Myxobolus arcticus and Parvicapsula minibicornis infections in sockeye salmon Oncorhynchus nerka following downstream migration in British Columbia.

Factors influencing the health of sockeye salmon Oncorhynchus nerka in British Columbia, Canada, are important for fisheries management and conservation. Juvenile salmon originating from the Fraser River were screened for 3 enzootic parasites (Myxobolus arcticus, Parvicapsula minibicornis, Ceratonova shasta) and the bacterium Renibacterium salmoninarum. Fish were collected from the Strait of Georgia in 2010, 2011 and 2012 and genotyped to stock of origin. Trends in infection status were estimated by year, spawning zone and catch area. The annual prevalences of P. minibicornis (n = 1448) were 23.3, 6.5 and 8.1%, and for M. arcticus (n = 1343), annual prevalences were 40.4, 66.3 and 27.4%, respectively. Logistic regression showed that P. minibicornis was most strongly associated with salmon from the lower Fraser River spawning zone and increased with distance caught from the mouth of the Fraser River. In contrast, infection with M. arcticus was most strongly associated with salmon from the middle Fraser River spawning zone, and there was no trend related to distance from the Fraser River. Neither R. salmoninarum nor C. shasta were detected. These observations are discussed in the context of salmon life history and pathogen biology.

Two Novel Microsporidia in Skeletal Muscle of Pike-Perch Sander lucioperca and Burbot Lota lota in Finland.

Two new species of Microsporidia were recognized in skeletal muscle of freshwater fishes from Finland. Myosporidium spraguei n. sp. from pike-perch Sander lucioperca occurred as mature spores within sporophorous vesicles (SPVs) within a xenoma. The ovoid spores were 3.8 µm long and 2.4 µm wide, based on transmission electron micrographs (TEM). The exospore and endospore were equally thick, the nucleus was monokaryotic and the polar filament was isofilar with 12 coils in a single rank, entirely adjacent to the prominent posterior vacuole. Small subunit (SSU) rDNA sequence confirmed the presence of M. spraguei n. sp. in burbot Lota lota . The second species, Microsporidium luciopercae n. sp., also from pike-perch, occurred within SPVs that occupied only a fraction of the volume of the otherwise intact myocyte; no xenoma was produced. Myocyte degeneration and necrosis occurred as mature spores dispersed into direct contact with the sarcoplasm. The ovoid spores were 4.6 µm long and 2.8 µm wide (based on TEM); they were monokaryotic and the polar filament was isofilar with 25 coils in a single rank in the posterior of the spore. The exospore was relatively thin with an irregular profile. Neither infection elicited an inflammatory response, although degenerate spores were observed within host cells, suggesting phagocytosis. Phylogenetic analysis of SSU sequences placed both organisms on distinct clades within the Marinosporidia.

Development of the salmon louse Lepeophtheirus salmonis and its effects on juvenile sockeye salmon Oncorhynchus nerka.

Responses of sockeye salmon Oncorhynchus nerka during infection with Lepeophtheirus salmonis were assessed in controlled laboratory trials. Juvenile salmon were exposed to 100 copepodids fish-1 (Trials 1 and 2) or 300 copepodids fish-1 (Trial 3) at mean weights of approximately 40, 80 and 135 g, respectively. Infections occurred on all salmon in all trials, and mean abundances (infection densities) ranged between 3.3 and 19.4 lice fish-1 (0.08 and 0.44 lice g-1 fish) in Trial 1, between 7.2 and 18.3 (0.09 and 0.22) in Trial 2 and between 19.5 and 60.7 (0.15 and 0.46) in Trial 3. A cumulative mortality of 24.4% occurred in Trial 3. At attachment sites on gills, we observed hyperplasia of basal epithelial cells and fusion of secondary lamellae occasionally associated with a cellular infiltrate. At attachment sites on fins, partial to complete skin erosion occurred, with limited evidence of hyperplasia or inflammation. Scale loss and abrasions coincided with pre-adult lice around 20 d post infection (dpi). Plasma osmolality was significantly elevated in exposed fish in Trials 1 (21 dpi), 2 (15 and 36 dpi) and 3 (20 dpi), whereas haematocrit was significantly depressed in exposed fish in Trials 1 (21 and 28 dpi) and 3 (20 dpi). Plasma cortisol was significantly elevated in exposed fish at 20 dpi (Trial 3). Physiological changes and mortality were related to the intensity of infection and became most prominent with pre-adult stages, suggesting patterns of infection and response in sockeye salmon similar to those reported for Atlantic and Chinook salmon.

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.

Differential expression of the pro-inflammatory cytokines IL-1beta-1, TNFalpha-1 and IL-8 in vaccinated pink (Oncorhynchus gorbuscha) and chum (Oncorhynchus keta) salmon juveniles.

Laboratory-reared pink and chum salmon juveniles (approximately 2g) received an intraperitoneal injection with a commercial, unadjuvanted Aeromonas salmonicida bacterin or sterile saline. Relative to elongation factor-1A, expression levels of genes encoding the proinflammatory cytokines interleukin-1beta-1 (IL-1beta), tumour necrosis factor-alpha-1 (TNFalpha) and interleukin-8 (IL-8) in pools of kidney and liver were examined 6- and 24-h after injection. Expression of IL-1beta was significantly elevated in pink and chum salmon by 6-h, and declined in pink salmon but not in chum salmon by 24-h. Similarly, expression of TNFalpha was significantly elevated in both species at 6h and only in chum salmon after 24-h. Expression of IL-8 was significantly elevated in both species at 6- and 24-h after injection. Expression of the three proinflammatory cytokine genes differed between salmon species both in the timing and magnitude of their expression. The significance of these differences with respect to immune function in these fish requires further research.

The effect of vaccination and sea water entry on immunocompetence and susceptibility to Kudoa thyrsites in Atlantic salmon (Salmo salar L.).

The effect of intraperitoneal (IP) vaccination and sea water entry (SWE) on the immunocompetence of Cascade Atlantic salmon was investigated. Smolts were IP injected with Aqua Health's Forte trade mark vaccine (Listonella (Vibrio) anguillarum, Listonella ordalii, Vibrio salmonicida and Aeromonas salmonicida) at four times (42, 238, 433 and 630 degree days, DD) prior to SWE and were examined for immunocompetence. Immune response measurements included mitogen-driven proliferation of peripheral blood leukocytes (PBL), head kidney leukocyte respiratory burst activity and alternative complement hemolytic titres and were measured 24h prior to SWE, 72 h post-SWE and again 3.5 weeks post-SWE. A 50% reduction in the number of PBL was observed 3 days post-vaccination. At this time LPS-driven proliferation was low (stimulation index, SI, 1.5-2.9) in all groups prior to SWE compared with that of PBL from freshwater-reared Atlantic salmon parr (6.7). By 72 h and 3.5 weeks post-SWE, the LPS-driven SI from unvaccinated salmon and those vaccinated 630 and 433 DD prior to SWE increased 3-fold. In contrast, SI from salmon vaccinated 42 and 238 DD prior to SWE remained low. A similar pattern was observed for cultured PBL stimulated with PHA, although unlike LPS-stimulated PBL, the SI of cells from parr and unvaccinated control smolts remained low following SWE but increased in fish vaccinated 433 and 630 DD prior to SWE. The respiratory burst activity of head kidney leukocytes was not affected by SWE but showed a transient 50% depression 3 days post-vaccination. The alternative complement activity (ACH50) was similar for all treatment groups prior to and at 72h post-SWE. By 3.5 weeks post-SWE, ACH50 values in salmon vaccinated 42 and 238 DD prior to SWE doubled to 874 and 860 U/ml, respectively. The prevalence and severity of Kudoa thyrsites infections, detected in all treatment groups approximately 2400 DD following SWE, were not significantly different among groups. Atlantic salmon parr should be IP vaccinated no earlier than 433 DD before SWE to avoid an enhanced risk of acquiring pathogens because of transient depression in some immune mechanisms.