Characterization of coho salmon (Oncorhynchus kisutch) insulin

Insulin has been isolated from islet tissue of coho salmon (Oncorhynchus kisutch) by gel filtration and HPLC and the complete amino acid sequence has been determined. The sequence differs from bovine insulin at 14 sites but all interchanges are conservative from the viewpoint of preservation of conformation. A comparison of insulin sequences from other fish is presented. Salmon insulin cross-reacts very weakly with antiserum to bovine insulin and vice versa. A completely homologous radioimmunoassay has been developed and used to estimate the insulin in salmon islet tissue and in plasma. The hypoglycemic effect of salmon insulin in salmon was more pronounced and persisted longer than that caused by identical doses of bovine insulin.     

Glacial biogeography of North American coho salmon (Oncorhynchus kisutch)

To study the glacial biogeography of coho we examined 20 microsatellite loci and mitochondrial DNA D-loop sequence in samples from Alaska to California. Microsatellite data divided our samples among five biogeographic regions: (1) Alaska and northern coastal British Columbia; (2) the Queen Charlotte Islands; (3) the mainland coast of British Columbia and northern Washington State; (4) the Thompson River; and (5) Oregon and California. The D-loop sequence data suggested three geographical regions: (1) Oregon and California; (2) the Thompson River; and (3) all the other sites north of the southern ice margin. Microsatellite data revealed no difference in the number of alleles in different regions, but mitochondrial DNA data revealed a cline of decreasing diversity from south to north. We suggest that the two signals presented by these different marker types illuminate two time frames in the history of this species. Endemic microsatellite diversity in Alaska and on the Queen Charlotte Islands provides evidence in favour of Fraser Glaciation refugia in these regions. The loss of mitochondrial variation from south to north suggests that one of the earlier, more extensive, Pleistocene glaciations eliminated coho from its northern range.     

A genetic linkage map for coho salmon (Oncorhynchus kisutch)

Construction of genetic linkage maps is an important first step for a variety of genomic applications, such as selective breeding in aquaculture, comparative studies of chromosomal evolution and identification of loci that have played key roles in the evolution of a species. Here we present a sex-specific linkage map for coho salmon. The map was constructed using 148 AFLP markers, 133 microsatellite loci and the phenotypic locus SEX . Twenty-four linkage groups spanning 287.4 cM were mapped in males, and 33 linkage groups spanning 429.7 cM were mapped in females. Several male linkage groups corresponded to two female linkage groups. The combination of linkage groups across both sexes appeared to characterize regions of 26 chromosomes. Two homeologous chromosomes were identified based on information from duplicated loci. Homologies between the coho and rainbow trout maps were examined. Eighty-six loci were found to form common linkage relationships between the two maps; these relationships provided evidence for whole-arm fissions, fusions and conservation of chromosomal regions in the evolution of these two species.     

Ontogeny of the olfacto-retinalis projection in coho salmon (Oncorhynchus kisutch)

Filling of nucleus olfacto-retinalis neurons by cobaltous lysine injections into one eye of coho salmon of different ages revealed that the terminal nerve projection to the retina is established when the fish leave their freshwater environment. At this time salmons go through a metamorphosis termed "smoltification." FMRF-amide-like immunoreactive neurons in the nucleus olfacto-retinalis already exist in pre-smolts before the retino-petal projection is established. Thus, for the first time in any vertebrate, a projection of the terminal nerve is shown to develop during an advanced ontogenetic state.     

Characterization of Phase I biotransformation enzymes in coho salmon (Oncorhynchus kisutch)

Wild stocks of Pacific salmon in the Northwestern United States have declined in recent years, and the major factors contributing to these losses include water pollution and loss of habitat. In salmon, sublethal chemical exposures may impact critical behaviors (such as homing, feeding, predator-avoidance) that are important for species survival. Therefore, understanding the potential for these species to biotransform organic compounds within sensitive target tissues such as liver, gills and olfactory region can help estimate or predict their susceptibility to pollutants. In this study, we used real-time quantitative polymerase chain reaction (Q-PCR), Western blotting, and catalytic assays to characterize the expression of Phase I biotransformation enzymes in coho salmon (Oncorhynchus kisutch), a sensitive species in the Pacific Northwest. Gene expression analysis using Q-PCR assays developed for coho genes revealed the presence of the predominant cytochrome P450 mRNAs (CYP1A, CYP2K1, CYP2M1, CYP3A27) in the olfactory rosettes and provided quantitative mRNA expression levels in coho liver and gills. Q-PCR analysis revealed relatively high expression of the major CYP isoforms in the liver and olfactory rosettes, which was generally confirmed by Western blotting. Extrahepatic CYP expression was generally higher in the olfactory rosettes as compared to the gills. Catalytic studies demonstrated functional CYP1A-dependent ethoxyresorufin-O-deethylase, CYP2-dependent pentoxyresorufin-O-dealkylase, CYP2K1-dependent testosterone 16beta-hydroxylase, and CYP3A27-dependent testosterone 6beta-hydroxylase activities in liver, but not at detectable levels in gills. In contrast, flavin-containing monooxygenase (FMO)-dependent thiourea S-oxidase activity was readily observed in the gills and was substantially higher than that observed in liver. Collectively, the results of this study suggest that the olfactory rosettes are important sites of extrahepatic biotransformation in coho salmon, and that tissue specific-differences in Phase I metabolism may lead to contrasting tissue-specific biotransformation capabilities in this species.     

Top-down and bottom-up control of life-history strategies in coho salmon (Oncorhynchus kisutch)

Sexual maturation profoundly affects population dynamics, but the degrees to which genetic, top-down, and bottom-up controls affect age at maturity are unclear. Salmonid fishes have plastic age at maturity, and we consider genetic and environmental effects on this trait by developing fitness functions for coho salmon (Oncorhynchus kisutch). The functions are based on size-specific survival and reproductive success, where reproductive success is the product of fecundity and ability to defend nests (females) or the product of sperm volume and ability to mate (males). We model genetic and bottom-up controls (e.g., food availability) with an environmentally explicit growth function and top-down control (predation mortality) with survival functions that consider both size-dependent and size-independent mortality. For females, we predict that early maturation rarely maximizes fitness, but males can maximize fitness by maturing early if they grow well in freshwater. We predict that early maturation is most affected by the bottom-up effects of resource distribution at sea, followed by bottom-up and genotypic effects in freshwater. Top-down processes are predicted to have strong effects on the likelihood of delayed maturation.     

Isolation and structures of coho salmon (Oncorhynchus kisutch) glucagon and glucagon-like peptide

Glucagon and glucagon-like peptide (GLP) containing 31 amino acids have been isolated from the principal islet of coho salmon (Oncorhynchus kisutch) by gel filtration of acid alcohol extracts followed by HPLC, and the complete amino acid sequence of both peptides has been determined. Salmon glucagon is a simple 29 residue peptide differing at 3 positions when compared to catfish glucagon and at 8 positions when compared to porcine glucagon. Salmon GLP differs at 6 positions when compared with the N-terminal 31 amino acids of the 34 amino acid catfish GLP. Both coho salmon glucagon and GLP cross-react weakly in our mammalian glucagon radioimmunoassay and therefore this technique could not be used to determine tissue content. Glucagon and GLP isolated amounted to 156 micrograms/g and 350 micrograms/g wet tissue, respectively.     

Gastric evacuation of single and multiple meals by piscivorous coho salmon,Oncorhynchus kisutch

Synopsis Gastric evacuation of multiple meals of recently emerged sockeye salmon, Oncorhynchus nerka, consumed by juvenile coho salmon, O. kisutch, were compared with that expected from a single meal evacuation rate model developed for the estimation of food consumption by coho in the field. Significant interaction occurred between meals consumed two hours apart (p<0.001). Evacuation of the first of two meals (29.9% of initial weight remaining) was significantly faster (p<0.05) than that of a single meal (36.1% of initial weight remaining) after four hours, whereas evacuation of the second meal (72.2% of initial weight remaining) was significantly slower (p<0.001) than that of single meals (50.6% of initial weight remaining) after two hours. The total weight of the multiple meal remaining after four (0.191 g), six (0.138 g) and eight (0.070 g) hours averaged within 6% of that predicted by the single meal evacuation model (0.186 g, 0.115 g and 0.073 g, respectively). These data suggest that the stomach evacuation model based on single meals is adequate for estimating the evacuation of prey consumed by continuously feeding coho salmon.     

Changes in intestinal fluid transport associated with smoltification and seawater adaptation in coho salmon, Oncorhynchus kisutch (Walbaum)

The intestinal role in osmoregulatory preadaptation of freshwater-adapted (FW) yearling coho salmon during smoltification was examined. Measurements of intestinal net fluid absorption (J_V) using an in vitro sac preparation showed that J_V of FW parr was significantly less than that of either FW or seawater-adapted (SW) smolts. From two to four weeks following the springtime thyroxin surge, J_V increases in FW coho to a level comparable with that observed for SW smolts. J_V remained elevated in FW smolts throughout most of the summer and then decreased in the autumn. Two months after the thyroxin peak, intestinal wet weight per unit serosal surface area increases. Coho stunts resulting from premature transfer into sea water had significantly higher J_V than that seen in SW smolts. The elevated J_V of stunts is only partially explicable on the basis of differences in body weight between stunts and smolts.
Our observations indicate that an increase in intestinal fluid absorption is a preadaptive change associated with smoltification occurring in concert with alterations in renal and branchial osmoregulatory mechanisms. The timing of the increase in J_V suggests a phase relationship to the thyroxin surge. This temporal relationship may help to explain the efficacy of using changes in plasma thyroxin levels to predict hatchery-fish release dates that may optimize eventual seawater survival.     

Changes in the lipid class and proximate compositions of coho salmon (Oncorhynchus kisutch) smolts infected with the nematode parasite Philonema agubernaculum

Coho salmon (Oncorhynchus kisutch) smolts infected with the nematode Philonema agubernaculum had 36% lower mean lipid content (1.4%) than nonparasitized coho salmon (2.2%) harvested simultaneously from the same outmigration. Lengths, weights, and condition factors, as well as protein and moisture content, did not differ significantly between the two groups. Lipid class compositions differed significantly between the parasitized and nonparasitized fish. None of the nematode-infected fish contained detectable triacylglycerols (TAG) or monoacylglycerols (MAG). In contrast, mean TAG and MAG contents of the nonparasitized fish totaled 5.5% of the extracted lipid. Infected smolts had lower cholesterol contents than did uninfected coho (17% for infected, 33% for uninfected). Parasitized fish had significantly higher levels of free fatty acids (mean of 57% for parasitized vs. 35% for nonparasitized) as well as the phospholipids phosphatidylethanolamine (PE) and phosphatidylcholine (PC). However, the PC/PE ratios for infected and noninfected coho did not differ significantly (2.2 for infected vs. 2.0 for uninfected). These differences suggest that the parasitic nematodes are either harvesting storage energy directly from the coho or are placing additional energetic demands on the fish to cope with the infection.     

The energetic equivalence of cover to juvenile coho salmon (Oncorhynchus kisutch): ideal free distribution theroy applied

Cover is often thought to be an important habitat characteristicfor juvenile stream salmonida. In addition to providing protectionfrom predators, cover may be associated with reduced food availability.Thus, an individual's use of cover is likely to reflect a trade-offbetween the conflicting demands of growth and survival. We measuredthe influence of cover on foraging-site selection in groupsof eight juvenile coho salmon (Oncorhynchus kisutch) by examiningtheir distribution across two stream channel patches, one providingaccess to cover but little food (the "poor" patch), the otherproviding more food but no cover (the "good" patch). Becausefish distributions in the absence of cover conformed to an idealfree distribution (IFD) for unequal competitors (i.e., the distributionof competitive abilities matched the distribution of food),we used IFD theory to quantify the energetic equivalence ofcover to the fish. In the presence of cover and a model avianpredator, use of the poor patch increased relative to the predictionsof the IFD model. Using this observed deviation from an IFD,we calculated how much extra food must be added to the goodpatch to return the distribution of fish to the previously observedIFD of unequal competitors. As predicted, adding this amountof food caused the fish to return to their previous distribution,demonstrating that IFD theory can be used to relate energy intakeand risk of predation in a common currency     

Gonadal morphology of normal and sex-reversed triploid and gynogenetic diploid coho salmon (Oncorhynchus kisutch)

In this study, the gonadal morphology of untreated and sex-reversed juvenile triploid and gynogenetic diploid coho salmon was compared with that of diploids. Testes of triploids were of the same size as those of diploids. Spermatogonia, however, were significantly bigger than those of diptoids in both diameter (P<0·001) and volume (P<0·01), suggesting that this characteristic can be a useful indicator of ploidy in the early stages of gonadal development. In females, induction of triploidy did not affect the lamellar structure of the ovaries but reduced their size considerably. Further, these ovaries had no oocytes. Treatment of triploids with oestrogen resulted in the feminization of genotypic males, which had ovaries similar to those found in tripioid females. However, gonads of triploid males partially sex-re versed into females were identified by their enlargement, the presence of remnants of the male vascular system, and by the appearance of ovarian lacunae and germinal and somatic cells typical of triploid females, Induction of gynogenesis resulted in 100% females, of which 34% had ovaries of reduced size with areas devoid of oocytes. However, and contrary to what has been found in cyprinids, no male germ cells were observed in these ovaries. This discrepancy may reflect differences, in the mechanisms of sex determination between salmonids and cyprinids. Treatment of gynogenetics with androgen increased the number of fish with abnormal ovaries but also resulted in the production of phenotypic-male gynogenetic diploids, of which 11% had testes indistinguishable from those of untreated control diploids.     

Observations on the inherent variability of measuring lysozyme activity in coho salmon (Oncorhynchus kisutch)

Lysozyme activity is a common measurement of innate immunity. It has also been used to investigate genetic variation and an animal's responses to factors such as stress, infections and variations in diet. This research demonstrates the inherent variation in lysozyme activity in unstimulated coho salmon (Oncorhynchus kisutch). The role of maternal contribution, early life stage development and fish mass are considered. Genetic variation within and between strains of coho was found to be significant at selected life stages. Our results indicate that strain differences in lysozyme activity are more accurately measured by comparing the genetic variation after the eyed stage, when maternal effects are reduced. A positive correlation between plasma/serum lysozyme activity and fish mass is reported here. In summary, this study shows the role of maternal, developmental stage and size in lysozyme activity in fish, and emphasizes the importance of considering such variables when measuring the variability of lysozyme activity in fish.     

Mass mortality in cultured coho salmon (Oncorhynchus kisutch) due to Saprolegnia parasitica coker.

Epizootics of saprolegniasis occurred in 20 to 60 g freshwater-cultured coho salmon (Oncorhynchus kisutch) in Miyagi Prefecture, Japan. Cotton-like mycelia occurred on the body surface of infected fish, especially around the head, the adipose fin and the caudal fin, and aseptate hyphae occurred in the lesions. The hyphae also penetrated into the muscle and blood vessels. The isolated fungus was identified by asexual morphological characteristics as Saprolegnia parasitica (syn. S. diclina Type 1), a known salmonid fish pathogen.     

Hepatic aryl hydrocarbon hydroxylase activities in coho salmon (Oncorhynchus kisutch) exposed to petroleum hydrocarbons

Coho salmon exposed to the water soluble fraction (WSF) of Cook Inlet crude oil for a maximum of 30 days showed a greater than three-fold increase in hepatic aryl hydrocarbon hydroxylase activity. The initial increase in enzyme activity appeared between 2 and 5 days of exposure and increased as a function of increased exposure time. Persistence of the induced enzyme activity was dependent on the length and the concentration of WSF exposure. Handling stress had no effect on the AHH activity, but starvation caused a decrease in the activity.     

Transient serotonin-immunoreactive neurons coincide with a critical period of neural development in coho salmon (Oncorhynchus kisutch)

Summary In coho salmon (Oncorhynchus kisutch), smolt transformation has been shown to be associated with sequential surges of neurotransmitters in the brain. In order to determine if the surge of serotonin (5-HT) is correlated with structural changes, we have used immunocytochemistry to observe changes in the serotonin immunoreactivity before, during and after the 5-HT surge. The following stages were studied: 12-month-old freshwater presmolts, 17-month-old freshwater presmolts, 18-month-old saltwater smolts, 19-month-old saltwater postsmolt, 24-month-old postsmolt, and adult spawners. In the 17-month-old samples, but not at any other stage, we found a set of transient (serotonin-immunoreactive) 5-HT-immunoreactive neurons in the lateral preoptic area, as well as a discrete population of 5-HT-immunoreactive neurons in the lateral part of the dorsal right habenular nucleus. In addition, a higher density of serotonergic fibers was found in the telencephalon at this stage compared to the following two stages. Since the transient 5-HT-immunoreactive structures presented here do not appear simultaneously with the 5-HT total brain concentration surge, we conclude that they are unlikely to be the source of the 5-HT surge, but are probably related to other developmental changes in the brain associated with smolt transformation.