Seasonal Variations in Clock‐Gene Expression in Atlantic Salmon (Salmo salar)

In homeothermic vertebrates inhabiting temperate latitudes, it is clear that the seasonal changes in daylength are decoded by the master circadian clock, which through secondary messengers (like pineal melatonin secretion) entrains rhythmic physiology to local conditions. In contrast, the entrainment and neuroendocrine regulation of rhythmic physiology in temperate teleosts is not as clear, primarily due to the lack of understanding of the clock gene system in these species. In this study, we analyzed the diel expression of the clock‐genes in brains of Atlantic salmon, a species that is both highly photoperiodic and displays robust clock‐controlled behavior. Atlantic salmon parr were acclimated to either long‐day (LD) or short‐day (SD) photoperiods for one month and thereafter sampled at 4 h intervals over a 24 h cycle. Clock, Bmal1, Per2, and Cry2 were all actively expressed in salmon brain homogenates and, with the exception of Per2, all displayed rhythmic expression under SD photoperiods that parallels that reported in zebrafish. Interestingly, daylength significantly altered the mRNA expression of all clock genes studied, with Clock, Bmal1, and Per2 all becoming arrhythmic under the LD compared to SD photoperiod, while Cry2 expression was phase delayed under LD. It is thus proposed that the clock‐gene system is actively expressed in Atlantic salmon, and, furthermore, as has been reported in homeothermic vertebrates, it appears that clock expression is daylength‐dependent.     

N‐Terminal Sequence and Main Characteristics of Atlantic Salmon (Salmo salar) Albumin

Abstract

Atlantic salmon (Salmo salar) serum albumin was purified from plasma and its N‐terminal sequence determined. Atlantic salmon albumin is the predominant plasma protein, negatively charged, at pH 8.6. Albumin was purified to >95% purity which yielded a single band on SDS‐PAGE and agarose gel electrophoresis. The molecular weight of the purified albumin was approximately 6,5 kDa. The N‐terminal sequence of Atlantic chinook salmon albumin was consistent with that predicted from its previously determined cDNA sequence and was identical to that of salmon (Oncorhynchus tshawytscha) albumin through the first 15 residues. However, the fact that the actual N‐terminus was different from that predicted from cDNA sequence indicates that Atlantic salmon albumin, like chinook salmon albumin, lacks a propeptide.     

Stress-Induced Reversion to Virulence of Infectious Pancreatic Necrosis Virus in Na?ve Fry of Atlantic Salmon (Salmo salar L.)

We have studied stress-induced reversion to virulence of infectious pancreatic necrosis virus (IPNV) in persistently infected Atlantic salmon (Salmo salar L.) fry. Naïve fry were persistently infected with a virulent strain (T₂₁₇A₂₂₁ of major structural virus protein 2, VP2) or a low virulent (T₂₁₇T₂₂₁) variant of IPNV. The fry were infected prior to immunocompetence as documented by lack of recombination activating gene-1, T-cell receptor and B-cell receptor mRNA expression at time of challenge. The fish were followed over 6 months and monitored monthly for presence of virus and viral genome mutations. No mutation was identified in the TA or TT group over the 6 months period post infection. Six months post infection TA and TT infected groups were subject to daily stress for 7 days and then sampled weekly for an additional period of 28 days post stress. Stress-responses were documented by down-regulation of mRNA expression of IFN-α1 and concomitant increase of replication levels of T₂₁₇T₂₂₁ infected fish at day 1 post stress. By 28 days post stress a T221A reversion was found in 3 of 6 fish in the T₂₁₇T₂₂₁ infected group. Sequencing of reverted isolates showed single nucleotide peaks on chromatograms for residue 221 for all three isolates and no mix of TA and TT strains. Replication fitness of reverted (TA) and non-reverted (TT) variants was studied in vitro under an antiviral state induced by recombinant IFN-α1. The T₂₁₇A₂₂₁ reverted variant replicated to levels 23-fold higher than the T₂₁₇T₂₂₁ strain in IFN-α1 treated cells. Finally, reverted TA strains were virulent when tested in an in vivo trial in susceptible salmon fry. In conclusion, these results indicate that stress plays a key role in viral replication in vivo and can facilitate conditions that will allow reversion from attenuated virus variants of IPNV.     

Variation in the timing of river entry of Atlantic salmon （Salmo salar L. ） in the Baltic

The timing of river entry in the Atlantic salmon is known to depend on genetic, demographic and environmental factors, but little is known about the relative magnitude of among population and among year variation and covariation in this respect in natural state Atlantic salmon rives. To investigate this, variability in the timing of river entry in three historical Finnish Atlantic salmon populations were analyzed using salmon trap data collected during 1870- 1902. The analyses reveled that 1 ） the timing of river entry differed substantially and consistently among the rivers, and that 2） variation among the rivers was much larger than variation among years. Annual variations were not explained by regional environmental conditions, whereas in one river the timing of the local flood peak was a significant predictor of the timing of river entry. Differences in the timing of salmon entry to geographically closely situated rivers suggests that a regionally fixed opening date for coastal fisheries might not be the best management strategy as it may lead to uneven exploitation of salmon populations from different rivers [ Current Zoology 55 （5） ： 342 - 349, 2009] .     

Fluctuating asymmetry in farmed Atlantic salmon (Salmo salar) juveniles: also a maternal matter?

Developmental stability reflects the degree to which phenotypic expression is unaffected by random accidents or developmental noise. Developmental stability may be measured by phenodeviance or fluctuating asymmetry (FA), and estimation of developmental stability has attracted substantial interest because it appears to represent a relatively simple method to identify sub lethal stress exposure and to assess animal welfare. As a part of a long-term study, the work presented here primarily aimed to investigate impacts on developmental instability in farmed salmon offspring ten months post hatch attributable to maternal cortisol administration prior to spawning and mild hyperthermia exerted during incubation. Main results show that maternal cortisol enhancement increased the level of FA in pectoral and pelvic fins, but did not affect the frequency of malformations in offspring. Mild hyperthermia during incubation increased weight and fork length and also increased pelvic fin FA. Malformed fish were heavier and longer than the normal ones, and pelvic fin asymmetry was positively related to condition factor. These results illustrate plausible lasting impacts on offspring development due to the maternal endocrinological state at spawning and indicate that developmental instability in farmed salmon juveniles may mirror aspects of the broodstock’s housing conditions.     

Cloning and sequence analysis of cDNAs encoding the MHC class II β chain in Atlantic salmon (Salmo salar)

Atlantic salmon (Salmo salar) cDNAs encoding the major histocompatibility complex (Mhc-Sasa) class II chain were isolated from a leucocyte library by a polymerase chain reaction (PCR) approach. Three different cDNAs (c144, c22, and c157) encoding the entire mature chain have been analyzed. Clone c144 differs from clone c157 in 12.6% of the nucleotides in the 1-encoding region. The corresponding differences between clones c144 and c22, and clones c22 and c157, are 10.3% and 5.2%, respectively. This variation is, at least in part, most likely attributable to allelism. The similarity indices between the highly conserved 2 domains from Atlantic salmon and corresponding sequences from humans (DQ), chicken (BL), carp (TLAII-1), and rainbow trout (O. M. No. 55) are 45%, 40%, 66%, and 97%, respectively. Variable residues in the 1 domains from Atlantic salmon correspond with polymorphic sites of 1 domains from higher vertebrates. The frequency of substitutions in the 1-encoding region exceeds that in the 3-untranslated (UT) region with several folds, indicating extensive 1 polymorphism in Atlantic salmon.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers (C 144) X 70165, (C 157) X 70166, and (C 22) X 70167.This study was supported by grants from the Norwegian Fisheries Research Council (NFFR) and the Norwegian Agricultural Research Council (NLVF).     

Changes in scale circulus spacings of an endangered Atlantic salmon Salmo salar population: evidence of a shift in marine migration?

Post‐smolt scale circulus spacing patterns for two Atlantic salmon Salmo salar populations from the Southern Upland (SU) of Nova Scotia, Canada, were compared with spacings from two endangered populations from the inner Bay of Fundy (iBoF) Nova Scotia and New Brunswick, to determine if growth patterns differed among these populations, and if growth patterns had changed as the abundance of these populations declined. An analysis of numbers of marine circuli from scales of post‐smolts and one‐sea‐winter adults of known age indicated that circuli were deposited at a rate of about one circulus per week in summer and slowed to one every 2 weeks in winter. During the summer and the autumn, mean circulus spacing in the iBoF populations, known to have occupied the outer Bay of Fundy during these seasons, was lower than in the SU populations, which are known to migrate to the North Atlantic. Similar circulus spacing patterns within SU populations is suggestive of a common marine distribution for these populations. In contrast, a cluster analysis revealed that within the geographically intermediate Big Salmon River (iBoF), some individuals exhibited wider spacing patterns that resemble the distant migrating SU populations, while others exhibited narrower spacing similar to other iBoF S. salar. Within the Big Salmon River, the relative abundance of the wider and the narrower spacing patterns varied in the earlier years, but all fish sampled since 1999, exhibited wider spacings similar to distant migrating SU S. salar. The apparent disappearance of the narrower pattern, characteristic of localized migration and indicative of historical iBoF populations, suggests that local migration may not presently be a successful strategy for these populations.     

Identification of a Low Digestibility δ-Conglutin in Yellow Lupin (Lupinus luteus L.) Seed Meal for Atlantic Salmon (Salmo salar L.) by Coupling 2D-PAGE and Mass Spectrometry

The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain δconglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein.     

Silver spoons in the rough: can environmental enrichment improve survival of hatchery Atlantic salmon Salmo salar in the wild?

This study tested the ‘silver spoon’ hypothesis which posits that individuals that develop under favourable conditions should enjoy a fitness advantage later in life because they are more likely to recognize and settle in high‐quality habitats. Atlantic salmon Salmo salar of two age classes (0+ and 1+ years) were reared in environmentally enriched or standard hatchery tanks for a short period (c. 10 weeks), were then released into a natural river and sampled on repeated occasions to test for silver‐spoon effects. Compared with controls, enriched fish had a 6·4% higher recapture rate and settled in higher velocity habitats when they were stocked as 0+ year fry, but not when they were stocked as 1+ year parr. The opportunity for selection was generally higher for environmentally enriched fish than for controls, and also higher for 0+ than for 1+ year fish. Selection favoured individuals with high condition factor, extensive fat reserves and longer than average pectoral fins in both age classes but favoured a small body size in 1+ year and a large body size in 0+ year releases. Stomach analysis showed that enriched fish ate more, and adapted quicker to natural prey than controls. These results provide support for silver‐spoon effects in fish and indicate that enrichment can improve post‐release performance in conservation programmes, but seemingly only if fish are not kept in captivity for too long.     

A SNP in the 5′ flanking region of the myostatin-1b gene is associated with harvest traits in Atlantic salmon (Salmo salar)

Background

Myostatin (MSTN) belongs to the transforming growth factor-β superfamily and is a potent negative regulator of skeletal muscle development and growth in mammals. Most teleost fish possess two MSTN paralogues. However, as a consequence of a recent whole genome-duplication event, salmonids have four: MSTN-1 (?1a and -1b) and MSTN-2 (?2a and -2b). Evidence suggests that teleost MSTN plays a role in the regulation of muscle growth. In the current study, the MSTN-1b gene was re-sequenced and screened for SNP markers in a commercial population of Atlantic salmon. After genotyping 4,800 progeny for the discovered SNPs, we investigated their association with eight harvest traits - four body-weight traits, two ratios of weight traits, flesh colour and fat percentage - using a mixed model association analysis.

Results

Three novel SNPs were discovered in the MSTN-1b gene of Atlantic salmon. One of the SNPs, located within the 5′ flanking region (g.1086C?>?T), had a significant association with harvest traits (p?<?0.05), specifically for: Harvest Weight (kg), Gutted Weight (kg), Deheaded Weight (kg) and Fillet Weight (kg). The haplotype-based association analysis was consistent with this result because the two haplotypes that showed a significant association with body-weight traits, hap4 and hap5 (p?<?0.05 and p?<?0.01, respectively), differ by a single substitution at the g.1086C?>?T locus. The alleles at g.1086C?>?T act in an additive manner and explain a small percentage of the genetic variation of these phenotypes.

Conclusions

The association analysis revealed that g.1086C?>?T had a significant association with all body-weight traits under study. Although the SNP explains a small percentage of the variance, our results indicate that a variation in the 5′ flanking region of the myostatin gene is associated with the genetic regulation of growth in Atlantic salmon.

     

Effects of sea lice (Lepeophtheirus salmonis Kröyer, 1837) infestation on macrophage functions in Atlantic salmon (Salmo salar L.)

Experiments were conducted to determine the effects of sea lice, Lepeophtheirus salmonis, on non-specific defence mechanisms in Atlantic salmon, Salmo salar, by experimentally infesting hatchery-reared 1 and 2 year old post-smolts, S1 and S2, with laboratory grown infective copepodids at moderate to high infection intensities ranging from 15-285 lice per fish. The effects of sea lice-induced stress were investigated by measuring the blood levels of cortisol and glucose as indicators of primary and secondary stress responses, and by changes in macrophage respiratory burst activity and phagocytosis as indicators of tertiary stress responses as well as non-specific defence mechanisms. Fish were sampled prior to sea lice infestation at day 0 and at days 3, 7, 14 and 21 post-infestation. Sea lice were at copepodid stage at day 3, at chalimus stages at days 7 and 14, and at pre-adult stage at day 21. Blood levels of cortisol and glucose were found to be significantly increased at day 21 in fish-infested with the highest levels. Macrophage respiratory burst and phagocytic activities were found to be significantly decreased only at day 21. These results indicate that sea lice do not suppress host defence mechanisms during the earlier stages of infestation. They do have effects on the development of chronic stress and on the host non-specific defence mechanisms soon after the lice reach the pre-adult stage.     

Bisucaberin – A dihydroxamate siderophore isolated from Vibrio salmonicida, an important pathogen of farmed Atlantic salmon (Salmo salar)

A siderophore of the bacterial fish pathogen, Vibrio salmonicida, was isolated from low-iron culture supernatant and structurally characterized as bisucaberin by FTICR- and FAB-MS, NMR and GC-MS analysis of the hydrolysis products. Although the cyclic dihydroxamate bisucaberin has previously been isolated from a marine bacterium, Alteromonas haloplanktis, its involvement in cold-water vibriosis of Atlantic salmon (Salmon salar) is novel. Bisucaberin production in iron-limited media was highest at temperatures around and below 10 °C, correlating well with temperatures at which outbreaks of cold-water vibriosis occur. Due to the very high stability constant of K = 32.2, bisucaberin is a most efficient iron scavenger which may contribute to the virulence of V. salmonicida in Atlantic salmon.     

Does catch and release affect the mating system and individual reproductive success of wild Atlantic salmon (Salmo salar L.)?

In this study, we documented the breeding system of a wild population of Atlantic salmon (Salmo salar L.) by genetically sampling every returning adult and assessed the determinants of individual fitness. We then quantified the impacts of catch and release (C&R) on mating and reproductive success. Both sexes showed high variance in individual reproductive success, and the estimated standardized variance was higher for males (2.86) than for females (0.73). We found a weak positive relationship between body size and fitness and observed that fitness was positively correlated with the number of mates, especially in males. Mature male parr sired 44% of the analysed offspring. The impact of C&R on the number of offspring was size dependent, as the reproductive success of larger fish was more impaired than smaller ones. Also, there was an interactive negative effect of water temperature and air exposure time on reproductive success of C&R salmon. This study improves our understanding of the complex reproductive biology of the Atlantic salmon and is the first to investigate the impact of C&R on reproductive success. Our study expands the management toolbox of appropriate C&R practices that promote conservation of salmon populations and limit negative impacts on mating and reproductive success.     

Does size matter? A test of size‐specific mortality in Atlantic salmon Salmo salar smolts tagged with acoustic transmitters

Mortality rates of wild Atlantic salmon Salmo salar smolts implanted with acoustic transmitters were assessed to determine if mortality was size dependent. The routinely accepted, but widely debated, ‘2% transmitter mass: body mass’ rule in biotelemetry was tested by extending the transmitter burden up to 12·7% of body mass in small [mean fork length (L_F) 138·3 mm, range 115–168 mm] downstream migrating S. salar smolts. Over the short timescale of emigration (range 11·9–44·5 days) through the lower river and estuary, mortality was not related to S. salar size, nor was a relationship found between mortality probability and transmitter mass: body mass or transmitter length: L_F ratios. This study provides further evidence that smolt migration studies can deviate from the ‘2% rule’ of thumb, to more appropriate study‐specific measures, which enables the use of fishes representative of the body size in natural populations without undue effects.     

Variation in the post-smolt growth pattern of wild one sea-winter salmon (Salmo salar L.), and its linkage to surface warming in the eastern North Atlantic Ocean

Variation in circulus spacing on the scales of wild Atlantic salmon is indicative of changes in body length growth rate. We analyzed scale circulus spacing during the post-smolt growth period for adult one sea-winter salmon (n = 1947) returning to Scotland over the period 1993–2011. The growth pattern of the scales was subjectively and visually categorized according to the occurrence and zonal sequence of three intercirculus spacing criteria (“Slow”, “Fast” and “Check” zones). We applied hierarchical time-series cluster analysis to the empirical circulus spacing data, followed by post hoc analysis of significant changes in growth patterns within the 20 identified clusters. Temporal changes in growth pattern frequencies showed significant correlation with sea surface temperature anomalies during the early months of the post-smolt growth season and throughout the Norwegian Sea. Since the turn of the millennium, we observed (a) a marked decrease in the occurrence of continuous Fast growth; (b) increased frequencies of fish showing an extended period of initially Slow growth; and (c) the occurrence of obvious growth Checks or hiatuses. These changes in post-smolt growth pattern were manifest also in decreases in the mean body length attained by the ocean midwinter, as sea surface temperatures have risen.     

Atlantic salmon (Salmo salar) protein hydrolysate in diets for weaning piglets ─ effect on growth performance,intestinal morphometry and microbiota composition

Salmon protein hydrolysates (SPH) from two different rest raw materials were evaluated in diets for weaning piglets. Four experimental diets were included in the study: a diet based on plant protein with soy protein as the main protein source (Diet PP), a diet based on fishmeal in exchange for soy protein (Diet FM) and two diets in which different SPH replaced fishmeal in the FM diet. The experimental diets were fed to piglets from the day of weaning until 32 d postweaning. In addition to the record of performance data, an intestinal sampling for mucosal morphometry and microbiota 16S rRNA gene sequencing were performed at day 11 on a subset of the animals. The duodenal villi absorption area was significantly larger in piglets receiving Diets SPH compared with Diet PP (p < 0.02). A significant positive correlation between duodenal villi height and average daily gain during the first 11 d postweaning was detected. Only small differences in intestinal microbiota community and no differences in growth performance were detected between the experimental diets. To conclude, SPH seem to be an interesting novel protein source in weanling piglets.