Biomedical subjects

F Nilsen

Publications and source records attributed to F Nilsen.

16 recordsLinked to original sources

PubMed2004

Characterization of nodavirus and viral encephalopathy and retinopathy in farmed turbot, Scophthalmus maximus (L.).

An outbreak of nodavirus infection in turbot larvae is described with respect to histopathology, immunohistochemistry, cell culture cultivation, RT-PCR amplification and sequence analysis of the capsid protein gene RNA2. Affected turbot developed classical signs of viral encephalopathy and retinopathy (VER) with abnormal swimming behaviour and high mortality levels. In the acute stage of infection, light microscopy revealed vacuolation of the central nervous system (CNS), with positive immunohistochemical staining for nodavirus. Later in the infection, CNS lesions appeared more chronic and contained clusters of cells immunopositive for nodavirus. Bacterial overgrowth in the intestines of the fish may have provoked or influenced the course of the nodavirus infection. We were unable to propagate the virus in cell culture. While RT-PCR using primers designed to detect Atlantic halibut nodavirus gave negative results, further testing with primers complementary to a more conserved region of RNA2 resulted in amplification of a product of the expected size. The entire RNA2 segment was cloned and sequenced. Sequence alignment showed that the turbot nodavirus (TNV) was different from previously described fish nodaviruses. In addition, phylogenetic analysis based on an 823 nt region of the sequence indicated that TNV clustered outside the four established fish nodavirus genotypes, suggesting a fifth genotype within the betanodaviruses.

Amino Acid Sequence↗

PubMed2002

Ovipleistophora gen. n., a new genus for Pleistophora mirandellae-like microsporidia.

Based on ultrastructural study and molecular analysis, a new genus, Ovipleistophora, is established for Pleistophora mirandellae-like microsporidia from roach and ruff oocytes. Unlike Pleistophora, Ovipleistophora has a thick additional envelope around the meront. This envelope breaks open to release the cells into the host cell cytoplasm. The cells, becoming multinuclear sporogonic plasmodia, already have a surface coat that transforms into the sporont wall and eventually into the sporophorous vesicle wall. The surface coat and its transformation differ from those of Pleistophora, but bear some resemblance to those of Trachipleistophora. In Trachipleistophora the sporonts, however, do not form plasmodia, as they do in Ovipleistophora and Pleistophora. Small subunit ribosomal DNA analysis supports the establishment of the new genus and assignment of P. mirandellae from 2 different fish hosts to the same species. The same small subunit ribosomal DNA analysis lends support for transferring P. ovariae into the genus Ovipleistophora.

Animals↗

PubMed2001

Diseases, prophylaxis and treatment of the Atlantic halibut Hippoglossus hippoglossus: a review.

After substantial investments in research, the Atlantic halibut Hippoglossus hippoglossus is now being cultivated commercially in Norway, Iceland, Scotland and Canada. As with other domesticated species, disease problems have been experienced. This review summarizes the current state of knowledge of diseases of the Atlantic halibut, and their diagnosis, prophylaxis and treatment. In economic terms, the most important losses have been suffered at the larval and juvenile stages. The most important infections are caused by nodaviruses, causative agents of Viral Encephalopathy and Retinopathy (VER), which are the major reason why Norway's production of halibut fry has been level since 1995. An aquatic birnavirus, Infectious Pancreatic Necrosis Virus, is also an important agent of mortality. Vibrio anguillarum, Flexibacter ovolyticus and atypical Aeromonas salmonicida are the major bacterial pathogens. The protozoan parasites recorded include Ichthyobodo sp., the microsporidium Enterocytozoon sp., Trichodina hippoglossi, and the metazoan pathogens include myxozoans, helminths, Entobdella hippoglossi, Lepeophtheirus hippoglossi and other parasitic copepods. Experimental vaccines have been tested against V anguillarum and atypical A. salmonicida, with good results. A recombinant vaccine against nodaviruses is under development. A few trials have been carried out on non-specific immunostimulants, but no such treatment is currently available. A number of efficacy and pharmacokinetic trials with various antibacterial agents have also been published.

Animals↗

PubMed2001

Strain variation, based on the hemagglutinin gene, in Norwegian ISA virus isolates collected from 1987 to 2001: indications of recombination.

Infectious salmon anemia (ISA) is caused by a virus that probably belongs to the Orthomyxoviridae and was first recorded in Norway in 1984. The disease has since spread along the Norwegian coast and has later been found in Canada, Scotland, the Faroe Islands, Chile, and the USA. This study presents sequence variation of the hemagglutinin gene from 37 ISA virus isolates, viz. one isolate from Scotland, one from Canada and 35 from Norway. The hemagglutinin gene contains a highly polymorphic region (HPR), which together with the rest of the gene sequence provides a good tool for studies of epizootics. The gene shows temporal and geographical sequence variation, where certain areas are dominated by distinct groups of isolates. Evidence of transmission of ISA virus isolates within and between regions is given. It is suggested that the hemagglutinin gene from different isolates may recombine. Possible recombination sites are found within the HPR and in the 5'-end flanking region close to the HPR.

Anemia↗

PubMed2001

Redescription of Microsporidium takedai (Awakura, 1974) as Kabatana takedai (Awakura, 1974) comb. n.

Ultrastructural study of the microsporidian Microsporidium takedai from the muscles of masu salmon Oncorhynchus masou proved that this species can be assigned to the genus Kabatana Lom, Dyková and Tonguthai, 2000. The parasites develop within disintegrated sarcoplasm without any delimiting boundary or cyst. Cylindrical multinucleate meronts proliferate by serial constrictions into uninucleate stages which repeat the process. Eventually, the uninucleate stages transform into uninucleate sporonts, which divide once to produce sporoblasts, thus functioning as sporoblast mother cells. Spores, with a subterminally located anchoring disc and 3 to 4 turns of the polar tube coil, average 3.3 by 1.9 microm in size. The exospore is divided into small fields; the endospore frequently makes small invaginations into the spore inside. Phylogenetic analysis using SSU rDNA sequence consistently placed Kabatana takedai in a group consisting of Microgemma sp., Spraguea lophii and Glugea americanus. The K. takedai could easily be separated from the other species in the same group by 2 inserts in the SSU rDNA sequence.

Animals↗

PubMed2001

Phylogenetic analysis of infectious salmon anaemia virus isolates from Norway, Canada and Scotland.

The sequences of gene segments 2 and 8 from 10 different isolates of infectious salmon anaemia virus (ISAV) sampled in Norway, Canada and Scotland between 1987 and 1999 were determined and compared. Pairwise comparisons revealed a high degree of homology between the European isolates, with identities of 98 to 100% for both genes examined. The Canadian isolate showed identities of 84 and 87 to 88% with the European isolates for the nucleotide sequence of segments 2 and 8, respectively. Phylogenetic analyses were performed to establish the interrelationship between the European virus isolates. The evolutionary rate based on 4 Norwegian isolates clustered together in the analysis of segment 2 was calculated to be 0.96 x 10(-3) nucleotides site(-1) yr(-1). On the basis of this mutation rate it was estimated that the Norwegian Glesvaer 90 and Canadian Bay of Fundy 97 isolates diverged around 1900, which coincides with transportation of salmonids between Europe and North America starting in the late nineteenth century.

Amino Acid Sequence↗

PubMed2001

Theloiania contejeani Henneguy, 1892: dimorphic life cycle and taxonomic affinities, as indicated by ultrastructural and molecular study.

Thelohania contejeani is a dimorphic species with two simultaneous routes for sporogony. In the first, diplokaryotic sporonts produce 8 uninucleate spores with 9-10 turns of polar tube, within a sporophorous vesicle wall. The episporontal space contains two kinds of tubules and a spongiform mass. In the second, single diplokaryotic sporonts produce small membrane-bound compartments in which they transform into mature diplokaryotic spores with 5-6 turns of polar tube. Analysis of the small subunit (SSU) rRNA revealed two copies of 16S-like rDNA, one of them 1,311 bp, the other 1,361 bp long, with an overall identity of 93%. The majority of sequence differences were located in a 120-bp stretch between positions 336 and 456, with only 40.5% identity between the sequences. Careful consideration suggests that the shorter sequence represents a pseudogene. According to the SSU rDNA sequence, T. contejeani is not closely related to any of the microsporidians where this sequence is available and could not be unambiguously placed in the 16S phylogenetic tree.

Animals↗

PubMed2001

Experimental infection of Atlantic salmon (Salmo salar) with marine Eubothrium sp. (Cestoda: Pseudophyllidea): observations on the life cycle, aspects of development and growth of the parasite.

The life cycle of marine Eubothrium sp. (Cestoda: Pseudophyllidea), from Atlantic salmon (Salmo salar L.) was experimentally completed in one year and included only one intermediate host (Acartia tonsa Dana) (Copepoda: Calanoida). Adult cestodes were collected from farmed salmon, and ripe eggs released by the cestodes were fed to Acartia tonsa. Ingested eggs hatched in the gut and the larvae developed in the haemocoel of the copepod for 15 days at 16 degrees C. A total of 170 seawater-reared salmon were exposed to infected copepods and the total prevalence of Eubothrium sp. in the salmon after infection was 95.3%, with a mean intensity of 15.0 (range 1-87). The infected salmon were kept in the laboratory where the growth of the cestodes was studied for eleven months. Mean length of the cestodes increased with time, but a large variation among the cestodes was observed. Growth and maturation of the cestodes were dependent on host size and the number of worms present in the intestine. No evidence of mortality of Eubothrium sp. was observed during the experimental period.

Animals↗

PubMed2000

Partial cloning of constitutive and inducible nitric oxide synthases and detailed neuronal expression of NOS mRNA in the cerebellum and optic tectum of adult Atlantic salmon (Salmo salar).

Studies of different species have implicated nitric oxide (NO) synthase (NOS) in various physiological and pathological events. Three major NOS isoforms are present in the brain of mammals; endothelial NOS (eNOS), neuronal NOS (nNOS) and inducible NOS (iNOS). Little is known about the significance of the presence of these proteins in the brain. We report the first investigation into the presence of nNOS and iNOS isoforms in a teleost, adult Atlantic salmon (Salmo salar). Complementary DNA was synthesized from cerebellum and thymus mRNA using RT-PCR techniques with primers against conserved regions of NOS. Cloning and sequencing revealed a partial gene sequence of 560 bp corresponding to mammalian nNOS from cerebellum cDNA. The predicted protein sequence of identified salmon nNOS possessed 85% identity to that of mammalian nNOS. Northern blot analysis of different tissues revealed expression in brain and heart, and indicated expression of three different nNOS mRNAs in the brain. In addition, a 389 bp sequence corresponding to iNOS was identified in thymus cDNA. Salmon iNOS is almost identical to rainbow trout iNOS (95%), but shows much less amino acid identity to goldfish (65%) and mammalian (52%) iNOS. Phylogenetically, all vertebrate nNOS and iNOS homologues are clustered separately. Expression studies by means of in situ hybridization revealed abundant nNOS mRNA transcripts in distinct neuronal populations throughout the Purkinje cell layer of the corpus cerebellum and the periventricular layer of the optic tectum. Our data show that adult Atlantic salmon possess a gene encoding an nNOS isoform and putative alternatively spliced forms, which are expressed in distinct neuronal populations in the cerebellum and optic tectum, and in yet unidentified cell types in the heart. The data suggest that the arising of different vertebrate NOS isoforms is an evolutionary old event. The well conserved sequences present in salmon and mammalian nNOS may reflect their importance in protein function, whereas interspecies distributional differences in cellular expression of nNOS and sequence differences of iNOS may reflect variations and specializations in routes of NO action in the vertebrate phylogeny.

Age Factors↗

PubMed2000

Small subunit ribosomal DNA phylogeny of microsporidia with particular reference to genera that infect fish.

Molecular data have proved useful in the study of microsporidia phylogeny. Previous studies have shown that there are several important differences between phylogenies based on rRNA and morphological data. In the present study, small subunit (SSU) rDNA sequences were obtained from 7 different fish-infecting microsporidia from 4 different genera (Glugea Thélohan, 1891, Loma Morrison and Sprague, 1981, Pleistophora Gurley, 1893, and Spraguea Weissenberg, 1976). The lengths of the SSU rDNA genes in these species were between 1,332 and 1,343 base pairs. Phylogenetic analysis was performed using parsimony, maximum likelihood, and Kimura 2-parameter with neighbor joining. The analyses revealed that the microsporidia could be divided into 3 major groups. With the exception of Nucleospora salmonis Hedrick, Groff, and Baxa, 1991, all the microsporidia infecting fishes occurred in the same group. The analysis showed that Pleistophora mirandellae Vaney and Conte, 1901 and Pleistophora aguillarum Hoshina, 1951 are not species of Pleistophora. Furthermore, the analysis showed that Loma is not a member of Glugeidae Thélohan, 1892.

Animals↗

PubMed1999

Three structurally and functionally conserved Hlx genes in zebrafish.

For the Hlx class, which includes homeodomains (HD) that are similar to Drosophila H2.0, few members have been identified in vertebrates. In this report, we describe three zebrafish genes, hlx1, hlx2 and hlx3, related to the murine Dbx genes. The proteins encoded by hlx1 and hlx2 have about the same sequence identity to Dbx1 (approximately 60%), suggesting that they derive from a duplication in the fish lineage. This is supported by similarities in the embryonic expression patterns and promoter sequence conservation. The zebrafish Hlx3 protein is related to murine Dbx2, but it is apparently too diverged to be orthologous. Our phylogenetic analysis of all the known HD sequences of the Hlx class also shows that it can be divided into at least two distinct families. All the Dbx-like genes have similar expression in the embryonic nervous system. However, the initial expression patterns of the zebrafish hlx genes are quite unique, suggesting that some functional divergence has occurred between fish and mammals.

Amino Acid Sequence↗

PubMed1999

Small subunit rDNA phylogeny of Bacillidium sp. (Microspora, Mrazekiidae) infecting oligochaets.

Small subunit (SSU) rDNA has been sequenced from a microsporidium, identified as a member of the genus Bacillidium obtained from an oligochaete. The length of the amplified PCR product was 1386 bp which is currently the longest microsporidium SSU sequence known. Phylogenetic analysis using 28 microsporidia SSU sequences, using 3 different tree-building methods indicated that Bacillidium sp. may be one of the earliest branches on the microsporidia tree. However, bootstrapping failed to give a high score (more than 50%) for the position of Bacillidium sp. The branch leading to Bacillidium sp. was long, indicating that this species is not closely related to any of the other microsporidia so far studied by means of rDNA.

Animals↗

PubMed1999

The putative polymerase sequence of infectious salmon anemia virus suggests a new genus within the Orthomyxoviridae.

The infectious salmon anemia virus (ISAV) is an orthomyxovirus-like virus infecting teleosts. The disease caused by this virus has had major economic consequences for the Atlantic salmon farming industry in Norway, Canada, and Scotland. In this work, we report the cloning and sequencing of an ISAV-specific cDNA comprising 2,245 bp with an open reading frame coding for a predicted protein with a calculated molecular weight of 80.5 kDa. The putative protein sequence shows the core polymerase motifs characteristic of all viral RNA-dependent RNA polymerases. Comparison of the conserved motifs with the corresponding regions of other segmented negative-stranded RNA viruses shows a closer relationship with members of the Orthomyxoviridae than with viruses in other families. The putative ISAV polymerase protein (PB1) has a length of 708 amino acids, a charge of +22 at neutral pH, and a pI of 9.9, which are consistent with the properties of the PB1 proteins of other members of the family. Calculations of the distances between the different PB1 proteins indicate that the ISAV is distantly related to the other members of the family but more closely related to the influenza viruses than to the Thogoto viruses. Based on these and previously published results, we propose that the ISAV comprises a new, fifth genus in the Orthomyxoviridae.

Amino Acid Sequence↗

PubMed1994

On the occurrence and morphology of Gyrodactylus pterygialis from saithe Pollachius virens in a Norwegian fjord.

A total of 308 Pollachius virens from a fjord near Bergen, western Norway, were examined for Gyrodactylus pterygialis over a 14-mo period; G. pterygialis showed the highest prevalence and density in winter/spring (November, January, March, and April pooled) but was absent in some months (August, September, and November). The morphology of the opisthaptoral hard parts of G. pterygialis were studied throughout the survey, and their variation is discussed with respect to host species, host locality, and different temperatures. On the basis of the present and previous descriptions, it is suggested that G. pterygialis is a parasite infecting gadine fishes of the genera Pollachius and Gadus, and that records from Pacific Eleginus and White Sea herring are distinct from G. pterygialis sensu Bychowsky and Poljansky, 1953.

Analysis of Variance↗

PubMedDate not supplied

Observations on an intranuclear microsporidian in lymphoblasts from farmed Atlantic halibut larvae (Hippoglossus hippoglossus L.).

An intranuclear microsporidian was observed in lymphoblasts from the kidney of farmed Atlantic halibut larvae (Hippoglossus hippoglossus). In addition to spores, intranuclear pre-spore stages were observed. Uninucleated stages with a slight thickening of the cell membrane were the most simple developmental stages observed. Multinucleated developmental stages contain electron-lucent vesicles and electron-dense discs. The spores are ovoid and measure 2.9 x 1.2 micron (mean). Sporophorous vesicles, diplokarya and mitochondria were not observed in any of the developmental stages. The developmental stages observed are similar to those of the family Enterocytozoonidae, genus Enterocytozoon. However, there are several differences between the present species and E. bieneusi from enterocytes in man. The relationship to the other species in the genus, E. salmonis can only be determined when all the developmental stages of the species from Atlantic halibut are identified.

Animals↗

PubMedDate not supplied

Molecular phylogeny of microsporidians with particular reference to species that infect the muscles of fish.

Ribosomal DNA from eight species of microsporidians infecting fish have been sequenced. Seven of these species infect the skeletal muscle of fish (Pleistophora spp.) and one species infects migratory mesenchyma cells (Glugea anomala). These sequences, in addition to other available microsporidian rDNA sequences from a broad range of host taxa, have been used in phylogenetic analysis. This analysis revealed that muscle-infecting microsporidians from fish are a polyphyletic group, indicating that characters supposed to be important in the classification of the genus Pleistophora have to be re-evaluated. One character that probably has a polyphyletic origin is the amorphous coat, which has been extensively used in the definition of this genus. Furthermore, our results showed that the insect parasitizing Pleistophora spp. are not related to the true pleistophorans parasitic in skeletal muscle of fish. Phylogenetic analysis of small subunit rDNA sequences revealed disagreements between the molecular phylogeny and classifications based upon ultrastructure. Many of the morphological characters claimed to be important in microsporidian classifications appeared to have arisen several times during evolution: for example, the diplokaryon and sporophorous vesicles.

Animals↗