Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Efficacy of bithionol as an oral treatment for amoebic gill disease in Atlantic salmon Salmo salar (L.)

This study examined the efficacy of bithionol as an oral treatment for Atlantic salmon Salmo salar affected by amoebic gill disease (AGD). The current commercial management strategy of AGD is a costly 3 h freshwater bath. It is labour intensive and the number of baths needed appears to be increasing; hence, there is an effort to identify alternative treatments. Efficacy was examined by feeding AGD-affected Atlantic salmon twice daily to satiation with bithionol, an antiprotozoal, at 25 mg kg^− 1 feed. Three seawater (35‰, 17 °C) re-circulation systems were used each consisting of three tanks containing 32 Atlantic salmon smolts with an average (± SEM) mass of 90.4 g (± 5.2). Three feeds were examined in the trial including bithionol, plain commercial control and oil coated commercial control. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L^− 1 and continued for 28 days post-exposure. Efficacy was determined by examining gross gill score and identifying percent lesioned gill filaments twice weekly for 4 weeks post-exposure. Bithionol when fed as a two-week prophylactic treatment at 25 mg kg^− 1 feed delayed the onset of AGD pathology and reduced the percent lesioned gill filaments by 53% and halved the gill score from 2 to 1 when compared with both the plain and oil controls during an experimental challenge. There were no palatability problems observed with mean feed intake of bithionol over the trial duration with fish consuming higher levels of the bithionol diet compared to both the oil and plain controls. This study demonstrated that bithionol at 25 mg kg^− 1 feed, when fed as a two-week prophylactic treatment for Neoparamoeba spp. exposure, delayed and reduced the intensity of AGD pathology and warrants further investigation as an alternative to the current freshwater bath treatment for AGD-affected Atlantic salmon.     

Impact of starvation and handling stress on rigor development and quality of Atlantic salmon (Salmon salar L)

Farmed Atlantic salmon (Salmo salar L) were fed to satiation or starved for 35 d, and subjected to acute pre-slaughter stress or careful handling before slaughtering. Rigor development, post-mortem energy metabolism and quality variations were analysed during 72 h cold storage. The body weight averaged 2.9 and 3.5 kg of the fed and starved salmon, respectively. The starved salmon had lower condition factor (0.99 vs. 1.11) and higher slaughter yield (92.6 vs. 87.5%), but the fat content was similar (11.8–12.9%) in the fed and starved salmon. The initial glycogen level was lower in the starved salmon (21 vs. 29 μmol g^− 1), whereas initial ATP levels (6.4–6.6 μmol g^− 1) were not affected by starvation. Pre-slaughter stress accelerated rigor development, stimulated lactate formation through post-mortem glycolysis, and increased the breakdown of ATP and CP. The effect of stress on rigor development was less pronounced in starved salmon, and a significant interaction was observed between nutritional status and stress. Rigor development was closely correlated to ATP breakdown, especially in fillets. Acute stress accelerated flesh softening during ice storage. After 72 h storage, the fillets of the starved salmon exposed to careful slaughter handling had the significantly firmest texture, whereas the most intense fillet colour was observed in the starved/stressed salmon. Liquid leakage from the muscle during cold storage was lower in the starved salmon, but pre-slaughter stress had no significant impact. It is concluded that a starvation period of five weeks can improve the resistant to acute stress prior to slaughtering of Atlantic salmon that in turn hampers rigor development.     

Effects of Cu-exchanged montmorillonite on growth performance, microbial ecology and intestinal morphology of Nile tilapia (Oreochromis niloticus)

A total of 360 Nile tilapia at an average initial body weight of 3.9 g were randomly allocated to 4 treatments, each of which had three replicates of 30 fish/tank and used to investigate the effects of Cu²⁺-exchanged montmorillonite (Cu-MMT) on growth performance, microbial ecology of the skin, gill and intestine, and intestinal morphology. The dietary treatments were: 1) basal diet, 2) basal diet + 1.5 g/kg MMT, 3) basal diet + 30 mg/kg copper as CuSO₄ (equivalent to the copper in the Cu-MMT treatment group), or 4) basal diet + 1.5 g/kg Cu-MMT. The results showed that supplementation with Cu-MMT significantly improved growth performance as compared to control and fish fed with Cu-MMT had higher growth performance than those fed with MMT or CuSO₄. Supplementation with Cu-MMT reduced (P < 0.05) the total intestinal aerobic bacterial counts and affected the composition of intestinal microflora with a tendency of Aeromonas, Vibrio, Pseudomonas, Flavobacterium, Acinetobacter, Alcaligence, Enterobacteriaceae decreasing as compared with control. Measurements of villus and microvillus heights at different sites of the intestinal mucosa indicated that dietary addition of MMT or Cu-MMT improved intestinal mucosal morphology. However, the microbial ecology of the skin and gill was not affected by the addition of CuSO₄, MMT or Cu-MMT. Supplementation with CuSO₄ had no (P > 0.05) effect on the growth performance, microbial ecology and intestinal morphology. The results showed that Cu-MMT exhibited antibacterial activity in vivo and protected intestinal mucosa from invasion of pathogenic bacterium and toxins, resulting in a positive effect on the growth performance.     

Environmental effects of the anti-sea lice (Copepoda: Caligidae) therapeutant emamectin benzoate under commercial use conditions in the marine environment

Sea lice infestation can pose a particular problem to intensive cage aquaculture of salmonids. It is most often treated by therapeutants added to the water or incorporated into fish feed. This study investigated the environmental effects of one such in-feed treatment, emamectin benzoate, under commercial use conditions. Atlantic salmon were fed the commercial preparation of emamectin benzoate at a production fish farm on the west coast of Scotland. Sediment dwelling fauna, large mobile fauna and sentinel organisms were monitored for potential impacts over a 1 year post-treatment period (23 September 1997 to 23 September 1998). Additionally, sediment and water samples, transplanted blue mussels and captured large fauna were analysed for accumulated emamectin benzoate and its desmethylamino metabolite to investigate the long term environmental fate of emamectin benzoate.

Results from analysis of sediments suggested that limited deposition of emamectin benzoate took place during and up to 1 week post-treatment, and may have continued up to 4 months of post-treatment. This was consistent with the release of the active ingredient from fish excretion and defecation after treatment. Quantifiable amounts of emamectin benzoate were found 10 m from the cages 12 months after treatment. However, clear reduction in overall sediment concentrations indicated that the deposited emamectin was degrading over time. Emamectin benzoate was not detected in water samples during the field trial. Quantifiable concentrations of emamectin benzoate were found in blue mussels deployed up to 100 m from the treatment cages at 1 week post treatment, but found only at 10 m from the cages at 1 month post-treatment. This suggests that while mussels were actively accumulating emamectin benzoate immediately after treatment by 1 month this had largely been depurated.

Macrobenthic faunal analysis indicated that there was no evidence that the occurrence of emamectin benzoate, or its desmethylamino metabolite, in sediments around fish farm cages after treatment had any toxic impacts on organisms in either water column or sediments.     

Effects of dietary estradiol-17β in juvenile shortnose sturgeon, Acipenser brevirostrum, Lesueur

Shortnose sturgeon, Acipenser brevirostrum, were fed estradiol-17β (E₂) in commercial salmon feed in two separate experiments, with the aim of creating all-female populations for aquaculture. In the first experiment, 5-month-old fish were fed 0 (control), 10, 50 and 100 mg E₂/kg feed during a 9-month trial starting in September. This experiment was confounded by copper contamination of the heated water source that appeared to cause E₂-dose-dependent mortality. As a result, a second, parallel experiment was conducted on the same year class of fish starting at age 7 months in November, but with hormone dosage reduced to 0, 10, 25 and 50 mg E₂/kg feed in a 7-month trial. In both experiments, control and 10 mg E₂/kg groups remained active and fed well, but the other fish were less active and fed less as E₂ dosage increased. Histological observations indicated that anatomical differentiation of the ovaries had begun in 3 of 5 individuals examined at the start of Experiment 2. In January, sub-samples of 6 fish from each group showed that E₂-treated fish had mottled livers, swollen kidneys and reduced gonad size. The number of hepatocytes and their staining characteristics were similar in both the control and 10 mg E₂/kg groups. At 25 mg E₂/kg and higher, the number of hepatocytes and the presence and quantity of yolk protein increased with the E₂ dosage. At final sampling in May, all of the treatment groups showed some individuals with these pathologies evident macroscopically, and their frequency and severity increased with E₂ dosage such that all individuals of the 100 mg E₂/kg group were affected. In general, fish grew better and had lower mortality in Experiment 2 (8.5%, as compared to 15.2% for Experiment 1) during the period of E₂ treatment. Final sampling revealed that all treated fish were apparently feminized in both experiments, based on anatomical differentiation of the gonads. Control groups, on the other hand, were 65 and 58% female in Experiments 1 and 2, respectively.     

Pigmentation, carotenoids, lipid peroxides and lipid composition of skin of red porgy (Pagrus pagrus) fed diets supplemented with different astaxanthin sources

A feeding experiment was carried out to determine the efficiency of different commercial sources, chemical forms and levels, of dietary astaxanthin, to appropriately pigment the red porgy (Pagrus pagrus) skin. According to this, total carotenoid content, profiles and chemical forms present in the skin were determined. In order to establish the potential for antioxidant protecting role of astaxanthin supplemented diets, peroxide levels and lipid composition of skin were also determined.

Red porgy alevins were fed six dietary treatments in triplicate; a basal diet (B) without carotenoids; two diets (N25 and N50) formulated to supply either 25 or 50 mg kg^− 1 of an esterified source of astaxanthin (Haematococcus pluvialis, NatuRose™); two diets (CP25 and CP50) with either 25 or 50 mg kg^− 1 of unesterified astaxanthin (Carophyll® Pink); and a positive control diet (B + S) proved as a successful pigmenting-diet in previous experiences (B + S, 88% basal diet:12% frozen shrimp) [Cejas, J., Almansa, E., Tejera, N., Jerez, S., Bolaños, A., Lorenzo, A., 2003. Effect of dietary supplementation with shrimp on skin pigmentation and lipid composition of red porgy (P. pagrus) alevins. Aquaculture 218, 457–469].

All fish fed carotenoid supplemented diets displayed a pink-coloured skin after 4 months of feeding in contrast to the greyish appearance displayed by fish fed the basal diet not supplemented with carotenoids (B). Furthermore, astaxanthin diesters were the major carotenoid in the skin of pink fish. A second carotenoid, tentatively identified as tunaxanthin diester, was also detected. The best results in terms of skin natural reddish hue, total carotenoid and astaxanthin contents were found by using the esterified forms of dietary astaxanthin (N25, N50 and B + S). Interestingly, the lowest levels of lipid peroxides were found in the fish fed these three treatments. However, no effect of treatment on lipid composition was found. In conclusion, red porgy alevins are able to efficiently utilise dietary natural or synthetic astaxanthin, and deposit this pigment in its esterified form to acquire an acceptable pink-coloured skin compared to that of the wild fish.     

The effect of artificial light treatment and depth on the infestation of the sea louse Lepeophtheirus salmonis on Atlantic salmon (Salmo salar L.) culture

Two field studies were carried out with farmed Atlantic salmon (Salmo salarL.) in sea cages to examine various effects of artificial light (AL) and the vertical distribution of salmon on lice infestation.

The use of AL light caused an overall increase in lice infestation in both experiments. The first study showed that salmon held at 0–4 m depth in cages developed higher infestation than salmon held at greater depths (4–8 and 8–12 m) under both natural light (NL) and AL. In the second study, salmon maintained in 14-m deep sea cages that were exposed to AL with different light intensities. The AL treatments resulted directly in different diel and seasonal patterns of vertical distribution of the salmon and also different temporal patterns in lice infestations. So indirectly the infestation pattern appeared to be correlated with median day-time swimming depth of the salmon.     

Effects of dietary canola oil level on growth, fatty acid composition and osmoregulatory ability of juvenile fall chinook salmon (Oncorhynchus tshawytscha)

This study assessed refined canola oil (CO) as a supplemental dietary lipid source for juvenile fall chinook salmon, Oncorhynchus tshawytscha, parr with respect to possible effects on their growth and osmoregulatory performance and body composition. Diets with equal protein ( 57%) and lipid ( 19%) content (dry weight basis) were supplemented with lipid from either anchovy oil (AO) or CO with AO so that CO comprised 0 (0CO), 11% (11CO), 22% (22CO), 33% (33CO), 43% (43CO) or 54% (54CO) of the dietary lipid content. Triplicate groups of juvenile chinook salmon were fed their prescribed diets for 104 days in freshwater (FW) and 31 days in seawater (SW) after a 4-day transition period. Dietary fatty acid compositions reflected the different proportions of AO and CO in the supplemental lipid. Diet treatment had no effect on fish growth, feed intake, feed efficiency, protein utilization, fish mortality or terminal whole body water and ash percentages. Whole body lipid percentages were higher in 11CO and 43CO fish than in 33CO fish and in 11CO fish versus 22CO fish. Whole body protein percentages were highest in 33CO, 43CO and 54CO fish and lowest in 0CO and 22CO fish. Terminal whole body fatty acid compositions were influenced strongly by the dietary fatty acid compositions. Haematocrit and muscle water percentages were not affected consistently and plasma Na⁺ and Cl⁻ concentrations were unaffected by diet treatment in FW or 24-h seawater challenges during FW residency. Also, diet treatment had no effect on the physiological parameters after SW residency. We conclude that dietary treatment had no effect on fish growth performance under our experimental conditions. Also, the dietary inclusion of CO neither facilitated nor impaired the transfer of chinook salmon parr to seawater. Thus, CO was found to be an excellent and cost-effective source of supplemental dietary lipid for culture of juvenile fall chinook salmon during freshwater residency.     

Effects of hook and release on Atlantic salmon in the River Alta, northern Norway

The purpose of the study was to collect information on angling procedures and the effects of hook and release on Atlantic salmon in the River Alta, northern Norway, covering both grilse and multi-sea-winter salmon in a non-artificial setting with real anglers. Information on the angling procedure, handling of the fish and the condition of the fish at release was collected for individual salmon in catch logs (n=543, mean body length 82 cm), whereas physiological stress was studied in a sub-sample (n=15, mean body length 77 cm). To study post-release behaviour, survival and recapture rates, salmon were tagged with radio transmitters (n=30, mean body length 83 cm) and anchor T-tags (n=353, mean body length 79 cm). To evaluate the effects of the hook and release programme on the salmon population, number of spawning redds were recorded from a helicopter in 6 years during 1989–2000.

The results showed that at water temperatures 10.0–14.5 °C, a high proportion of the radio tagged salmon (97%) survived hook and release and stayed in known spawning areas during the spawning period. However, the behaviour after release seemed to be affected by hook and release. Only a small proportion (4%) of the anchor T-tagged salmon was caught more than once within the same season. Increased playing time, increased number of runs during the angling event, hooking in the throat, bleeding at the hook wound, increased handling time, air exposure and water temperature were factors that affected hooked and released Atlantic salmon negatively, either indicated by a poor condition at release, increased stress levels or unnatural behaviour after release. Number of spawning redds were more than doubled after the introduction of compulsory release of all angled salmon in Sautso (the upper 16% of the watershed inhabited by salmon) in 1998, which indicates that hook and release can be an effective management tool to enhance declining Atlantic salmon populations.     

Safety and efficacy of emamectin benzoate administered in-feed to Atlantic salmon, Salmo salar L., smolts in freshwater, as a preventative treatment against infestations of sea lice, Lepeophtheirus salmonis (Krøyer)

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg⁻¹ fish day⁻¹ for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg⁻¹ day⁻¹, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg⁻¹ day⁻¹. Among smolts that received 272 μg kg⁻¹ day⁻¹, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified.

In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg⁻¹ day⁻¹ (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P<0.001) on treated fish than on controls. When fish challenged at day 109 were sampled at day 128, efficacy was 44.3%, but survival of chalimus to adult lice on treated fish was lower, and at day 159, efficacy had increased to 73%. These results demonstrate that treatment of salmon smolts with emamectin benzoate in freshwater was well tolerated and highly effective in preventing sea lice infestation following transfer of fish to seawater.     

Growth and parr-smolt transformation of Atlantic salmon (Salmo salar L.) under different light intensities and subsequent survival and growth in seawater

Groups of potential Atlantic salmon (Salmo salar L.) smolts were reared under a simulated natural photoperiod from November 1987 to May 1988. Three different light intensities were used: 27, 335 or 715 lux. Growth rate was not significantly influenced by differences in light intensity. All groups developed the silvery external appearance, increased hypo-osmoregulatory ability and decreased condition coefficient associated with smolting. Smolts from all groups were successfully acclimatized to seawater in early May. The subsequent seven months of rearing in seawater did not reveal any differences in post-smolt performance of the groups. The incidence of early maturation during the first autumn was similar in all groups. We conclude that within the limits of the light sources used in the present experiment, light intensity had no significant effect on the growth and smolting in juvenile Atlantic salmon.     

Immunization of grouper, Epinephelus coioides, confers protection against a protozoan parasite, Cryptocaryon irritans

The present study aimed to determine whether protection is conferred by immunization of grouper, Epinephelus coioides, against a protozoan parasite, Cryptocaryon irritans. The immunization of E. coioides was carried out by a low level exposure of fish to live C. irritans theronts from predetermined number of tomonts and by an intraperitoneal injection of a vaccine consisting of formalin-killed C. irritans theronts.

Mucus titers detected by ELISA were significantly higher in fingerling and adult grouper subjected to the low level of exposure to C. irritans theronts at 3-week post-exposure compared to fish that had no previous exposure. In addition, significantly smaller tomonts were produced from adult grouper after three successive exposures than the tomonts produced after a single exposure to the parasite.

In the vaccine-immunization experiment, no mortality was monitored in fish that received high dose vaccine (100 μg/fish), while 40% cumulative mortality and 100% cumulative mortality were recorded in low dose group (10 μg/fish) and control group (PBS-injected), respectively. In the succeeding replicate, the vaccine-immunized group (high dose) had 37.5% cumulative mortality and 100% cumulative mortality for the control. In addition, a total of 1830 tomonts were collected at 5-day post-challenge from the control group while none from the vaccine-immunized group. Significantly fewer trophonts and tomonts were enumerated at 5-day and 7-day post-challenge, respectively, in the vaccine-immunized group than the control.

Results suggest that a protective immunity has been conferred on the immunized grouper as indicated by high antibody titers in the mucus of C. irritans-exposed fish and higher survival and fewer parasites in vaccine-immunized fish than the control groups. The conferred immunity played a major role in preventing or limiting the adhesion, invasion, and development of C. irritans theronts on the skin of the immunized grouper.     

In vitro survival and the effect of water chemistry and oxidative chemical treatments on isolated gill amoebae from AGD-affected Atlantic salmon

Amoebic gill disease (AGD) of cultured salmonids in Tasmania is caused by the amphizoic parasitic amoeba Neoparamoeba pemaquidensis. The freshwater tolerance of amoebae isolated from the gills of AGD-affected salmon (predominantly N. pemaquidensis) was tested in vitro using a trypan blue exclusion assay. Amoebae exposed to water containing high concentrations of Ca²⁺ or Mg²⁺ (200 mg l⁻¹) showed high levels of survival up to 3 h of exposure. Exposure to water containing elevated Na⁺, choline chloride or water at different pH all had no significant survival of amoebae. Exposure of amoebae to different concentrations of chlorine dioxide, chloramine-T or hydrogen peroxide in artificially hard water demonstrated that chloramine-T and hydrogen peroxide were the most efficacious at killing amoebae in vitro. This work suggests that the hardness of freshwater may be an important factor for the survival of marine amoebae (predominantly N. pemaquidensis) on the gills of AGD-affected salmon and have significant implications with regard to the efficacy of freshwater bathing practices for the control of AGD on farms. Additionally, chloramine-T and hydrogen peroxide appear to be efficacious at killing marine gill amoebae in vitro and may be useful for the control of AGD in farmed Atlantic salmon.     

Short-term storage and cryopreservation of milt from Atlantic salmon and sea trout

Experiments on short-term preservation of sperm were performed with Atlantic salmon (Salmo salar). Fertility was maintained for up to 10 days when 2 mm thick samples were stored at 0° C under an oxygen atmosphere in the presence of antibiotics (125 IU penicillin and 125 μg streptomycin per ml sperm). Fertility was completely lost after 24 days. Sperm stored without antibiotics fertilized 100% of eggs after 6 days.

Cryopreservation was carried out with milt from Atlantic salmon and sea trout (Salmo trutta). Semen mixed with extender was frozen on dry ice (pellets) with subsequent storage in liquid nitrogen. Sperm pellets were thawed in a 0.12-M NaHCO₃-solution (10° C) before insemination. The suitability of an extender as previously described by Stoss and Holtz and of a 0.3-M glucose solution with the addition of 10% DMSO, was tested on two different batches of sperm and eggs in Atlantic salmon and sea trout. In addition, the extender earlier reported by Mounib and an aqueous solution of 10% DMSO were only used in Atlantic salmon with one batch of gametes.

Insemination with cryopreserved Atlantic salmon sperm resulted in 36 to 91.3% eyed eggs (control = 100). The differences were caused by the type of extender and the batch of gametes employed. The very simple extender consisting of 0.3 M glucose and 10% DMSO only was the most successful. Results with cryopreserved sea trout sperm ranged between 38.6–54.8% eyed eggs, showing no difference between treatments.     

Growth, osmoregulation and sexual maturation of underyearling Atlantic salmon smolt Salmo salar L. exposed to different intensities of continuous light in sea cages

Atlantic salmon smolts are usually transferred to sea water during early spring, but photoperiod treatment can be used to produce underyearling (0+) smolts for transfer to sea water in late autumn, 7–8 months earlier than usual. This study investigated the effect of exposure to additional continuous light (LL) of different intensities on 0+ smolts after transfer to sea water. 0+ smolts transferred to sea cages in mid-October were exposed to natural light (NL) or LL of one of three intensities (low, medium or high) until January, whereupon they experienced NL until harvest in December. Fish in all groups displayed a normal ability to hypoosmoregulate, assessed by monitoring plasma chloride concentrations after transfer to sea water. No consistent differences in length, weight or condition were found between LL groups. The LL groups had greater growth in length than the NL group during the time of LL exposure, followed by an increase in weight when exposed to NL. This resulted in fish of larger size than the NL group. By August, the fish in the NL group had caught up with the LL groups in respect of length, weight and condition factors. This suggests that the LL treatment led to increased winter growth and phase-shifted a seasonal pattern of growth. The incidence of sexual maturation was low (< 1%), with no differences between groups.     

An experimental study of the susceptibility of Atlantic salmon fry, Salmo salar L., to viral haemorrhagic septicaemia

Abstract. Infection trials using two serotypes of VHS viruses (type 1 and 23/75) demonstrated that Atlantic salmon fry were susceptible to the disease when injected intraperitoneally (i.p.) with 10³ pfu of virus/fish but resistant to infection by a bath method when exposed for 3 h in water containing 5 × 10⁴ pfu of virus/ml. In the i.p.-infected fish, mortality reached 78 and 67% within 13 days with VHSV₁ nad 23/75 serotypes, respectively. High virus yields were recovered from infected fish and virus shedding was demonstrated by the onset of VHS in rainbow trout kept in the outflow water from the aquaria containing infected salmon. Neither mortality nor virus shedding occurred in salmon infected by the water route but virus multiplication was demonstrated in 2 of 60 fish with VHSV₁ and 3 of 60 fish with virus 23/75. On day 79 post-infection the sera from surviving salmon of both i.p. and bath infection trials exhibited good neutralizing titres (around 1000) against the homologous viruses.     

Involvement of trypsin and chymotrypsin activities in Atlantic cod (Gadus morhua) embryogenesis

Proteases play a key role in yolk formation and degradation during embryogenesis of marine fish. This study presents the first clear data on the involvement of trypsin and chymotrypsin activities in the embryogenesis of Atlantic cod (Gadus morhua). Both enzyme activities were shown to be present in unfertilized eggs followed by a significant decline (P < 0.01) in trypsin activity during the first 4 days post fertilization (dpf). Thereafter, the trypsin- and chymotrypsin activities increased to a maximum around day 9 pf. A decline in trypsin and chymotrypsin activities was observed from day 10 pf with minimal activity just prior to first feeding (day 15 pf). Western blot analysis, using polyclonal antibodies raised to Atlantic cod trypsins I and Y, mostly coincided with the trypsin activity profile. The novel trypsin Y was previously shown to have both trypsin- and chymotrypsin-like activities. Thus, some of the chymotrypsin activity observed in the samples may be originated from trypsin Y. The low trypsin and chymotrypsin activities just prior to first feeding (13–15 dpf) may indicate insufficient digestive function as trypsin has been shown to be a suitable short-term indicator reflecting the nutritional quality of marine fish larvae.     

Atlantic salmon, Salmo salar, utilizes wax ester-rich oil from Calanus finmarchicus effectively

Against a background of decreasing availability of fish oils for use in aquaculture, the present study was undertaken to examine whether a wax ester-rich oil derived from the calanoid copepod Calanus finmarchicus, could be used effectively by Atlantic salmon when supplied in their diet. Individually tagged Atlantic salmon of initial weight around 500 g were divided into replicate tanks of two dietary groups and fed either a fish oil supplemented diet, or an experimental diet coated with Calanus oil. Wax esters accounted for 37.5% of the lipids in the Calanus oil diet but were absent from the fish oil diet in which triacylglycerols (TAG) were the major lipid class. Over the feeding period (140 days) the salmon fed fish oil displayed a greater increase in length, but there was no significant difference between the two groups in weight gained. The specific growth rates (0.75) and the feed conversion ratio of fish fed the two diets were similar throughout the study. No differences were observed in the apparent digestibility coefficients (ADC) of fish fed Calanus oil or fish oil. The ADC of fatty acids decreased with chain length and increased with unsaturation. Long-chain alcohol utilization showed a similar tendency although there was a notable difference in that saturated long-chain alcohols were utilized better than the comparable fatty acid homologue. In fecal lipid of fish fed Calanus oil, the content of 16:0 alcohol decreased in both the free long-chain alcohol and wax ester fractions, while the corresponding fatty acid increased in the feces of both dietary groups of fish. In contrast, the proportion of the 22:1n−11 alcohol increased in both fecal wax esters and free long-chain alcohol fractions whereas 22:1n−11 fatty acid displayed no accumulation. The observed patterns of fatty acid and long-chain alcohol compositions in fecal lipid compared to those of the initial dietary lipid are consistent with the digestive lipases of salmon preferentially hydrolyzing esters containing polyunsaturated fatty acid (PUFA) moieties. The wax esters of Calanus oil contained substantial amounts of the n−3 PUFA, 20:5n−3 and 22:6n−3, that were effectively deposited in muscle and liver tissues. No major differences were seen in either lipid content/lipid classes or in gross fatty acid composition of these tissues between the two dietary groups. It is concluded that that Atlantic salmon in seawater can effectively utilize diets in which a major lipid component is derived from zooplankton rich in wax ester without any detrimental change in growth or body lipid composition. This finding gives support to the use of lipid from zooplankton from high latitudes as an alternative or as a supplement to fish oil and a provider of long-chain n−3 PUFA in diets for use in salmon aquaculture.     

Urea in feeds for sea water farmed Atlantic salmon: effect on growth, carcass quality and outbreaks of winter ulcer

The main objective of the present study was to evaluate whether dietary supplementation of urea might reduce the incidence of winter ulcer in sea water-farmed Atlantic salmon. Salmon destined to be S0 smolt were fed with a urea-supplemented diet (0 or 20 g kg⁻¹ urea) over an 8-week period prior to sea water transfer and were then fed supplemented diet (0, 5, 10 or 20 g kg⁻¹ urea) during the first and second winters in the sea. During the first winter positive relationships between dietary urea and plasma urea and between plasma urea and plasma osmolality were observed. Further, plasma osmolality displayed a negative relationship to mortality. Of the salmon that died during the first winter in the sea 90% had one or more skin ulcers. Both during the first and second winter there were fewer salmon with ulcer among fish fed with the diets supplemented with urea. Salmon fed with 20 g kg⁻¹ urea tended to have higher percentage water in the muscle. Mortality and incidence of salmon with ulcer seemed to relate to plasma osmolality amongst fish fed on diets that differed in levels of urea supplementation, suggesting that an osmotic imbalance may contribute to the development of winter ulcer in farmed salmon. Salmon fed with 20 g kg⁻¹ urea showed significantly greater body weight during the second winter in sea. Fish killed without prior starvation had significantly higher level of muscle urea in the 20 g kg⁻¹ urea group compared with fish fed with the unsupplemented diet. However, a 13-day starvation period reduced urea content in the muscle to the level of the control. No effects of dietary urea supplementation on the sensory quality of market size Atlantic salmon were observed.