A thicker chorion gives ova of Atlantic salmon (Salmo salar L.) the upper hand against Saprolegnia infections

Since the ban of malachite green in the fish farming industry, finding alternative ways of controlling Saprolegnia infections has become of utmost importance. Much effort has been made to elucidate the mechanisms by which Saprolegnia invades fish eggs. Little is known about the defence mechanisms of the hosts, making some eggs more prone to infection than others. One clue might lie in the composition of the eggs. As the immune system in the embryos is not developed yet, the difference in infection levels could be explained by factors influenced by the mother herself, by either transferring passive immunity, influencing the physical aspects of the eggs or both. One of the physical aspects that could be influenced by the female is the chorion, the extracellular coat surrounding the fish egg, which is in fact the first major barrier to be overcome by Saprolegnia spp. Our results suggest that a thicker chorion in eggs from Atlantic salmon gives a better protection against Saprolegnia spp. In addition to the identification of differences in sensitivity of eggs in a fish farm set‐up, we were able to confirm these results in a laboratory‐controlled challenge experiment.     

Pathogenicity of Saprolegnia spp. to Atlantic salmon, Salmo salar L., eggs

Live and dead Atlantic salmon eyed eggs were challenged with eight different Saprolegnia isolates, selected because of their varied origins, known morphological characteristics and growth/germination pattern. Some isolates were also tested for pathogenicity to Atlantic salmon parr. Challenge of eggs was performed by exposure to spores in suspension or by co-incubation of live eggs with infected dead eggs. The phenotypic characteristics of the isolates were evaluated in relation to their observed pathogenicity from the challenge experiment, to identify possible virulence factors leading to egg-infection by Saprolegnia. The results from the experiments confirm that live eggs are refractory to infection with Saprolegnia spores in suspension and that an infection of live eggs can only occur from an infection nucleus represented by dead eggs or debris. It was observed that strains pathogenic to salmon parr were not particularly infective towards eggs, and the isolates that gave the highest infection rates to eggs were species considered to be saprotrophs.     

Modelling salmon lice,Lepeophtheirus salmonis,reproduction on farmed Atlantic salmon,Salmo salar L

The aim of this study was to model sea lice levels and the effect on reproduction by a stochastic simulation model and to evaluate the uncertainty of lice estimates based upon counts. Two empirical data sets were examined to parameterize the models. An overall fit of the data to the Poisson distribution was found and thus was used as the base of the stochastic models. In the model, salmon lice reproduction is not linear with the number of adult females and at low lice loads a smaller proportion of the adult female lice will reproduce. Depending on the variance structure, it was estimated that between 40% and 60% of the adult female lice will reproduce at an abundance of 0.5 adult females per fish. Lice counts, especially when examining few fish at low lice loads, are uncertain and at a true abundance of 0.1 one may count between 0 and 5 lice when examining 10 fish. Understanding the dynamics of sea lice reproduction is a key factor in the development of sustainable control strategies.     

In vitro activity of chemicals and commercial products against Saprolegnia parasitica and Saprolegnia delica strains

Oomycetes of the genus Saprolegnia are responsible for severe economic losses in freshwater aquaculture. Following the ban of malachite green in food fish production, the demand for new treatments pushes towards the selection of more safe and environment‐friendly products. In the present work, in vitro activity of ten chemicals and three commercial products was tested on different strains of Saprolegnia, using malachite green as reference compound. The compounds were screened in agar and in water to assess the minimum inhibitory concentration (MIC) and the minimum lethal concentration (MLC), respectively. Two strains of Saprolegnia parasitica and one isolate of Saprolegnia delica were tested in triplicate per each concentration. Among tested chemicals, benzoic acid showed the lowest MIC (100 ppm) followed by acetic acid, iodoacetic acid and copper sulphate (250 ppm). Sodium percarbonate was not effective at any tested concentration. Among commercial products, Virkon^?S was effective in inhibiting the growth of the mycelium (MIC = MLC = 1,000 ppm). Actidrox® and Detarox® AP showed MIC = 5,000 and 1,000 ppm, respectively, while MLCs were 10‐fold lower than MICs, possibly due to a higher activity of these products in water. Similarly, a higher effectiveness in water was observed also for iodoacetic acid.     

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.     

Genetic variation in Norwegian piscine myocarditis virus in Atlantic salmon,Salmo salar L.

Cardiomyopathy syndrome (CMS) in Atlantic salmon, Salmo salar L., is a severe cardiac disease characterized by a necrotizing myocarditis involving the atrium and the spongious part of the ventricle. The disease is caused by piscine myocarditis virus (PMCV), a double‐stranded RNA virus likely belonging to the family Totiviridae. The objective of this study was to evaluate the genetic variation in Norwegian PMCV isolates focusing on the putative structural proteins encoded by open reading frames (ORFs) 1 and 3. The virus isolates were sampled from a total of 36 farms along the Norwegian coastline. This study represents the first investigation of PMCV genome variation and shows that Norwegian isolates are highly similar, with the most divergent isolates sharing 98.6% nucleotide identity. Interestingly, amino acid sequence diversity within ORF3 is approximately threefold higher than for ORF1. While phylogenetic analysis based on concatenated nucleotide data covering ORF1 and ORF3 revealed four main clusters, the maximum sequence variation of 1.4% at the nucleotide level suggests that all Norwegian isolates belong to a single genogroup. Substantial sequence variation within farms was also observed, which may complicate future molecular epidemiological investigations. The genetic homogeneity among the Norwegian isolates might facilitate development of both diagnostic tools and an efficient vaccine against CMS in the future.     

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.     

Comparative virulence of Infectious salmon anaemia virus isolates in Atlantic salmon, Salmo salar L.

Infectious salmon anaemia virus (ISAV) surveillance in the Bay of Fundy has identified the existence of a large number of genetically distinct ISAV isolates which appear to be of variable virulence. Genetically distinct isolates are currently being designated based on sequencing of the hyper polymorphic region (HPR) of genomic segment 6, which encodes the haemagglutinin–esterase protein, but it has been difficult to elucidate a clear association between these molecular variations and variations in virulence. This has hampered the establishment of proactive management decisions regarding infected fish, and ISAV infections, regardless of type, must be treated as one. Field data of ISAV infections is difficult to collect and to compare between infections because of a wide range of confounding factors including time of year, fish stock, cage site location, mitigating factors and stressors. An important tool in determining the relationship between molecular differences and virulence comes from analysis of quarantine studies. The goal of this study was to compare the virulence, by co-habitation and intraperitoneal injection, of four regionally common and recent ISAV isolates in a controlled environment. We found significant differences in mortality between ISAV molecular isolates, and present data showing that survival of ISAV infection confers significant resistance to re-infection with a different ISAV isolate. These findings, if borne out in field studies, will significantly alter the way ISAV infections are managed in the Bay of Fundy and elsewhere.     

Saprolegnia diclina IIIA and S. parasitica employ different infection strategies when colonizing eggs of Atlantic salmon,Salmo salar L.

Here, we address the morphological changes of eyed eggs of Atlantic salmon, Salmo salar L. infected with Saprolegnia from a commercial hatchery and after experimental infection. Eyed eggs infected with Saprolegnia spp. from 10 Atlantic salmon females were obtained. Egg pathology was investigated by light and scanning electron microscopy. Eggs from six of ten females were infected with S. parasitica, and two females had infections with S. diclina clade IIIA; two Saprolegnia isolates remained unidentified. Light microscopy showed S. diclina infection resulted in the chorion in some areas being completely destroyed, whereas eggs infected with S. parasitica had an apparently intact chorion with hyphae growing within or beneath the chorion. The same contrasting pathology was found in experimentally infected eggs. Scanning electron microscopy revealed that S. parasitica grew on the egg surface and hyphae were found penetrating the chorion of the egg, and re‐emerging on the surface away from the infection site. The two Saprolegnia species employ different infection strategies when colonizing salmon eggs. Saprolegnia diclina infection results in chorion destruction, while S. parasitica penetrates intact chorion. We discuss the possibility these infection mechanisms representing a necrotrophic (S. diclina) vs. a facultative biotrophic strategy (S. parasitica).     

Feed intake of Atlantic salmon parr Salmo salar L. in relation to temperature and feed composition

The design and testing of a feed intake monitoring system based on feed waste collection is described. The system was used in a study of feed intake and growth of Atlantic salmon Salmo salar L. parr in relation to temperature (2 °C or 8 °C) and feed composition (21% fat, 50% protein and 22.5 MJ kg^?1 or 34% fat, 40% protein and 24.8 MJ kg^?1). Fish had lower feed intake and slower growth at the lower temperature, but temperature‐corrected growth (TGC) was better in fish held at the lower temperature. There was an increase in TGC over time at low temperature, probably as a result of long‐term thermal acclimation. Feed conversion for an increase in body size from 19 to 38 g was better for the fish held at low temperature. Feed consumption of low‐fat feed was higher than that of high‐fat feed, and growth of the fish fed the low‐fat feed was better at the higher temperature. These data are consistent with the ideas that the fish compensated for differences in feed energy densities to maintain energy and nutrient intakes, and that lipostatic factors may be operating to regulate feed intake and growth.     

An estimation of dietary iron requirement of Atlantic salmon, Salmo salar L., parr

A study was conducted to determine the dietary iron requirement of fingerling Atlantic salmon Salmo salar L. During the first 4 weeks of the experiment, fish with an initial weight of 5 g were fed a casein–gelatine-based purified diet which contained 11 mg iron kg^?1. Thereafter duplicate tanks (200 fish in each) were fed the casein–gelatine purified diets containing supplemental iron levels of 0, 10, 20, 30, 40, 60, 100, 200 or 400 mg iron kg^?1 (added as FeSO_?4* 7H₂O) for 12 weeks. Weight gain, body length and mortality were monitored. Liver iron and ascorbic acid concentration were analysed in addition to whole-body iron, manganese and zinc concentration. Several haematological parameters were also measured. There were no significant differences in weight gain and survival of salmon fed diets containing different iron levels. Haematological values, hepatic and whole-body iron concentrations were, however, significantly affected by the dietary iron content. Liver vitamin C concentration decreased with increasing dietary iron levels. Dietary supplementation with iron significantly reduced whole-body manganese, but no effect of dietary iron on whole-body zinc was found. Based on haematology and hepatic iron concentration, the iron requirement of Atlantic salmon was determined to be between 60 and 100 mg iron kg¹.     

Investigating factors contributing to reduced embryo survival in farm‐raised Atlantic salmon,Salmo salar L.

Embryo mortality of Atlantic salmon, Salmo salar, has been increasing for more than a decade in the State of Maine, a leading producer of this species in the United States. Increasing embryo mortality not only creates a financial bottleneck for farms but also prevents the sale of surplus eggs as an additional source of revenue. Blood and egg samples were collected at three Maine Atlantic salmon farms from female broodstock at the time of spawning over a 2‐year period. Correlative factors for reduced embryo survival were investigated by measuring egg and maternal plasma concentrations of 17β‐estradiol (E2), 11‐ketotestosterone (11‐KT), testosterone and calcium, as well as maternal hepatic ethoxyresorufin‐O‐deethylase (EROD) activity and fork length. Significant positive correlations were found between maternal plasma concentrations of E2 and 11‐KT and embryo survival. Interestingly, there was no correlation with egg concentrations of sex steroids and embryo survival, suggesting that embryo mortality does not likely rest with the maternal deposition of sex steroids into the egg, but with another hormone regulated process related to egg assembly, ovulation or post‐ovulatory ageing.     

The responses of hatchery-reared Atlantic salmon, Salmo salar L., parr to pelletted and wild prey

Abstract. Juvenile hatchery-reared Atlantic salmon, Salmo salar L., were offered choices of pelletted or four types of wild prey (chironomid larvae, ephemeropteran nymphs Ecdyanurus and Baetis and tnchopteran larvae Hydropsyche) of the same particle widths, in a test flume Preference for wild foods increased over 16 trials (1·5 h), and was greatest for Ecdyonurta and Hydropsyche, which had prey width fish fork length (PFR) ratios of 0·019 and 0·022 respectively, Pelletted food was rejected sooner when Ecdyonurus nymphs were offered as the alternative food than when chironomid larvae were (P < 0·05). Pret'erence (P < 0·05) for Ecdyonurus was achieved after 11 trials and for chironomids after 15. The time required to capture the first prey item was two to three times as long for pellets as for wild prey and did not change with experience for any prey type. Mean capture distance did not change with experience. It is concluded that acceptability of wild prey should not limit performance of pellet-fed salmon released into the wild.     

Reproductive strategies in Atlantic salmon, Salmo salar L.

Abstract. Life-history strategies are means by which animals solve the problems of successful reproduction in varying environments. Their development patterns are consequences of responses to the opportunities the environment offers them. Understanding them requires an understanding of the way they evolved, their ontogenetic development, their physiological control, and their adaptive value. The present paper views the salmonids as marine fishes, which have radiated into fresh water through using river beds as protected spawning grounds. It also takes the view that the maturation process has priority over somatic growth in fish, and that it has already been initiated by the time of first feeding. Its completion is environmentally dependent, and can be arrested annually. Whether or not it will be arrested depends on the status of the energy stores of the individual at particular critical times of year. This mechanism has adaptive value both for immediate reproductive success — adequate energy to provision the next generation — and for later overwinter survival, ensuring that if energy stores are inadequate for reproduction they are spared. Atlantic salmon show variation in their reproductive patterns, and examples are given from laboratory and aquaculture experiments to demonstrate some environmental controls which result in these variations. A hypothetical model is presented to account for the operation of these controls.     

Nocardiosis in tank-reared Atlantic salmon, Salmo salar L.

Nocardiosis is an infection caused by aerobic Gram-positive, branching, filamentous rods of the genus Nocardia . The organism is resident in both soil and plants ( Austin & Austin 1993 ; Frerichs 1993 ), and is closely related to Mycobacterium spp. Nocardiosis caused by Nocardia asteroides and N. seriolae (previously N. kampachi ) has been reported in several finfish species, both freshwater and marine. The first reported incidence was recorded by Valdez & Conroy (1963) in neon tetras, Hyphessobrycon innesi (Myers), and subsequently in other fish species, some of commercial significance including rainbow trout, Oncorhynchus mykiss (Walbaum) ( Snieszko, Bullock, Dunbar & Pettijohn 1964 ), brook trout, Salvelinus fontinalis (Mitchill), ( Campbell & MacKelvie 1968 ) and yellowtail, Seriola quinqueradiata (Temminck & Schlegel; Kubota, Kariya, Nakamura & Kira 1968 ). Despite two of these incidences occurring in salmonids, infection by Nocardia spp. in Atlantic salmon, Salmo salar L., has not previously been reported.     

The parameters of energetic status of the wild Atlantic salmon (Salmo salar L.) spermatozoa

The energetic status of fresh sperm of wild Atlantic salmon (Salmo salar L.) was presented in selected males (n=10) with motility rate ≥80%. Purine and pyridine nucleotides: adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), guanosine triphosphate, guanosine diphosphate, guanosine monophosphate, inosine monophosphate, nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate; nucleosides: adenosine, guanosine and inosine; bases: hypoxanthine, xanthine and uric acid were determined using the high‐performance liquid chromatography method. The average value of adenylate energy charge in the group was 0.77 ± 0.10, and the mean total adenine nucleotide content (TAN) was 65.1 ± 18.3 pmol × 10^?6 spermatozoa. The mean concentrations of ATP, ADP and AMP were 43.5, 11.8 and 9.9 pmol × 10^?6 spermatozoa respectively. The concentrations of the other energetic nucleotides studied were lower. In all males studied (n=23) with a motility rate from 0% to 95%, no statistically significant correlation between the per cent of motile sperm and ATP concentration was found (R_s=0.35), whereas the correlation between the per cent of motile sperm and ADP was statistically significant (R_s=0.60). A negative correlation was found between hypoxanthine and the per cent of motile spermatozoa (R_s=?0.44). Our results suggest that hypoxanthine is the final product of salmon spermatozoa of adenine nucleotide catabolism.     

Haemorrhagic kidney syndrome of Atlantic salmon, Salmo salar L.

This report describes a new syndrome affecting farmed Atlantic salmon on the Canadian east coast that has resulted in increased morbidity and mortality in affected stocks. The major pathological findings are apparent only microscopically and include renal interstitial haemorrhage and acute tubular necrosis and tubular casting. As a result, the disease has become known as haemorrhagic kidney syndrome (HKS). Affected fish are lethargic and anorectic, and lack external lesions. Clinically, HKS fish are anaemic, hypoproteinaemic and hyperosmolalic, with increased serum concentrations of sodium and chloride. At necropsy, internal changes ranged from apparently normal to include one or several of the following: swelling and/or patchy reddening of the kidney, pale gills, exophthalmos, serosanguinous ascites, darkening of the posterior intestine and splenomegaly. Ultrastructurally, viral inclusions were found in the cytoplasm of erythrocytes of HKS fish, and there were unusual electron‐dense inclusions within the tips of renal tubular microvilli of HKS fish. The significance and relevance of the ultrastructural findings to HKS are unknown. Virus isolation was attempted using CHSE, RTG‐2, FH‐10, BB and EPC cell lines; no virus was isolated. Bacteriological analysis failed to reveal significant pathogens. Analysis of tissues for heavy metals and pesticides was negative. Assays for clostridial toxins, lipopolysaccharide and verotoxins were negative. The aetiology of HKS remains unresolved.     

Development and detection of phosphorus deficiency in Atlantic salmon, Salmo salar L., parr and post-smolts

A pathological condition was induced in Atlantic salmon, Salmo salar L., by feeding a diet severely deficient in phosphorus. Parallel results were obtained in two consecutive experiments, one in fresh water and one in sea water. The deficiency condition was first manifest as a reduction in whole-body calcium (Ca) and phosphorus (P) levels and the development of abnormally soft and malformed bones. In later stages, growth was severely impaired and there was elevated mortality compared with fish fed a diet with sufficient phosphorus.
Two semipurified casein-based test diets were used in the experiments. The low-P diet contained only the P from casein, 3.5 g kg⁻¹ or 0.35%, whereas the reference diet was supplemented with P and contained 9.0 g kg⁻¹ or 0.9%, The diets were fed to salmon parr in fresh water (initial weight 4.7 g) and to post-smolts in sea water (initial weight 113 g) for 12 and 15 weeks, respectively. In both experiments, the fish grew adequately for a prolonged period, whereas the whole-body Ca and P levels declined almost immediately. Within a few weeks, the whole-body P content was reduced to ≊ 65% of the initial value, and Ca to 40%. The reductions in mineral content were most severe in the mineral-rich tissues, bones and scales. On gross examination, all bony structures were abnormally soft. The ribs were wrinkly, and the spine displayed scoliotic changes. These changes were correlated to the reduction in bone mineralization, and were manifest at the time when reductions in growth rate could be noted.
After the deficient fish were fed adequate levels of P for 9 weeks, their whole-body mineral levels returned to normal. The bone lesions did not heal completely during the same period of time.