Plasma protein changes in Atlantic salmon, Salmo salar L., with infectious salmon anaemia

Moderate to severe anaemia and hypoproteinaemia were reported in a Canadian outbreak of 'haemorrhagic kidney syndrome' in Atlantic salmon, later shown to be caused by a variant of infectious salmon anaemia virus (ISAV). The progressive anaemia associated with ISA has been previously reported, but hypoproteinaemia in salmon infected with European isolates of ISA virus has not been well documented. The present study showed a very significant positive correlation between decreasing haematocrit values and total plasma protein concentrations in Atlantic salmon infected with two Canadian and two Norwegian ISA viral isolates. However, variations in the concentration of individual plasma proteins, typical of acute phase responses in higher vertebrates, were not observed.     

Comparing the behavioural thermoregulation response to heat stress by Atlantic salmon parr (Salmo salar) in two rivers

Climate change is expected to increase the frequency and magnitude of extreme thermal events in rivers. The Little Southwest Miramichi River (LSWM) and the Ouelle River (OR) are two Atlantic salmon (Salmo salar) rivers located in eastern Canada, where in recent years, water temperatures have exceeded known thermal limits (~23°C). Once temperature surpasses this threshold, juvenile salmon exploit thermal heterogeneity to behaviourally thermoregulate, forming aggregations in coolwater refuges. This study aimed to determine whether the behavioural thermoregulation response is universal across rivers, arising from common thermal cues. We detailed the temperature and discharge patterns of two geographically distinct rivers from 2010 to 2012 and compared these with aggregation onset temperature. PIT telemetry and snorkelling were used to confirm the presence of aggregations. Mean daily maximum temperature in 2010 was significantly greater in the OR versus the LSWM (p = 0.005), but not in other years (p = 0.090–0.353). Aggregations occurred on 14 and 9 occasions in the OR and LSWM respectively. Temperature at onset of aggregation was significantly greater in the OR (T_onset = 28.3°C) than in the LSWM (T_onset = 27.3°C; p = 0.049). Logistic regression models varied by river and were able to predict the probability of aggregation based on the preceding number of hours >23°C (R² = 0.61 & 0.65; P₅₀ = 27.4°C & 28.9°C; in the OR and LSWM respectively). These results imply the preceding local thermal regime may influence behaviour and indicate a degree of phenotypic plasticity, illustrating a need for localised management strategies.     

Fluoride retention of Atlantic salmon (Salmo salar) fed krill meal

The present experiment was performed to study how fluoride from krill meal enriched muscle, whole fish and bone of adult Atlantic salmon (Salmo salar) reared in sea water. Atlantic salmon (mean weight 0.5 kg) were divided into four triplicate groups and fed a commercial fish meal based diets with 0, 100, 200 and 300 g krill kg^?1 feed, respectively, for 12 weeks. The fluoride concentrations in the experimental feeds were analysed to be 18, 132, 235 and 358 mg kg^?1, respectively. Growth, mortality and feed efficiency were recorded through the experiment. Fluoride concentration was measured in muscle, whole‐body, and bone initially and after 12 weeks of feeding. The fluoride concentrations in the samples were determined by alkali fusion and fluoride ion‐selective electrode. Growth, mortality and feed efficiency ratio were not affected by the dietary treatments. The results showed that fluoride concentration in muscle, whole body and bone were not affected by the dietary fluoride level. The fluoride concentration in the tissues showed great variation among replicates of the group given the same diet. Fillets of the fish varied between 0.3 and 1.4 mg fluoride kg^?1 wet weight, whereas the whole‐body concentration of fluoride varied between 3.3 and 6.1 mg kg^?1 wet weight and the fluoride bone concentration varied between 5.8 and 7.2 mg kg^?1 fresh weight. These results suggest that Atlantic salmon are highly tolerant of dietary fluoride given as krill meal with concentration of fluoride up to 350 mg kg^?1 diet, and that accumulation of fluoride from feeding diets containing krill meal does not lead to tissue accumulation in the fish, at least over a short period of time.     

Transport of alpha-tocopherol in Atlantic salmon (Salmo salar) during vitellogenesis

The transport of α-tocopherol was studied during vitellogenesis in Atlantic salmon that were fed diets with two levels of α-tocopherol. α-Tocopherol levels were measured in the flesh, liver, ovary and serum, and in the serum the α-tocopherol levels in the very low density lipoprotein (VLDL), low density lipoprotein (LDL), high density lipoprotein (HDL) and very high density lipoprotein (VHDL or vitellogenin) were also measured. Atlantic salmon store α-tocopherol mainly in their flesh because the muscle mass comprises 50% or more of live weight. During vitellogenesis the α-tocopherol content declined to about 10% of the level prior to maturation. The relative range of level of α-tocopherol in the lipoproteins was: HDL> LDL> VLDL> VHDL, irrespective of dietary levels of α-tocopherol. From the recent knowledge on lipid transport during vitellogenesis and the present data, we hypothesize that α-tocopherol is transported from peripheral tissues to liver by HDL and further transported from liver to ovary by LDL. Vitellogenin appears to play a minor role in the transportation of vitamin E to the ovary.     

Storage of lipids in the myosepta of Atlantic salmon (Salmo salar)

Storage of lipids in the myosepta of Atlantic salmon (Salmo salar) was determined by dissection of myosepta from both white and dark muscle, by analyzing lipid classes of myosepta, and by analysis of muscle tissues with and without myosepta. By using Iatroscan-TLC/FID it was found that triglyceride was the only major lipid class present in the myosepta. In a sample pooled from three fish, 39.1% of white muscle lipids were found to be stored in the associated myosepta. In the dark muscle up to 62.4% of the total lipids were located in the associated myosepta.After removal of the myosepta, the ratio of the total lipid content of dark muscle to white muscle decreased from 5.25 to 3.66. The proportion of polar lipids increased in both white muscle and dark muscle after removal of the myosepta; however the ratio of phosphatidyl choline (PC) to phosphatidyl ethanolamine (PE) in both muscle types remained constant. This suggests that neutral lipids accounted for most of the lipids in the myosepta. It is concluded that this type of connective tissue, not the actual muscle fibres, stores most of the muscle lipids in Atlantic salmon.     

Signal crayfish (Pacifastacus leniusculus) predation upon Atlantic salmon (Salmo salar) eggs

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Seawater transmission and infection dynamics of pilchard orthomyxovirus (POMV) in Atlantic salmon (Salmo salar)

The Tasmanian salmon industry had remained relatively free of major viral diseases until the emergence of pilchard orthomyxovirus (POMV). Originally isolated from wild pilchards, POMV is of concern to the industry as it can cause high mortality in farmed salmon (Salmo salar). Field observations suggest the virus can spread from pen to pen and between farms, but evidence of passive transmission in sea water was unclear. Our aim was to establish whether direct contact between infected and naïve fish was required for transmission, and to examine viral infection dynamics. Atlantic salmon post‐smolts were challenged with POMV by either direct exposure via cohabitation or indirect exposure via virus‐contaminated sea water. POMV was transmissible in sea water and direct contact between fish was not required for infection. Head kidney and heart presented the highest viral loads in early stages of infection. POMV survivors presented low viral loads in most tissues, but these remained relatively high in gills. A consistent feature was the infiltration of viral‐infected melanomacrophages in different tissues, suggesting an important role of these in the immune response to POMV. Understanding POMV transmission and host–pathogen interactions is key for the development of improved surveillance tools, transmission models and ultimately for disease prevention.     

Protein-nitrogen flux and protein growth efficiency of individual Atlantic salmon (Salmo salar L.)

Protein-nitrogen flux (the proportions of consumed and absorbed protein-nitrogen partitioned into protein synthesis and growth) was examined in Atlantic salmon, Salmo salar L. Salmon were held in groups and fed high or low rations or starved. Individual food consumption rates were measured using radiography. Fish varied widely in protein growth efficiency (protein growth divided by protein consumption), but this did not correlate with consumption rate, digestive capacity (as measured by absorption efficiency, trypsin levels and pyloric caecal size) or feeding hierarchy rank. Protein synthesis rates, measured in whole-animals, were linearly correlated with protein consumption and assimilation. There was a significant correlation between protein growth efficiency and the efficiency of retention of synthesised proteins. The capacity for protein synthesis and RNA activity were positively correlated with rates of food consumption and growth but were not correlated with protein growth efficiency. It was concluded that individual differences in protein growth efficiency related to differences in synthesis retention efficiency, but not to differences in the capacity for protein synthesis, RNA activity, digestive capacity or feeding hierarchy rank.     

Control of phosphorus homeostasis of Atlantic salmon (Salmo salar) in fresh water

Studies were conducted to determine the absorption, excretion and requirement of dietary phosphorus (P) by Atlantic salmon (Salmo salar). Triplicate groups of salmon parr, initial weight 15 ± 0.5±g , were fed, diets containing 4, 5, 6, 7, 8, 9, 11 and 13 mg P and 20 KJ of digestible energy (DE) per±g of diet (dry matter basis, DMB) to satiation for 16 weeks. The basal diet containing 4 mg P g^-1 (0.15 mg available P per KJ DE) was supplemented with graded levels of calcium phosphate, Ca(H₂PO₄)²H₂O to formulate the eight experimental diets. The fish were reared in fresh water at a temperature of 15 °C on a 12 h photoperiod. Vertebrae ash increased from 316 to 516 mg g^-1 fat-free dry matter with an increase in dietary P content. P requirement was estimated by using a four-parameter sigmoidal equation. The data suggests that a diet of 0.28 mg available P per KJ DE is needed for Atlantic salmon. Phosphate and calcium levels in plasma and bone increased, whereas levels of magnesium and liver cholecalciferol decreased, with an increase in dietary P.Phosphate excretion in urine and apparent availability of P were determined in deficient and replete fish. In deficient fish, the urine phosphate concentration was 0.10 mmol L^-1 before feeding and 0.25 mmol l^-1 after feeding, whereas in replete fish these concentrations were 1.09 and 5.11 mmol l^-1, respectively. The increase in urine phosphate concentration was higher in replete fish than in deficient fish, however, the apparent absorption of P was found to be significantly lower in replete fish than in deficient fish. These results suggest that similarly to terrestrial vertebrates, P homeostasis in Atlantic salmon is controlled by absorption in the intestine, conservation in the kidney and storage in the bones.     

Hydrolysed fish protein concentrate (FPC) reduces viscera mass in Atlantic salmon (Salmo salar) fed plant‐protein‐based diets

Atlantic salmon fed diets devoid of fishmeal but added 0.5 g  kg^?1 fish protein concentrate (FPC) showed reduced growth and lipid deposition without affecting protein accretion as compared to fish fed a fishmeal‐based control diet. The aim of the current study was to assess whether higher inclusion of FPC improved the growth and lipid deposition of Atlantic salmon (initial body weight 380 g) fed high plant protein diets. Quadruplicate groups of fish were fed diets containing 200 g kg^?1 fishmeal of which was replaced with FPC (150, 112, 75, 38 and 0 g kg^?1) for a period of 79 days. The rest of the diet protein was a mixture of plant proteins. The lipid source used was fish oil. A fishmeal‐based diet was included as a positive control for growth performance. None of the test diets differed from the positive control‐fed fish in voluntary feed intake, growth performance or nutrient accretion. Thus, the test diets were found appropriate to assess the effect of FPC inclusion. Replacement of fishmeal with increasing concentration of FPC did not affect voluntary feed intake (P = 0.56), but growth performance decreased (P = 0.02) resulting in an increased feed conversion ratio (P = 0.003). Viscerosomatic index decreased as diet FPC inclusion increased (P = 0.012) without affecting the dress out weight (P = 0.08). Thus, the apparently improved growth in fish fed the diets with the low FPC inclusion was because of a higher visceral mass. Possible reasons for the reduced visceral mass following addition of FPC to high plant protein diets are discussed.     

Apoptosis inhibition of Atlantic salmon (Salmo salar) peritoneal macrophages by Piscirickettsia salmonis

To improve the understanding of the piscirickettsiosis pathogenesis, the in vivo apoptosis modulation of peritoneal macrophages and lymphocytes was studied in juvenile Salmo salar intraperitoneally injected with Piscirickettsia salmonis. Five fish were sampled at post‐exposure days 1, 5, 8 (preclinical), 20 (clinical) and 40 (post‐clinical period of the disease), and the leucocytes of their coelomic washings were analysed by flow cytometry (using the JC‐1 cationic dye), TUNEL and cytology to detect apoptotic cells. A selective and temporal pattern of apoptosis modulation by P. salmonis infection was observed. Apoptosis in lymphocytes was not affected, whereas it was inhibited in macrophages but only during the preclinical stage of the induced piscirickettsiosis. Hence, it is postulated that P. salmonis inhibits macrophage apoptosis at the beginning of the disease development to survive, multiply and probably be transported inside these phagocytes; once this process is complete, macrophage apoptosis is no longer inhibited, thus facilitating the exit of the bacteria from the infected cells for continuing their life cycle.     

Selective segregation in intraspecific competition between juvenile Atlantic salmon (Salmo salar) and brown trout (Salmo trutta)

Interactive segregation has been suggested as the ruling competition mechanism determining niche and niche segregation between juvenile Atlantic salmon (Salmo salar) and brown trout (Salmo trutta). Results from allopatry–sympatry observations of habitat use in both nature and in experiments were contrary to predictions derived from the interactive segregation hypothesis. Habitat use parameters under natural conditions such as distance to shore for Atlantic salmon parr were nearly identical in allopatric (mean ± SD; 3.2 ± 1.4 m) and sympatric (3.3 ± 1.4 m) situations. Occupied water depths largely reflected available water, but water depths <15 cm were avoided by salmon parr. Under experimental conditions, habitat use of allopatric salmon was density independent and salmon size had only minor effects, with smaller fish being more likely to occur in the shallow. Habitat use of salmon in sympatry with trout did not differ from allopatric salmon habitat use, and only salmon size had minor effects on depth choice – occurrence of trout or fish density had no effect. Allopatric trout was in general more frequent in the shallow habitat than salmon. Habitat use of sympatric trout was affected by the occurrence of salmon and trout size, resulting in a higher use of the shallow habitats for small trout. To conclude, selective segregation has a dominant role in salmon habitat use (not affected by trout occurrence), whereas a mixed situation occurs in trout habitat use with elements of interactive segregation when competing with Atlantic salmon (affected by salmon occurrence).     

Effects of dietary (n-3) and (n-6) polyunsaturated fatty acid ratio on the immune response of Atlantic salmon, Salmo salar L.

To examine the influence of the dietary ratio of (n-3) to (n-6) polyunsaturated fatty acids (PUFAs) on the immune system of Atlantic salmon, Salmo salar L., two dietary trials were carried out in which parr were maintained on diets containing either fish oil [(n-3)/(n-6) PUFA = 5.2] or sunflower oil [(n-3)/(n-6) PUFA = 0.3] and assessed for differences in immunological parameters. There were no significant differences in blood cell counts, differential leucocyte counts or haematocrit values between dietary groups, and while no apparent differences were observed in the non-specific immune parameters measured, there was a significantly higher number of B cells responding to Aeromonas salmonicida, in the kidney and spleen of vaccinated fish maintained on high (n-3)/(n-6) PUFAs diets. There was also a significant difference (P≤ 0.01) between the dietary groups in trial 1 and trial 2 when non-vaccinated fish were challenged with Aeromonas salmonicida and Vibrio anguillarum, respectively, with the (n-6) group succumbing to the bacterium before the (n-3) group. The results suggest that Atlantic salmon fed diets with a low ratio of (n-3)/(n-6) PUFA may be less resistant to infection than those fed diets containing lipid with a high (n-3)/(n-6) PUFA ratio.     

Expression of insulin-like growth factor-I in the gastrointestinal tract of Atlantic salmon (Salmo salar L.)

The expression of insulin-like growth factor (IGF-I) in the gut of Atlantic salmon (Salmo salar L.) smolt was investigated at the cellular level using in situ hybridization. A hybridization protocol was adapted for use on formalin fixed, paraffin embedded tissues and was based on a digoxigenin-labelled oligoprobe that recognised a conserved region of IGF-I mRNA. The specificity of the anti-sense IGF-I probe and the performance of the hybridization protocol were evaluated using a sense IGF-I probe, an Ep stein-Barr virus probe and an insulin probe cocktail on serial sections of fish gut, mouse thyroid and human lymph node tissue. In the gut of Atlantic salmon smolt, IGF-I was found to be expressed in single epithelial cells or small clusters of epithelial cells in the pyloric ampulla and in the pyloric caeca. Expression was not detected in the lamina propria of the gut or in epithelial cells of the stomach, midgut or hindgut. The restriction of IGF-I expression to the pyloric ampulla and pyloric caeca is consistent with the role of these gut segments in osmoregulation and the scattered epithelial expression supports the putative autocrine/paracrine mechanism of action of IGF-I in osmoregulation.     

Effect of growth hormone on muscle protein synthesis in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar)

This paper reports on the effect of administration of mammalian growth hormone (GH) on muscle protein synthesis as measured in white muscle using the phenylalanine flooding technique. The effect of exogenous GH was compared with that of insulin and prolactin, and with endogenous GH.The rate of protein synthesis in white muscle of rainbow trout 6 h after the injection of bovine GH or bovine insulin was twice (2.6 and 2.9% d^–1) that of the control saline-injected fish (1.2% d^–1). A metabolic effect of GH, as observed with insulin, is suspected.The rates of change in body weight and body length and the fractional rate of protein synthesis in muscle of rainbow trout were enhanced by mammalian GH administration. The effect of GH on muscle RNA/protein ratios was not significant. An opposite effect of antibodies against salmon GH (Lebailet al. 1989) on growth rate and muscle protein synthesis rate was found in rainbow trout. It is suggested that the effects of exogenous and endogenous GH on capacity and efficiency of muscle protein synthesis were similar.The long-term effects of mammalian GH on presmolt Atlantic salmon was also tested. The same trends were found with ovine prolactin supplementation in Atlantic salmon but not as high as those observed with ovine GH.     

Dietary phosphorus requirement of juvenile Atlantic salmon, Salmo salar L.

The objective of this study was to determine the dietary phosphorus (P) requirement of juvenile Atlantic salmon, Salmon salar L. Triplicate groups of fish (mean initial weight 1.4 g) were fed semipurified, casein-gelatine-based diets containing one of five levels of P (4, 8, 10, 15 and 25 g kg⁻¹) from Ca(H₂PO₄)₂·H₂O, or a commercial feed (17 g kg⁻¹ P) for 9 weeks. Weight gains did not differ significantly among treatment groups fed the experimental diets but were slightly less than gains in fish fed the commercial feed. Feed efficiency (wet weight gain/dry feed consumed) was similar in all groups, averaging 1.45. Availability of dietary P, estimated from apparent retention and apparent digestibility, was 86%. Whole-body P concentrations declined in fish fed diets containing less than 10 g kg⁻¹ P. Fitting a logistic curve to dietary P vs. whole-body P concentrations indicated that a minimum of 11 g kg⁻¹ dietary P (9 g kg⁻¹ digestible P) was required by juvenile Atlantic salmon to maintain whole-body P concentrations at initial levels. Calculation of a dietary requirement using a simple factorial model which incorporated measurements of P availability, feed efficiency and normal whole-body P concentration indicated that the dietary requirement was approximately 10 g kg⁻¹. The dietary requirement established in this study (10–11 g kg⁻¹) is higher than previously reported for Atlantic salmon or other fishes. Possible reasons for the wide range of reported dietary P requirements in fishes are discussed.     

Infectious pancreatic necrosis in Atlantic salmon, Salmo salar L

A clinical and histopathological review was carried out of 21 outbreaks of acute infectious pancreatic necrosis (IPN) in Scottish Atlantic salmon, Salmo salar L., farms (13 marine and eight fresh water) during 1991-2004. A distinctive syndrome was evident in both post-smolts in sea water and fry in fresh water, where liver lesions, which had not previously been associated with IPN, became a consistent finding in addition to the more typical pancreatic and intestinal changes. Initial cases were described in post-smolts in Shetland, but by the end of the period of investigation this type of pathology had extended down the West coast of Scotland and into Ireland. Limited viral strain analysis suggested that similar strains were involved in both fresh water and sea water and that these differed from earlier isolates from rainbow trout, Oncorhynchus mykiss (Walbaum). In fresh water, recovered fish frequently developed a greatly distended intestine associated with accumulation of undigested food. In sea water, after the initial, often significant (50% or more), losses, there were many fish which failed to grow and became chronically emaciated and prone to sea louse infection. Although use of transfer diets containing immune enhancers and the selection of IPN resistant broodstock has reduced losses the disease remains a serious cause of economic loss.