Atlantic salmon Salmo salar and ballan wrasse Labrus bergylta display different susceptibility to clonal strains of Paramoeba perurans.

Amoebic gill disease (AGD), caused by the marine amoeba Paramoeba perurans, is an important disease of farmed Atlantic salmon Salmo salar L. in Norway. The use of wrasse as cleaner fish in salmon net pens raises questions about interspecies transmission of pathogens such as P. perurans. In this study, cohabitant transmission of clonal isolates of P. perurans between Atlantic salmon and ballan wrasse Labrus bergylta Ascanius was examined, using isolates originating from both salmon and wrasse. The challenges resulted in AGD in both species, although less severely in wrasse. The amoeba isolate originating from ballan wrasse was more virulent than that originating from salmon, suggesting P. perurans strain-related virulence differences. The isolate originating from salmon showed limited proliferation in bath-challenged wrasse and salmon, and limited transfer to cohabitants. Our results support previous observations suggesting that salmon may be more susceptible to P. perurans and AGD than ballan wrasse. Treatment of P. perurans infection in wrasse is challenging, as it is a strictly marine fish species. In this study, brackish water (<15‰ seawater) treatment of AGD affected salmon and wrasse was examined. Both salmon and wrasse were treated for short periods (3 h and 24 h), and treatment of wrasse over longer periods (3-5 d) was also examined. Short exposure to brackish water was not enough to remove P. perurans, although the 24 h treatment reduced amoeba levels. It was not possible to culture or detect P. perurans from wrasse exposed to brackish water for 3 d, suggesting that this treatment would be effective in controlling the parasite.

Dietary histidine requirement to reduce the risk and severity of cataracts is higher than the requirement for growth in Atlantic salmon smolts, independently of the dietary lipid source.

The present study was carried out to investigate whether the dietary histidine requirement to reduce cataract development is higher than that for growth in Atlantic salmon smolts (Salmo salar L.) after seawater transfer and whether dietary vegetable oils contribute to cataractogenesis. Duplicate groups of salmon smolts were fed ten experimental diets with either fish oil (FO) or a vegetable oil (VO) mix replacing 70 % FO and histidine at five target levels (10, 12, 14, 16 and 18 g His/kg diet) for 13 weeks after seawater transfer. The VO diet-fed fish exhibited somewhat inferior growth and feed intakes compared with the FO diet-fed fish, irrespective of the dietary histidine concentration. Both cataract prevalence and severity were negatively correlated with the dietary histidine concentration, while lens N-acetyl-histidine (NAH) concentrations were positively correlated with it. The fatty acid profiles of muscle, heart and lens reflected that of the dietary oils to a descending degree and did not affect the observed cataract development. Muscle, heart and brain histidine concentrations reflected dietary histidine concentrations, while the corresponding tissue imidazole (anserine, carnosine and NAH) concentrations appeared to saturate differently with time. The expression level of liver histidase was not affected by the dietary histidine concentration, while the liver antioxidant response was affected in the VO diet-fed fish on a transcriptional level. The lowest severity of cataracts could be achieved by feeding 13·4 g His/kg feed, independently of the dietary lipid source. However, the present study also suggests that the dietary histidine requirement to minimise the risk of cataract development is 14·4 g His/kg feed.

Effects of chronic and periodic exposures to ammonia on the eye health in juvenile Atlantic halibut (Hippoglossus hippoglossus).

The effects of chronic and periodic peaks of un-ionised ammonia (UIA-N) exposure on eye health and cataract formation in juvenile Atlantic halibut, Hippoglossus hippoglossus, were examined. Fish with mean initial weight 51.7 g (SD 13.2) were exposed to five treatments consisting of control group and three groups (ChronicLow, ChronicMedium and ChronicHigh,) chronically exposed with UIA-N of 0.06, 0.12 to 0.17 mg/l, respectively, for 62 days at 11.9°C, pH 8.0 and salinity 34‰. Furthermore, a fifth group (HighPulse) was exposed to the same high levels as ChronicHigh for a short daily period (peak of 15 mg/l 30 min after exposure, 10 mg/l 1 h after exposure and 1.2 mg/l 3 h after exposure). In the subsequent period of the experimental study (from day 63 until day 100), no ammonia was added to the water. Mean weights were significantly lower in groups exposed to chronically high ambient ammonia concentrations compared to corresponding control group throughout the experimental period. The sampled fish exhibited signs of mild cataract formation, although the results showed no clear evidence that the ammonia treatments contributed to differences. Minor differences were found in measured muscle free amino acids, which could be used to explain potential changes in buffering capacity. The eye histidine status differed significantly at day 62, and osmotic differences in the eye lenses (measured as differences in N-acetyl histidine) were found in all group exposed to chronic levels of ammonia.

Ghrelin is involved in voluntary anorexia in Atlantic salmon raised at elevated sea temperatures.

Due to global and local climate changes, farmed salmon may experience periods of elevated sea temperatures. An experiment was conducted to examine endocrine and dietary effects of high sea temperatures in adult (2.0 kg) and sexually immature Atlantic salmon, Salmo salar L. Groups of salmon were exposed to 19 °C while others were kept as controls at 14 °C. The experiment lasted for 56 days, and fish were given iso-nitrogenous diets with either a normal (335 g kg(-1); L34) or a lower lipid level (298 g kg(-1); L30). Fish held at 19 °C had a reduction in the daily feed intake, growth and feed utilization of more than 50% compared to the controls. Fish at 19 °C retained little ingested fat, and high maintenance cost lead to depleted endogenous energy body reserves. Circulating ghrelin concentration and stomach ghrelin-1 and hypothalamus growth hormone secretagogue receptor 1a-like receptor (GHSR1a-LR) mRNA levels were significantly reduced in salmon at 19 °C. An increasing number of fish kept at 19 °C had empty gastrointestinal tract after 21 days (11-67%) and 56 days (56-100%), with the highest numbers in fish fed the L34 diet. We suggest that lower circulating ghrelin during negative energy homeostasis induce down-regulation of GHSR1a-LR, neuropeptide Y, and anorexigenic factors at transcriptional levels in the hypothalamus, which over time lead to a voluntary anorexia development in adult salmon held at 19 °C. Reduction of feed intake and growth may be an important coping strategy for salmon during elevated temperatures.

Susceptibility of Atlantic salmon lenses to hydrogen peroxide oxidation ex vivo after being fed diets with vegetable oil and methylmercury.

The development of cataract in farmed Atlantic salmon (Salmo salar L.) has been related to changes in feed composition resulting in sub-optimal lens nutrition. The present study was performed to investigate the ability of Atlantic salmon lenses to withstand oxidative stress ex vivo, with focus on the nutritional lipid history and exposure to methylmercury (MeHg) as a relevant dietary contaminant. Since dietary histidine has been shown to have a mitigating effect on the prevalence of cataract in farmed salmon, the antioxidative abilities of histidine and NAH, a major imidazole in the salmon lens, was also investigated ex vivo. Lenses from Atlantic salmon prefed diets based on either fish oil (FO) or vegetable oil (VO) as lipid source, with or without addition of 5 mg MeHgkg(-1) feed, were cultured for 96 h in normal medium (control), medium added 5 mM H(2)O(2) or in histidine enriched medium. Lipid class composition of the lenses was not affected by the dietary lipids; while VO fed fish had a decrease in lens n-3/n-6 fatty acid ratio due to minor but significant increase in the concentration of 18:2 n-6 and 20:4 n-6, and decrease in 20:5 n-3 fatty acids compared to FO fed fish. The lenses accumulated mercury in response to dietary levels, but neither the oxidative status nor any physiological responses were affected. The cultured lenses responded to H(2)O(2) exposure with loss of transparency, accumulation of auto-fluorescent compounds, volume increase and reduced glutathione concentration similarly and irrespective of the dietary history. Lenses extracted histidine from the media, and synthesised NAH during the culture period. The innate antioxidative defence system appeared to be influenced both by the dietary lipid history and histidine enrichment on a transcriptional level. Catalase and SPARC were expressed higher in lenses from FO fed fish, and glutaredoxin showed elevated expression levels in FO lenses cultured in histidine enriched medium, suggesting that histidine is related to the innate antioxidant defence in salmon lenses. Further, the concentration of NAH was significantly reduced in oxidatively stressed lenses. Based on the results from this study it is suggested that NAH has a novel role as antioxidant in the Atlantic salmon lens.

Lack of evidence for vertical transmission of SAV 3 using gametes of Atlantic salmon, Salmo salar L., exposed by natural and experimental routes.

Pancreas disease (PD) is an important cause of losses in farmed salmonids in Norway, the United Kingdom and Ireland. As the spread of salmonid alphavirus (SAV), the causal agent, to naïve populations is of major concern to the farming industry, it is important to uncover the transmission routes of the virus. This study was conducted to investigate the potential for vertical transmission of SAV subtype 3. Progeny of broodstock with signs of late-stage PD and persistent RT-PCR signals for SAV were followed from fertilization to smoltification in an experimental facility. Fertilized ova were either not disinfected or taken through one of three different disinfection regimes. Also, ova and milt from uninfected broodfish from a different population were exposed to a cell-cultured strain of SAV 3 immediately before fertilization to simulate a viraemic phase in parent fish. A group of uninfected controls were also included in the study. Fertilized ova from bath exposed and negative control groups were double disinfected. Following fertilization, experimental fish went through a normal freshwater phase. However, fry were stressed at first feeding to enhance replication of possibly latent virus. Smoltification was induced by an artificial light regime, and experimental fish were followed to the late smoltification phase. Selected samples were investigated by real-time RT-PCR for SAV, by histology for evidence of PD and by serology for neutralising antibodies against SAV. All analysed samples of progeny were negative. This result shows that SAV 3 is not readily transmitted vertically from parents to offspring. Additional negative PCR results from salmon sampled in commercial hatcheries support these findings. Also, recent studies have shown that risk factors for the horizontal transmission route explain the vast majority of PD outbreaks in Norway. It is concluded that if it happens at all, vertical transmission is of minor importance in the spread of SAV 3.

Dietary histidine supplementation prevents cataract development in adult Atlantic salmon, Salmo salar L., in seawater.

The aim of the present study was to investigate the cataract preventive effect of dietary histidine regimes in adult Atlantic salmon (Salmo salar L.) in seawater, both through manipulating the dietary histidine level and feeding period. Mean body weight of individually tagged Atlantic salmon at the start of the experiment was 1662 (sd 333) g. Low prevalence of mild cataracts were recorded in the beginning of June. Three fishmeal and fish oil-based extruded diets (crude protein: 375 g/kg and fat: 342 g/kg), differing only in histidine content (low (L): 9.3, medium (M): 12.8 and high (H): 17.2 g histidine/kg diets), were fed to duplicate net pens in seawater. The experimental period was divided into three seasons (June-July; July-September; September-October), each starting and ending with individual cataract examination, assessment of somatic data, and sampling of lens and muscle tissues for analysis of histidine and histidine derivatives. In July and September, a part of the population fed L- and H-histidine feeds were transferred (crossed over) to respective series of replicate net pens fed L-, M- and H-histidine diets (i.e. eleven experimental feeding groups at trial conclusion). The fish doubled their body weight from June to October, with no systematic effects on weight gain of dietary histidine feeding regimes. Development of severe cataracts was observed between July and September. The cataract severity was directly related to the dietary histidine level fed during the first and second periods. Feeding histidine-supplemented diets (M or H) in the first period from June to July mitigated later cataract outbreaks. The status of selected free imidazoles in muscle and lens tissues reflected the dietary histidine feeding regimes, relative to both feed concentration and feeding duration. The study shows the risk for cataract development for adult Atlantic salmon, 1 year after the transfer of salmon smolts from freshwater to seawater, which to a major extent can be prevented by histidine supplementation just before and during the early phase of cataract development.

Pancreas disease (PD) in sea-reared Atlantic salmon, Salmo salar L., in Norway; a prospective, longitudinal study of disease development and agreement between diagnostic test results.

A prospective longitudinal study was performed on three cages at each of three Norwegian Atlantic salmon seawater sites that experienced outbreaks of pancreas disease (PD). Once salmonid alphavirus (SAV) ribonucleic acid (RNA) was detected by real-time RT-PCR (Rt RT-PCR) at a site, it became detected in all studied cages and was persistently found until the end of the study period up to 19 months after first detection. SAV-specific antibodies were detected at all sites until the end of the study period and were also found at a high prevalence in broodfish at the time of stripping. No evidence of increased viral activity was detected in these broodfish. One site tested negative over several months prior to the first detection of SAV by Rt RT-PCR and SAV-specific antibody, which occurred 1 month prior to clinical manifestations of PD. Moribund fish or thin fish/runts that were sampled after the first PD diagnosis had almost twice the risk of testing positive by one or more diagnostic tests compared to that of randomly selected apparently healthy individuals. This paper describes the first detailed investigation of the disease development of PD at site and cage level in Norway, as well as an assessment of the performance and agreement of the commonly used diagnostic tests.

Salmonid alphavirus (SAV) and pancreas disease (PD) in Atlantic salmon, Salmo salar L., in freshwater and seawater sites in Norway from 2006 to 2008.

A cohort study was initiated in the spring of 2006 to investigate epidemiological aspects and pathogenesis of salmonid alphavirus (SAV) subtype 3 infections and pancreas disease (PD). The aims were to assess involvement of the freshwater production phase, the extent and frequency of subclinical infections and to follow PD-affected populations throughout the entire seawater production cycle, as well as investigate possible risk factors for PD outbreaks. Fish groups from 46 different Atlantic salmon freshwater sites in six counties were sampled once prior to seawater transfer and followed onto their seawater sites. A total of 51 Atlantic salmon seawater sites were included, and fish groups were sampled three times during the seawater production phase. SAV subtype 3 was not identified by real-time RT-PCR from samples collected in the freshwater phase, nor were any SAV-neutralizing antibodies or histopathological changes consistent with PD. In the seawater phase, SAV was detected in samples from 23 of 36 (63.9%) studied sites located within the endemic region. No SAV subtype 3 was detected in samples from seawater sites located outside the endemic region. The cumulative incidence of PD during the production cycle amongst sites with SAV detected was 87% (20 of 23 sites). Average fish weight at time of PD diagnosis ranged from 461 to 5978 g, because of a wide variation in the timing of disease occurrence throughout the production cycle. Mortality levels following a PD diagnosis varied greatly between populations. The mean percentage mortality was 6.9% (+/-7.06) (range 0.7-26.9), while the mean duration of increased mortality following PD diagnosis was 2.8 months (+/-1.11) (range 1-6).

Histidine nutrition and genotype affect cataract development in Atlantic salmon, Salmo salar L.

The aim of this study was to investigate effects of dietary levels of histidine (His) and iron (Fe) on cataract development in two strains of Atlantic salmon monitored through parr-smolt transformation. Three experimental diets were fed: (i) a control diet (CD) with 110 mg kg(-1) Fe and 11.7 g kg(-1) His; (ii) CD supplemented with crystalline His to a level of 18 g kg(-1) (HD); and (iii) HD with added iron up to 220 mg kg(-1) (HID). A cross-over design, with two feeding periods was used. A 6-week freshwater (FW) period was followed by a 20-week period, of which the first three were in FW and the following 17 weeks in sea water (SW). Fish were sampled for weighing, cataract assessment and tissue analysis at five time points. Cataracts developed in all groups in SW, but scores were lower in those fed high His diets (P < 0.05). This effect was most pronounced when HD or HID was given in SW, but was also observed when these diets were given in FW only. Histidine supplementation had a positive effect on growth performance and feed conversion ratio (P < 0.05), whereas this did not occur when iron was added. Groups fed HD or HID had higher lens levels of His and N-acetyl histidine (NAH), the latter showing a marked increase post-smoltification (P < 0.05). The HD or HID groups also showed higher muscle concentrations of the His dipeptide anserine (P < 0.05). There was a strong genetic influence on cataract development in the CD groups (P < 0.001), not associated with tissue levels of His or NAH. The role of His and His-related compounds in cataractogenesis is discussed in relation to tissue buffering, osmoregulation and antioxidation.

Light scattering in normal and cataractous lenses of farmed Atlantic salmon (Salmo salar): a slit lamp and Scheimpflug photographic study.

To investigate normal light scattering and cataract formation, the anterior eye segments of farmed Atlantic salmon (Salmo salar) reared in fresh water and sea water were documented in vivo for the first time with a Topcon SL-45 Scheimpflug camera. A total of 40 fish from the fresh-water-rearing period, obtained from 2 groups of identical age but showing a different growth rate, and 24 fish from the sea-water-rearing period, sampled from 2 groups with identical age but being fed different food brands, were included in this study. The fish were anaesthetized before examination. Due to the naturally wide pupil, no mydriatic compound was applied. All fish were removed from the water for photography, which was performed for each eye in 0 degrees = vertical slit position. Images were recorded on Kodak Tmax 400 black-and-white film. Microdensitometric image analysis of all negatives was performed using a Joyce-Loebl online microdensitometer. In spite of the virtual absence of an anterior chamber gap between cornea and lens and very little light scattering in the normal fish lens, a small number of distinct layers could be reproducibly identified in the lens. While there was little abnormal light scattering which could point to cataract development in young fish from the fresh water period, the evaluation of the lenses from the 2 sea water groups showed the presence of specific forms of cataract especially in the cortical and supranuclear layers. There were significant differences between the groups fed different food brands at the sea water site. In conclusion, Scheimpflug photography proved to be applicable to eye research in fish in vivo. It is suggested that this method should be employed for reproducible documentation as an extension to slit lamp monitoring in experimental research to reveal causative factors for cataracts in farmed fish.

Cataract development in Atlantic salmon (Salmo salar L) in fresh water.

Irreversible bilateral cataracts were diagnosed by slit-lamp biomicroscopy in 178 of 200 farm-raised Atlantic salmon (Salmo salar L) fed a standard diet over a five-month period. Initial changes were anterior polar opacities, progressing to involve both the anterior and posterior cortex before changes in the lens nucleus were seen. The lens changes were recorded and given scores according to the severity of the cataracts. At each of 3 samplings, after 2, 4 and 5 months, 200 fish were measured, weighed and examined by slit-lamp biomicroscopy. At all 3 samplings, there was a significant correlation between body length and both cataract incidence and cataract severity. There was also a significant correlation between body weight and cataract incidence and severity for the 2 last samplings. There was a significant correlation between K-factor as a measure of the shape of the fish, and both cataract incidence and severity, at all 3 samplings. Evaluation of specific growth rate in the periods between the examinations showed that the rapidly-growing fish were most susceptible to cataract formation. After cataract developed, however, the growth rate slowed. Follow-up examination of severely affected fish 3 months after transfer to sea water showed a normal cortical zone in the periphery of the lens in 24 out of 28 fish.