under field conditions. Previously published results from a laboratory evaluation indicated that the IFAT had a much higher sensitivity than did histology. One hundred seventy-seven moribund chinook salmon from 3 farms located in British Columbia were sampled. Sensitivity, specificity and their respective quality indices were estimated for the IFAT relative to histology. The IFAT was shown to be unreliable, particularly with respect to sensitivity. Cohen's kappa was also calculated and revealed that the agreement between the 2 tests was no better than random. In contrast to previously published results the IFAT did not perform better than histology in the presence of bacterial kidney disease. The results emphasize the importance of evaluating tests in the field conditions in which they are to be used. The possible reasons for the shortcomings of the IFAT are discussed.

Production of monoclonal antibodies specific for antigens derived from tissue of chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia.

Two distinct monoclonal antibodies (MAB) were prepared for testing with kidney, spleen, and retrobulbar tissue imprints made from chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia. (PL). Hybridomas were prepared from mice immunized with whole and lysed cells purified from renal or retrobulbar PL-positive tissues, which had been obtained from naturally and experimentally infected fish from British Columbia, Canada. The MAB reacted with at least 4 morphologically different cell types; fluorescence was associated with the plasma membrane and cytoplasm. The MAB also reacted with kidney imprints made from chinook salmon affected with a PL-like lymphoproliferative disease in California, indicating that these 2 diseases might be caused by a similar agent. The MAB did not react with any of the kidney or spleen imprints made from wild chinook salmon collected from a river in Ontario, Canada.