Vibrio tasmaniensis sp. nov., isolated from Atlantic salmon (Salmo salar L.)

We describe the polyphasic characterization of four Vibrio isolates which formed a tight AFLP group in a former study. The group was closely related to V. cyclitrophicus, V. lentus and V. splendidus (98.2-98.9% similarity) on the basis of the 16S rDNA sequence analysis, but by DNA-DNA hybridisation experiments it had at maximum 61% DNA similarity towards V. splendidus. Thus, we propose that the isolates represent a new Vibrio species i.e. V. tasmaniensis (LMG 20012T; EMBL under the accession numbers AJ316192; mol% G+C of DNA of the type strain is 44.7). Useful phenotypical features for discrimination of V. tasmaniensis from other Vibrio species include gelatinase and beta-galactosidase activity, fatty acid composition (particularly 14:0), utilisation and fermentation of different compounds (e.g. sucrose, melibiose and D-galactose) as sole carbon source.     

Ichthyobodo hippoglossi n. sp. (Kinetoplastea: Prokinetoplastida: Ichthyobodonidae fam. nov.), an ectoparasitic flagellate infecting farmed Atlantic halibut Hippoglossus hippoglossus

Diseased Atlantic halibut Hippoglossus hippoglossus juveniles from a hatchery in western Norway showed gill and skin infections with an Ichthyobodo species. Genus Ichthyobodo contains a single valid species, I. necator, a parasite originally described from the skin and fins of a salmonid fish in freshwater. Many studies have identified this species from other hosts, but recent molecular evidence suggests that many Ichthyobodo spp. occur in both fresh- and seawater. We redescribe I. necator from Atlantic salmon Salmo salar skin infections in Norway and compare the morphology of I. necator with the form from halibut. A scheme to standardise the measurements of Ichthyobodo cells is presented. Morphologically, the Ichthyobodo species from the skin and gills of halibut differs from I. necator from salmon skin by shape (in air dried stained smears), by a low number of variably sized kinetoplasts and by a long flagellar pocket. There is also a clear increase in the number of kinetoplasts in L necator with increasing cell size (area), a pattern absent from Ichthyobodo sp. from halibut. The 2 forms are also clearly separated by their small subunit (ssu) rDNA sequences; alignments of partial ssu sequences showed 93.5 % similarity. Consequently, Ichthyobodo sp. from halibut is considered a new species, and is named I. hippoglossi n. sp. Its closest relative is Ichthyobodo sp. IV from another marine fish, the Atlantic cod Gadus morhua. A family, Ichthyobodonidae fam. nov. in the order Prokinetoplastida Vickerman, 2004, is erected to encompass Ichthyobodo spp.     

First feeding of Atlantic salmon (Salmo salar L.) under different photoperiods

Juvenile Atlantic salmon ( Salmo salar L. ) were first fed under four different static photoperiods (LD 24:0, LD 21:3, LD 18:6, LD 12:12) in two experiments. Under LD 24:0, continuous feeding or feeding 12 hours per day did not affect growth rate. In both experiments the roup on continuous light (LD 24:0) had highest growth rate. Growth was reduced on reduced dayfength. It is suggested that the difference in growth between long day (LD 21:3) and continuous light (LD 24:0) was caused by a growth inhibition during the dark period.

Zusammenfasung

Erste Nahrung des Atlantischen Lachses (Salmo salar L .)
Jungfische des atlantischen Lachses ( Salmo ralar L. ) wurden unter vier verschiedenen Photoperioden in zwei Experimenten erstmals gefüttert (LD 24:0; LD 21:3; LD 18:6; LD 12:12). Bei kontinuierlicher Fütterung (LD 24:0) oder Fütterung über 12 Stunden pro Tag, war die Wachsturnsrate nicht beeinflußt. In beiden Exerimenten hatte die Gruppe, die unter durchgehender Beleuchtung aufgezogen wurde, die höchste Wacgstumsrate. Das Wachsturn war bei kürzerer Tageslänge reduziert. Aus den Daten wird geschlossen, daß die Differenz im Wachstum zwischen dem Langtag (LD 21:3) und der durchgehenden Beleuchtung (LD 24:0) durch eine Wachtumshemmung während der Dunkelzeit verursacht wurde.     

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.     

Cloning and characterization of alpha-amylase from Atlantic salmon (Salmo salar L.)

Amylase has a lower activity in carnivorous fish species, particularly in Atlantic salmon. We report the first cloning of a salmonid alpha-amylase cDNA from Atlantic salmon, a major species in aquaculture. By amino acid alignment of several species, we identified a seven amino acid deletion in one of the large loops of the enzyme in relatively close proximity to the active site, that could impair substrate binding. We also found the signal peptide to be less hydrophobic compared to other species. This may affect import into ER during protein synthesis. Active site residues were shown to be conserved. Amylase mRNA expression was shown in pancreatic tissue, liver, and in the heart. Using blocked p-nitrophenyl-maltoheptaoside as a substrate, we measured a low amylase activity in Atlantic salmon intestinal content, which was about half of the activity measured in Atlantic cod, whereas activity measured in rainbow trout was fourteen times higher. Amylase activities in all three species showed similar degree of reduction in hydrolytic activity in a dose-response trial with a wheat amylase inhibitor preparation. This indicates similar specific activity per amylase molecule.     

Neoparamoeba perurans n. sp., an agent of amoebic gill disease of Atlantic salmon (Salmo salar)

Amoebic gill disease (AGD) is a potentially fatal disease of some marine fish. Two amphizoic amoebae Neoparamoeba pemaquidensis and Neoparamoeba branchiphila have been cultured from AGD-affected fish, yet it is not known if one or both are aetiological agents. Here, we PCR amplified the 18S rRNA gene of non-cultured, gill-derived (NCGD) amoebae from AGD-affected Atlantic salmon (Salmo salar) using N. pemaquidensis and N. branchiphila-specific oligonucleotides. Variability in PCR amplification led to comparisons of 18S rRNA and 28S rRNA gene sequences from NCGD and clonal cultured, gill-derived (CCGD) N. pemaquidensis and N. branchiphila. Phylogenetic analyses inferred from either 18S or 28S rRNA gene sequences unambiguously segregated a lineage consisting of NCGD amoebae from other members of the genus Neoparamoeba. Species-specific oligonucleotide probes that hybridise 18S rRNA were designed, validated and used to probe gill tissue from AGD-affected Atlantic salmon. The NCGD amoebae-specific probe bound AGD-associated amoebae while neither N. pemaquidensis nor N. branchiphila were associated with AGD-lesions. Together, these data indicate that NCGD amoebae are a new species, designated Neoparamoeba perurans n.sp. and this is the predominant aetiological agent of AGD of Atlantic salmon cultured in Tasmania, Australia.     

Movement rhythms in juvenile Atlantic salmon, Salmo salar L.

Nocturnal downstrean migration of juvenile Atlantic salmon is usually interpreted as increased locomotor activity. The frequency of downstream passages of 0–1 + salmon in an endless stream channel was greater by night than by day in both smoking and non-smolting fish in autumn and spring. Movement increased at dusk, and decreased after dawn. Mature male 1 + fish moved slightly less than immatures in October, but significantly more in November. Total movement frequency was lower at full moon than at other moon phases, and movement was reduced when the moon was up. Under turbid conditions by day, the threshold water velocity inducing nett downstream movement was 8.2 cm s⁻¹, and the relative velocity of fish swimming downstream was never more than one third that of fish holding station at the normal maximal flow of 25–30 cm s⁻¹.
At the end of their first growing season in October, fish which had been offered food continuously through 24 h did not differ in size from those fed by day only, but the latter were significantly larger than those offered food only at night.
We conclude that: (1) the fish fed actively by day, and not by night; (2) station-holding represented activity, and downstream nocturnal movement represented relative inactivity (displacement) which occurred on loss of visual orientation, hence migration resulted from reduced activity; (3) lack of displacement in early autumn has adaptive value for maturing fish, but not for non-spawners.     

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.     

Arsenobetaine in Atlantic salmon (Salmo salar L.): influence of seawater adaptation

Glycine betaine has been suggested to improve the maintenance of ionic and osmotic homeostasis during seawater adaptation in teleost fish. Arsenobetaine may also behave as an osmolyte, due to its structural similarity to glycine betaine. The influence of seawater adaptation on intestinal uptake and muscle accumulation of arsenobetaine in the teleost Atlantic salmon (Salmo salar L.) was investigated. Atlantic salmon (freshwater and seawater adapted) were given a single oral dose of arsenobetaine, which was absorbed over the intestine within 6 h after exposure. Seawater adapted Atlantic salmon had significantly higher levels of accumulated arsenobetaine in blood compared to the freshwater adapted salmon. However, seawater adaptation had no effect on the levels of accumulated arsenobetaine in muscle tissue. Similar retention of the administered dose was found in muscle tissue in both freshwater and seawater adapted salmon, with 49+/-6% and 50+/-10% retention after 144 h, respectively. Results indicate that muscle retention was not influenced by salinity in seawater adapting teleosts.     

Infestations by Gyrodactylus sp. of Atlantic salmon, Salmo salar L., in Norwegian rivers

Young Atlantic salmon, Salmo salar , brown trout, Salmo trutta , and Arctic char, Salvelinus alpinus , from eight rivers in North and Mid Norway were examined for Gyrodactylus. The fish were collected from 1975 to 1980. A Gyrodactylus salaris type was observed infecting salmon from six of these rivers. No trout or char were infected. A high frequency and intensity of infection of salmon were observed in all but one of the rivers surveyed. In the Saltdalselva, only one specimen of Gyrodactylus infecting one fish was observed. No subsequent mortality of salmon was observed in this river while there were signs of a high mortality of salmon in the other rivers. The salmon parr were more frequently attacked than the fry, the mortality of salmon seemed to have happened in most of the rivers one year after the first observations of Gyrodactylus were made. The mortality of salmon and the high infection rate of Gyrodactylus in these rivers appears unique, and as far as known to the authors there are no other described cases of mortality due to Gyodactylus in Atlantic salmon in natural waters. The reasons for the outbreak of Gyrodactylus in these rivers are not known. Two theories are discussed: one that the fish were weakened by environmental factors and the other that Gyrodactylus was introduced from some of the infected salmon hatcheries in Scandinavia.     

Periodicity in Atlantic salmon Salmo salar L. smolt migration

Behavioural and physiological mechanisms postulated for the control of downstream migration of Atlantic salmon smolts are reviewed briefly, and some new evidence is presented for their refusal to undergo sustained swimming. Although these mechanisms imply passive displacement as the primary means of emigration, it is likely that active components must also exist as the rates of travel of smolts through loch systems are only slightly slower than those recorded for river systems. The timing of these movements within 24 h periods is reviewed and it is shown that the predominantly nocturnal emigration pattern is evident on occasions in alevin, fry and parr stages also. Thus at migration the diel periodicity probably represents a seasonal locomotor rhythm which, under changed behavioural and physiological circumstances, results in downstream displacement.     

Energetics in Atlantic salmon (Salmo salar L.) parr fed elevated dietary cadmium

Atlantic salmon (Salmo salar L.) parr were reared for 4 months on experimental diets supplemented with Cd (0.5, 5, 25, 125, or 250 mg Cd x kg(-1)) to assess the long-term energetic changes based on the digestibility and biochemical deposition of the major dietary nutrients and to evaluate a maximum tolerable dietary toxicant concentration. Growth did not differ significantly (P>0.05) from the control groups. The biochemical composition of the carcass, but not the viscera, was negatively affected by dietary Cd exposure. The significant decreases in protein, lipid, and glycogen concentrations in the carcass (P<0.05, 25 mg x kg(-1) compared to control groups) caused a reduction in calculated whole-body energy content in fish fed 125 mg x kg(-1)compared to control groups. This reduction in calculated whole-body energy content was explained by a concurrent significant disturbance to the gastrointestinal function (measured as reduced digestibility). Only at the highest dietary Cd exposure (250 mg x kg(-1)), increased metabolic costs to cope with Cd toxicity was thought to contribute significantly to the reduction in carcass energy content. The most important factor effecting calculated total energetics was nutrient digestibility. Based on the logarithmic effective median concentration for reduced calculated energy digestibility (dietary Cd concentration corresponding to 50% reduction, EC50), the maximum tolerable dietary Cd concentration is 11 mg x kg(-1) diet.     

Phagocytosis by B-cells and neutrophils in Atlantic salmon (Salmo salar L.) and Atlantic cod (Gadus morhua L.)

Phagocytosis by fish cells has mostly been studied using adherent leucocytes, excluding suspended cells such as the majority of B-cells and neutrophils, but a recent study describes professional phagocytosis of latex beads and bacteria by B-cells from rainbow trout. In the present study, phagocytosis by B-cells and neutrophils from salmon and cod was studied. Leucocytes were isolated from peripheral blood (PBL) and head kidney (HKL). By flow cytometry analyses, proportions of MAb labelled cell populations with internalized fluorescent beads, as well as the number of beads within each cell, could be determined. Phagocytic capacity and ability were demonstrated in B-cells and neutrophils from salmon and cod. In salmon, B-cells had higher phagocytic ability than neutrophils in HKL, but not in PBL. For cod the phagocytic ability of B-cells were lower than for neutrophils in both HKL and PBL, but the phagocytic capacity of cod B-cells were higher than for neutrophils in both HKL and PBL. For salmon B-cells the phagocytic capacity was lower than or similar to neutrophils in HKL and PBL. The total phagocytic ability of leucocytes was different in the species studied. The highest phagocytic ability was observed in cod, showing similar values for PBL and HKL. Salmon PBL displayed about twice the phagocytic ability of cod PBL. There seemed to be some major differences between the two fish species concerning phagocytosis. In salmon, a rather large proportion of phagocytic leucocytes were phagocytic B-cells, indicating that B-cells may have an important function in particle clearance in this species. In cod, phagocytic leucocytes in HKL and PBL were mostly neutrophils, and only a small proportion of B-cells were phagocytic, supporting the more prominent role of innate immune functions in cod neutrophils.     

Biochemical genetics of the Atlantic salmon Salmo salar L.

In recent years, blood group and protein polymorphisms in Atlantic salmon have been investigated extensively with a view, primarily, to their use in identifying individuals of different spawning populations present in high seas fisheries. Erythrocyte antigens, haemoglobins, serum proteins and various tissue enzymes—mainly esterases and de-hydrogenases—have been studied by electrophoretic and immunological techniques. These studies are reviewed here for the first time.
Many of the protein systems exhibit multiple components and this fact, together with cytological evidence, indicates the occurrence of tetraploidy in the course of Salmonid evolution. The significance of a tetraploid origin in the evolution and ecological adaptation of Salmonids is discussed briefly.
Some protein systems studied exhibit phylogenetic variation, and analyses of phenotype ratios and allele frequencies indicate that the populations of different river systems are genetically distinct. Allele frequencies have not yet been shown to be stable from generation to generation however, and some of the factors likely to affect allele frequencies are discussed.
Different spawning populations can not be identified in high seas fisheries using these protein characters, although it may be possible to identify the continent of origin (N. America or Europe) of some individuals. Indeed, it has been proposed independently by two groups that North American and European populations of Atlantic salmon be assigned to different sub-species viz. S. s. americanus and S. s. europaeus respectively. The contradictory evidence on which these taxa are proposed is discussed, together with the evidence for other population groups proposed in the European part of the salmon's range. The possible role and future direction of studies on the biochemical genetics of salmon are outlined.     

Migration of smolts of Atlantic salmon (Salmo salar L.) and sea trout (Salmo trutta L.) in a chalkstream

Synopsis Migration of smolts in the River Piddle, Dorset, was studied over three years in relation to factors that could influence downstream movement. The river originates mostly from groundwater springs, resulting in stable flows and low turbidity except in very rainy weither. Fish were intercepted at the tidal limit in a fixed trap-net, and measurements of water temperature, discharge, turbidity, barometric pressure, rainfall and solar radiation taken nearby. Slightly increased turbidity and discharge following heavy rain initiated major movements during two nights of the total of 55 days studied. At other times large-scale movements rock place during sunny warm afternoons. Both solar radiation and water temperature were correlated with intensity and timing of movement. The pattern of migration is different from that reported on other rivers, reflecting the relatively stable flow regime of the chalkstream.