Proteomic analysis of inviable salmonid hybrids between female masu salmon Oncorhynchus masou masou and male rainbow trout Oncorhynchus mykiss during early embryogenesis

Early embryos of inviable hybrids between female masu salmon Oncorhynchus masou masou and male rainbow trout Oncorhynchus mykiss at 9, 12, 15 and 20 days after fertilization were examined for protein expression profiles. A total of 44 proteins, mostly down-regulated products of house-keeping genes and those involved in nucleic acid metabolism or chromatin replication, were identified in hybrid embryos by mass spectrometry analysis and protein database searching. The identified down-regulated proteins may be responsible for the inviability in the hybrids.     

Species-specific releaser effect of urine from ovulated female masu salmon and rainbow trout

MT (17α-methyltestosterone)-treated immature masu salmon Oncorhychus masou and rainbow trout Oncorhynchus mykiss , exposed to the urine of ovulated females in their non-reproductive season, made significantly more entries into the channel, of a Y-maze preference trough, conditioned by the urine of conspecific females than by heterospecifics. Immature, non-treated controls in both species showed no preference for the urine of ovulated females. These results suggest that, for both species, the releaser pheromones that attract males in the urine are species-specific. Therefore, utilization of species-specific chemical signals may be one of the mechanisms of the reproductive isolation in Oncorhynchus species.     

Differential dispersal patterns of male and female masu salmon fry

Behavioural experiments using artificial and natural stream channels were undertaken to determine whether there were differences in dispersal between newly emerged male and female masu salmon. Eyed eggs from a cross with wild spawners were planted in the middle pool of an artificial channel. After emergence, more males than females moved into an upstream trap, while fewer males moved downstream. In a natural stream, eyed eggs were marked with alizarin complexone to distinguish them from naturally spawned eggs and these were planted into artificial redds. More newly emerged male fry remained at the planted site than female fry. In contrast, more females moved downstream than males. These results imply that differences in dispersal patterns between male and female masu salmon fry are genetically controlled.     

Recombinant genotypes in backcrosses of male Atlantic salmon × brown trout hybrids to female Atlantic salmon

When male hybrids of Atlantic salmon × brown trout were backcrossed to female Atlantic salmon, approximately 1% of diploid progeny hatched. These were shown to exhibit recombinant genotypes when examined electrophoreticalty at five enzyme loci. This is the first confirmation of genie recombination in backcrosses of these species. Triploidization greatly increases the proportion of backcross progeny which hatch.     

DNA homologies between the rainbow trout, chum salmon and coho salmon

The thermal stabilities of hybrid duplexes between the DNAs from three salmonid fish species were monitored as measures of DNA homology. The chum salmon, Oncorhynchus keta, and coho salmon, Oncorhynchus kisutch, had more DNA homology with each other than either had with the rainbow trout, Salmo gairdnerii. Morphological, ecological and protein similarities between the coho salmon and the rainbow trout may be due to parallel or convergent evolution.     

Urine of ovulated female masu salmon attracts immature male parr treated with methyltestosterone

Methyltestosterone-treated immature male masu salmon Oncorhynchus masou parr were attracted to both the urine of ovulated females and the ether soluble basic substances extracted from the urine, but not to immature female urine. It is suggested that the male response to the sex attractant (releaser pheromone) in the urine is under the control of androgens.     

Contrasting seasonal modulation of the stress response in male and female rainbow trout

Between June and September the magnitude of the plasma cortisol response of maturing male and female rainbow trout to confinement was indistinguishable. A progressive increase in confinement-induced cortisol levels in both sexes occurred during May to September, associated with the seasonal rise in water temperature. Between September and January a reduction of >50% in the magnitude of the cortisol response to confinement in male fish (but no decline in females) coincided with declining water temperature and significant increases in plasma 11-ketotestosterone and elevated plasma testosterone levels. Plasma oestradiol-17β levels were significantly greater in females than males throughout the study period and this difference was maximal between September and January. However, plasma testosterone was also elevated in females during this period and levels overall were higher than those in male fish. Previous studies have shown oestradiol-17β and testosterone to have diametrically opposed effects on stress responsiveness in trout, with the former enhancing, and the latter suppressing, the cortisol response to a stressor. The relative roles of androgens, estrogen and water temperature in modulating the stress responsiveness of rainbow trout are discussed.     

Absence of developmental incompatibility in hybrids between rainbow trout and two subspecies of cutthroat trout

We examined the developmental rate of hybrids between rainbow trout (Salmo gairdneri) and two subspecies of cutthroat trout: westslope cutthroat trout (Salmo clarki lewisi) and Yellowstone cutthroat trout (Salmo clarki bouvieri). These taxa show considerable genetic divergence at 42 structural loci encoding enzymes; the mean Nei's d between the rainbow trout and the two species of cutthroat trout is 0.22. We used four measures of developmental rate: time of hatching and yolk resorption, rate of increase in activity of four enzymes, and time of initial detection of seven isozyme loci. The two cutthroat trout subspecies reached hatching and yolk resorption earlier than rainbow trout. Cutthroat trout had higher relative enzyme activities than rainbow trout from deposition of eye pigment to hatching. There was no difference in the rate of increase in enzyme activity or time of initial expression of these loci between these species. Hybrids showed developmental rates intermediate or similar to that of the parental species using all measures. Our results indicate an absence of regulatory and developmental incompatibility between these taxa.This research was supported by NSF Grants ISP-8011449 and BSR-8300039. M.M.F. was supported by a postgraduate scholarship from the Natural Sciences and Engineering Research Council of Canada.     

The existence of natural hybrids between the European trout and the Atlantic salmon

Natural hybrids between Atlantic salmon ( Salmo salar L.) and European trout ( S. trutta L.) have been detected in a sample of 4431 fish which appeared to be Atlantic salmon. The methods of analysis for serum protein, serum esterase and serum lactic dehydrogenase are given. The possible occurrence of hybrids having the appearance of European trout and the effect of introgression on salmon stocks are discussed.     

The sockeye salmon neo-Y chromosome is a fusion between linkage groups orthologous to the coho Y chromosome and the long arm of rainbow trout chromosome 2

Unlike other Pacific salmon, sockeye salmon (Oncorhynchus nerka) have an X(1)X(2)Y sex chromosome system, with females having a diploid chromosome number of 2n = 58 and males 2n = 57 in all populations examined. To determine the origin of the sockeye Y chromosome, we mapped microsatellite loci from the rainbow trout (O. mykiss; OMY) genetic map, including those found on the Y chromosomes of related species, in kokanee (i.e. non-anadromous sockeye) crosses. Results showed that 3 microsatellite loci from the long arm of rainbow trout chromosome 8 (OMY8q), linked to SEX (the sex-determining locus) in coho salmon (O. kisutch), are also closely linked to SEX in the kokanee crosses. We also found that 3 microsatellite loci from OMY2q are linked to those markers from OMY8q and SEX in kokanee, with both linkage groups fused to form the neo-Y. These results were confirmed by physical mapping of BAC clones containing microsatellite loci from OMY8q and OMY2q to kokanee chromosomes using fluorescence in situ hybridization. The fusion of OMY2q to the ancestral Y may have resolved sexual conflict and, in turn, may have played a large role in the divergence of sockeye from a shared ancestor with coho.     

Carbohydrate energy reserves and effects of food deprivation in male and female rainbow trout

We investigated the effects of nutritional state on carbohydrate, lipid, and protein stores in the heart, liver, and white skeletal muscle of male and female rainbow trout. For fed animals we also partitioned glycogen into fractions based on acid solubility. Fish (10-14 months-old, ~400-500 g) were held at 14 °C and either fed (1% of body weight, every other day) or deprived of food for 14 days. Under fed conditions, glycogen was increased 54% in ventricles from males compared with females, and elevated in the liver (87%) and white muscle (70%) in sexually-maturing versus immature males. Acid soluble glycogen predominated over the acid insoluble fraction in all tissues and was similar between sexes. Food deprivation 1) selectively reduced glycogen and free glucose in male ventricles by ~30%, and 2) did not change glycogen in the liver or white muscle, or triglyceride, protein or water levels in any tissues for both sexes. These data highlight sex differences in teleost cardiac stores and the metabolism of carbohydrates, and contrast with mammals where cardiac glycogen increases during fasting and acid insoluble glycogen is a significant fraction. Increased glycogen in the hearts of male rainbow trout appears to pre-empt sex-specific cardiac growth while storage of acid soluble glycogen may reflect a novel strategy for efficient synthesis and mobilization of glycogen in fishes.     

Natural hybrids between Atlantic salmon, Salmo salar L., and trout, Salmo trutta L., in juvenile salmonid populations in south-west England

Electrophoretic analysis of juvenile salmonids from south-west England revealed the presence of natural hybrids between Atlantic salmon, Salmo salar L., and brown trout, Salmo rrurra L., within three of five rivers. Amongst 409 juvenile 'salmon', 1.2% were hybrids, and of 150 juvenile 'trout' sampled, 2.0% were natural hybrids. In one river, similar hybridization levels in 'salmon' parr samples were found in two successive years. Levels of hybridization are high when compared with most similar studies, especially others within the United Kingdom. Possible factors influencing this are discussed.     

Comparison of the frequency of the growth hormone pseudogene between juvenile and adult female masu salmon Oncorhynchus masou

The proportions of individual masu salmon Oncorhynchus masou whose genotypic and phenotypic sex differed were compared among juvenile and adult fish in three rivers, and genotypically male but phenotypically female individuals were observed 6–16% more often among adults than among juveniles. This suggests that during the transition from juvenile to adult, survival rates of genotypically male but phenotypically female individuals are higher than those of normal females. In contrast, genotypically female but phenotypically male individuals were only found in the juvenile period, which suggests that they exhibit a decreased survival rate in comparison with normal males.     

New subtype of salmonid alphavirus (SAV), Togaviridae, from Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss in Norway

In Europe, 2 closely related alphaviruses (Togaviridae) are regarded as the causative agents of sleeping disease (SD) and salmon pancreas disease (SPD): SD virus (SDV) has been isolated from rainbow trout Oncorhynchus mykiss in France and the UK, while SPD virus (SPDV) has been isolated from salmon Salmo salar in Ireland and the UK. Farmed salmonids in western Norway also suffer from a disease called pancreas disease (PD), and this disease is also believed to be caused by an alphavirus. However, this virus has not yet been characterised at the molecular level. We have cultured a Norwegian salmonid alphavirus from moribund fishes diagnosed with cardiac myopathy syndrome (CMS) and fishes diagnosed with PD. The virus has also been found in salmon suffering from haemorrhagic smolt syndrome in the fresh water phase. The genomic organisation of the Norwegian salmonid alphavirus is identical to that in SPDV and SDV, and the nucleotide sequence similarity to the other 2 alphaviruses is 91.6 and 92.9%, respectively. Based on the pathological changes, host species and the nucleotide sequence, we suggest naming this virus Norwegian salmonid alphavirus (NSAV). Together with SPDV and SDV it constitutes a third subtype of salmonid alphavirus (SAV) species within the genus Alphavirus, family Togaviridae.     

Interpopulation variation in reproductive traits of female masu salmon, Oncorhynchus masou

Several life history models predict that larger eggs and lower fecundity should be favored in a low-growth environment. We applied the model of Sibly et al. to seven Japanese populations of masu salmon ( Oncorhynchus masou ) in order to test the predictions that populations in which individual growth rate is low are characterized by larger eggs and lower fecundity. Two populations, Shumarinai and Shikaribetsu, had the lowest growth rates of individuals, largest egg sizes and lowest fecundities of all populations examined. In contrast, the Shiribetsu, Chitose and Uono populations had highest growth rates of individuals, smallest egg sizes and highest fecundities. The Shibetsu and Toya populations were intermediate between these two groups. The gonadsomatic index (GSI) of the Shumarinai and Shikaribetsu populations was smaller than that of all other populations. Correlation analysis indicated that populations with lower individual growth rates had larger eggs and lower fecundities. The results were consistent with the predictions of the Sibly et al. model: increased egg size which results in decreased fecundity is probably an adaptation to low-growth environments. Therefore, in masu salmon, growth differences among populations may explain interpopulation variation in egg size and fecundity.     

Responses of immature male masu salmon parr to the urine of mature males

Responses of male masu salmon Oncorhynchus masou parr to mature male urine were investigated in a Y-maze trough. Immature masu salmon made fewer entries into the channel conditioned by mature male urine than that with control water. This phenomenon may suggest that immature males avoid sexually active males in the spawning season.     

A releaser pheromone that attracts males in the urine of mature female masu salmon

A releaser pheromone of masu salmon Oncorhynchus masou was investigated using Y-maze behavioural experiments. During the reproductive season, urine of mature females contains a releaser pheromone which acts as a sex attractant for spermiating mature male parr. The releaser pheromone in mature female urine is one or more low molecular weight substances (less than 10 000) soluble in ether under basic conditions. The attractant was not present in either the coelomic fluid of ovulated females nor in neutral or acidic extracts of female urine which contain free steroids and F-type prostaglandins, respectively.