Variation in the susceptibility of sockeye salmon Oncorhynchus nerka to infectious haemopoietic necrosis virus

Haemoglobin concentrations, haematocrit values, red blood cell counts, red blood cell diameter, erythrocyte sedimentation rate and plasma haemoglobin concentration were measured on the air-breathing mud eel Amphipnous cuchia .     

Susceptibility of captive adult winter-run Chinook salmon Oncorhynchus tshawytscha to waterborne exposures with infectious hematopoietic necrosis virus (IHNV)

Sexually mature female Chinook salmon Oncorhynchus tshawytscha with no prior history of exposure to infectious hematopoietic necrosis virus (IHNV) were susceptible to experimental infection induced by additions of virus to the water. The resulting infections resembled those observed among naturally infected hatchery and wild populations of Chinook salmon. Virus was detected as early as 4 d post-exposure (p.e.) and subsequently in all virus-exposed fish that died or that were examined at 14 d p.e. when the study was terminated. The greatest concentrations of virus, up to 10(8) plaque-forming units (pfu) ml(-1), were found in the ovarian fluid at 13 to 14 d p.e., but the virus was also found in high concentrations in the gill, kidney/spleen and plasma. In contrast, the virus was not recovered from unexposed control adult salmon that died or were sampled at the end of the study. Despite detecting concentrations of IHNV in excess of 10(7) pfu g(-1) of tissue, no specific microscopic lesions were found in IHNV-exposed compared to unexposed control salmon. The results of this initial study suggest that virus in the spawning environment, either from adult salmon or other sources, may contribute to its rapid spread among adult Chinook salmon, thereby considerably increasing the prevalence of IHNV infection in both wild and hatchery populations of adult Chinook salmon.     

Evidence for a carrier state of infectious hematopoietic necrosis virus in chinook salmon Oncorhynchus tshawytscha.

In British Columbia, Canada, infectious hematopoietic necrosis virus (IHNV) is prevalent in wild sockeye salmon Oncorhynchus nerka and has caused disease in seawater net-pen reared Atlantic salmon Salmo salar. In this study, chinook salmon Oncorhynchus tshawytscha experimentally exposed to an isolate of IHNV found in British Columbia became carriers of the virus. When Atlantic salmon were cohabited with these virus-exposed chinook salmon, IHNV was isolated from the Atlantic salmon. Identification of chinook salmon populations that have been exposed to IHNV may be difficult, as virus isolation was successful only in fish that were concurrently infected with either Renibacterium salmoninarum or Piscirickettisia salmonis. Also, IHNV-specific antibodies were detected in only 2 of the 70 fish experimentally exposed to the virus. Two samples collected from chinook salmon exposed to IHNV while at a salt water net-pen site had a seroprevalence of 19 and 22%; however, the inconsistencies between our laboratory and field data suggest that further research is required before we can rely on serological analysis for identifying potential carrier populations. Because of the difficulty in determining the exposure status of populations of chinook salmon, especially if there is no concurrent disease, it may be prudent not to cohabit Atlantic salmon with chinook salmon on a farm if there is any possibility that the latter have been exposed to the virus.     

Retinal projections in sockeye salmon smolts (Oncorhynchus nerka)

Summary The retinal projections in 2-year-old salmon smolt (Oncorhynchus nerka) are significantly different from those observed in other teleosts examined to date in that the projections are more extensive. Very noticeable are extensive projections to most of the dorsal thalamus, to all layers of the optic tectum, and into the periaqueductal gray of the torus semicircularis. The salmon smolt has bilateral retinal projections to the diencephalon and pretectum. A small retinal projection to the lateral habenular nucleus has not been described previously. Although these findings suggest striking differences in retinal projections among teleosts, this variation may relate to age differences since the previously studied teleosts were adults.     

Multigenerational outbreeding effects in Chinook salmon (Oncorhynchus tshawytscha)

Outbreeding, mating between genetically divergent individuals, may result in negative fitness consequences for offspring via outbreeding depression. Outbreeding effects are of notable concern in salmonid research as outbreeding can have major implications for salmon aquaculture and conservation management. We therefore quantified outbreeding effects in two generations (F₁ hybrids and F₂ backcrossed hybrids) of Chinook salmon (Oncorhynchus tshawytscha) derived from captively-reared purebred lines that had been selectively bred for differential performance based on disease resistance and growth rate. Parental lines were crossed in 2009 to create purebred and reciprocal hybrid crosses (n = 53 families), and in 2010 parental and hybrid crosses were crossed to create purebred and backcrossed hybrid crosses (n = 66 families). Although we found significant genetic divergence between the parental lines (F_ST = 0.130), reciprocal F₁ hybrids showed no evidence of outbreeding depression (hybrid breakdown) or favorable heterosis for weight, length, condition or survival. The F₂ backcrossed hybrids showed no outbreeding depression for a suite of fitness related traits measured from egg to sexually mature adult life stages. Our study contributes to the current knowledge of outbreeding effects in salmonids and supports the need for more research to better comprehend the mechanisms driving outbreeding depression.     

Assignment of sockeye salmon (Oncorhynchus nerka) to spawning sites using DNA markers

Randomly amplified polymorphic DNA (RAPD) markers were used to assign individual adult sockeye salmon to their spawning sites using a genotype assignment test. Six primers were selected for use by screening bulked DNA samples for markers missing in fish from one or more of 5 sites in British Columbia or Alaska. Of 73 markers scored, 54 showed variation between or within sites among the sampled fish. Thirty-seven of the variable markers were not detected in any fish from one or more sites; 18 variable markers were detected in all fish from one or more other sites. Thus 25% of markers scored were found in all fish of some sites and in no fish of some other sites. An assignment test placed all 70 fish tested into their correct populations. Principal coordinate analysis of genetic variation produced clusters of fish corresponding to each sampling site. No sex-specific RAPD markers were detected among more than 1300 screened.     

Compensatory swimming in the kokanee and sockeye salmon Oncorhynchus nerka (Walbaum)

When negatively buoyant, such as by increased pressure or loss of swimbladder gas, kokanee and sockeye salmon ( Oncorhynchus nerka ) attempt to swim upward by increased use of the pectoral fins. This response is termed compensatory swimming. Prior to initial filling of the swimbladder, sockeye fry showed no behavioural response to pressures above atmospheric. Following air-gulping at the surface and bladder inflation, kokanee and sockeye fry responded to increased pressure by assuming a more vertical position and by beating the pectoral fins more rapidly. In young sockeye this response occurred over the pressure range of atmospheric to 20 lb/in², and the effect of this behaviour would be to distribute these fish in the upper 14 m of the lacustrine environment. Fingerling kokanee showed a more gradual increase in compensatory swimming over the range of pressure equivalent to depths of 0–50 m. The behaviour of yearling kokanee would tend to concentrate these fish in the upper 30 m. Sockeye older than 1 year responded to negative buoyancy with increased horizontal swimming whilst planing upward on the pectoral fins. Depth distribution postulated on the basis of pressure-induced compensatory swimming is consistent with the known vertical distribution of kokanee and sockeye salmon.     

Morphological changes in senescing adult male sockeye salmon (Oncorhynchus nerka Walbaum)

Mature male sockeye salmon ( Oncorhynchus nerka Walbaum) were measured, tagged and released into a gravel holding pond adjacent to the spawning site, then remeasured when dead for possible changes in shape or size. Body length, depth, snout length and caudal peduncle thickness decreased at the end of life, 1–2 weeks later.     

Microsatellite DNA variation in sockeye salmon Oncorhynchus nerka (Walbaum, 1792) populations of eastern Kamchatka

The allelic variations of eight microsatellite loci (Ots107, Oki1a, Oki1b, One104, One109, OtsG68, OtsG85, and Oki6) were examined in sockeye salmon Oncorhynchus nerka (Walbaum) populations of eastern Kamchatka. Samples from river basins of the Karaginskiy and Olyutorskiy districts in the northern part of the investigated region are characterized by a smaller number of allelic variants and lower heterozygosity, compared to samples from the Kamchatka River basin. A fit to the Hardy-Weinberg distribution was found at most of the loci that were studied in all samples. The average differentiation between populations (st) over the eight loci was 2.93%, which corresponds to the statistical significance level. The most highly polymorphic loci proved to be the least differentiating between the analyzed samples.     

Population differences in Chinook salmon (Oncorhynchus tshawytscha) DNA methylation: Genetic drift and environmental factors

Local adaptation and phenotypic differences among populations have been reported in many species, though most studies focus on either neutral or adaptive genetic differentiation. With the discovery of DNA methylation, questions have arisen about its contribution to individual variation in and among natural populations. Previous studies have identified differences in methylation among populations of organisms, although most to date have been in plants and model animal species. Here we obtained eyed eggs from eight populations of Chinook salmon (Oncorhynchus tshawytscha) and assayed DNA methylation at 23 genes involved in development, immune function, stress response, and metabolism using a gene‐targeted PCR‐based assay for next‐generation sequencing. Evidence for population differences in methylation was found at eight out of 23 gene loci after controlling for developmental timing in each individual. However, we found no correlation between freshwater environmental parameters and methylation variation among populations at those eight genes. A weak correlation was identified between pairwise DNA methylation dissimilarity among populations and pairwise F _ST based on 15 microsatellite loci, indicating weak effects of genetic drift or geographic distance on methylation. The weak correlation was primarily driven by two genes, GTIIBS and Nkef. However, single‐gene Mantel tests comparing methylation and pairwise F _ST were not significant after Bonferroni correction. Thus, population differences in DNA methylation are more likely related to unmeasured oceanic environmental conditions, local adaptation, and/or genetic drift. DNA methylation is an additional mechanism that contributes to among population variation, with potential influences on organism phenotype, adaptive potential, and population resilience.     

Observations on saprolegniasis of adult sockeye salmon, Oncorhynchus nerka (Walbaum)

Some observations on the mycology and pathology of saprolegniasis of maturing sockeye salmon are reported. Saprolegnia spp. were isolated from all lesions examined. Some isolates did not produce oogonia and could not be identified further. The other isolates have affinities to the S. diclina-S. parasitica complex. Observations on the gross and histo-pathology of the lesions show the fungi can be active pathogens. It is hypothesized that increased levels of plasma corticosteroids and depletion of ascorbic acid reserves in the fish increase the probability that Saprolegnia spp. will initiate infections, either alone or concurrently with other opportunistic parasites.     

Seasonal and interannual variations of sockeye salmon (Oncorhynchus nerka) microsatellite DNA in two Kamchatka lake-river systems

Seasonal and interannual variations in the sockeye salmon populations from two lake-river systems of the East and West Kamchatka were studied. Stability of allele and genotypic frequencies of six microsatellite DNA loci in the adjacent generations and spawning populations of the sockeye salmon of the Bol'shaya River was confirmed experimentally. The pairwise intersample differentiation (F(st)) of the local sockeye salmon populations from the southwestern Kamchatka coast (Ozernaya and Bol'shaya Rivers)was almost 7 times higher than the corresponding values for the spawning populations of the Bol'shaya River sockeye salmon of the adjacent years; 15 times, for the adjacent Bol'shaya River sockeye salmon generations; and four times, for the seasonal races within the Kamchatka River.     

Effects of rearing temperature on immune functions in sockeye salmon (Oncorhynchus nerka)

To determine if the defences of sockeye salmon (Oncorhynchus nerka) raised in captivity are affected by the rearing temperature or their life-cycle stage, various indices of the humoral and cellular immune functions were measured in fish reared at either 8 or 12 degrees C for their entire life-cycle. Measures of humoral immunity included the commonly used haematological parameters, as well as measurements of complement, and lysozyme activity. Cellular assays quantified the ability of macrophages from the anterior kidney to phagocytise Staphylococcus aureus cells, or the activities of certain bactericidal systems of those cells. The T-dependent antibody response to a recombinant 57 kDa protein of Renibacterium salmoninarum was used to quantify the specific immune response. Fish were sampled during the spring and fall of their second, third and fourth years, corresponding to a period that began just before smolting and ended at sexual maturation. Fish reared at 8 degrees C tended to have a greater percentage of phagocytic kidney macrophages during the first 2 years of sampling than the fish reared at 12 degrees C. During the last half of the study the complement activity of the fish reared at 8 degrees C was greater than that of the 12 degrees C fish. Conversely, a greater proportion of the blood leucocytes were lymphocytes in fish reared at 12 degrees C compared to the fish reared at 8 degrees C. Fish reared at 12 degrees C also produced a greater antibody response than those reared at 8 degrees C. Results suggested that the immune apparatus of sockeye salmon reared at 8 degrees C relied more heavily on the non-specific immune response, while the specific immune response was used to a greater extent when the fish were reared at 12 degrees C. Although a seasonal effect was not detected in any of the indices measured, varying effects were observed in some measurements during sexual maturation of fish in both temperature groups. At that time there were dramatic decreases in complement activity and lymphocyte numbers. This study was unique in its scope because it was the first quantitative assessment of salmon immune functions for an entire life-cycle.     

Juvenile habitat partitioning and relative productivity in allochronically isolated sockeye salmon (Oncorhynchus nerka)

Allochronic divergence, like spatial isolation, may contribute to population diversity and adaptation, however the challenges for tracking habitat utilization in shared environments are far greater. Adult Klukshu River (Yukon, Canada) sockeye salmon, Oncorhynchus nerka, return as genetically distinct "early" and "late" runs. Early and late adult spawning populations (1999 and 2000) and their subsequent fry (sampled at 7 sites in 2000 and at 8 sites in 2001 throughout Klukshu Lake and River) were genotyped at eight microsatellite loci. Bayesian assignment was used to determine the spatial distribution of early versus late fry; although intermixed, the distribution of fry significantly differed in Klukshu Lake and in the Klukshu River in 2001, based on crosstab analyses. Late-run fry predominated in Klukshu Lake at all sites, while early-run fry were most common in the north and south of Klukshu Lake and in Klukshu River. Early-run spawners had significantly higher relative productivity (early life survival) than late-run fish (2.9 times more fry produced per early-run adult in 2000, and 9.2 times more in 2001). This study demonstrates spatial habitat partitioning and differences in the contribution of allochronically isolated populations to fry abundance, and highlights annual variability that likely contributes to recruitment variation.     

Some aspects of pathogenesis of infectious hematopoietic necrosis (IHN)

The histopathogenesis of infectious haematopoietic necrosis (IHN) virus infection was studied by exposing juvenile sockeye salmon ( Oncorhynchus nerka ) to the IHN virus. Fish samples were taken every 24 h for histological examination and for determination of virus concentration. A close correlation was found between histopathological changes and virus concentration. The most significant changes occurred 4 days after exposure. The haematopocitic tissue of the kidney was the most extensively involved but minor degenerative changes were seen in the liver, pancreas, and in the granular cells of the digestive tract. On the 4th day, maximum tissue concentration of virus was reached and the mortality increased. By the 5th day, 90% of the samples showed extensive pathological changes in the kidney, together with variable changes in spleen, liver, pancreas, and gut. Similarities in the histopathogenesis of IHN, Oregon sockeye disease (OSD), Sacramento River chinook disease (SRCD) and viral haemorrhagic septicaemia (VHS), are discussed.     

The invasion of Patagonia by Chinook salmon (Oncorhynchus tshawytscha): inferences from mitochondrial DNA patterns

The Chinook salmon Oncorhynchus tshawytscha, which was introduced deliberately in Chile four decades ago for sport fishing and aquaculture, represents a rare example of a successful translocation of an anadromous Pacific salmon into the southern Hemisphere, offering a unique opportunity to examine the role of introduction history and genetic variability in invasion success. We used historical information and mitochondrial displacement loop sequences (D-loop) from seven colonized sites in Chile and Argentina and from native and naturalized Chinook salmon populations to determine population sources and to examine levels of genetic diversity associated with the invasion. The analysis revealed that the Chinook salmon invasion in Patagonia originated from multiple population sources from northwestern North America and New Zealand, and admixed in the invaded range generating genetically diverse populations. Genetic analyses further indicated that the colonization of new populations ahead of the invasion front appear to have occurred by noncontiguous dispersal. Dispersal patterns coincided with ocean circulation patterns dominated by the West Wind Drift and the Cape Horn Currents. We conclude that admixture following multiple introductions, as well as long-distance dispersal events may have facilitated the successful invasion and rapid dispersal of Chinook salmon into Patagonia.     

Environmental adaptation in Chinook salmon (Oncorhynchus tshawytscha) throughout their North American range

Landscape genomics is a rapidly growing field with recent advances in both genotyping efficiency and statistical analyses that provide insight towards local adaptation of populations under varying environmental and selective pressure. Chinook salmon (Oncorhynchus tshawytscha) are a broadly distributed Pacific salmon species, occupying a diversity of habitats throughout the northeastern Pacific with pronounced variation in environmental and climate features but little is understood regarding local adaptation in this species. We used a multivariate method, redundancy analysis (RDA), to identify polygenic correlations between 19 703 SNP loci and a suite of environmental variables in 46 collections of Chinook salmon (1956 total individuals) distributed throughout much of its North American range. Models in RDA were conducted on both rangewide and regional scales by hierarchical partitioning of the populations into three distinct genetic lineages. Our results indicate that between 5.8 and 21.8% of genomic variation can be accounted for by environmental features, and 566 putatively adaptive loci were identified as targets of environmental adaptation. The most influential drivers of adaptive divergence included precipitation in the driest quarter of the year (Rangewide and North Coastal Lineage, anova P = 0.002 and 0.01, respectively), precipitation in the wettest quarter of the year (Interior Columbia River Stream‐Type Lineage, anova P = 0.03), variation in mean diurnal range in temperature (South Coastal Lineage, anova P = 0.005), and migration distance (Rangewide, anova P = 0.001). Our results indicate that environmental features are strong drivers of adaptive genomic divergence in this species, and provide a foundation to investigate how Chinook salmon might respond to global environmental change.