Morphological and genetic comparison between naturally produced smolts of Atlantic salmon,brown trout and their hybrids

Morphological characters were compared between smolts (standard length 110–220 mm) of Atlantic salmon Salmo salar L., anadromous brown trout Salmo trutta L. and their hybrids captured in River Driva, western Norway. Morphological discrimination between S. salar, S. trutta and their hybrids was correct for 93% of the individuals (correct form was determined genetically). Morphological discrimination of hybrids was incorrect with 9.1% (n = 7) classified as S. salar and 1.3% (n = 1) as S. trutta, suggesting that the hybrids were morphologically more similar to S. salar than S. trutta despite a bias towards S. trutta mothers (7 of 8 cases). The largest morphological differences were found in head (notably maxilla length) and caudal peduncle morphology. Hybrids between S. salar and S. trutta had the longest pectoral and pelvic fins. This indicates that precise discrimination based on morphological traits can be made between S. salar, S. trutta and their hybrids.     

Aeromonas salmonicida infects Atlantic salmon (Salmo salar) erythrocytes

Aeromonas salmonicida subsp. salmonicida (hereafter A. salmonicida) is the aetiological agent of furunculosis in marine and freshwater fish. Once A. salmonicida invade the fish host through skin, gut or gills, it spreads and colonizes the head kidney, liver, spleen and brain. A. salmonicida infects leucocytes and exhibits an extracellular phase in the blood of the host; however, it is unknown whether A. salmonicida have an intraerythrocytic phase. Here, we evaluate whether A. salmonicida infects Atlantic salmon (Salmo salar) erythrocytes in vitro and in vivo. A. salmonicida did not kill primary S. salar erythrocytes, even in the presence of high bacterial loads, but A. salmonicida invaded the S. salar erythrocytes in the absence of evident haemolysis. Naïve Atlantic salmon smolts intraperitoneally infected with A. salmonicida showed bacteraemia 5 days post‐infection and the presence of intraerythrocytic A. salmonicida. Our results reveal a novel intraerythrocytic phase during A. salmonicida infection.     

Stress response of Salmo salar (Linnaeus 1758) facing low abundance infestation of Caligus rogercresseyi (Boxshall & Bravo 2000), an object in the tank,and handling

This study looks at how low infestation loads of adult Caligus rogercresseyi and other stressors affect the physiology of Salmo salar. Experimental fish groups were with (infested) or without (control) exposure to the parasite. The parasite cohort was followed for 78 days post‐infestation (dpi), and only adult lice were observed. Additional stressors were applied at 60 and 75 dpi. The analysis included measurements of fish physiology and weight. Low‐level infestations by adult C. rogercresseyi for more than 50 dpi induced moderate stress in S. salar as well as a high energy demand and increased small skin mucous cells. Threshold lice loads were identified, and above those loads, a high stress response was observed. Additional stressors altered fish physiology, inducing downregulation of the cortisol response after the first stressor and upregulation after the second stressor, but infested fish responded more strongly. Parasitism by C. rogercresseyi is energetically demanding, affecting the primary and secondary responses (e.g. cortisol and glucose levels), as well as the tertiary response (fish weight).     

Impacts of parasites on marine survival of Atlantic salmon: a meta‐analysis

Parasites can, in theory, have large impacts on the survival of fish populations. One method to evaluate such impacts on anadromous species is to apply manipulative field experiments in which parallel groups of antiparasitically treated and non‐treated fish are simultaneously released and then subsequently recaptured as returning adults. A systematic review and meta‐analysis on all such Norwegian studies on Salmo salar provided a data set for the time period 1996 to 2011 on 118 release groups comprising 657 624 fish released and 3989 recaptured. The overall risk ratio (RR) was estimated to be 1.18 (95% CI: 1.07–1.30). The effect varied strongly between groups, (Higgins I² = 40.1%). Over 70% of this heterogeneity could be explained by the release location, time period and baseline survival. The most important predictor variable was baseline survival. In groups with low recapture in the control group (low baseline survival), the effect of treatment was high (RR = 1.7), while in groups with high recapture in the control group (high baseline survival), there was no effect of treatment (RR ~ 1.00). The most prevalent parasite in the region affected by the drugs administered was Lepeophtheirus salmonis. Hence, the meta‐analysis supports the hypothesis that L. salmonis contributes to the mortality of S. salar during outward migration. However, the effect of treatment was not consistent, but was evidently strongly modulated by other risk factors. The results suggest that the population‐level effects of parasites cannot be estimated independently of other factors affecting the marine survival of S. salar.     

Serum enolase: a non‐destructive biomarker of white skeletal myopathy during pancreas disease (PD) in Atlantic salmon Salmo salar L.

Diseases which cause skeletal muscle myopathy are some of the most economically damaging diseases in Atlantic salmon, Salmo salar L., aquaculture. Despite this, there are limited means of assessing fish health non‐destructively. Previous investigation of the serum proteome of Atlantic salmon, Salmo salar L., during pancreas disease (PD) has identified proteins in serum that have potential as biomarkers of the disease. Amongst these proteins, the enzyme enolase was selected as the most viable for use as a biomarker of muscle myopathy associated with PD. Western blot and immunoassay (ELISA) validated enolase as a biomarker for PD, whilst immunohistochemistry identified white muscle as the source of enolase. Enolase was shown to be a specific marker for white muscle myopathy in salmon, rising in serum concentration significantly correlating with pathological damage to the tissue.     

Relationship between sea lice levels on sea trout and fish farm activity in western Scotland

The relationship between aquaculture and infestations of sea lice on sea trout, Salmo trutta L., is controversial. Here, the association between sea lice infestations on wild sea trout and characteristics of local Atlantic salmon, Salmo salar L., farms were investigated using data collected on the Scottish west coast. The proportion of sea trout with louse burdens above a critical level was positively related to the fork length of the sea trout and the mean weight of salmon on the nearest fish farm, and negatively related to the distance to that farm. The distance to the nearest fish farm did not influence the probability of infestations above the critical level beyond 31 km although there was considerable uncertainty around this cut‐off distance (95% limits: 13–149 km). The results support a link between Atlantic salmon farms and sea lice burdens on sea trout in the west of Scotland and provide the type of information required for marine spatial planning.     

Gaping and loss of fillet firmness in farmed salmon (Salmo salar L.) closely correlated with post‐slaughter cleaning of the abdominal cavity

This study analysed the effect of cleaning intensity of the abdominal cavity and storage temperature from slaughter to the end of processing on the quality of farmed salmon (Salmo salar L.) fillets. These two parameters were manipulated in an experimental setup using in total thirty salmon with an average weight of 4.2 kg. The experiment was designed to imitate realistic scenarios in a normal production process in the Faroe Islands. The salmon stored at low temperatures had an average muscle temperature of 4.65°C, whereas the salmon stored at ambient temperature had an average muscle temperature of 11.27°C. After the salmon were gutted to remove all viscera except the kidney, the abdominal cavity was either rinsed lightly or meticulously cleansed of kidneys, all blood and bodily fluids. A wide range of quality and production parameters were measured either straight after cleaning or after the salmon had been stored in chipped ice at 1.5°C for 7 days. All measured parameters were analysed for possible correlations by principal component analysis (PCA). Blood and remains left in the abdominal cavity were shown to have a significant negative effect on fillet firmness (P < 0.01) and gaping (P < 0.01). The different storage temperatures between slaughter and gutting, tested in this experiment, did not significantly affect fillet firmness or gaping. However, the fillet colour showed significant negative correlation (P < 0.01) with the storage temperatures applied.     

Muscle fiber types,growth and development in the whole myotome of cultured Pacific bluefin tuna Thunnus orientalis

Following the successful development of Pacific bluefin tuna (PBT) aquaculture, it is of considerable importance to determine the muscle fiber types and their growth patterns for future development. Muscle fiber profiles of dorsal ordinary, lateral ordinary (LO) and dark muscles and their growth patterns in PBT from 3.0 to 54.3 kg body weight were studied. Muscle fibers were histochemically stained for NADH-diaphorase and myosin adenosine triphosphatase activity (mATPase), and immunohistochemically stained with S-58 slow-muscle myosin antibody. All muscle fibers in dorsal and LO muscles showed low NADH-diaphorase activity, and acid-labile (pH 4.0 or 4.3) and alkali-stable mATPase activity. In LO muscle adjacent to dark muscle, three intensities of mATPase activity were observed after acid pre-incubation at pH 4.5 or 5.0, and the activity was related to the muscle fiber diameter. In dark muscle, all small and some large fibers stained intensely for NADH-diaphorase activity, related to their high aerobic metabolism. The high-active fibers with NADH-diaphorase in dark muscle were positive for S-58 antibody. Some large fibers in dark muscle showed intermediate NADH-diaphorase activity and high mATPase activity after alkali pre-incubations. These are fast-twitch oxido-glycolytic fibers in dark muscle and transformed to red muscle fibers with increasing body weight.     

Riverine and fjord migration of wild and hatchery‐reared Atlantic salmon smolts

Migration timing, speed, survival and effects of environmental parameters on migration, between wild and hatchery produced Atlantic salmon, Salmo salar L., smolts in the River Lærdalselva were studied. Hatchery‐reared (n = 40) and wild pre‐smolts (n = 40) were tagged with acoustic tags, and an array of receivers along the migration route was deployed. In all, 77 and 85% of the fish from the two groups, respectively, were recorded as migrating smolts, that is, predation rate and/or numbers of fish opting to remain in the river were low. Hatchery‐reared smolts showed a migration pattern, speed and migration route similar to wild smolts, even though the time period between river release and onset of migration was relatively short. Both groups of smolt showed high migration speed through both the river and the fjord compared with other studies.     

Effects of temperature and feed composition on essential fatty acid (n-3 and n-6) retention in Atlantic salmon (Salmo salar L.) parr

Retentions of total n-3 and n-6 essential fatty acids (EFAs) were assessed in Atlantic salmon (Salmo salar L.) parr held at 8 °C and 2 °C until they increased in weight from ca. 19 g to 38 g. Feeds contained sandeel oil or a rapeseed:linseed oil blend at 21 and 34% dietary fat. EFA retention efficiencies [(g EFA gained g EFA ingested-1) × 100] were estimated by the 'mass balance method' from measurements of feed intake, changes in biomass for each tank of fish, and fatty acid compositions of the feeds and fish. The n-3 EFA retentions were higher (overall mean 71%) across feed treatments and temperatures than the n-6 EFA retentions (overall mean 63%). Retentions of the n-3 fatty acids were higher in the fish given the feeds with the lower fat content (77% vs. 65%), implying improved retention with reduced n-3 EFA availability. n-3 EFA retention tended to be higher at 2 °C than at 8 °C, although this was not consistent across feeds. At low temperature there was very high retention of the n-3 EFAs in feeds containing sandeel oil (80%). Such high retention may represent an adaptation response to low temperature. Lower n-6 EFA retentions imply that more n-6 fatty acids were metabolized than n-3 EFAs. Feed oil influenced retention of the n-6 fatty acids, retention being lower for the salmon parr given the feeds containing sandeel oil (56% vs. 71%). This could indicate a higher tissue deposition of n-6 fatty acids when they are freely available via the diet. Abbreviations: AA – arachidonic acid (C20:4 n-6); DHA – docosahexaenoic acid (C22:6 n-3); EFA – essential fatty acid; EPA – eicosapentaenoic acid (C20:5 n-3); HUFA – highly-unsaturated fatty acid (\ge4 double bonds); MUFA – monounsaturated fatty acid (1 double bond); PL – phospholipid; PUFA – poly-unsaturated fatty acids (\ge2 double bonds); SFA – saturated fatty acid (no double bond); TAG – triacylglycerol. This revised version was published online in July 2006 with corrections to the Cover Date.     

Para-nitrophenol hydroxylation by fish liver microsomes: kinetics and effect of selective cytochrome P450 inhibitors

The study investigated the kinetics of p-nitrophenol hydroxylase (PNPH) in hepatic microsomes obtained from Atlantic salmon (Salmo salar). The selective inhibitors for some major mammalian cytochrome P450 (CYP450) were used to investigate the potential inhibitory effect on enzymes involved in p-nitrophenol hydroxylation. The following inhibitors were used: α-naphtoflavone (CYP1A), ellipticine (CYP1A1), furafylline (CYP1A2), 8-methoxypsoralen (8MOP, CYP2A6), 4-methylpyrazole (4MP, CYP2A6/2E1), diallyl sulfide (DAS, CYP2E1), and ketoconazole (CYP3A4). Additionally, the natural steroids 17-beta-oestraiol (E2) and testosterone were investigated as potential inhibitors of PNPH activity. It was found that formation of 4-nitrocatechol from p-nitrophenol followed monophasic kinetics with K _m  = 0.17 ± 0.03 mM and V _max = 21.8 ± 1.05 pmol/min/mg. PNPH activity was competitively inhibited by diallyle sulfide with the K _i value of 285.1 ± 94.2 μM μM and uncompetitively by ellipticine with K _i value of 65.7 ± 7.8 μM. Moreover, E2 showed an ability to reduce PNPH activity through the mechanism-based inhibition mode. Our results suggest that hepatic microsomes from Atlantic salmon possess CYP2E1-like activity. However, specific isoform-mediated PNPH activity should be identified.     

Drift diving: A quick and accurate method for assessment of anadromous salmonid spawning populations

The accuracy of drift diving surveys of Atlantic salmon, Salmo salar L. and sea trout, Salmo trutta L., was evaluated by comparing the abundance and size distribution with catches in a fish trap over 6 years in the River Etneelva, western Norway. The population count from drift diving accounted for on average 96.3% of the salmon in the trap after accounting for the catches during fishing. Size structure registered during drift diving corresponded with trap catches of salmon, but the number of small salmon (<3 kg) appeared to be somewhat underestimated, while large salmon (>7 kg) were overestimated in drift diving. For sea trout, the match between drift diving counts and trap registrations was poorer (average 76.3%), but may have been affected by the surveys being performed too late with regards to sea trout spawning time. The study illustrates the utility of drift counting for estimating the entire population of anadromous salmonids in a river.     

Anesthesia induces stress in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>), Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) and Atlantic halibut (<Emphasis Type="Italic">Hippoglossus hippoglossus</Emphasis>)

Stress in response to anesthesia with benzocaine, MS-222, metomidate and isoeugenol was studied in Atlantic salmon (Salmo salar), Atlantic halibut (Hippoglossus hippoglossus), and Atlantic cod (Gadus morhua) with no concomitant stress from handling or confinement in association with anesthesia or sampling. All of the anesthetics tested induced a stress response in all species, displayed by a release of cortisol to the water. MS-222 anesthesia elicited the highest cortisol release rates, reaching maximum levels 0.5 h post-exposure and returning to basal levels after 3–4 h. Benzocaine anesthesia caused a bimodal response where the initial peak in cortisol release rate was followed by a second increase lasting towards the end of the trial (6 h). This bimodality was more profound in Atlantic salmon than in Atlantic halibut and Atlantic cod. Metomidate anesthesia induced the lowest release of cortisol of the agents tested in both Atlantic halibut and Atlantic cod, but resulted in a bimodal response in Atlantic salmon where the initial increase in cortisol release was followed by a larger increase peaking at 2–2.5 h post exposure before returning to basal levels after 5 h. The stress induced in Atlantic salmon by isoeugenol anesthesia resembled that of MS-222, but did not reach the same elevated level. Overall, the cortisol release was most profound in Atlantic salmon followed by Atlantic halibut and Atlantic cod.     

Ochroconis globalis infecting Atlantic salmon (Salmo salar), with a review of Ochroconis species in cold‐blooded animals

Necropsy examination of an adult Atlantic salmon (Salmo salar) from the Dalälven River in Sweden revealed numerous large, white nodules, with spherical cysts and granulomata in kidney and liver. Histopathology showed dark, septate, thin‐walled hyphae. The aetiologic agent was found to be an Ochroconis species (Venturiales) that differed from known fish‐associated species of the genus. Molecular phylogenetic studies of the culture (strain UIII09 = CBS 135766) demonstrated that Ochroconis globalis was concerned. The isolate proved to be susceptible to all investigated antifungals, as it is known for another Ochroconis species. The role of Ochroconis in opportunism of cold‐blooded animals was discussed, and the diagnostic methods using DNA sequences for routine identification of the fungus were proposed.     

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.     

Inactivation of Piscine orthoreovirus

Piscine orthoreovirus infects various salmonid fish species, and the infection is associated with diseases such as heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon (Salmo salar). There are no vaccines available or genetically selected resistant hosts that can efficiently control piscine orthoreovirus (PRV) infection. Currently, the only prophylactic measure against PRV is general biosecurity measures aiming to break the transmission cycle. Methods to eradicate infectious virus from contaminated facilities are desirable, but the knowledge on how to inactivate PRV is lacking. A major bottleneck for inactivation studies is the lack of ability to propagate PRV in cell culture. Therefore, in this study we developed an in vivo model for detection of infectious PRV particles after treatment of the virus with inactivation tools such as heat, pH, iodine, UV and commercially available disinfectants. The results show that standard iodine treatment is efficient in inactivation of the virus, and similarly are high and low pH extremes and treatment with Virocid, a commercially available disinfectant. A UV dose of at least 50 mJ/cm² is required for inactivation, and the virus has high resistance against heat treatment.     

Serological diversity in Flavobacterium psychrophilum: A critical update using isolates retrieved from Chilean salmon farms

Chile is currently the second largest producer of farmed salmon worldwide, but Flavobacterium psychrophilum, as one of the most detrimental pathogens, is responsible for major losses during the freshwater culturing step in salmonid fish farms. An antigenic study conducted 10 years ago reported four serological groups using 20 F. psychrophilum Chilean strains. To reduce disease outbreaks and to develop vaccine candidates, antigenic knowledge needs to be regularly updated using a significant number of additional recent F. psychrophilum isolates. The present study aimed at investigating the serological diversity of 118 F. psychrophilum isolates collected between 2006 and 2018 from farmed Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss) and coho salmon (Oncorhynchus kisutch). The current study supports an expansion of the known antigenic groups in Chile from 4 to 14. However, the use of the slide-agglutination technique for serotyping is costly, is labour-intensive and requires significant technical expertise. Addressing these points, the mPCR-based procedure was a very useful tool for serotyping the collected Chilean F. psychrophilum isolates. This technique revealed the presence of diverse mPCR serotypes (i.e. types 0, 1, 2 and 4). Therefore, mPCR should be employed to select the bacterial strain(s) for vaccine development and to conduct follow-up, selective breeding or epidemiological surveillance in Chilean fish farms. Given the presented findings, changes to Chilean fish-farming practices are vital for ensuring the continued productivity and well-being of farmed salmonids.