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Biomedical subjects

G Olivier

Publications and source records attributed to G Olivier.

At least 19 recordsLinked to original sources

Molecular detection and characterization of nodavirus in several marine fish species from the northeastern Atlantic.

Nodaviruses (NNV) are responsible for causing disease outbreaks mainly in hatchery-reared larvae and juveniles of a wide variety of fishes throughout the world. This disease has seriously limited the culture of marine fishes over the last decade. In the Atlantic provinces of Canada, disease caused by a nodavirus was first reported in juvenile Atlantic cod being reared in Nova Scotia, in 1999. More recently, disease outbreaks caused by nodavirus have been identified in hatchery-reared Atlantic cod and haddock in Newfoundland and New Brunswick, respectively, and along the east coast of the USA. The presence of NNV in wild Atlantic cod adults and wild winter flounder has also been reported. Nodaviruses were isolated from cultured Atlantic cod and haddock, as well as from wild winter flounder from a variety of geographical localities, and their virus coat (capsid) protein genes were partially sequenced. An analysis of the data indicates that all of the nodaviruses isolated from eastern North America were closely related to one another, but that they were distinct from the European isolates already sequenced. Regardless of host species, isolates from close geographical localities were more similar than those from distant geographical areas. At the protein level, differences in coat protein sequences were seen only for strains isolated from Atlantic cod originating from Newfoundland. Our results suggest that NNV may have been present in the Atlantic off Canada and on the east coast of the USA for some time, and has evolved to form a monophyletic group, distinct from other isolates found in cold-water species. Non-lethal methods for detection of NNV are necessary to develop management strategies for this disease, and would be an asset to diagnosticians and producers. Based on the results of this study, new primers were designed and developed for an improved RT-PCR assay able to detect North Atlantic nodaviruses in ovarian fluids, eggs and other tissues. The application of this test to field samples is discussed.

Amino Acid Sequence↗

Characterization of attenuated Renibacterium salmoninarum strains and their use as live vaccines.

Two nutritionally mutant strains of Renibacterium salmoninarum (Rs) were isolated that grew on tryticase soy agar (Rs TSA1) or brain heart infusion agar (Rs BHI1). These 2 strains could be continuously cultured on these media, whereas typical R. salmoninarum would only grow on KDM-2 agar. We determined no other phenotypic difference that could be used to distinguish them from wild-type R. salmoninarum. Both strains were found to be avirulent when 5 x 10(6) bacteria were intraperitoneally (i.p.) injected into Atlantic salmon. Rs TSA1, Rs BHI1, and Rs MT-239 (a R. salmoninarum strain previously shown to be attenuated) were tested as live vaccines in 2 separate trials. The best protection was seen with Rs TSA1. Vaccinated Atlantic salmon had relative percent survival (RPS) of 50 at 74 d post-challenge in Trial 1 and 76 at 60 d post-challenge in Trial 2. In both trials, 100% of the control salmon died from bacterial kidney disease (BKD) (within 40 d for Trial 1 and 50 d for Trial 2) after i.p. challenge with 5 x 10(6) live cells of the virulent isolate Rs Margaree.

Animals↗

Generation of oculomotor images during tasks requiring visual recognition of polygons.

This paper concerns the contribution of mentally simulated ocular exploration to generation of a visual mental image. In Exp. 1, repeated exploration of the outlines of an irregular decagon allowed an incidental learning of the shape. Analyses showed subjects memorized their ocular movements rather than the polygon. In Exp. 2, exploration of a reversible figure such as a Necker cube varied in opposite directions. Then, both perspective possibilities are presented. The perspective the subjects recognized depended on the way they explored the ambiguous figure. In both experiments, during recognition the subjects recalled a visual mental image of the polygon they compared with the different polygons proposed for recognition. To interpret the data, hypotheses concerning common processes underlying both motor intention of ocular movements and generation of a visual image are suggested.

Adult↗

Genetic diversity of the fish pathogen Aeromonas salmonicida demonstrated by random amplified polymorphic DNA and pulsed-field gel electrophoresis analyses.

The current taxonomy of Aeromonas salmonicida includes 4 subspecies. A. salmonicida subsp. salmonicida is associated with salmonid furunculosis, and A. salmonicida subsp. achromogenes, A. salmonicida subsp. masoucida, and A. salmonicida subsp. smithia are strains that show variation in some biochemical properties. This classification does not readily encompass isolates from a wide range of fish hosts currently described as atypical A. salmonicida. This study examined 17 typical strains, 39 atypical strains and 3 type A. salmonicida subspecies strains for genetic similarity using the random amplified polymophic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) techniques. On the basis of RAPD- and PFGE-derived profiles, similarity matrices and dendrograms were constructed. The results showed that species A. salmonicida constituted a genetically heterogeneous group of strains, encompassing within an homogeneous or clonal lineage comprised solely of typical strains and the A. salmonicida subsp. salmonicida type strain.

Aeromonas↗

Co-culture of Aeromonas salmonicida and host cells in intraperitoneal implants is associated with enhanced bacterial survival.

An experimental procedure that we named "in vivo co-culture technology" allowed us to study the interactions between Aeromonas salmonicida and host cells, inside semipermeable chambers implanted in the peritoneal cavity of Atlantic salmon. Intraperitoneal implants containing bacteria and host cells, or bacteria and lysed cells, consistently yielded higher numbers of viable bacteria than implants containing bacteria only. Electron microscopy confirmed that 30 min after chamber inoculation, numerous bacteria were already internalized by exudate cells, and that at 3 h, destruction of these cells was evident. Thus, the rapid invasion and (or) the A. salmonicida-mediated lysis of host cells may constitute a survival strategy in vivo. The co-culture of bacteria with exudate peritoneal cells may be applicable to the in vivo study of other pathogens.

Aeromonas↗

Host cell invasion and intracellular residence by Aeromonas salmonicida: role of the S-layer.

Virulent strains of the fish pathogen Aeromonas salmonicida, which have surface S-layers (S+), efficiently adhere to, enter, and survive within macrophages. Here we report that S+ bacteria were 10- to 20-fold more adherent to non-phagocytic fish cell lines than S-layer-negative (S-) mutants. When reconstituted with exogenous S-layers, these S- mutants regained adherence. As well, latex beads coated with purified S-layers were more adherent to fish cell lines than uncoated beads, or beads coated with disorganized S-layers, suggesting that purified S-layers were sufficient to mediate high levels of adherence, and that this process relied on S-layer structure. Gentamicin protection assays and electron microscopy indicated that both S+ and S- A. salmonicida invaded non-phagocytic fish cells. In addition, these fish cells were unable to internalize S-layer-coated beads, clearly suggesting that the S-layer is not an invasion factor. Lipopolysaccharide (which is partially exposed in S+ bacteria) appeared to mediate invasion. Surprisingly, A. salmonicida did not show net growth inside fish cells cultured in the presence of gentamicin, as determined by viable bacterial cell counts. On the contrary, bacterial viability sharply decreased after cell infection. We thus concluded that the S-layer is an adhesin that promotes but does not mediate invasion of non-phagocytic fish cell lines. These cell lines should prove useful in studies aimed at characterizing the invasion mechanisms of A. salmonicida, but of limited value in studying the intracellular residence and replication of this invasive bacterium in vitro.

Aeromonas↗

Motor dimension of visual mental image transformation processes.

This paper concerns the influence of perceptual motor skills on the transformation processes during mental visual imaging. We first administered a visual recognition task to subjects, during which they scanned and rotated visual mental images. Then, we measured motor skills in perceptual situations. The main result is that both mental scanning and mental rotation processes are quicker when they simulate a perceptual motor skill of the subject. Furthermore, mental rotation seems to be canceled when the hemisphere activated by perception has to control the behavioral response. These results suggest that visual mental images are transformed partly via motor processes.

Adolescent↗

The cell surface of Aeromonas salmonicida determines in vitro survival in cultured brook trout (Salvelinus fontinalis) peritoneal macrophages.

Aeromonas salmonicida strains phenotypically differing in their A-layer, lipopolysaccharide, and macrophage cytotoxicity were tested in vitro for survivability in brook trout (Salvelinus fontinalis) serum with or without antibodies, and in vivo following intraperitoneal injection. The ability of brook trout peritoneal macrophages to phagocytize and kill the different phenotypes was investigated in an in vitro assay. The virulent strain, A. salmonicida 80204, which has the full complement of known virulence factors, as well as the recently described macrophage cytotoxin, was resistant in vitro to both the bactericidal activity of normal and immune serum, and to brook trout peritoneal macrophages. A. salmonicida SS-70.1, which possesses the A-layer but lacks the cytotoxin, was resistant to the bactericidal activity of normal and immune serum but was avirulent and killed by macrophages. Phenotypes lacking the A-layer, regardless of whether or not they possessed the macrophage cytotoxin were avirulent, susceptible to normal and immune serum and the bactericidal activity of peritoneal macrophages. A. salmonicida virulence expression requires both the A-layer and the macrophage cytotoxin.

Aeromonas↗

Characteristics of 'atypical', cytochrome oxidase-negative Aeromonas salmonicida isolated from ulcerated flounders (Platichthys flesus (L.)).

'Atypical', cytochrome oxidase-negative variants of the fish pathogen Aeromonas salmonicida, isolated from ulcerated flounder (Platichthys flesus), were studied using different methods. Two of the strains possessed a protein that corresponded to the A-layer protein of Aer. salmonicida. The strains reacted with antibodies against the A-layer and monoclonal antibodies against the O-antigen of typical Aer. salmonicida. These tests confirm that the isolates from flounder should be classified as Aer. salmonicida. Analysis of the fatty acids showed that the isolates were rather homogeneous but the values of the guanine plus cytosine content of the DNA of the bacteria varied too much for any conclusion to be drawn on their taxonomic location. The strains examined exhibited several biochemical characters that differed from those of the type strains of Aer. salmonicida subsp. salmonicida and Aer. salmonicida subsp. achromogenes. The results suggest that these 'atypical', cytochrome oxidase-negative variants may form a new subspecies of Aer. salmonicida.

Aeromonas↗

Interaction between Aeromonas salmonicida and peritoneal macrophages of brook trout (Salvelinus fontinalis).

The phagocytic and bactericidal properties of peritoneal macrophages obtained from brook trout (Salvelinus fontinalis), injected with either glycogen or a modified Freund's complete adjuvant (MFCA), were evaluated against an avirulent and a virulent strain of Aeromonas salmonicida. Avirulent bacteria were effectively phagocytized and killed by macrophages obtained from fish injected with both irritants. With glycogen-elicited macrophages, no enhancement of killing was observed following opsonization of avirulent bacteria with specific antibodies. A killing index (K.I.) of 38 was obtained, compared to a K.I. of 39 for unopsonized bacteria. When avirulent bacteria were opsonized with complement, the K.I. was increased to 67. Virulent bacteria were less susceptible to the phagocytic and the bactericidal activities of glycogen-elicited macrophages, even after opsonization with antibodies and/or complement, K.I. of 9 to 15. In contrast, MFCA-elicited macrophages showed increased phagocytic and bactericidal activities against both strains. The K.I. of unopsonized virulent bacteria was increased to 47 and 46 compared to K.I. of 4 and 7 obtained with glycogen-elicited macrophages.

Aeromonas↗

Immunity to Aeromonas salmonicida in coho salmon (Oncorhynchus kisutch) induced by modified Freund's complete adjuvant: its non-specific nature and the probable role of macrophages in the phenomenon.

Juvenile coho salmon (Oncorhynchus kisutch), vaccinated with one intraperitoneal injection of formalin-killed virulent Aeromonas salmonicida cells suspended in saline, showed increased protection against approximately one LD60 of homologous challenge administered at 30 days post-vaccination. Under similar conditions, coho vaccinated with a modified complete Freund's adjuvant (MFCA) alone were also equally protected. When measured against a more severe A. salmonicida challenge of approximately one LD95, the strength of the MFCA-induced protection was found to exceed that produced by the homologous bacterin administered in saline or incomplete adjuvant, and the protection was still evident at 90 days post-treatment. Other more precise measurements indicated the LD50 for MFCA-treated coho to be up to 450 times that for saline-treated coho. Two other tested adjuvants, levamisole and MDP (N-acetyl-muramyl-L-alanyl-D-isoglutamine), administered in a modified Freund's incomplete adjuvant, also enhanced anti-A. salmonicida immunity but to a lesser degree. The active factor in MFCA was a killed Mycobacterium butyricum preparation, and the anti-A. salmonicida immunity it induced was non-specific because the immunity extended to two other serologically distinct fish pathogens tested: A. hydrophila (LD50 increase of 5.3-fold) and Vibrio ordalii (LD50 increase of 560-fold). Macrophages are believed to account for the M. butyricum-induced anti-A. salmonicida immunity because the immunity was a) non-specific, b) very rapid in onset (it was measurable by 4 days), and c) influenced by particulate preparations, known to affect macrophage function and immunity in mammals. The possible benefits of adjuvant-induced non-specific immunity in cultured fish are discussed.

Aeromonas↗

Serogrouping of motile Aeromonas species isolated from healthy and moribund fish.

A total of 195 strains of motile Aeromonas isolated from fish were characterized as Aeromonas hydrophila and Aeromonas sobria. In view of the frequency of isolation and the importance of motile Aeromonas species as fish pathogens, a serological classificaton of these organisms was attempted. Antisera were prepared in rabbits against formalinized whole cell suspensions and against boiled cells of 12 different isolates. Seventy-six strains could be grouped by tube agglutination with whole cells as antigen and anti-whole cell antiserum. However, only 39 strains were typable with anti-O serum. Differentiation was made between heat-stable antigens and heat-labile antigens which did not block the O agglutination reaction. The same heat-labile antigen could be associated with different heat-stable particulate antigens, and a relationship was observed between the heat-stable particulate antigens and the virulence of A. hydrophila for fish. In addition to these two types of antigen, motile Aeromonas possessed heat-stable soluble antigens which could be detected by indirect hemagglutination. One strain seemed to possess various heat-stable soluble antigens; so far, however, it does not appear to be feasible to use these antigens for serology. Finally, we also observed cross-reactions between some A. hydrophila and A. sobria strains.

Aeromonas↗

A toxigenic profile of Aeromonas hydrophila and Aeromonas sobria isolated from fish.

Forty strains of motile Aeromonas were isolated from healthy and diseased fish. These strains were identified as A. hydrophila or a A. sobria. It was found that only strains of A. hydrophila produced a dermonecrotic factor and two zones of hemolysis on blood agar. All the strains of A. sobria tested and 72% of the A. hydrophila were enterotoxigenic. Finally, we observed that only A. hydrophila strains could regularly produce hemolysin at 10 degrees C.

Aeromonas↗