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.     

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.     

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.     

Vibrio cortegadensis sp. nov., isolated from clams

A group of four strains isolated from clams (Venerupis decussata and Venerupis philippinarum) in Galicia (NW Spain) were subjected to a polyphasic characterization, based on the phenotypic characteristics, the analysis of chemotaxonomic features, the sequencing of the 16S rRNA and five housekeeping (atpA, pyrH, recA, rpoA and rpoD) genes, as well as DNA–DNA hybridization (DDH). The analysis of the phenotypic and chemotaxonomic characteristics and the results of a phylogenetic study, based on the 16S rRNA gene sequence analysis and multilocus sequence analysis, clearly indicated that these strains belong to the genus Vibrio and were allocated between the Splendidus and Anguillarum clades showing a close relationship with the type strains of Vibrio tapetis (98.8 %), Vibrio pomeroyi (98.0 %) and Vibrio crassostreae (97.9 %). DNA–DNA hybridization results confirmed that these isolates constitute a new species. The name Vibrio cortegadensis sp. nov. is proposed with C 16.17^T (=CECT 7227^T=LMG 27474^T) as the type strain.     

Cranial nodules in farmed Atlantic salmon, Salmo salar L., fry

Cranial nodules are described from Atlantic salmon, Salmo salar fry hatched from certain egg batches. Conspicuous, smooth, mostly unilateral nodules were recorded on the cranium. Histologically, the cerebellum appeared normal, but was displaced dorsally. Karyorhectic malpighian cells were absent from the epidermis and the meninges appeared normal. There was no apparent contact of the cerebellum with the water, or evidence of fungal, bacterial or parasitic infection. Mortality of up to 15% as recorded. The aetiology of this condition is currently unknown.     

Cytogenetic studies of Atlantic salmon, Salmo salar L., in Scotland

Chromosome numbers for Atlantic salmon, Salmo salar L., from ten populations in Scotland were ascertained. The majority of fish had 2n = 58, NF = 74 karyotypes, and no polymorphisms between populations were found. The findings suggest that Atlantic salmon in Scotland are cytogenetically homogeneous.     

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.     

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.     

Antigenic and molecular characterization of Vibrio ordalii strains isolated from Atlantic salmon Salmo salar in Chile

Biochemical, serological and molecular properties of a group of 14 Vibrio ordalii strains isolated from cultured Atlantic salmon Salmo salar in Chile in recent years were studied. The characteristics of isolates were compared with the type strain V. ordalii ATCC 33509T. The Chilean V. ordalii represented a biochemically homogenous group; however, some minor differences with the type strain were observed. The serological relationships among isolates, as well as the study of their antigenic determinant (LPS) revealed a strong reaction with antisera raised against Atlantic salmon strains and the antiserum raised against Listonella anguillarum serotype O2. However, LPS electrophoretic patterns were completely different from the V. ordalii type strain, regardless of the serum employed, suggesting the possibility that the Chilean strains constitute a new serological subgroup within this bacterial species. Genetic analyses by PFGE, RAPD, REP-PCR and ERIC-PCR demonstrated that all V. ordalii strains were genetically homogenous, displaying similar DNA patterns, regardless of the techniques used. Moreover, the analysis of DNA banding patterns generated by ERIC-PCR and REP-PCR also clearly separated the type strain from the Chilean strains. This is the first report of characterization of V. ordalii strains from the Southeastern Pacific area, the results of which should facilitate the development of vaccines for protecting cultured Atlantic salmon against vibriosis in this area.     

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.     

Growth control of Saprolegnia from UDN-infected Atlantic salmon Salmo salar L.

Nine antiseptic compounds, at concentrations ranging from O.1 to 0.0001 %, were tested in vitro for their ability to inhibit the growth of Saprolegnia . This fungus was isolated from suspected UDN-infected Atlantic salmon. Malachite green was the most effective antiseptic. Fungal growth was completely inhibited at all the concentrations tested.