Estimation of infectious dose and viral shedding rates for infectious pancreatic necrosis virus in Atlantic salmon,Salmo salar L., post‐smolts

Infectious dose and shedding rates are important parameters to estimate in order to understand the transmission of infectious pancreatic necrosis virus (IPNV). Bath challenge of Atlantic salmon post‐smolts was selected as the route of experimental infection as this mimics a major natural route of exposure to IPNV infection. Doses ranging from 10² to 10^?4 50% end‐point tissue culture infectious dose (TCID₅₀) mL^?1 sea water were used to estimate the minimum infectious dose for a Scottish isolate of IPNV. The minimum dose required to induce infection in Atlantic salmon post‐smolts was <10^?1 TCID₅₀ mL^?1 by bath immersion (4 h at 10 °C). The peak shedding rate for IPNV following intraperitoneal challenge using post‐smolts was estimated to be 6.8 × 10³ TCID₅₀ h^?1 kg^?1 and occurred 11 days post‐challenge. This information may be incorporated into mathematical models to increase the understanding of the dispersal of IPNV from marine salmon sites.     

Suspected myocardial necrosis in farmed Atlantic salmon, Salmo salar L.: a field case

Arteriosclerosis of the coronary artery has been described as a 'fact of life' for Pacific and Atlantic salmonids due to the high prevalence in spawning fish. The lesions are believed to be the result of overstretching of the highly distensible bulbus arteriosus whereby the endothelium of the main coronary vessel becomes mechanically damaged and a smooth muscle proliferation ensues with resultant partial occlusion of the vessel. The physiological significance for the function of the heart has yet to be demonstrated, but experimental studies show that, for example, swimming performance is compromised in fish in which the coronary artery has been ligated. This paper describes a case of myocardial necrosis in harvest-size Atlantic salmon during and after transportation to the slaughterhouse. Mortality during this process reached 10% in some of the transports and affected fish showed characteristic signs of congestive cardiac failure. Histology revealed extensive myointimal proliferation in the coronary artery and patchy necrosis of the compact ventricular myocardium. Several unfavourable factors such as high water temperature, skeletal malformations and crowding all probably contributed to extra cardiac workload. To the best of our knowledge, this is the first reported field case showing a link between coronary lesions and severe cardiac pathology.     

Effect of hyperoxygenation and low water flow on the primary stress response and susceptibility of Atlantic salmon Salmo salar L. to experimental challenge with IPN virus

Intensive salmon smolt production normally includes reduced water flow and hyperoxygenation (added oxygen) of remaining water. There is little information on how different water quality parameters influence the fish health and the susceptibility to infectious diseases. The current experiment was carried out to evaluate if the combination of hyperoxygenation and reduced water flow (hyperoxic) can act as a chronic stressor to salmon in freshwater (FW) in such a way that it increases the susceptibility to IPN virus (IPNV) following seawater transfer. In FW, after 22 days of hyperoxic exposure plasma ion, TBARS and cortisol were measured. The cortisol levels were significantly (p = 0.011) higher in the hyperoxic group compared to controls maintained under normal oxygen saturation and water flow (normoxic), indicating chronic stress. Hyperoxygenation in FW caused decreased plasma [Cl⁻] compared to the normoxic group (p = 0.037), while [K⁺] tended to be higher in the hyperoxic group (p = 0.088). No significant differences were observed in plasma [Na⁺], total osmolality, TBARS or hematocrit, but there was a tendency towards a lower hct in the hyperoxic compared to the normoxic group. In SW the mortality was higher in the hyperoxic group challenged with IPNV (34%) compared to the normoxic group challenged with IPNV (20%) (p = 0.02), and no mortality was observed in the PBS injected fish. The challenged fish showed an overall increase in plasma cortisol day 8, 10, 12 and 14 post-challenge (p = 0.015, p = 0.000, p = 0.046 and p = 0.022 respectively). After SW transfer and challenge, plasma [K⁺] was elevated in both challenged groups, but no consistent trends were found for plasma [Cl⁻], [Na⁺] or total osmolality during the SW phase. There were no significant differences in the gene expression level of IFN 1, Mx and IL 1β prior to challenge, suggesting that the basic expression level of these genes were not affected by hyperoxygenation. IPNV was detected in kidney and pylorus, by immunohistochemistry, cell culture, and RT-PCR in head kidney. This experiment indicates that chronic stress induced by a combination of low water flow and hyperoxygenation increases the susceptibility to IPNV challenge.     

Adiposity and growth of post-smolt Atlantic salmon Salmo salar L.

The long‐term adipose homeostasis seen in mammals gives rise to a ‘lipostatic’ model in which signals produced in proportion to fat stores serve to regulate energy intake. An extension of this predicts an impact of these signals on growth; downregulation of feeding in animals with increased adiposity should result in reduced growth. This was tested by monitoring fat deposition and growth in Atlantic salmon Salmo salar L. provided with feeds that differed in fat content. Salmon parr (mass c. 20 g) were fed either high‐ (H: 34%) or low‐fat (L: 22%) feeds, based on either fish (F) or vegetable (V) oils for 6 months to create groups of fish that differed in adiposity (10–12% and 5–7% body fat) at parr–smolt transformation (mass c. 130 g). Fish fed the high‐fat feeds deposited more body fat, and this was confirmed by measurement of fat concentrations in the fillet, viscera and remaining carcass. The fish were then grown‐on in sea water (c. 35 g L^?1, 8 °C, 24L:0D) for 14 weeks while being fed either high‐ or low‐fat feed formulated with fish oil to give the following treatments: HF→ H, HF→ L, LF→ L, LF→ H, HV→ H, HV→ L, LV→ L, LV→ H. Although fish exposed to the various feed treatments did not differ markedly in growth rate (SGR range 1–1.14% day^?1) over the 14 weeks of rearing in sea water, the results were in general agreement with predictions from the ‘lipostatic’ model, i.e. fish with the greatest fat reserves after the parr–smolt transformation grew more slowly than fish that were ‘leaner’ at this time. This suggests that adiposity, or ‘fatness’, may exert a negative feedback on feeding in salmon, thereby having an influence upon growth.     

Further development of bithionol therapy as a treatment for amoebic gill disease in Atlantic salmon, Salmo salar L.

This study examined the efficacy of bithionol as a prophylactic or therapeutic oral treatment for Atlantic salmon (AS), Salmo salar , affected by amoebic gill disease (AGD). Furthermore, it explored the interaction of bithionol oral therapy with the current standard treatment (a freshwater bath for at least 3 h). The efficacy of three medicated feeds was determined in the trial by feeding AGD-affected AS at 1% body weight (BW) day⁻¹ either oil coated commercial feed (control) or prophylactic and therapeutic bithionol at 25 mg kg⁻¹ feed. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L⁻¹ and continued for 49 days post-exposure (PE). Bithionol when fed as a 2-week prophylactic or therapeutic treatment at 25 mg kg⁻¹ feed delayed the onset of AGD pathology and reduced the percentage of gill filaments with lesions. Administration of a 3-h freshwater bath at 28 days PE significantly reduced amoeba numbers to a similar level across all treatments; in contrast, gross gill score and percent lesioned filaments were reduced to different extents, the control having a significantly higher score than both bithionol treatments. Following the freshwater bath, clinical signs of AGD increased at a similar level across all treatments, albeit controls were significantly higher than the bithionol treatments immediately following freshwater treatment. This study demonstrated that bithionol at 25 mg kg⁻¹ feed, when fed as a 2-week prophylactic or a therapeutic treatment, delayed and reduced the intensity of AGD pathology and warrants further investigation as a treatment for AGD-affected AS.     

Incidence of hybridization between Atlantic salmon, Salmo salar L., and brown trout, Salmo trutta L., in Ireland

The substantial growth of the farmed salmon industry in Europe since the 1970s has highlighted concerns regarding the genetic impact of escaped farmed salmon on wild salmonid stocks. High incidences of salmon × trout hybrids have been recorded in rivers situated near intensive salmon farming in Norway and Scotland, which may be indicative of a breakdown in reproductive isolation between salmon, Salmo salar L., and brown trout, Salmo trutta L. In the present study, salmonid fry and 0+ parr were collected from rivers in western Ireland. Allozyme and minisatellite DNA analysis were carried out on fry to determine the frequency of F1 hybrids from 10 rivers located within 38 km of salmon farms and three rivers at least 80 km from salmon farms. A total of 49 hybrids were recorded from 4135 salmonid fry (frequency = 1.2%). Mitochondrial DNA analysis showed that all hybrids arose from Atlantic salmon female × brown trout male crosses. Hybrid parr were recorded from one of the low-risk rivers (1.0%), but were present in seven out of the 10 catchments located within 38 km of salmon farms, with frequencies ranging from 0.7% to 3.1%. The results of the present survey, which represents the first extensive record of the levels of salmon-trout hybridization in Ireland, are discussed in relation to the continued growth of salmon farming in this country.     

Allozyme heterozygosity and development in Atlantic salmon, Salmo salar

The development of Atlantic salmon (Salmo salar) parr was monitored during the first year in a fresh-water hatchery environment to test whether multilocus heterozygosity (computed for six allozyme loci: _s AAT-4^*, IDDH-1^*, IDDH-2^*, IDPH-3^*, _s MDH-3,4^* and_m MEP-2^*) is associated with growth (body mass, fork length) and maturity status (mature male parr). The length frequency distribution of population was unimodal in September, but was clearly bimodal by December. UMG fish tended to be more heterozygous and had higher growth rates than LMG fish. The heterozygosity of UMG fish and mature male parr (MMP) did not differ significantly but MMP showed lower growth rates than UMG. There were positive correlations between the number of heterozygotic loci and the fork length, weight or growth rates of individuals.     

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.     

Target spawning requirements for Atlantic salmon, Salmo salar L., in Newfoundland rivers

Abstract In recent years, there has been increasing pressure from user groups in Newfoundland and Labrador, Canada, to manage Atlantic salmon, Salmo salar L., on a river-specific basis. The requirement to maximize benefits to users while ensuring that conservation goals are met has resulted in an initiative to develop river-specific target spawning requirements. In Newfoundland, anadromous juvenile Atlantic salmon make widespread use of lacustrine habitat for rearing. Depending on the river, lacustrine habitat can account for a substantial proportion of total production. Therefore, target spawning requirements for a number of rivers in several management areas were derived in terms of the contribution of both riverine and lacustrine habitats. The approach adopted, and derivations of associated parameter values, are reviewed, and limitations and research needs are discussed.     

Digestibility of diets with precooked rye or wheat by Atlantic salmon, Salmo salar, L.

Four extruded diets, differing in grain supplementation, were fed to triplicate groups of Atlantic salmon ( Salmo salar , L.) with a mean weight of 0.5 kg in a digestibility trial. The diets contained 100 or 150 g kg⁻¹ of precooked rye or wheat, replacing fish meal. There were no significant differences in apparent digestibility coefficients (ADC) of protein, fat or starch which could be related to dietary carbohydrate source or inclusion level. The ADC of dry matter and energy were higher for the diets with wheat than for those with rye, and higher for the diets with wheat than for those with rye, and higher for the diets with 100 g precooked grain kg⁻¹ than for those with 150 g kg⁻¹. The mean ADC of starch was 52% for rye, and 53% for wheat.     

Presence of anti-Neoparamoeba sp. antibodies in Tasmanian cultured Atlantic salmon, Salmo salar L

Previous studies have indicated that when Atlantic salmon, Salmo salar L., are exposed to Neoparamoeba sp. the fish produce anti-Neoparamoeba sp. antibodies. It appears unlikely that these antibodies elicit any specific protection against amoebic gill disease (AGD) as fish with demonstrable activities have been affected by AGD. Experiments were conducted on Atlantic salmon cultured throughout Tasmania to assess the natural production of antibodies towards Neoparamoeba sp. Fish were sampled from areas where AGD was prevalent and from areas where there had been no reported cases. An enzyme-linked immunosorbent assay (ELISA) was used to measure anti-Neoparamoeba sp. antibody activities in serum. All fish from sea water had antibody activities greater than the negative control fish, including fish from areas with no reported cases of AGD. Time trial samples indicated that time after transfer to sea water did not appear to be a significant (P > 0.05) factor in antibody activity, however location was (P < 0.05). There was no agreement (corrected kappa value, 0.16) between the ELISA result and the isolation of Neoparamoeba sp. from the gills of the same fish. The results suggest that Atlantic salmon in seawater culture in Tasmania produce anti-Neoparamoeba sp. antibodies regardless of infection history, suggesting the presence of Neoparamoeba sp. in the environment.     

Occurrence of gyrodactylids on wild Atlantic salmon, Salmo salar L., in Danish rivers

Three hundred and one parr of wild salmon, Salmo salar L., from three river systems in Denmark were examined in December 2005 and August 2006 for gyrodactylid infections. The species determination was done by morphological analyses of the opisthaptoral hard parts and molecular analyses of the internal transcribed spacer (ITS) region using PCR restriction fragment length polymorphism (RFLP) and sequencing. Five species of the genus Gyrodactylus were found: Gyrodactylus derjavini, Gyrodactylus teuchis, Gyrodactylus truttae, Gyrodactylus arcuatus and Gyrodactylus salaris. The parasite species G. derjavini dominated over G. teuchis and G. truttae and only one specimen of G. arcuatus and one specimen of G. salaris were recovered. This specimen of G. salaris had the same mutation in the ITS-region as a previously described non-pathogenic Danish G. salaris variant. The prevalence of gyrodactylids on the salmon ranged from 0% to 100% but the mean abundance never exceeded 12.8.     

Genetic characterization of Streptococcus phocae strains isolated from Atlantic salmon, Salmo salar L., in Chile

Streptococcus phocae is a beta-haemolytic bacterium frequently involved in disease outbreaks in seals causing pneumonia or respiratory infection. Since 1999, this pathogen has been isolated from diseased Atlantic salmon, Salmo salar , causing serious economic losses in the salmon industry in Chile. In this study, we used different molecular typing methods, such as pulsed-field gel electrophoresis (PFGE), randomly amplified polymorphic DNA (RAPD), enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), repetitive extragenic palindromic PCR (REP-PCR) and restriction of 16S–23S rDNA intergenic spacer regions to evaluate the genetic diversity in S. phocae . Thirty-four strains isolated in different years were analysed. The S. phocae type strain ATCC 51973^T was included for comparative purposes. The results demonstrated genetic homogeneity within the S. phocae strains isolated in Chile over several years, suggesting the existence of clonal relationships among S. phocae isolated from Atlantic salmon. The type strain ATCC 51973^T presented a different genetic pattern with the PFGE, RAPD, ERIC-PCR and REP-PCR methods. However, the fingerprint patterns of two seal isolates were distinct from those of the type strain.     

Content of synthetic astaxanthin in escaped farmed Atlantic salmon, Salmo salar L., ascending Norwegian rivers

Abstract  Isomeric ratios of astaxanthin in eggs and alevins of Atlantic salmon, Salmo salar L., have proven useful in identifying female spawners of farmed origin, but the method underestimates the proportion of fish of farmed origin. The rate of underestimation was studied by analysing astaxanthin content in tissue of 55 farmed Atlantic salmon ascending two Norwegian rivers in the autumn of 1991. The astaxanthin content fell into two distinct classes. Fifty-one per cent of the adult escaped salmon had isomeric ratios similar to salmon fed synthetic astaxanthin, whereas all the remaining fish had ratios similar to wild fish. Discriminant analysis classified 96.4% of the fish with known astaxanthin content into the correct astaxanthin class on basis of tail-fin erosion, length, weight and gill-cover damage. This discriminant function was used to estimate the astaxanthin classification of 1017 farmed salmon caught in nine rivers during 1989–1991. The classification success varied among years from 52 to 64%. Corresponding numbers for females and males were 45–48% and 54–70%, respectively. Thus, estimates of spawning rates of farmed female salmon via astaxanthin content in eggs or alevins from redds should be adjusted accordingly. The observed isomeric ratios of astaxanthin in the escaped farmed salmon and the relationship with morphology indicates that a significant proportion of the escapees ascending rivers have spent more than 1 year in the wild after escape.     

Occurrence of canthaxanthin in Atlantic salmon, Salmo salar L., fry in Irish rivers as an indicator of escaped farmed salmon

The rapid growth of Atlantic salmon, Salmo salar L., culture in north-western Europe has given rise to concerns regarding the biological consequences of fish farm escapes on wild salmonid populations. Canthaxanthin, a carotenoid pigment additive to farmed salmon feed which is passed from females to their progeny, may be used as an indicator of the numbers of escaped farmed salmon which spawn in the wild. In the present study, thin-layer chromatography and high-performance liquid chromatography (HPLC) were used to screen emergent Atlantic salmon fry sampled from seven river catchments in Ireland for canthaxanthin. The incidence of fry containing canthaxanthin at greater than trace levels (<5% of total carotenoid pigment) was 0–4%, with an average of 1.7%, among the seven rivers sampled, indicating that the progeny of farmed salmon were present at similarly low frequencies. Canthaxanthin was detected at trace levels in an unexpectedly high proportion (35%) of salmon fry. Canthaxanthin was present at levels exceeding trace amounts in 24% of 21 non-anadromous brown trout, Salmo trutta L., sampled from six Irish rivers and present at trace levels in a further 57% of the fish, indicating that dietary canthaxanthin is freely available to salmonids in Irish rivers. The widespread presence of trace levels in salmon fry may be attributable, at least in part, to the increased sensitivity of the HPLC methods and to rapid dietary uptake during early post-emergence feeding.     

Development of a method to assess binding of astaxanthin to Atlantic salmon Salmo salar L. muscle proteins

Several methods were examined to characterize the binding between astaxanthin and salmon muscle protein(s) in order to provide tools for evaluation of the role of muscle proteins on astaxanthin retention in Atlantic salmon Salmo salar L. flesh. The methods included gel filtration chromatography, displacement of a hydrophobic probe and ultrafiltration. With gel filtration chromatography, aggregation of astaxanthin under the experimental conditions was a major problem for the separation of bound astaxanthin from free astaxanthin because the apparent molecular weight of aggregated astaxanthin or astaxanthin micelles was in the range of protein–astaxanthin complexes. Displacement of the fluorescent probe 8‐anilino‐1‐naphthalenesulphonate (ANS) was not effective as astaxanthin quenched the fluorophore so that displacement could not be observed. An ultrafiltration method was developed using 200‐mM sodium cholate for dispersion of astaxanthin aggregates. This allowed unbound astaxanthin to be separated from bound astaxanthin using a 30‐kDa filter. After salmon muscle proteins were solubilized in different fractions by sequential extraction using low ionic strength solutions, the astaxanthin binding of different fractions was assessed using the ultrafiltration method. The significant difference (P<0.05) observed in the astaxanthin binding of the various fractions suggests an application of this assay to detect differences in affinity of proteins for astaxanthin. The results also suggest that proteins other than actomyosin or actin can bind astaxanthin in Atlantic salmon flesh. This method can be used for the identification of astaxanthin‐binding proteins in salmon flesh and other tissues.     

Lake-use by juvenile Atlantic salmon (Salmo salar L.) and other salmonids in northern Norway

Abstract– The utilization of lakes, and inlet and outlet streams by juvenile Atlantic salmon ( Salmo salar L.), brown trout ( Salmo trutta L.) and Arctic charr ( Salvelinus alpinus (L.), were investigated in 16 watercourses northern Norway, all known to inhabit salmon stocks. In lakes, fish were caught by small mesh size gill nets, while in rivers fish were caught electrofishing. In the shallow littoral (0-3 m depth) there were juvenile salmon in 15 of 19 investigated lakes, juvenile trout in 17 and juvenile charr in seven. Trout dominated significantly in numbers in the shallow littoral of seven lakes, while salmon and charr dominated in three lakes each. When trout and salmon were frequent in the shallow littoral, charr was usually not present in this habitat, but were found in the profundal zone in most of the lakes. Atlantic salmon parr utilized both shallow and deep lakes, and used both stones and macrophytic vegetation as shelter. The utilization of lakes by salmon parr seemed to be closely related to utilization of small inlet streams for spawning. In most inlet and outlet streams salmon dominated over trout in numbers, while charr were absent. This is the first documentation of lake-use by naturally occurring salmon parr in Scandinavia.