Deep-diving of Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>) during their marine feeding migrations

Data from seven data storage tags recovered from Atlantic salmon marked as smolts were analyzed for depth movements and patterns of deep diving during the marine migration. The salmon mostly stayed at the surface and showed diurnal activity especially from autumn until spring. During the first months at sea the salmon stayed at shallower depths (<100 m). The salmon took short deep dives (>100 m), that were rare or absent during the first summer at sea but increased in frequency and duration especially in late winter. The maximum depth of the dives varied from 419 to 1187 m. Most of dives were short, (<5 h) but could last up to 33 h. The duration of dives increased in late winter until spring and the overall depth and maximum depth per dive increased exponentially over time. The initiation of the dives was more common in evenings and at night, suggesting nocturnal diving. We hypothesized that deep diving is related to feeding of salmon as mesopelagic fish can be important food for salmon during winter.     

Tolbutamide hydroxylation by hepatic microsomes from Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.)

Metabolic transformations of two substrates for human cytochrome P450 (CYP450) 2C9, tolbutamide and diclofenac, were investigated in hepatic microsomes from Atlantic salmon (Salmo salar L.). Tolbutamide hydroxylation followed Michaelis–Menten kinetics. Mean apparent Michaelis–Menten constant (K_m) and maximum reaction velocity (V_max) values for 4-hydroxytolbutamide (TBOH) formation were 0.09 ± 0.031 mM and 49.5 ± 6.03 pmol/min/mg, respectively. Addition of sulfaphenazole, an inhibitor for mammalian CYP2C9, in a range from 1 to 200 μM decreased formation of TBOH in a concentration-dependent manner, but not to 50%. Neither fluconazole, an inhibitor of human CYP2C9, nor ketoconazole, inhibitor of CYP1A and CYP3A in fish, affected TBOH formation. In contrast ellipticine, an inhibitor of CYP1A in fish inhibited TBOH formation with the IC₅₀ value of 12.1 μM. The rate of TBOH formation was competitively inhibited by 100 μM of sesamin in the incubations, but the degree of inhibition did not increase with increased sesamin concentration. Ethoxyresorufin hydroxylase (EROD) activity was inhibited by tolbutamide in a non-competitive manner (inhibition constant K_i = 218 μM). Our data suggest that tolbutamide is metabolized by salmon microsomes with formation of TBOH. CYP1A might be involved in this reaction as suggested by decreased TBOH formation in the presence of ellipticine and decreased EROD activity in the presence of tolbutamide. Incubation of diclofenac with the microsomes yielded no metabolite formation, suggesting that salmon does not possess diclofenac-metabolizing activity.     

Damaging effect of the fish pathogen <Emphasis Type="Italic">Aeromonas salmonicida</Emphasis> ssp. <Emphasis Type="Italic">salmonicida</Emphasis> on intestinal enterocytes of Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.)

In fish, bacterial pathogens can enter the host by one or more of three different routes: (a) skin, (b) gills and (c) gastrointestinal tract. Bacteria can cross the gastrointestinal lining in three different ways. In undamaged tissue, bacteria can translocate by transcellular or paracellular routes. Alternatively, bacteria can damage the intestinal lining with extracellular enzymes or toxins before entering. Using an in vitro (Ussing chamber) model, this paper describes intestinal cell damage in Atlantic salmon (Salmo salar L.) caused by the fish pathogen Aeromonas salmonicida ssp. salmonicida, the causative agent of furunculosis. The in vitro method clearly demonstrated substantial detachment of enterocytes from anterior region of the intestine (foregut) upon exposure to the pathogen. In the hindgut (posterior part of the intestine), little detachment was observed but cellular damage involved microvilli, desmosomes and tight junctions. Based on these findings, we suggest that A. salmonicida may obtain entry to the fish by seriously damaging the intestinal lining. Translocation of bacteria through the foregut (rather than the hindgut) is a more likely infection route for A. salmonicida infections in Atlantic salmon.Financial support from the Commission of the European Communities, quality of Life and Management of Living Resources programme, project Q5RT-2000-31656 Gastrointestinal Functions and Food Intake Regulation in Salmonids: Impact of Dietary vegetable Lipids (GUTINTEGRITY) and from Magnus Bergvalls Stiftelse for KS, is acknowledged.This work does not represent the opinion of the European Community, which is thus not responsible for any use of the data presented.     

Etho-biochemical mechanisms of early differentiation in juveniles of the Atlantic salmon <Emphasis Type="Italic">Salmo salar</Emphasis>

The social status of hatchery-reared juveniles of Atlantic salmon Salmo salar at the age of 11–19 months in conditions of lack of shelters was investigated experimentally. It was demonstrated that biochemical differentiation in such juveniles begins at the age of 13 months—one month before the first differences in the social status of fish and four months prior to the beginning of smoltification. The dominants occupy bottom areas with shelters and extrude the subordinates to the water column. The subordinates undergo smoltification or die from aggression of the dominants. Concentrations of dopamine, noradrenalin, and of their metabolites in the brain of fish in the beginning of smoltification were higher in the dominants and, by the end of smoltification, were higher in smolts. The social behavior of fish is considered to be one of the ethological mechanisms of differentiation of salmon juveniles into smolts and parr.     

Development of ethologo-physiological differentiation between parrs and smolts of the Atlantic salmon <Emphasis Type="Italic">Salmo salar</Emphasis>

Dynamics of the level of monoamines and their metabolites in the brain of juveniles of the Atlantic salmon Salmo salar is investigated in the initial period of differentiation of the progeny into parrs and smolts. The first differences in the level of metabolism of noradrenaline (NA) and 5-HT arise in the optical tectum of the brain before the beginning of smolting. Appearance of the first signs of smolting in juveniles is accompanied by the increase of differences in the level of activity of HA-and 5-HT-ergic systems in the forebrain and hypothalamus. The first differences in the activity of the DAergic system between parrs and smolts were observed in the period of visible differences in fish coloration. In the same period, considerable differences in the level of adrenaline and noradrenaline in the fish blood were recorded. In experiments on a prolonged exposure of fish to a current of different intensity, it was shown that behavior and physiological response of the fish organism are different: in parrs, the stressogenous response is developed, they stop feeding, weight loss takes place, and they cannot withstand the water flow; smolts adapt to the current using heterogeneity of water movements.     

Adaptation of cultured salmon <Emphasis Type="Italic">Salmo salar</Emphasis> (Salmonidae) yearlings to natural environments

The process of adaptation of the yearling Atlantic salmon Salmo salar cultured under the farming conditions in the Umba fish hatchery to the natural environments has been studied during a 1-year period after releasing them into the river. The significant differences in a range of the morphophysiological parameters between the hatchery-reared juveniles and their wild-origin peers, which were revealed immediately before the release into the river and 1 month later, are gradually reduced. The index of body condition, the liver index, and the body cavity fat content are normalized 4 months later; 13 months later, the other analyzed parameters (the hemoglobin concentration and the indices of heart, gills, and pectoral fins) are brought back into the normal ranges. The behavioral patterns of the hatchery-reared and wild-origin juveniles remain different during the whole observation period, since the behavioral habits acquired under the farming conditions, when compared to the developmental abnormalities, more commonly affect the juvenile fish survival in the natural environments.     

Characterization of Na,K-ATPase genes in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>) and comparative genomic organization with rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

A combination of molecular and in silico approaches was employed to assemble a survey of Na, K-ATPase genes contained in the ancestrally tetraploid genome of the Atlantic salmon (Salmo salar). Molecular characterization of genomic clones coding for the subunit revealed two single genes (1a and 2) and two pairs of presumably homeologous genes (1b/i-ii and 1c/i-ii). Each of the six genes showed high sequence similarity to isoforms previously isolated from rainbow trout and extensive structural differences relative to putative orthologs in the human genome. In silico analysis of expressed sequence tag (EST) collections indicated that at least five (1a, 1b, 1c, 2, and 3) and four (1a, 1b, 2, and 3b) subunit isoforms are expressed in Atlantic salmon. Meiotic linkage analysis further showed that Na, K-ATPase genes are dispersed throughout the salmon genome, with the exception of two multigene clusters on linkage groups AS-22 and AS-28. Duplicate gene copies for the isoform 1b were assigned to linkage groups with multiple homeologous anchors (AS-22 and AS-23), while 2 duplicates suggested a new homeologous affinity between AS-05 and AS-28. In addition, the comparison of linkage arrangements with rainbow trout also showed that the genomic organization of Na, K-ATPase genes is consistent with the evolutionary conservation of syntenic chromosome regions between these species.Electronic Supplementary Material Supplementary material is available for this article at .     

Fatty acid composition of sprat (<Emphasis Type="Italic">Sprattus sprattus</Emphasis>) and herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>) in the Baltic Sea as potential prey for salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

Sprat (Sprattus sprattus) and small herring (Clupea harengus) are the dominant prey fish of Atlantic salmon (Salmo salar) in the Baltic Sea. If the fatty acid (FA) proportions of sprat and herring differ, the dietary history of ascending salmon could be determined from their FA profiles. Therefore, we investigated the FA composition of several age groups of whole sprat and small herring, caught from the three main feeding areas of salmon in autumn and spring. Oleic acid (18:1n-9) was the most prevalent FA in sprat and characteristic of this species. In herring, palmitic acid (16:0) was the most common FA, but herring lipid was characterized by n-6 polyunsaturated FAs, and moreover, by palmitoleic acid (16:1n-7) and vaccenic acid (18:1n-7). Due to the higher lipid content of sprat, the concentrations of all other FAs, excluding these, were higher in sprat than in herring. The concentration of docosahexaenoic acid (DHA, 22:6n-3) increased with an increase in the lipid content and was consequently highest in the youngest specimens, being in young sprat almost double that of young herring, and 2.6 times higher in the sprat biomass than in that of herring. As a result of a decrease in the DHA concentration with age, the ratio thiamine/DHA increased with respect to age in both species, and was lower in sprat than in herring. It is concluded that an abundance of DHA in the diet of salmon most likely increases oxidative stress because of the susceptibility of DHA to peroxidation, and thus decreases thiamine resources of fasting, prespawning salmon. Because the FA composition of sprat and herring differs, and the relative abundancies of prey fish differ between the feeding areas of salmon, the feeding area of ascending salmon can most probably be derived by comparing their FA profiles.     

Somatosensory evoked potentials in the telencephalon of Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>) following galvanic stimulation of the tail

Electric activity in the brain which is time-locked to a given stimulation of the somatosensory system can be recorded as a somatosensory evoked potential (SEP). We investigated whether a galvanic stimulation of the tail base in Atlantic salmon (Salmo salar) would elicit a SEP in the telencephalon. The telencephalon is central in learning and memory, and activity here may be a prerequisite for processing of external stimuli on a cognitive or emotional level. Anaesthetized salmon (n = 11) were subjected to craniotomy and a recording electrode was inserted into the telencephalon. The fish were given stimulations of four intensities, i.e., 2, 5, 10 and 20 mA. A SEP was elicited in the contralateral dorsal telencephalon for all intensities. This result agrees with findings in other fish species. Furthermore, there was a significant difference between the maximum peak amplitude and mean amplitude of the SEP elicited by putative non-noxious (2 mA) and putative noxious (20 mA) stimulation intensities (P < 0.01). The stronger stimulation intensities also tend to introduce longer-latencies components in the SEP. The results added to the body of literature indicates that the exteroceptive senses are represented by processing within the telencephalon of the fish.     

Influence of shelters and food resources on smoltification of juveniles of the Atlantic salmon <Emphasis Type="Italic">Salmo salar</Emphasis>

Influence of feeding conditions and of the presence of a various number of shelters in basins on development of differentiation of juveniles of Salmo salar at the age 1+ and formation of smolts were studied under experimental conditions. The experiments had been carried out on groups of ten fish. The influence of investigated factors turned out to be significant. Under normal feeding four times a day, the quantity of formed smolts depends on the number of shelters: the more shelters that are available, the fewer smolts appear in groups. The presence of one shelter is a strong stimulant enhancing aggressive activity towards conspecific individuals, resulting in destruction of some fish in the group. Under feeding once a day, i.e., under food shortage, more fish perished, and the number of smolts decreased as the fish did not accumulate a sufficient energy potential for the morphophysiological transformation. One of the potential mechanisms of development of differentiation of fish juveniles into parrs and future smolts is discussed, based on domination of some fish possessing the individual territory and exclusion of subdominant specimens into open water as a result of social interaction.     

An extended panel of single nucleotide polymorphisms in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) mitochondrial DNA

A total of 6,324 base pairs, distributed on eight different regions of the mitochondrial DNA (mtDNA) were re-sequenced for 32 specimens of Norwegian farmed and wild Atlantic salmon. A total of 16 new and three previously reported single nucleotide polymorphisms (SNPs) were detected. These SNPs were validated using 240 farm and 119 wild Norwegian salmon. Alignments of sequences obtained in this study and previously published sequences revealed another 15 previously unreported SNPs. The panel of mitochondrial SNPs detected in the present study will, in combination with previously identified SNPs, prove useful in designing efficient assays for analyzes of mtDNA variation.     

Feeding of wild and hatchery reared Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) smolts during downstream migration

In general, hatchery salmonid smolts experience higher mortality during migration than wild smolts, which is suggested to be due to domestication effects and that hatchery fish lack experience of the natural environment. However, possible differences in feeding during smolt migration between hatchery and wild smolts have rarely been addressed. We compared the number of feeding smolts and stomach fullness among wild Atlantic salmon smolts, hatchery-reared smolts released as 1-year-old parr, and hatchery-reared smolts released as 2-year-old smolts during their descent to sea in River Tornionjoki. In addition, estimations of prey selection among the smolt groups were conducted. A high proportion of wild smolts and smolts stocked as parr actively fed during the smolt migration. A lower proportion of smolts stocked as smolts was feeding and their stomach fullness were much reduced in comparison with the two other groups. The study also indicated that the feeding of migrating smolts is selective rather than opportunistic. In conclusion, this study suggests that stocked 2-year-old smolts may enter sea with an inferior foraging behaviour and it is a possibility that this may contribute to the observed low post-smolt survival in the Baltic Sea.     

Reduced anti-predator responses in multi-generational hybrids of farmed and wild Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.)

Cultured organisms undergo genetically-based behavioural changes that may reduce their ability to survive in the wild. This has raised concerns that interbreeding between escaped cultured and wild organisms will generate hybrids exhibiting maladaptive behaviours which may ultimately reduce the fitness of the wild counterpart. We compared anti-predator responses in Atlantic salmon (Salmo salar) from two wild North American populations, the major farmed strain used in regional aquaculture, and their wild-farmed hybrids (F₁, F₂, and wild backcross). Anti-predator responses of fry (age 0+ parr) were measured under common environmental conditions, using a model of a natural predator (belted kingfisher, Ceryle alcyon). Farmed fry exhibited significantly reduced anti-predator responses relative to fry from both wild populations. The anti-predator responses of wild-farmed hybrid fry were intermediate to those of the parental populations (pure farmed or wild). The magnitude by which wild-farmed hybrids differed in anti-predator responses from pure wild fish also depended on the wild population. These results suggest that: (1) the observed behavioural differences have a genetic basis; (2) wild-farmed hybrids have, on average, reduced anti-predator responses relative to wild fish; and that (3) the effects of wild-farmed interbreeding on anti-predator responses will differ between wild populations. Our study is consistent with the general hypothesis that continual farmed-wild interbreeding may have detrimental effects on the fitness of wild organisms.     

Unique haplotypes of co-segregating major histocompatibility class II <Emphasis Type="Italic">A</Emphasis> and class II <Emphasis Type="Italic">B</Emphasis> alleles in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>) give rise to diverse class II genotypes

Sequence-based typing of a breeding population (G1) consisting of 84 Atlantic salmon individuals revealed the presence of 7 Sasa-DAA and 7 Sasa-DAB expressed alleles. Subsequent typing of 1,182 individuals belonging to 33 families showed that Sasa-DAA and Sasa-DAB segregate as haplotypes. In total seven unique haplotypes were established, with frequencies in the population studied ranging from 0.01 to 0.49. Each haplotype is characterized by a unique minisatellite marker size embedded in the 3' untranslated region of the Sasa-DAA gene. These data corroborate the fact that Atlantic salmon express a single class II locus, consisting of tightly linked class II A and class B genes. The seven haplotypes give rise to 15 genotypes with frequencies varying between 0.01 and 0.23; 21 class II homozygous individuals were present in the G1 population. We also studied the frequency distribution in another breeding population (G4, n=374) using the minisatellite marker. Only one new marker size was present, suggesting the presence of one new class II haplotype. The marker frequency distribution in the G4 population differed markedly from the G1 population. The genomic organizations of two Sasa-DAA and Sasa-DAB alleles were determined, and supported the notion that these alleles belong to the same locus. In contrast to other studies of salmonid class II sequences, phylogenetic analyses of brown trout and Atlantic class II A and class II B sequences provided support for trans-species polymorphism.     

Lipid status of fingerlings of the Atlantic salmon <Emphasis Type="Italic">Salmo salar</Emphasis> from different microbiotopes of the Varzuga River

Mechanisms of formation of phenotypic groups of fingerlings of the Atlantic salmon are investigated, related to diversity of embryos and subsequent start possibilities of dispersion of larvae to microbiotopes differing in their life conditions. The fingerlings of salmon, which after hatching and dissolution of the yolk sac moved from the mainstream of the Varzuga River to mouths of its tributaries, had an increased level of triacylglycerols and a higher growth rate than the juveniles remaining in the coastal zone of the river. The revealed differences between the compared groups of fingerlings by the spectrum of stock and structural lipids are mainly connected with distinctions of feeding of juveniles of the same age. The found stable differences by lipid spectra in fingerlings of salmon from the investigated biotopes of the Varzuga in July, August, and October are considered as biochemical prerequisites of the origin of different phenotypic groups of juveniles. Subdivision of fingerlings into phenotypic groups may further on influence the oncoming of the smoltification time of juveniles at the age 2+, 3+, and 4+. Accordingly, this is reflected in formation of the complex age structure (by the number of years spent in the river and in the sea) of the Varzuga population of Atlantic salmon.     

Molecular faunistics of accidental infections of <Emphasis Type="Italic">Gyrodactylus</Emphasis> Nordmann, 1832 (Monogenea) parasitic on salmon <Emphasis Type="Italic">Salmo salar</Emphasis> L. and brown trout <Emphasis Type="Italic">Salmo trutta</Emphasis> L. in NW Russia

Salmon Salmo salar L. and brown trout S. trutta L. juveniles were examined for the presence of accidental monogenean ectoparasitic species of Gyrodactylus Nordmann, 1832 in the Baltic and White Sea basins of Russian Karelia in order to estimate the frequency of host-switching attempts on an ecological timescale. To collect phylogeographical information and for exact species identification, the parasites were characterised by nuclear internal transcribed spacer sequences of rDNA (ITS) and, for some species, also by their mitochondrial DNA (CO1 gene) sequences. Four accidental Gyrodactylus species were observed on salmon and brown trout. A few specimens of G. aphyae Malmberg, 1957, the normal host of which is the Eurasian minnow Phoxinus phoxinus (L.), were observed on lake salmon from the Rivers Kurzhma (Lake Kuito, White Sea basin) and Vidlitsa (Lake Ladoga, Baltic basin). G. lucii Kulakovskaya, 1952, a parasite of the northern pike Esox lucius L., was observed on salmon in the Kurzhma. In the River Vidlitsa, two specimens of G. papernai Ergens & Bychowsky, 1967, normally on stone loach Barbatula barbatula (L.), were found on salmon. On anadromous White Sea salmon in the River Pulonga in Chupa Bay, a few salmon parr carried small colonies of G. arcuatus Bychowsky, 1933, which were shown to have originated from the local three-spined stickleback Gasterosteus aculeatus L. consumed as prey. No specimens of Gyrodactylus salaris Malmberg, 1957 were observed, although the Pulonga is the nearest salmon spawning river to the River Keret', which is heavily infected with introduced G. salaris. In the River Satulinoja, Lake Ladoga, three specimens of G. lotae Gusev, 1953, from burbot Lota lota (L.), were collected from a single brown trout S. trutta. All nonspecific gyrodactylid infections on salmonids were judged to be temporary, because only a few specimens were observed on each of the small number of infected fishes. The prevalence of endemic G. salaris was also low, only 1% (Nfish = 296) in Lake Onega and 0.7% (Nfish = 255) in Lake Ladoga, while brown trout specific Gyrodactylus species were not observed on any of the 429 trout examined from the Ladoga basin. The host-specific and unspecific burden of Gyrodactylus spp. on these 'glacial relict' populations of salmon and brown trout was very low, suggesting a generalised resistance against the co-evolved freshwater parasite community, or some kind of 'vaccination' effect. These hypotheses deserve further testing.