Pancreas disease in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in Norway.

The present paper describes, for the first time, clinical signs and pathological findings of pancreas disease (PD) in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in sea water in Norway. Similarities and differences with reports of PD from Ireland and Scotland are discussed. Samples of 68 rainbow trout from disease outbreaks on 14 farms and from 155 Atlantic salmon from outbreaks on 20 farms collected from 1996 to 2004 were included in the present study. The histopathological findings of PD in Atlantic salmon and rainbow trout in sea water were similar. Acute PD, characterized by acute necrosis of exocrine pancreatic tissues, was detected in nine Atlantic salmon and three rainbow trout. Salmonid alphavirus (SAV) was identified in acute pancreatic necroses by immunohistochemistry. Most fish showed severe loss of exocrine pancreatic tissue combined with chronic myositis. Myocarditis was often but not consistently found. Kidneys from 40% and 64% of the rainbow trout and Atlantic salmon, respectively, had cells along the sinusoids that were packed with cytoplasmic eosinophilic granules. These cells resembled hypertrophied endothelial cells or elongated mast cell analogues. Histochemical staining properties and electron microscopy of these cells are presented. SAV was identified by RT-PCR and neutralizing antibodies against SAV were detected in blood samples.

Granulomatous uveitis associated with vaccination in the atlantic salmon.

This study addressed histologic and immunopathologic changes in ocular tissues and investigated the distribution of major histocompatibility class II (MHC class II)-positive cells in Atlantic salmon (Salmo salar) suffering from severe postvaccination disease. Twenty-nine fish with generalized inflammation, probably a result of vaccination, were investigated. One individual that had escaped vaccination was included in the study. Material was investigated by cultivation methods for fungi and bacteria. Histology using conventional staining procedures and immunohistochemistry with antisera against MHC class II beta chain were performed. No growth was observed from the cultivation investigations. Histology revealed occlusion of the lumen in the larger choroid vessels and in the choriocapillaris, inflammatory infiltrations and loss of structure in the choroid rete, and, in some cases, aggregations of multinucleated giant cells (MGC) and Splendore-Hoeppli material. Immunohistochemistry demonstrated massive MHC class II+ cellular infiltrations in the uveal tract. Such infiltrations were also seen in the ventral ciliary cleft, a condition that is associated with glaucoma. Immunoreactive cells included dendritelike cells, epithelioid cells, and MGCs. The endothelia of smaller vessels were frequently MHC class II+, and immunoreactive infiltrations were seen in the optic nerve in several individuals. No pathologic changes were detected in the unvaccinated individual. In conclusion, generalized inflammatory reactions in fish may lead to severe ocular inflammation, occlusion of uveal vessels, and perivascular changes with MHC class II+ upregulation in cells in the uveal tract and optic nerve.

Vaccination induces major histocompatibility complex class II expression in the Atlantic salmon eye.

The aim of the study was to investigate the presence, distribution and density of major histocompatibility complex (MHC) class II+ cells in the ocular tissues of the Atlantic salmon, Salmo salar, prior to and following vaccination. Eyes were collected 14 days prior to and at 4, 11, 25 and 39 days and 4 months subsequent to vaccination with a commercial fish vaccine. A quantitative analysis was performed in sections on the number of immunopositive cells in the retinal layers. In all groups, MHC class II+ cells were detected in the area of the limbus but not in the central parts of the cornea. In the uvea, immunopositive cells were present in unvaccinated and vaccinated fish. Abundant immunopositive cells were identified in the choroid rete (or choroid gland) in all groups as well as in the ventral ciliary cleft, where macrophage-like MHC class II+ cells were seen. Quantitative histology of the retina revealed a significant increase in MHC class II+ cells in the outer plexiform layer (OPL) and the inner nuclear layer (INL) 4 days following vaccination. Positive cells were detected in all layers of the retina with the exception of the photoreceptor layer.

Production of rabbit antisera against recombinant MHC class II beta chain and identification of immunoreactive cells in Atlantic salmon (Salmo salar).

In the present work, rabbit antisera recognising the Atlantic salmon (Salmo salar) MHC class II beta chain polypeptide were produced and used in immunoblotting, immunohistochemistry and immunogold electron microscopy. A construct encoding the beta1 and beta2 domains fused to the E. coli protein thioredoxin was used to express the recombinant MHC class II beta chain. Immunoblotting revealed a band of approximately 30kDa in total protein samples from head kidney, spleen, gills, thymus and blood leukocytes, while being absent in muscle. The distribution of MHC class II positive cells was immunohistochemically demonstrated in Atlantic salmon epithelial and haemopoietic tissues. Ultrastructural demonstration of immunoreactive organelles in mid-kidney cells was performed by immunogold electron microscopy. The results indicate expression in lymphocytes, macrophages, epithelial cells and endritic-like cells. This is the first study to address morphological MHC class II expression in a fish species.

Subclinical herpesvirus infection in farmed turbot Scophthalmus maximus.

Subclinical infections with a herpesvirus were detected by light and electron microscopic examination of juvenile turbot collected during a national surveillance programme. Virions detected in the epidermis of skin and in the epithelium of gills had a morphology consistent with those of Herpesvirus scophthalmi described from turbot in the United Kingdom and Denmark. This is the first report of herpesvirus infection in turbot in Norway.

Redescription of Neospora caninum and its differentiation from related coccidia.

Neospora caninum is a protozoan parasite of animals, which before 1984 was misidentified as Toxoplasma gondii. Infection by this parasite is a major cause of abortion in cattle and causes paralysis in dogs. Since the original description of N. caninum in 1988, considerable progress has been made in the understanding of its life cycle, biology, genetics and diagnosis. In this article, the authors redescribe the parasite, distinguish it from related coccidia, and provide accession numbers to its type specimens deposited in museums.

The anatomy of the oesophagus, stomach and intestine in common wolffish (Anarhichas lupus L.): a basis for diagnostic work and research.

The alimentary canal of laboratory-reared common wolffish (Anarhichas lupus L.) was studied using light and electron microscopy. In the oesophagus, a simple columnar microvillous epithelium with transport characteristics was observed in addition to the main striated squamous epithelium. An osmoregulatory function is proposed for the simple columnar epithelium, which was supported by wide, thin-walled vessels. In the stomach, a separate type of neck cells was observed leading into the acinar gastric glands, which morphologically consist of one cell type: chief cells. The intestine was divided into a proximal and distal segment by an intestinal valve. Pyloric caeca were not present. We propose that shallow, crypt-like structures in the intestinal mucosa are the sites of epithelial-cell proliferation in juveniles and adults. The length of microvilli decreased from approximately 4 microns in the cranial part of the proximal intestine, to 1.5 microns in the distal intestine. In the distal intestine, rod-shaped bacteria were observed between microvilli. An extensive system of thin-walled vessels was observed in the submucosa of juvenile and adult wolffish stomach and intestine. Eosinophilic granular cells were numerous in the perivascular connective tissue in the gastric and intestinal submucosa of adults and juveniles, but were not observed in larvae.

Genetic markers linked to neuronal ceroid lipofuscinosis in English setter dogs.

The neuronal ceroid lipofuscinoses (NCL) are a group of fatal autosomal recessive neurodegenerative diseases occurring in human and some domesticated animal species. A canine form of the disease (CNCL) has been extensively studied in a Norwegian colony of inbred English setters since 1960. A resource family developed for genetic mapping and comprising 170 individuals was typed for 103 genetic markers. Linkage analysis showed three genetic markers to be linked to the disease locus with the closest marker at a distance of about 3 CM. Two other loci were linked with these markers making a linkage group of five genetic markers. The linkage group spanned a distance of 54 CM. Two genes for human forms of the disease, CLN2 and CLN3, have been identified and mapped to human chromosome 11p15 and 16p12, respectively. The present study did not indicate any linkage between CNCL and the canine CLN3 homologue or to homologues of markers for genes that map close to human CLN2.

An outbreak of cataract with lens rupture and nuclear extrusion in wolf-fish (Anarhicas spp.).

Eight spotted (Anarhicas minor Olafsen) and five common wolf-fish (Anarhicas Lupus L), developed cataracts shortly after an episode of increased water temperature and decreased salinity 5 years prior to examination. On clinical examination, the cataracts were mostly bilateral and complete, and a majority of the lenses were lobulated. Inflammatory reaction was, apart from one eye with severe inflammation, limited to iris atrophy. Of the 14 eyes collected for pathomorphological examination, eight had lens rupture with extrusion of the nucleus to the posterior chamber, two showed partly dislocated nuclei with posterior protrusion and two lenses were morgagnian. A multilayered squamous epithelium with abundant desmosomes had developed on the surface of seven of the extruded nuclei. The main cause of the cataracts was considered to be the rapid decrease in water salinity, causing osmotic changes within the eyes with secondary swelling of lens fibers and rupture of the lens capsules.

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.

Identification and characterization of Neospora caninum tachyzoite antigens useful for diagnosis of neosporosis.

The purpose of the present study was to identify antigens of the protozoan Neospora caninum that could be useful for the diagnosis of neosporosis in domestic animals. As revealed by immunoblotting, immune sera from a wide range of animal species exhibited a similar recognition pattern of four major and several minor N. caninum antigens. In contrast to preinoculation sera, all tested immune sera recognized nonreduced immunodominant 17-, 29-, 30-, and 27-kDa antigens. A 46-kDa protein which showed faint recognition by preimmune sera also exhibited a strong response by immune sera. Immunolocalization of the four immunodominant N. caninum antigens was investigated by immunogold electron microscopy using monospecific polyclonal antisera. The 17-kDa antigen appears to be associated with the body part of the rhoptries, while the 29- and 30-kDa antigens were associated with the dense granules, network, and limiting membrane of the parasitophorous vacuole. Studies were also conducted to compare antibody responses to N. caninum and the related protozoan Toxoplasma gondii. Although N. caninum and T. gondii (RH strain) tachyzoites shared a few cross-reacting antigens, the immunodominant antigens of both parasites were not recognized by heterologous sera. Also, immunogold staining with rabbit anti-Neospora hyperimmune serum exhibited almost no labeling of external membranes of Neospora tachyzoites compared with the very marked labeling seen when Toxoplasma tachyzoites (RH strain) were incubated with rabbit anti-Toxoplasma hyperimmune serum. These unique antigenic differences should be useful in developing a diagnostic assay for N. caninum.

Extraglomerular lesions in kidneys of mink with encephalitozoonosis.

Extraglomerular renal lesions were studied by light and electron microscopy in 13 farmed mink which showed cataractous eyes associated with spontaneous encephalitozoonosis. The extraglomerular renal lesions consisted of multiple renal cysts, multifocal-to-coalescing interstitial nephritis and vasculitis. Tubular cysts of varying size were present in the corticomedullary junction and medulla. The inflammatory infiltrates were composed mostly of lymphocytes and plasma cells and usually accompanied an interstitial fibrosis. Vasculitis, perivasculitis and sclerotic arteries were frequently seen.

Experimental pestivirus infections in pregnant goats.

Fifty pregnant goats, inoculated intramuscularly at different gestational stages with a non-cytopathic ovine pestivirus or a cytopathic bovine pestivirus, all developed pestivirus-neutralizing antibodies within 5 weeks of inoculation. The incidence of reproductive failure was similar for the two agents. Parturition at term with only healthy kids occurred in 13 (26 per cent) of the goats. Viable kids were not born to any of the 17 goats inoculated at about day 40 of gestation. Three of the 17 delivered dead or weak kids, seven aborted and three of seven which were necropsied during pregnancy had markedly underdeveloped and autolysed or mummified fetuses in utero, while four were barren. When inoculated at around the 60th day of gestation, two of 18 animals gave birth to only healthy kids, 12 to dead and/or weak kids, two aborted and, at necropsy, a small, decomposed fetus was found in one goat while one other was barren. In this group, one kid was ataxic and seven others had body tremors characteristic of border disease. One of the latter kids was viable. Of 15 goats inoculated at around day 100 of gestation, 11 gave birth to healthy kids only, three to dead and/or weak kids and one aborted. In 23 progeny, histological changes in the central nervous system (CNS) consisted mainly of cerebral white matter necrosis, cerebellar dysplasia, hypercellular areas in white matter and lymphocytic perivascular cuffings. All seven weak-born kids with signs of border disease had CNS lesions, particularly cerebellar dysplasia and/or hypercellular areas. Non-cytopathic pestivirus was isolated from tissues from all eight progeny examined in the 40-day inoculation group, from tissues and/or serum from 10 of 23 progeny in the 60-day group, and from four of 24 in the 100-day group. Persistent infection was demonstrated in a healthy kid, in a viable shaker and in two other kids which appeared normal at birth. Examination of offspring before ingestion of colostrum revealed pestivirus antibodies in one kid in each of the 40- and 60-day inoculation groups and in five kids in the 100-day group.

Outbreaks of border disease in goats induced by a pestivirus-contaminated orf vaccine, with virus transmission to sheep and cattle.

Five herds with a total of 276 female goats experienced severe outbreaks of reproductive failure due to a pestivirus-contaminated experimental orf vaccine given early in the breeding season. The reproductive failures comprised barrenness in 42 goats, abortion in 53 and, in 118, the birth of dead or weak kids. The incidence of female goats with such failures was 82 per cent overall, herd incidence rates ranging from 79 to 96 per cent. No progeny showed characteristic signs of border disease (BD). Microscopic lesions in the CNS were moderate, mostly characterized by hypercellularity, cell disorganization and decreased myelin content. Non-cytopathic strains of pestivirus were demonstrated in tissue or serum from two weak-born and two apparently healthy kids. Neutralizing antibodies against a bovine pestivirus were found in all 254 goats examined about 4 months after vaccination. At the end of the breeding season, all kids were removed and 74 young kids were introduced from presumably normal herds. Approximately 8 months later, 64 (86 per cent) of the bought-in kids had pestivirus antibodies. Two years after the outbreaks, such antibodies were found in all but three of 127 vaccinated goats still alive, and another 2 years later, in all 22 vaccinated goats in one of the herds. Sheep also were kept on the same five farms and cattle on one. Sheep in two of the flocks showed reproductive failure probably caused by pestivirus transmitted from infected goat offspring. A pair of twin lambs showed nervous signs and brain lesions characteristic of BD, while 17 other lambs had a very low growth rate. Virus was not isolated from any lamb. The prevalence rates of ewes with pestivirus antibodies in the two affected flocks were 33 and 72 per cent, respectively. One of nine cows aborted a pestivirus-infected foetus, and all were antibody-positive.

Neuropathology and host-parasite relationship of acute experimental toxoplasmosis of the blue fox (Alopex lagopus).

The neuropathology and host-parasite relationship of experimental infection with the RH-strain of Toxoplasma gondii were studied in 27 blue foxes (Alopex lagopus) aged 0 to 23 days, using light microscopy, including immunohistochemical staining, and transmission and scanning electron microscopy. All cases displayed multifocal necrotic lesions with numerous parasitic tachyzoites in the brain and spinal cord. The gray matter and the meninges were most seriously affected. Although a wide variety of cell types were parasitized, neurons and astrocytes seemed to be the main target cells. Individual parasitophorous vacuoles usually contained only a few tachyzoites, with rosette formations as a prominent feature. The present ultrastructural study supports the theory that the parasites actively invade the host cells by mechanisms that are different from those of phagocytosis. This is apparently the first report indicating that the formation of the network of tubular structures within the parasitophorous vacuole of T. gondii is associated with a transient, sack-like formation in the posterior end of the tachyzoites.

Experimental pestivirus infections in newborn goat kids.

Two experiments were carried out in which 37 healthy newborn goat kids were inoculated with a non-cytopathic ovine (BDV) or a cytopathic bovine pestivirus (BVDV) by intramuscular or intracerebral injection. No kids showed signs of disease or gross lesions which could be attributed to these viruses, but inoculated kids had lower mean growth rates than the controls. Significant histological changes in the CNS of 14 kids were restricted largely to the white matter and consisted mainly of hypercellular foci comprising microglial/histiocytic cells and mild perivascular infiltration by mononuclear cells. Varying degrees of infiltration of the myocardium by lymphocytes and plasma cells were observed. All kids remained negative for neutralizing antibodies against pestivirus until 2 to 3 weeks after infection. Titres increased during the following weeks. Pestiviruses were recovered from kids necropsied 10 days after inoculation, but not from any kids killed 20 days after inoculation or later. Non-cytopathic virus was isolated from various tissues of four kids that had received BDV and three kids that had been given BVDV. Cytopathic viruses were not recovered from any kids. Mean white blood cell counts in all kids were within the normal range at 4 and 8 weeks after inoculation. The lymphocyte response to stimulation by phytohaemagglutinin was significantly increased on both sampling occasions in the BDV-inoculated kids, while in the BVDV-inoculated animals, a similar increase was seen only at 8 weeks.

Spongy degeneration of white matter in the central nervous system of silver foxes (Vulpes vulpes).

A disorder of central nervous white matter in Norwegian-bred silver foxes is described from the case histories of 21 clinically affected foxes. The main presenting sign of this disorder was caudal limb ataxia, which appeared between 2 1/2 and 4 months of age and progressed over the next 4-8 weeks. Only four affected foxes were allowed to live beyond this period, but they showed moderate to marked improvement. Light microscopic examination of specimens from 16 affected foxes necropsied between 3 1/2 and 6 1/2 months of age revealed lesions that were restricted to the white matter of brain and spinal cord. The lesions were characterized by a symmetrical spongy change with vacuoles of varying sizes and included significant myelin deficiency. There was a relative preservation of axons and nerve cells and no significant inflammation or vascular reaction. An astrocytic hypertrophy was usually associated with the spongy change. Ultrastructural examination of central nervous tissue from two, perfusion-fixed, 6-month-old foxes showed intramyelin vacuoles resulting from splitting of the myelin lamellae at the intraperiod line and was interpreted as indicating myelin edema. Expanded extracellular spaces and watery astrocytic processes also contributed to the vacuolar appearance. Astrocytic processes in affected areas were hypertrophic and contained abundant filaments. Although the 16 silver foxes had severe clinical signs, their lesions had features in common with the juvenile form of Canavan's disease in children and a spongy degeneration reported in Labrador Retrievers; however, the clinical course in the foxes was not uniformly progressive.