High Gyrodactylus salaris infection rate in triploid Atlantic salmon Salmo salar

We describe an unusually high infection rate of Gyrodactylus salaris Malmberg in juvenile Atlantic salmon Salmo salar L. of Baltic Sea origin, which are generally believed to be more resistant to G. salaris than East Atlantic salmon populations. Based on analyses of mitochondrial (complete cytochrome oxidase 1 [CO1] gene, 1548 bp) and nuclear (ADNAM1, 435 bp; internal transcribed spacer [ITS] rDNA region, 1232 bp) DNA fragments, the closest relatives of the characterized Estonian G. salaris strain were parasites found off the Swedish west coast and in Raasakka hatchery, Iijoki (Baltic Sea, Finland). Analyses of 14 microsatellite loci of the host S. salarrevealed that approximately 40% of studied fish were triploids. We subsequently identified triploid Atlantic salmon of Baltic origin as more susceptible to G. salaris infection than their diploid counterparts, possibly due to compromised complement-dependent immune pathways in triploid salmon. This is in accordance with earlier studies that have shown elevated susceptibility of triploids to various viral or bacterial pathogens, and represents one of the first reports of increased susceptibility of triploid salmonid fish to an ectoparasite. However, further experimental work is needed to determine whether triploid Atlantic salmon is generally more susceptible to G. salaris compared to their diploid counterparts, irrespective of the particular triploidization method and population of origin.     

Dexamethasone treatment affects skin mucous cell density in Gyrodactylus derjavini infected Salmo salar

Atlantic salmon, Salmo salar, is normally rather refractive to infection with the ectoparasitic monogenean Gyrodactylus derjavini but dexamethasone treatment of the host increases the susceptibility. The causative mechanisms were elucidated in this work. Groups of Atlantic salmon were treated by intra-peritoneal dexamethasone injections and subsequently infected with G. derjavini. It was shown that both the infection level and the mucous cell density of caudal and pelvic fins were affected by the treatment. Significantly higher mucous cell densities were found on infected and treated fish whereas non-infected and treated fish showed no significant elevation of cell density. This suggests that mucous cell discharge elicited by infection is inhibited by the drug. The association with elevated parasite counts in these fish can be explained either by decreased anti-parasitic mucus action or by parasite predilection for intact mucous cells.     

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus

Atlantic salmon Salmo salar L. (Salmonidae) were experimentally infected with Spironucleus barkhanus (Diplomonadida: Hexamitidae). Parasites were found in the blood 1 to 8 wk after infection, after which they disappeared from the blood and were found mainly in the internal organs (e.g. spleen and liver), eye socket or muscles. Mortality (38 out of 40 infected fish) occurred when fish had lesions in internal organs and/or on the body surface. Uninfected fish cohabiting with infected fish became infected after 4 wk, indicating direct transmission. There was no difference in susceptibility to spironucleosis between 3 different families of Atlantic salmon. All families developed the disease with a similar pattern of parasitaemia in the blood, similar clinical signs and gross pathology, and with very high mortality (29 out of 30). Clinical signs of systemic spironucleosis may include anemia, skin blisters, muscle ulcerations or unilateral exophthalmia. Gross pathologies include hemorrhaging of internal organs, splenomegaly or deformed (globulated) spleen, or granulomatous lesions in the spleen and liver.     

Infestations of Atlantic salmon, Salmo salar, by Gyrodactylus salaris in Norwegian rivers

Over the 6 years 1980–85, 212 Norwegian rivers have been examined for occurrence of Gyrodactylus salaris: it was found in 26 rivers and six salmon hatcheries scattered throughout the country from Troms county in the north to Sogn og Fjordane in western Norway. The distribution of G. salaris is connected with the stocking of fish from infected salmon hatcheries. The populations of salmon parr have been drastically reduced in the infected rivers. In later years catches of ascending salmon in these rivers have also sharply declined: in 1984 salmon fishery losses were estimated at 250–500 t.
Gyrodactylus salaris is most probably a recent introduction to Norwegian rivers. A primary aim is to exterminate this parasite from all infected rivers and hatcheries: so far this has been accomplished in one river and one hatchery.     

Identification of genetic markers associated with Gyrodactylus salaris resistance in Atlantic salmon Salmo salar

Gyrodactylus salaris Malmberg, 1957 is a freshwater monogenean ectoparasite of salmonids, first recorded in Norway in 1975 and responsible for extensive epizootics in wild Atlantic salmon Salmo salar L. The susceptibility of different populations of Atlantic salmon to G. salaris infection differs markedly, with fish from the Baltic being characterised as relatively resistant whereas those from Norway or Scotland are known to be (extremely) susceptible. Resistance to Gyrodactylus infection in salmonids has been found to be heritable and a polygenic mechanism of control has been hypothesised. The current study utilises a 'Quantitative trait loci' (QTL) screening approach in order to identify molecular markers linked to QTL influencing G. salaris resistance in B1 backcrosses of Baltic and Scottish salmon. Infection patterns in these fish exhibited 3 distinct types; susceptible (exponential parasite growth), responding (parasite load builds before dropping) and resistant (parasite load never increases). B1 backcross fish were screened at 39 microsatellite markers and single marker-trait associations were examined using general linear modelling. We identified 10 genomic regions associated with heterogeneity in both innate and acquired resistance, explaining up to 27.3% of the total variation in parasite loads. We found that both innate and acquired parasite resistance in Atlantic salmon are under polygenic control, and that salmon would be well suited to a selection programme designed to quickly increase resistance to G. salaris in wild or farmed stocks.     

The ultrastructure of hypersymbionts on the monogenean Gyrodactylus salaris infecting Atlantic salmon Salmo salar

There is increasing pressure to develop alternative control strategies against the pathogen Gyrodactylus salaris, which has devastated wild Atlantic salmon Salmo salar in Norway. Hyperparasitism is one option for biological control and electron microscopy has revealed two ectosymbionts associated with G. salaris: unidentified rod-shaped bacteria, and the protist, Ichthyobodo necator. No endosymbionts were detected. The flagellate I. necator occurred only occasionally on fish suffering costiosis, whereas bacterial infections on the tegument of G. salaris were observed throughout the year, but at variable densities. Bacteria were seldom observed attached to fish epidermis, even when individuals of G. salaris on the same host were heavily infected. Wounds on salmon epidermis caused by the feeding activity of bacteria-infected G. salaris did not appear to be infected with bacteria. On heavily infected gyrodactylids, bacteria were most abundant anteriorly on the cephalic lobes, including the sensory structures, but no damaged tissue was detected by transmission electron microscopy in the region of bacterial adherence. Furthermore, transmission and survival of infected G. salaris on wild salmon did not appear to be influenced by the bacterial infection. The lack of structural damage and impact on G. salaris biology indicates that these bacteria are not a potential agent for control of gyrodactylosis. However, this may not be the case for all gyrodactylid-bacterial interactions and a review of bacterial infections of platyhelminths is presented.     

Infestations by Gyrodactylus sp. of Atlantic salmon, Salmo salar L., in Norwegian rivers

Young Atlantic salmon, Salmo salar , brown trout, Salmo trutta , and Arctic char, Salvelinus alpinus , from eight rivers in North and Mid Norway were examined for Gyrodactylus. The fish were collected from 1975 to 1980. A Gyrodactylus salaris type was observed infecting salmon from six of these rivers. No trout or char were infected. A high frequency and intensity of infection of salmon were observed in all but one of the rivers surveyed. In the Saltdalselva, only one specimen of Gyrodactylus infecting one fish was observed. No subsequent mortality of salmon was observed in this river while there were signs of a high mortality of salmon in the other rivers. The salmon parr were more frequently attacked than the fry, the mortality of salmon seemed to have happened in most of the rivers one year after the first observations of Gyrodactylus were made. The mortality of salmon and the high infection rate of Gyrodactylus in these rivers appears unique, and as far as known to the authors there are no other described cases of mortality due to Gyodactylus in Atlantic salmon in natural waters. The reasons for the outbreak of Gyrodactylus in these rivers are not known. Two theories are discussed: one that the fish were weakened by environmental factors and the other that Gyrodactylus was introduced from some of the infected salmon hatcheries in Scandinavia.     

Susceptibility of Baltic and East Atlantic salmon Salmo salar stocks to Gyrodactylus salaris (Monogenea)

The susceptibility of a Baltic salmon stock Salmo salar (Indals?lv, central Sweden) to Norwegian Gyrodactylus salaris (Figga strain, central Norway) was experimentally tested and compared with previously obtained results on East Atlantic salmon (Lierelva, SE Norway). Contrary to expectation, the Baltic salmon, which had no prior exposure to this parasite strain, appeared almost as susceptible as the Norwegian salmon parr that naturally experience G. salaris-induced mortality. Individually isolated salmon of both stocks sustained G. salaris infections with little evidence of innate resistance. A few individuals of the Indals?lv stock controlled their infection from the beginning, but overall there was considerable heterogeneity in the course of infection in both stocks. On individual hosts, G. salaris growth rates declined steadily throughout the infection, a trend which was particularly marked amongst the Lierelva stock. On shoaling Lierelva fish, there was some evidence of reduced parasite population growth towards the end of the infection; this was not apparent in Indals?lv fishes. These results reflect a growing awareness that not all Baltic salmon may be resistant to Norwegian G. salaris, and that Norwegian and Baltic G. salaris strains may differ in virulence. Consequently, management decisions concerning this parasite-host system should be based upon the actual, and tested, susceptibility of stocks under consideration and not upon identification of stocks as either Atlantic or Baltic.     

Diseases of farmed Atlantic salmon Salmo salar associated with infections by the microsporidian Paranucleospora theridion

The microsporidian Paranucleospora theridion was discovered in Atlantic salmon Salmo salar suffering from proliferative gill disease in a marine farm in western Norway in 2008. The parasite develops in cells of the reticuloendothelial system, cells important for normal immune function. The aim of this study was to see if P. theridion could play a part in some of the diseases with unclear causes in salmon production in Norway, i.e. proliferative gill disease (PGI), pancreas disease (PD), heart and skeletal muscle inflammation (HSMI) and cardiomyopathy syndrome (CMS). P. theridion was present in all areas with salmon farming in Norway, but high prevalence and densities of the parasite in salmon and salmon lice were only seen in southern Norway. This region is also the main area for PGI and PD in Norway. Quantification of pathogens associated with PGI, PD, HSMI and CMS diagnoses showed that P. theridion levels are high in southern Norway, and may therefore play a role in susceptibility and disease development. However, among the different diagnoses, fish with PGI are particularly heavily infected with P. theridion. Therefore, P. theridion appears as a possible primary agent in cases with high mortality in connection with PGI in western Norway.     

Experimental infection models and susceptibility of Atlantic salmon Salmo salar to a Scottish isolate of infectious salmon anaemia virus.

Infection models for both fresh and seawater salmon were established using a Scottish isolate of infectious salmon anaemia virus (ISAV). Modes of infection were intra-peritoneal injection, cohabitation and immersion exposure, and a range of doses was tested. Development of these models using a Scottish isolate of ISAV provided an approximation of the minimum infective dose leading to mortality under different infection regimens. The models also allow prediction of the time to first mortality and an estimation of expected total mortality following the various routes of infection. Such knowledge is important to the development of anti-ISAV vaccines and to future studies aimed at understanding the biology of ISAV in general.     

Comparative susceptibility of Atlantic salmon (Salmo salar) to the enteric redmouth bacterium and Aeromonas salmonicida

The bacterium causing enteric redmouth (ERM) and Aeromonas salmonicida were found to be equally pathogenic for fingerling Atlantic salmon (Salmo salar). Injection of 5 x 10(5) cells of ERM or A. salmonicida killed all salmon within 96 h. After a 30 min exposure to water-borne cells of the two test bacteria about one-half of the test salmon died within 14 days. Both ERM and A. salmonicida were transmitted horizontally. Results indicate that efforts should be made to prevent introduction of ERM into watersheds where Atlantic salmon occur.     

Liver ultrastructure of juvenile Atlantic salmon (Salmo salar)

Light and transmission electron microscopy of the liver of juvenile Atlantic salmon (Salmo salar) reveals a tubular arrangement of parenchymal cells, with biliary passages typically located at the center of tubules. Hepatocytes generally contain a single nucleus surrounded by a cuff of rough endoplasmic reticulum (RER), with many round to elongate mitochondria associated with the perinuclear RER. Whereas glycogen deposits are common and usually lie at the cell periphery, parenchymal cells seldom contain lipid droplets. Golgi complexes and heterogeneous dense bodies also occur in many hepatocytes, often in close proximity to bile canaliculi. Numerous microvilli from hepatocytes extend into the subendothelial space of Disse, which is also the location of stellate fat-storing cells. Interhepatocytic macrophages, sometimes containing prominent phagolysosomes and residual bodies, are common in the liver. The intrahepatic biliary system consists of intercellular canaliculi, bile pre-ductules, ductules, and ducts. In contrast to some other teleosts, the liver of the Atlantic salmon contains no intracellular bile canaliculi or Kupffer cells. The hepatic endothelium, arterioles, and perivenous regions are also described.     

Phylogenetic position of a paramyxovirus from Atlantic salmon Salmo salar

A paramyxovirus has been isolated from Atlantic salmon Salmo salar suffering from epitheliocystis. This virus does not cause any mortality when used to challenge disease-free salmon, but has been associated with 2 cases of mortality in salmon farms in Norway. Atlantic salmon paramyxovirus (ASPV) has been suggested as a name for the virus. The ASP virus is a slow-growing virus in cell cultures (rainbow trout gill cells: RTgill-W1). Little is known about its importance and its phylogenetic position is uncertain. Hence, the need for a fast and sensitive diagnostic method for studying the prevalence of this virus in salmon farms and for more basic knowledge about its identity were the motivation for this study. A partial nucleotide sequence (816 bp) from the large protein (L protein) gene of the ASP virus has been sequenced from 2 different isolates. The putative amino acid sequence has been compared with the L protein of other paramyxoviruses. This sequence gives strong support to a relationship between the ASP virus and members of the subfamily Paramyxovirinae, genus Respirovirus.     

Identification of olfactory receptor genes in Atlantic salmon Salmo salar

It has been hypothesized that salmonids use olfactory cues to return to their natal rivers and streams. The key components of the molecular pathways involved in imprinting and homing, however, are still unknown. Aquatic chemical cues are received through the nares and into the nasal cavity that contains a single olfactory organ, the olfactory rosette. The olfactory rosette contains sensory neurons, each of which is thought to express only one olfactory receptor. If odorants are involved in salmonid homing migration then olfactory receptors should play a critical role in the dissipation of information from the environment to the fish. Therefore, to understand the molecular basis for imprinting and homing in Atlantic salmon Salmo salar it is important to identify and characterize the repertoire of olfactory receptors in this species. The first public assembly of the S. salar genome was searched for genes encoding three of the superfamilies of fish olfactory receptors: V2R-like (olfc), V1R-like (ora) and main olfactory receptor (mor). A further six ora genes were added to ora1 and ora2, which had been described previously. In addition, 48 putative mors were identified, 24 of which appear to be functional based on their gene structures and predicted amino-acid sequences. Phylogenetic analyses were then used to compare these S. salar olfactory receptor genes with those of zebrafish Danio rerio, two pufferfish species Takifugu rubripes and Tetraodon nigroviridis, medaka Oryzias latipes and three-spined stickleback Gasterosteus aculeatus.     

Experimentally induced marine flexibacteriosis in Atlantic salmon smolts Salmo salar. I. Pathogenicity

Tenacibaculum maritimum causes marine flexibacteriosis in many cultured fish species, including Atlantic salmon Salmo salar in Tasmania, Australia. Several aspects of the pathogenicity of this bacterium were investigated in naive Atlantic salmon smolts using different isolates, growth conditions and doses to produce a model of infection. We found that T. maritimum is pathogenic to Atlantic salmon using either marine Shieh's or marine Ordal's culture medium. The use of aeration in broth culture produced a dose effect in challenge due to a 'clumping' of the bacteria during culture. The virulence of a strain appears to be connected with this 'clumping'; the more adherent the cells, the more pathogenic the strain. Differences in virulence between 3 strains was apparent, with 1 of the strains (89/4747) being non-pathogenic and unable to produce disease in the host. The 2 other strains (89/4762, 00/3280) were highly virulent, resulting in 100% mortalities within 3 d. A reproducible model of infection has been established in the present study using strain 89/4762. Results from the present study provide a better insight into the nature of the disease.     

Soybean meal‐induced uptake block in Atlantic salmon Salmo salar distal enterocytes

Soybean meal‐induced enteritis was characterized by the disappearance of the supranuclear vacuoles (SNV) in the enterocytes of the distal intestine of Atlantic salmon Salmo salar. The loss of SNV was correlated with an endocytosis uptake block as shown with ferritin as a marker molecule.     

Naked DNA vaccination of Atlantic salmon Salmo salar against IHNV

A naked plasmid DNA encoding the glycoprotein (pCMV4-G) of a 1976 isolate of infectious hematopoietic necrosis virus (IHNV) obtained from steelhead Oncorhynchus mykiss was used to vaccinate Atlantic salmon Salmo salar against IHNV. Eight weeks post-vaccination the fish were challenged with a strain of IHNV originally isolated from farmed Atlantic salmon undergoing an epizootic. Fish injected with the glycoprotein-encoding plasmid were significantly (p < 0.05) protected against IHNV by both immersion and cohabitation challenge. Survivors of the first challenges were pooled and re-challenged by immersion 12 wk after the initial challenge. Significant (p < 0.05) protection was observed in all of the previously challenged groups including those receiving the complete vaccine. Fish injected with the glycoprotein-encoding plasmid produced low levels of virus-neutralizing antibodies prior to the first challenge. Neutralizing antibodies increased in all groups after exposure to the IHNV. Passive transfer of pooled sera from pCMV4-G vaccinates and IHN survivors provided relative survivals of 40 to 100% compared to fish injected with sera collected from fish immunized with control vaccines or left unhandled. In this study, DNA vaccination effectively protected Atlantic salmon smolts against challenges with IHNV.