Piscine reovirus (PRV) in wild Atlantic salmon,Salmo salar L., and sea‐trout,Salmo trutta L., in Norway

This is the first comprehensive study on the occurrence and distribution of piscine reovirus (PRV) in Atlantic salmon, Salmo salar L., caught in Norwegian rivers. PRV is a newly discovered reovirus associated with heart and skeletal muscle inflammation (HSMI), a serious and commercially important disease affecting farmed Atlantic salmon in Norway. A cross‐sectional survey based on real‐time RT‐PCR screening of head kidney samples from wild, cultivated and escaped farmed Atlantic salmon caught from 2007 to 2009 in Norwegian rivers has been conducted. In addition, anadromous trout (sea‐trout), Salmo trutta L., caught from 2007 to 2010, and anadromous Arctic char, Salvelinus alpinus (L.), caught from 2007 to 2009, were tested. PRV was detected in Atlantic salmon from all counties included in the study and in 31 of 36 examined rivers. PRV was also detected in sea‐trout but not in anadromous Arctic char. In this study, the mean proportion of PRV positives was 13.4% in wild Atlantic salmon, 24.0% in salmon released for stock enhancement purposes and 55.2% in escaped farmed salmon. Histopathological examination of hearts from 21 PRV‐positive wild and one cultivated salmon (Ct values ranging from 17.0 to 39.8) revealed no HSMI‐related lesions. Thus, it seems that PRV is widespread in Atlantic salmon returning to Norwegian rivers, and that the virus can be present in high titres without causing lesions traditionally associated with HSMI.     

Bacterial infections of Chinook salmon, Oncorhynchus tshawytscha (Walbaum), returning to gamete collecting weirs in Michigan

Herein, we describe the prevalence of bacterial infections in Chinook salmon, Oncorhynchus tshawytscha (Walbaum), returning to spawn in two tributaries within the Lake Michigan watershed. Ten bacterial genera, including Renibacterium, Aeromonas, Carnobacterium, Serratia, Proteus, Pseudomonas, Hafnia, Salmonella, Shewanella and Morganella, were detected in the kidneys of Chinook salmon (n = 480) using culture, serological and molecular analyses. Among these, Aeromonas salmonicida was detected at a prevalence of ～15%. Analyses revealed significant interactions between location/time of collection and gender for these infections, whereby overall infection prevalence increased greatly later in the spawning run and was significantly higher in females. Renibacterium salmoninarum was detected in fish kidneys at an overall prevalence of >25%. Logistic regression analyses revealed that R. salmoninarum prevalence differed significantly by location/time of collection and gender, with a higher likelihood of infection later in the spawning season and in females vs. males. Chi‐square analyses quantifying non‐independence of infection by multiple pathogens revealed a significant association between R. salmoninarum and motile aeromonad infections. Additionally, greater numbers of fish were found to be co‐infected by multiple bacterial species than would be expected by chance alone. The findings of this study suggest a potential synergism between bacteria infecting spawning Chinook salmon.     

Pancreas disease in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in Norway

The present paper describes, for the first time, clinical signs and pathological findings of pancreas disease (PD) in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in sea water in Norway. Similarities and differences with reports of PD from Ireland and Scotland are discussed. Samples of 68 rainbow trout from disease outbreaks on 14 farms and from 155 Atlantic salmon from outbreaks on 20 farms collected from 1996 to 2004 were included in the present study. The histopathological findings of PD in Atlantic salmon and rainbow trout in sea water were similar. Acute PD, characterized by acute necrosis of exocrine pancreatic tissues, was detected in nine Atlantic salmon and three rainbow trout. Salmonid alphavirus (SAV) was identified in acute pancreatic necroses by immunohistochemistry. Most fish showed severe loss of exocrine pancreatic tissue combined with chronic myositis. Myocarditis was often but not consistently found. Kidneys from 40% and 64% of the rainbow trout and Atlantic salmon, respectively, had cells along the sinusoids that were packed with cytoplasmic eosinophilic granules. These cells resembled hypertrophied endothelial cells or elongated mast cell analogues. Histochemical staining properties and electron microscopy of these cells are presented. SAV was identified by RT-PCR and neutralizing antibodies against SAV were detected in blood samples.     

Laboratory evaluation of sample collection methods (organs vs swabs) for Tasmanian salmon reovirus detection in farmed Atlantic salmon,Salmo salar L.

The use of swabs relative to organs as a sample collection method for the detection of Tasmanian salmon reovirus (TSRV) in farmed Tasmanian Atlantic salmon, Salmo salar L., was evaluated by RT‐qPCR. Evaluation of individual and pooled sample collection (organs vs swabs) was carried out to determine the sensitivity of the collection methods and the effect of pooling of samples for the detection of TSRV. Detection of TSRV in individual samples was as sensitive when organs were sampled compared to swabs, and in pooled samples, organs demonstrated a sensitivity of one 10‐fold dilution higher than sampling of pooled swabs. Storage of swabs at 4 °C for t = 24 h demonstrated results similar to those at t = 0. Advantages of using swabs as a preferred sample collection method for the detection of TSRV compared to organ samples are evident from these experimental trials.     

The use of a real-time PCR primer/probe set to observe infectivity of Yersinia ruckeri in Chinook salmon, Oncorhynchus tshawytscha (Walbaum), and steelhead trout, Oncorhynchus mykiss (Walbaum)

Yersinia ruckeri is the causative agent of enteric redmouth disease (ERM), a common pathogen affecting aquaculture facilities and implicated in large losses of cultured fish. Fisheries scientists continue to gain a greater understanding of the disease and the pathogen by investigating methods of identification and pre- and post-infection treatment. In this study, a real-time PCR probe set for Y. ruckeri was developed to detect daily changes in the bacterial load during pathogen challenges. Two species of fish, Chinook salmon, Oncorhynchus tshawytscha, and steelhead trout, Oncorhynchus mykiss, were exposed to two strains of Y. ruckeri (Hag and SC) during bath challenges. A subset of fish was killed daily for 14 days, and the kidney tissue was biopsied to enumerate copies of pathogen DNA per gram of tissue. While Chinook exposed to either the Hag or SC strains exhibited similar pathogen loads, those exposed to the Hag strain displayed higher mortality (～66%) than fish exposed to the SC strain (～24% mortality). Steelhead exposed to the Hag strain exhibited a greater pathogen load and higher mortality (～42%) than those exposed to the SC strain (<1% mortality). Steelhead challenged with either strain showed lower pathogen loads than Chinook. The study illustrates the efficacy of the probe set to enumerate Y. ruckeri bacterial growth in the kidneys of fish. Also, strains of Y. ruckeri display species-specific growth patterns that result in differential mortality and pathogen load.     

Monitoring infection with Piscine myocarditis virus and development of cardiomyopathy syndrome in farmed Atlantic salmon (Salmo salar L.) in Norway

An epidemiological study was carried out in Norway in 2015–2018, investigating the development of infection with Piscine myocarditis virus (PMCV) and development of cardiomyopathy syndrome (CMS) in farmed Atlantic salmon. Cohorts from 12 sites were followed and sampled every month or every other month from sea transfer to slaughter. PMCV was detected at all sites and in all sampled cages, and fish in six sites developed clinical CMS. The initial infection happened between 1 and 7 months post‐sea transfer, and the median time from infection with PMCV until outbreak of CMS was 6.5 months. Generally, fish from sites with CMS had higher viral titre and a higher prevalence of PMCV, compared to sites that did not develop clinical CMS. The virus persisted until the point of slaughter at most (11 out of 12) of the sites. The detection of PMCV in all sites suggests that PMCV is more widespread than previously known. Screening for PMCV as a tool to monitor impending outbreaks of CMS must be supported by observations of the health status of the fish and risk factors for development of disease.     

Cross-sectional study of histopathology and piscine orthoreovirus during a marine production cycle of farmed Atlantic salmon (Salmo salar L.) in British Columbia,Canada

Two cohorts of farmed Atlantic salmon, Salmo salar L., in British Columbia, Canada, were sampled for histopathology (nine organs) and piscine orthoreovirus (PRV-1) PCR after seawater entry at 2, 4, 6, 8, 10, 13, 16 and 19 months (20 fish per cohort per date). One cohort—from a PRV+ hatchery—remained PRV+ throughout the study (sample prevalence 80%–100%). In an adjacent pen, the other cohort—from a PRV− hatchery—was 0% PRV+ at 78 days, 30% PRV+ at 128 days and ≥95% PRV+ thereafter. Among sample cohorts that were ≥80% PRV+, median Ct values were nominally less among fish sourced from the PRV− hatchery (28.7–33.3) than the PRV+ hatchery (30.8–35.2). No microscopic lesions were associated with PRV Ct value (minimum = 25.6). About 3% of fish in both cohorts had moderate inflammatory heart lesions; among these fish, only one had skeletal muscle inflammation (mild), and PRV Ct values were similar to unaffected cohorts sampled the same day. Also, among 16 moribund or freshly dead fish sampled opportunistically during the study, 14 were PRV+, and none had significant inflammatory heart lesions. These data support the hypothesis that British Columbia PRV-1 does not contribute to mortality.     

Preliminary evaluation of purified lignin and hemicellulose as prebiotics candidates for Atlantic Salmon,Salmo salar L.

A study was conducted to investigate the potential of purified lignin and hemicellulose as prebiotics in diets for Atlantic salmon, Salmo salar L., postsmolt in seawater (30 ppt) at 14.9 ± 1.2°C. Triplicate groups of fish (initial individual body mass of 101.6 ± 1.2 g) were fed either the fishmeal-dense (32% of fishmeal) control diet (A) or one of the nine diets (15% fishmeal) supplemented with lignin:hemicellulose ratios (% diet:% diet) of 0.05:0 (B), 0.25:0 (C), 0.50:0 (D), 1.00:0 (E), 0:0.05 (F), 0:0.25 (G), 0:0.50 (H), 0.05:0.25 (I), and 0.25:0.50 (J) for 12 weeks in a complete randomized experimental design. At the end of the experiment, the thermal-unit growth coefficient (TGC) was significantly lower in salmon fed diets with 0.50% and 1.00% purified lignin than those fed the control diet, while there was no significant difference between the other treatments and the control diet (P < 0.05). The feed efficiency (FE) was significantly lower in salmon fed diets with 0.50% and 1.00% purified lignin and 0.05% hemicellulose than those fed the control diet, while there was no significant difference between the other treatments and the control diet (P < 0.05). The hepatosomatic index (HSI), intestinal villus length, and apparent digestibility coefficient (ADC) of nutrients in diets were similar (P > 0.05) among all the diets. This study demonstrated that inclusions of up to 0.25% of dietary purified lignin alone, and 0.25%–0.50% of dietary hemicellulose alone or in combination with dietary lignin can be considered as candidate prebiotics in Atlantic salmon nutrition.     

Climate effects on size‐at‐age and growth rate of Chinook Salmon (Oncorhynchus tshawytscha) in the Fraser River,Canada

Decline in size‐at‐age of Chinook Salmon (Oncorhynchus tshawytscha) has been observed for many populations across the entire Northeast Pacific Ocean, and identifying external drivers of this decline is important for sustainable management of these ecologically, economically, and culturally valuable resources. We assessed size‐at‐age of 96,939 Chinook Salmon sampled in the Fraser River watershed (Canada) from 1969 to 2017. A broad decline in size‐at‐age was confirmed across all population aggregates of Fraser River Chinook Salmon, in particular since year 2000. By developing a novel probability‐based approach to calculate age‐ and year‐specific growth rates for Fraser River Chinook Salmon and relating growth rates to environmental conditions in specific years through a machine learning method (boosted regression trees), we were able to disentangle multi‐year effects on size‐at‐age and thus identify environmental factors that were most related to the observed size‐at‐age of Chinook Salmon. Among 10 selected environmental variables, ocean salinity at Entrance Island in spring, the Aleutian Low Pressure Index and the North Pacific Current Bifurcation Index were consistently identified as important contributors for four of the seven age and population aggregate combinations. These top environmental contributors could be incorporated into future stock assessment and forecast models to improve Chinook Salmon fisheries management under climate change.     

Astaxanthin deposition in the flesh of Atlantic Salmon, Salmo salar L., in relation to dietary astaxanthin concentration and feeding period

Atlantic salmon, Salmo salar L., were fed nine experimental diets containing from 0 to 200 mg astaxanthin per kg^?1 for six time periods, ranging from 3 to 21 months, in sea cages at Matre Aquaculture Research Station, Matredal, Norway. The sampled fish had an initial mean weight of 115 g and reached a weight of 3.2 kg at the termination of the experiment. Every third month, 10 fish from each dose and time group were sampled and the astaxanthin concentration in the flesh determined. The amount of astaxanthin in the flesh ranged from 0.7 to 8.9 mg kg^?1 at the termination of the experiment. This paper discusses deposition of astaxanthin in the flesh of Atlantic salmon in relation to dietary carotenoid levels in the 0–200 mg kg^?1 range and feeding times of 3–21 months. Under the conditions of this experiment, no significant effect on astaxanthin deposition rate could be achieved by increasing the astaxanthin level above 60 mg kg dry feed^?1. Atlantic salmon should be fed astaxanthin-supplemented diets during the whole seawater stage in order to obtain maximal astaxanthin level in the flesh.     

Susceptibility of Chinook salmon,Oncorhynchus tshawytscha (Walbaum), and rainbow trout,Salmo gairdneri Richardson,to infection with Vibrio anguillarum following sublethal copper exposure*

Abstract. Chinook salmon, Oncorhynchus tshawytscha (Walbaum), and rainbow trout, Salmo gairdneri Richardson, were used to determine if sublethal copper exposure would increase their susceptibility to Vibrio anguillarum infection. Fish were pretreated with copper at fractional levels of the 96 h copper LC₅₀ before exposure to the pathogen. Mortality by vibriosis was greater among fish exposed to 9% of the copper LC₅₀ for 96 h than unexposed fish. Peak susceptibility to vibriosis depended in part on the interaction of exposure time and copper concentration. The higher copper concentrations produced peak susceptibility to infection in shorter time periods. After the peak of susceptibility, sensitivity to infection declined to near control levels in those fish where exposure was continued. Rainbow trout stressed by copper required about 50% fewer pathogens to induce a fatal infection than non-copper exposed fish.     

Morphological and physiological characteristics of Saprolegnia spp. strains pathogenic to Atlantic salmon, Salmo salar L

Seventeen strains of Saprolegnia spp. were examined for morphological and physiological characteristics, and seven were examined for their pathogenicity to Atlantic salmon, Salmo salar L. Two of the Saprolegnia strains tested caused 89 and 31% cumulative mortality in challenged salmonids and were significantly more pathogenic than the other strains tested. The positive control (Saprolegnia parasitica ATCC 90213) caused 18% mortality, but this was not significantly higher than non-pathogenic strains (0-3% cumulative mortality). All the pathogenic Saprolegnia strains and two non-pathogenic strains had secondary cysts with long, hooked hairs, a characteristic which is claimed to be typical of S. parasitica. This characteristic is apparently necessary, but does not in itself determine the ability to cause mortality in Atlantic salmon. However, all the pathogenic Saprolegnia strains in the present study showed a significantly higher initial growth rate of cysts in sterilized tap water than did non-pathogenic strains. The results of the present study suggest that initial growth rate of germinating cysts in pure water, together with the presence of long hooked hairs on the secondary cysts, may be indicators of pathogenicity of Saprolegnia strains to Atlantic salmon.     

Mortality related to pancreas disease in Norwegian farmed salmonid fish,Salmo salar L. and Oncorhynchus mykiss (Walbaum)

High mortality during the salmonid seawater phase is a continuous problem for the salmonid aquaculture industry, although the mortality levels show a large variation both in farms with a disease history and those without. We wanted to examine the mortality patterns in farms with pancreas disease (PD) and compare it to farms without this diagnosis. Further, we wished to investigate the factors influencing the maximum mortality in both groups. We examined data from all salmonid farms in Norway stocked after January 2003 and slaughtered before December 2007. In total, 1884 cohorts were included, and 150 of these were diagnosed with PD. We found that season accounted for more of the variation in mortality than water temperature in PD‐positive cohorts and that infection pressure influenced the mortality in non‐PD cohorts, suggesting outbreaks of disease that are not diagnosed. We also found that the mortality in PD cohorts decreased significantly from 2003 to 2007, suggesting that increased knowledge about PD and targeted actions have been effective. Our study further suggests that crude mortality figures may be of limited use when wanting to examine a particular disease and risk factors for increased mortality. We suggest farmers and legislation should turn to a more modern approach with cause‐specific mortality records.     

Reduction of brown trout, Salmo trutta L., salmon, S. salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), smolt bycatches in eel pound nets

Two types of bycatch reduction devices (BRDs) were tested in the Randers Fjord, Denmark, aimed at deflecting the surface-oriented smolts of brown trout, Salmo trutta L., salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), before they became trapped, while retaining the catches of eel, Anguilla anguilla (L.). The methods tested were: a) placing a floating guard net between the wings; and b) submerging the pot net (i.e. last enclosure before fyke net) 55–100 cm below mean sea level. After each haul the pound net-setups were changed from the control to one of the types of BRD, and vice versa. Both methods significantly reduced the bycatch of brown trout smolts, while catches of legal-sized eels were not affected. Submerging the pot net reduced smolt catches of all species; mean reductions were 91.1% for brown trout, 74.5% for rainbow trout, and 86.1% for salmon.     

Growth, feed utilization and health of Atlantic salmon Salmo salar L. fed genetically modified compared to non-modified commercial hybrid soybeans

The present paper represents a part of a major scientific effort aiming to reveal possible effects, nutritional or health related, of genetically modified (GM) feed ingredients for Atlantic salmon. For 3 months groups of post‐smolt Atlantic salmon were treated with diets holding 130 g kg¹ of the protein as soybean, one which contained 800 g kg^?1 GM type RR (Roundup Ready^?), and compared with a standard counterpart (commercial hybrid, not isogenic line) analysed to be non‐GM (nGM), and again compared with a standard fishmeal diet without soybean protein. All diets were composed to be within the category ‘compositional equivalent’ and held exactly the same proximate compositions, starch and sugar levels, above requirements for methionine and lysine, equal fatty acid profiles, vitamin, mineral and pigment contents. There was, however, a slight difference in levels of anti‐nutrients between the three diets. The various dietary treatments resulted in more than tripling of fish weight in all groups. In addition no significant differences in feed utilization, whole body, liver and muscle proximate compositions, and no significant differences in muscle fatty acid profiles were measured. The diet without soybean of either type resulted in greater retention of lipid, but equal retention of protein (protein productive value). The relative sizes of liver, kidney, head–kidney and brain were the same in all dietary groups, while the relative size of the spleen showed significant differences between fish fed the genetically modified soy diet compared with fish fed the nGM soybeans. Fish fed the soy diets of either type also showed a somewhat reduced mean erythrocyte cell volume. All other haematological values were equal between diet groups. The detoxification system, measured as glutathione peroxidase (GPx) and lysozyme activities, showed equal values for all groups when measured in plasma and liver or head–kidney. The distal part of the intestine showed reduced sizes as an effect of soybean additions, without any differences between the GM and nGM type. Our results showed high growth, no mortality, haematological values within normal ranges, and efficient and equal responses in the detoxification system. This was a first indication that up to 130 g kg^?1 RR‐soybean protein can safely be used in diets for Atlantic salmon. However, there is still a need to elucidate higher inclusion levels of GM feed ingredients, and why spleen index was reduced, and if this was a long‐ or short‐term effect.     

Feasibility of using rare earth elements (REEs) to mark and identify escaped farmed Atlantic salmon Salmo salar L.

A series of rare earth elements (REEs) were evaluated as potential markers in scales of Atlantic salmon 1⁺ and 0⁺ smolt by addition of these elements to the feed. Chlorides of 5 REEs were tested, alone or in combination: lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd) and dysprosium (Dy). The labelled‐diets contained 125 or 250 mg REE kg^?1 feed and were administered for up to 10 weeks. Fish scales were collected and analysed by inductively coupled plasma‐mass spectrometry (ICP‐MS) before start, 5 and 10 weeks after labelling started and finally 2 or 4 months after marking. The results demonstrated that the five tested elements were clearly incorporated into the scales of the fish fed the supplemented diets. Uptake and concentrations increased gradually during the feeding period, and no principal differences were found between the two smolt types. Combining 2 REEs in the same feed did not affect the incorporation of either of them. After a ‘dilution’‐period of 2–4 months the concentrations of all markers (except La) in the scales of the treated‐fish were still significantly higher than those of the untreated‐fish. The background concentrations of the tested markers and some other elements were also measured and found to give interesting information.     

Growth performance and organ development in Atlantic salmon, Salmo salar L. parr fed genetically modified (GM) soybean and maize

Although the inclusion of genetically modified (GM) plants in diets fed to fish is a contentious issue, there are few empirical data. The present study addressed nutritional value and potential risks of four maize types (two traditional and two GM maize varieties) and two soy types (one traditional and one Roundup Ready^® soy) included at moderate levels in diets fed to Atlantic salmon parr (initial mean weight ± SD; 0.21 ± 0.02 g) during the first 8 months of feeding (March to October), which included the parr‐smolt transformation. The GM‐maize varieties [Dekalb 1 (D1) and Pioneer 1 (P1)] were hybrids of traditional maize variants [Dekalb 2 (D2) and Pioneer 2 (P2)] and the GM maize MON810^®. Four maize diets, two soy diets and one Standard fishmeal‐based diet were formulated and fed to fish in triplicate. The maize diets were formulated with 121 g kg^?1 GM maize (P1 and D1) or 121 g kg^?1 of the traditional untransformed line (nGM‐maize, P2 and D2) and the soy diets were formulated with 125 g kg^?1 GM soy or 125 g kg^?1 of the traditional untransformed line (nGM‐soy), all of equivalent nutrient composition. All diets supported good growth and showed no evidence of diet‐related mortality. Based on samplings every 6th week, growth was within the normal range and at conclusion of the study body weight did not differ among any of the treatments (range 101–116 g). Besides minor differences on heptatosomatic index (HSI), plasma triacylglycerol (TAG) values and thermal growth coefficient (TGC), body composition, relative organ weights, plasma nutrient concentrations and enzyme activities did not vary among treatments at any sampling. The present findings indicate that the inclusion of GM plants at the given level in salmonid diets poses little, or no, adverse risk to the health of first feeding Atlantic salmon parr and promote normal growth. The paper presents the production related data of this feeding study. Results regarding structure and function of intestinal segments and intestinal organs are presented in Sanden et al.     

Detection and antigenic characterization of salmonid alphavirus isolates from sera obtained from farmed Atlantic salmon, Salmo salar L., and farmed rainbow trout, Oncorhynchus mykiss (Walbaum)

A simple method of detecting the presence of the salmonid alphaviruses (SAVs), salmon pancreas disease virus (SPDV) and sleeping disease virus (SDV), from serum samples is described. Using a 96-well tissue-culture plate format, test sera are diluted in medium and added to chinook salmon embryo (CHSE-214) cells. After incubation for 3 days at 15 degrees C, plates are fixed and stained using a monoclonal antibody (mAb)-based immunoperoxidase (IPX) detection system, and virus-infected cells are observed microscopically by white light. Application of this screening test, which is now used routinely in our laboratory in conjunction with an IPX-based virus neutralization (IPX-VN) test for detecting antibodies to SAVs, has resulted in the recovery of 12 additional isolates from salmon sera and four additional isolates from trout sera. A low level of antigenic variation was detected when these SAV isolates were investigated by indirect immunofluorescence using a panel of mAbs raised to reference SPDV and SDV isolates.