The production of gynogenetic Atlantic salmon,Salmo salar L.

The production of Atlantic salmon gynogenomes by the combined use of a novel method for sperm irradiation and differently timed high hydrostatic pressure shocks is described. Sperm solutions were exposed to UV irradiation in a temperature-controlled flow-through device. Eggs fertilised with such sperm were exposed to shocks of 9500 psi at 30 min or approximately 7 h after fertilisation in order to produce meiotic and mitotic gynogenomes respectively. Yields of meiotic gynogenomes were generally high (up to 95%); those of mitotic gynogenomes were lower (range 2–20%). Analyses of the offspring by ploidy status and fingerprinting confirmed their gynogenetic origin. Small numbers of mitotic gynogenetic fish were grown on for 2 years in fresh and salt water. S1/S2 ratios were lower in gynogenetic fish and mean age at maturity was greater. Of the presumptive gynogenetic fish subjected to destructive sampling (n = 87) all were female.     

Major histocompatibility polymorphism associated with resistance towards amoebic gill disease in Atlantic salmon (Salmo salar L.)

The association between major histocompatibility (MH) polymorphism and the severity of infection by amoebic gill disease (AGD) was investigated across 30 full sibling families of Atlantic salmon. Individuals were challenged with AGD for 19days and then their severity of infection scored by histopathological examination of the gills. Fish were then genotyped for the MH class I (Sasa-UBA) and MH class II alpha (Sasa-DAA) genes using polymorphic repeats embedded within the 3' untranslated regions of the Sasa-UBA and Sasa-DAA genes. High variation in the severity of infection was observed across the sample material, ranging from 0% to 85% gill filaments infected. In total, seven Sasa-DAA-3UTR and ten Sasa-UBA-3UTR marker alleles were identified across the 30 families. A significant association between the marker allele Sasa-DAA-3UTR 239 and a reduction in AGD severity was detected. There was also a significant association found between AGD severity and the presence of two Sasa-DAA-3UTR genotypes. While the associations between MH allele/genotypes and AGD severity reported herein may be statistically significant, the small sample sizes observed for some alleles and genotypes means these associations should be considered as suggestive and future research is required to verify their biological significance.     

Post-thawed motility and fertility from Atlantic salmon (Salmo salar L.) sperm frozen with four cryodiluents in straws or pellets

The cryopreservation of salmonid sperm is a complex process involving the interplay of many factors. Although cryopreservation protocols can be evaluated through a range of responses at various stages in the process, the number of progeny is the ultimate indicator of success. We compared reproductive success from freezing Atlantic salmon (Salmo salar L.) sperm using the eight combinations of (1) the penetrating cryoprotectants, 10% dimethyl sulfoxide (DMSO) or methanol (MeOH); (2) the nonpenetrating cryoprotectants glucose (0.3 M) or sucrose (0.6 M), and freezing in 0.1 mL pellets or 0.25 mL straws. All cryodiluents were supplemented with 10% (v/v) of hen's egg yolk. Response variables were the percentage and degree of motility of thawed and activated sperm using computer assisted sperm analysis (CASA), and rates of eyed embryos, hatch and egg sac larvae. Growth rates of alevins were assessed to two months post hatch. Atlantic salmon milt cryopreserved in straws had higher spermatozoa motility and fertilization success than milt cryopreserved in pellets (P < 0.05). Type of sugar tested did not significantly affect the response variables. In the MeOH treatment, thawed spermatozoa achieved higher speed and a higher fertilization rate evaluated at the eyed embryo stage than spermatozoa subjected to the DMSO treatment. Higher mortality rate (especially before hatching) of MeOH offspring than DMSO offspring led to equal numbers of progeny for the two treatments from the swimming stage to the end of the study. Moreover, during feeding fish from the MeOH group produced significantly lower weight larvae than the DMSO and control groups. Even so, the weight of the MeOH group was satisfactory. Length and the condition factors did not differ significantly among the larvae groups. Significant positive correlations were found between fertilization success (measured in number of eyed eggs) and both motility (rs = 0.81), and velocity (rs = 0.49). Freezing in straws gave betters results than freezing in pellets for cryopreservation of salmon milt; whereas type of sugar tested (glucose vs sucrose) did not have significant effects. Penetrating cryoprotectants DMSO and MeOH differed in their effect on post-thawed sperm velocity, fertilization rate and mortality rate of progeny, suggesting the need for further research on the influence of these cryoprotectants on frozen sperm and and post-fertilization devopmental processes.     

First access to territorial space and exposure to strong predation pressure: A conflict in early emerging Atlantic salmon (Salmo salar L.) fry

Summary This work aimed to elucidate conflicting factors that may explain the narrow and synchronous emergence in salmonids. Fry are highly vulnerable to predation and stand a better chance of surviving if they emerge synchronously. On the other hand, fry that leave the gravel first should increase their chance of obtaining one of the limited number of feeding territories. The risk involved in early emergence for Atlantic salmon fry was evaluated by exposing them to predatorySalmo trutta. Yolk sac alevins were incubated in an artificial redd in order to catch them by their time of emergence. Early-, peak- and late-emerging fry were then successively marked and transferred to flume tanks in which the predators were either present from the start or not introduced until all fry had been added. When the predators were initially present, the predation pressure differed depending on the time of fry emergence, resulting in survival rates of 11.6, 44.9 and 51% in early-, peak- and late-emerging fry, respectively. By assuming that the predation rate of the three emergence groups was dependent both on time of emergence and fry density the survival rates were calculated to be 7.5, 41.1 and 53.5% in groups I, II and III, respectively. These figures corresponded well to the observed rates. When the predators were added after completed emergence the resulting survival rates were 56.9, 39.7 and 25.2% in early-, peak- and late-emerging fry, respectively. Thus, predation after complete emergence gave a survival probability that varied across the three emergence-date groups, despite being exposed to a predator during the same number of days. The presence of fish predators in combination with a limited territorial space seems to make both early and late emergence hazardous and favours synchronous swimming movements.     

Diel activity pattern of juvenile Atlantic salmon (Salmo salar) in early and late winter

Radiotelemetry was used to investigate the diel activity pattern of juvenile Atlantic salmon (Salmo salar) in early and late winter. Fish were active throughout the diel cycle. However, there was significantly less daytime than nighttime movement and movement declined significantly with increasing fork length. Maximizing winter growth rate, through an overall increase in foraging activity, may reduce the risk of starvation in smaller fish. The results of the present study provide evidence that the activity patterns of juvenile salmonids are quite complex and support the suggestion that individual variation in activity patterns are, at least, partially driven by body size.     

Spatial and temporal variation of genetic diversity and estimation of effective population sizes in Atlantic salmon (Salmo salar, L.) populations from Asturias (Northern Spain) using microsatellites

Rivers in Asturias (northern Spain) constitute the southern limit of the distribution of Atlantic salmon (Salmo salar L.) in Europe, a biological resource facing one of the more serious challenges for conservation today. In this work, eight microsatellite loci have been used to analyse samples collected in 1993 and 1999 from four Asturian rivers (Esva, Narcea, Sella, and Cares), obtaining information about the temporal and the spatial genetic variation in these populations and, in addition, estimations of their effective population sizes. The temporal analysis revealed a general decrease in all the estimated genetic variability parameters when samples from 1993 (mean A ₍₁₉₉₃₎ = 6.47, mean H _O(1993) = 0.472, mean H _E(1993) = 0.530) were compared with those obtained in 1999 (mean A ₍₁₉₉₉₎ = 6.16, mean H _O(1999) = 0.460, mean H _E(1999) = 0.490). This reduction was particularly notable for the case of the Esva river. Our results pointed to a pattern of spatial genetic differentiation inside the Asturian region (F _{ST (1993)} = 0.016 P < 0.01; F _{ST (1999)} = 0.023 P < 0.01). Using the standard Temporal Method we found estimates of N _e^{^} _(Esva) = 75.1 (33.2–267.2); N _e^{^} _(Cares) = 96.6 (40.0–507.5), N _e^{^} _(Sella) = 106.5 (39.1–9396.4) and N _e^{^} _(Narcea) = 113.9 (42.0–3693.3). The use of likelihood-based methods for the N _e^{^} estimations improved the results (smaller CIs) for the Esva and Cares rivers (N _e^{^} _(Esva) = 63.9 (32.3–165.3); N _e^{^} _(Cares) = 76.4 (38.8–202.0) using a Maximum likelihood approach) and suggested the presence of larger populations for the Sella and Narcea rivers (N _e^{^}≈200). These results showed that the Asturian Atlantic salmon populations (in particular Esva and Cares river populations) could be close to the conservation genetic borderline for avoiding inbreeding depression although we discuss some implications of the analysis of temporal genetic change in populations with overlapping generations.     

Osmoregulation in immature atlantic salmon (Salmo salar L.) following transfer from sea-water to fresh water

• 1.1. Immature Atlantic salmon post-smolts weighting approximately 150 g were transferred abruptly to fresh water (FW) after 5 months in sea water (SW).
• 2.2. Losses of ions and gain of body water are reversed after 3 days with about 10–12 days taken for complete FW adaptation.
• 3.3. Immediately on transfer from SW to FW, immature salmon take up sodium at 45 μmol/kg/hr, about one-third the rate observed in maturing salmon on their spawning migration.
• 4.4. The sodium uptake rate increases to that of maturing salmon after 2 days in freshwater. Differences in the osmoregulatory ability of immature and maturing salmon are discussed.

     

Intestinal water transport in juvenile Atlantic salmon (Salmo salar L.) during smolting and following transfer to seawater.

1. The rate of mucosal to serosal water movement was measured in vitro in non-everted midgut segments in Atlantic salmon (Salmo salar L.) during parr-smolt transformation (February-July) and following transfer of smolts to seawater in May. 2. The rate increased significantly during smolting from 5.61 microliters/cm2/hr in the parr (February) to 11.03 microliters/cm2/hr in smolts in May. 3. Measured at intervals over a period of 20 days in seawater, the rate of water transport was not significantly different from that found in the freshwater-adapted smolts (11.20 microliters/cm2/hr). 4. Intestinal water transport is sodium-linked and inhibited by ouabain but is not stimulated by cortisol.     

Initial mortality of radio-tagged Atlantic salmon (Salmo salar L.) smolts following release downstream of a hydropower station

The fate of radio-tagged hatchery-reared salmon smolts (Salmo salar L.) was investigated in the tailrace of a hydropower station in the Danish River Gudenaa during 2 years. Seventeen and 27 smolts were tagged and released in late May 1996 and during April 1999, respectively. Out of the total of 44 smolts, only two were recorded to leave the river and enter the estuary. In both years of study, electrofishing was used to sample tagged smolts. In 1996, these attempts were unsuccessful, while in 1999, 19 (70% of total) transmitters were retrieved in the stomachs of pikeperch (Stizostedion lucioperca (L.)) and pike (Esox lucius L.), and in grey heron (Ardea cinerea L.) nests. The present study demonstrates that the physical condition of the watershed, with respect to spawning and rearing, may not be the only crucial determining factors of the success of reintroduction of Atlantic salmon. Hydropower plants not only block the free movement of migratory fish, hence preventing the utilisation of spawning and rearing areas, but also form the basis of areas with increased mortality.     

Genetic variation in haemolytic activity in Atlantic salmon (Salmo salar L.)

Spontaneous and antibody-dependent haemolytic activity against sheep red blood cells (SRBC) has been studied in serum from Atlantic salmon, Salmo salar L. Considerable increase in haemolytic activity was detected when SRBC were sensitized with haemolysin produced in Atlantic salmon. Haemolytic activity was sensitive both to divalent cations and to heat treatment. Significant reduction in haemolytic activity was detected after absorption with SRBC, indicating the presence of natural antibody against SRBC in Atlantic salmon serum. Persistent haemolytic activity of unsensitized SRBC in serum absorbed to remove natural antibodies was found, which suggests the activation of the alternative complement pathway, while the observed increased haemolytic activity in the presence of specific antibody against SRBC suggests activation of the classical complement pathway. Spontaneous and antibody-dependent haemolytic activity were both analysed in absorbed and non-absorbed sera in a family material of Atlantic salmon. The material consisted of 574 fish belonging to 57 fullsib groups within 20 paternal halfsib groups. Fish with signs of sexual maturity generally showed reduced haemolytic activity. Statistically significant effect of sire on the spontaneous haemolytic activity of both absorbed and nonabsorbed serum, and on antibody-dependent activity, provides evidence of significant additive genetic variation in both the alternative and the classical complement activation in Atlantic salmon. Neither on a phenotypical nor on a family basis were the two traits statistically correlated. The estimated heritabilities were 0.2–0.3 with a standard error of approximately 0.1.     

Mate choice for major histocompatibility complex genetic divergence as a bet-hedging strategy in the Atlantic salmon (Salmo salar)

Major histocompatibility complex (MHC)-dependent mating preferences have been observed across vertebrate taxa and these preferences are expected to promote offspring disease resistance and ultimately, viability. However, little empirical evidence linking MHC-dependent mate choice and fitness is available, particularly in wild populations. Here, we explore the adaptive potential of previously observed patterns of MHC-dependent mate choice in a wild population of Atlantic salmon (Salmo salar) in Québec, Canada, by examining the relationship between MHC genetic variation and adult reproductive success and offspring survival over 3 years of study. While Atlantic salmon choose their mates in order to increase MHC diversity in offspring, adult reproductive success was in fact maximized between pairs exhibiting an intermediate level of MHC dissimilarity. Moreover, patterns of offspring survival between years 0+ and 1+, and 1+ and 2+ and population genetic structure at the MHC locus relative to microsatellite loci indicate that strong temporal variation in selection is likely to be operating on the MHC. We interpret MHC-dependent mate choice for diversity as a likely bet-hedging strategy that maximizes parental fitness in the face of temporally variable and unpredictable natural selection pressures.     

Effect of dimethylglycine and trimethylglycine (Betaine) on the response of Atlantic salmon (Salmo salar L.) smolts to experimental Vibrio anguillarum infection

Atlantic salmon (Salmo salar L.) were fed on a control diet or experimental diets containing betaine (15 mg g^-1) or dimethylglycine (DMG, I mg g^-1 or 5 mg g^-1). After 10 weeks of feeding, resistance to infection was assessed following inoculation with Vibrio anguillarum. Total blood and differential leucocyte counts were made, and plasma lysozyme and ceruloplasmin were assayed as non-specific humoral factors. The mortality during the bacterial exposure was of the same magnitude in all feeding groups. Betaine or DMG had no effect on the 'basal' levels of plasma total protein, lysozyme or ceruloplasmin, but 3 days postinjection the lysozyme and ceruloplasmin levels were higher in the control group compared with the experimental groups. In both DMG groups, the lymphocyte response took place 1-2 weeks earlier than in the control or betaine supplemented group indicating that DMG has an immunomodulating effect on salmon.     

Effects of salmon lice infection and salmon lice protection on fjord migrating Atlantic salmon and brown trout post-smolts

Effects of artificial salmon lice infection and pharmaceutical salmon lice prophylaxis on survival and rate of progression of Atlantic salmon (n = 72) and brown trout post-smolts (n = 72) during their fjord migration, were studied by telemetry. The infected groups were artificially exposed to infective salmon lice larvae in the laboratory immediately before release in the inner part of the fjord to simulate a naturally high infection pressure. Groups of infected Atlantic salmon (n = 20) and brown trout (n = 12) were also retained in the hatchery to control the infection intensity and lice development during the study period. Neither salmon lice infection nor pharmaceutical prophylaxis had any effects on survival and rate of progression of fjord migrating Atlantic salmon post-smolts compared to control fish. Atlantic salmon spent on average only 151.2 h (maximum 207.3 h) in passing the 80 km fjord system and had, thus, entered the ocean when the more pathogenic pre-adult and adult lice stages developed. The brown trout, in comparison to Atlantic salmon, remained to a larger extent than Atlantic salmon in the inner part of the fjord system. No effect of salmon lice infection, or protection, was found in brown trout during the first weeks of their fjord migration. Brown trout will, to a larger extent than Atlantic salmon, stay in the fjord areas when salmon lice infections reach the more pathogenic pre-adult and adult stages. In contrast to Atlantic salmon, they will thereby possess the practical capability of returning to freshwater when encountering severe salmon lice attacks.     

大西洋鲑杀鲑气单胞菌无色亚种的分离鉴定和致病性研究

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Colocalization of neuropeptide Y (NPY)-like and FMRFamide-like immunoreactivities in the brain of the Atlantic salmon (Salmo salar)

Summary The colocalization of the peptides neuropeptide Y (NPY) and Phe-Met-Arg-Phe-NH₂ (FMRFamide) in the brain of the Atlantic salmon was investigated with double immunofluorescence labeling and peroxidase-antiperoxidase immunocytochemical techniques. Colocalization of NPY-like and FMRE amide-like immunoreactivities was observed in neuronal cell bodies and fibers in four brain regions: in the lateral and commissural nuclei of the area ventralis telencephali, in the nucleus ventromedialis thalami, in the laminar nucleus of the mesencephalic tegmentum, and in a group of small neurons situated among the large catecholaminergic neurons in the isthmal region of the brainstem. All cell bodies in these nuclei were immunoreactive to both NPY and FMRF. We consistently observed larger numbers of FMRF-immunoreactive than NPY-immunoreactive fibers. In the nucleus ventromedialis thalami NPY- and FMRFamide-like immunoreactivities were colocalized in cerebrospinal fluid (CSF)-contacting neurons. NPY-immunoreactive, but not FMRF-immunoreactive, neurons were found in the stratum periventriculare of the optic tectum, and at the ventral border of the nucleus habenularis (adjacent to the nucleus dorsolateralis thalami). Neurons belonging to the nucleus of the nervus terminalis were FMRF-immunoreactive but not NPY-immunoreactive. The differential labeling indicates, as do our cross-absorption experiments, that the NPY and FMRFamide antisera recognize different epitopes. Thus, it is probable that NPY-like and FMRFamide-like substances occur in the same neurons in some brain regions.     

The effects of handling on flounder (Platichthys flesus L.) and Atlantic salmon (Salmo salar L.)

The effect of 9 min of net confinement on two marine teleost species, the flounder and the Atlantic salmon, was investigated in order to gauge how different species respond to the same stress stimulus. Net confinement in both species induced significant elevations in plasma cortisol, glucose, lactate, osmolality, and monovalent ion levels; the responses to net confinement in salmon were generally of a greater magnitude. In both species, handling induced significant alterations in PFFA levels though there were marked species differences in the qualitative and quantitative aspects of this response. Plasma protein levels were significantly elevated only in the flounders. In general, the two species responded similarly to the net confinement with differences only in the absolute levels and durations of the responses.     

Histological changes in intestine of Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) following in vitro exposure to pathogenic and probiotic bacterial strains

Furunculosis and vibriosis are diseases that cause severe economic losses in the fish-farming industry. The foregut of the Atlantic salmon (Salmo salar L.) was exposed in vitro to two fish pathogens, Aeromonas salmonicida (causative agent of furunculosis) and Vibrio anguillarum (causative agent of vibriosis), and to one probiotic strain, Carnobacterium divergens, at 6 × 10⁴ or 6 × 10⁶ viable bacteria per milliliter. Histological changes following bacterial exposure were assessed by light and electron microscopy. Control samples (foregut exposed to Ringer’s solution only) and samples exposed only to C. divergens had a similar appearance to intact intestinal mucosal epithelium, with no signs of damage. However, exposure of the foregut to the pathogenic bacteria resulted in damaged epithelial cells, cell debris in the lumen, and disorganization of the microvilli. Co-incubation of the foregut with a pathogen and C. divergens did not reverse the damaging effects caused by the pathogen, although these were alleviated when probiotic bacteria were used. Based on these results, we suggest that the probiotic bacterium, C. divergens, is able to prevent, to some extent, pathogen-induced damage in the Atlantic salmon foregut. I. Salinas thanks Fundacion Seneca for a PhD studentship.