Differentiation of Bacteroides nodosus biotypes and colony variants in relation to their virulence and immunoprotective properties in sheep

Skerman, T. M.; Erasmuson, Sylvia; Every, D.

doi:10.1128/iai.32.2.788-795.1981

Cited by 42 publications

(14 citation statements)

References 29 publications

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“…Colonies of 14 strains (198, 226, 291. 305, 309, 310, 312, 331, 332, 333, 334, 335, 336 and 337) were examined with a stereomicroscope, on hoof agar medium (Thomas 1958) containing either 2% or 4% agar$, by the procedures of Stewart (1975), Thorley (1976) and Skerman et al(1981) and on blood Eugonagare (Gradin and Schmitz 1977) containing 0.2% yeast extract and 10% defibrinated bovine blood. Strains 334 and 336 were also grown on TAS medium (Skerman 1975) containing 4% agar$.…”

Section: Colony Morphology and Electron Microscopymentioning

confidence: 99%

The pathogenicity and cultural characteristics of virulent, intermediate and benign strains of Bacteroides nodosus causing ovine foot‐rot

et al. 1986

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The relationship between the cultural and biochemical characteristics of 22 strains of Bacteroides nodosus and their virulence for sheep was examined. Virulent, intermediate and benign strains were recognised. Although there was some relationship between virulence and colony morphology on hoof medium with 4% agar, colonies of one virulent and 4 intermediate strains resembled those of benign strains. However, on hoof medium with 2% agar and on blood Euonagar, colonies of this virulent and one intermediate strain differed from each other and the other 3 intermediate strains, which in turn differed from the benign. The degree of piliation, as assessed by electron microscopy, was not a reliable indicator of virulence in strains not possessing a beaded colony type. Together, the results of colony morphology and proteolytic tests such as zymogram, degrading proteinase and elastin-agar tests allowed better discrimination of virulent and benign strains. Intermediate strains generally possessed virulent protease activity. In strains with benign zymogram patterns, activity bands 2 and 3 were more labile than in strains with virulent patterns. The addition of CaCl2 to the culture medium resulted in greater stability of proteolytic activity, particularly with benign strains, and prevented the disappearance of protease activity in the band 5 position in virulent, intermediate and benign strains during prolonged incubation. There were slight differences in the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) patterns of outer membrane proteins of some benign strains but those of intermediate category resembled virulent strains. There was some relationship between the apparent Mr of the pilin monomer on SDS-PAGE gels and serogroup specificity.

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Section: Colony Morphology and Electron Microscopymentioning

confidence: 99%

The pathogenicity and cultural characteristics of virulent, intermediate and benign strains of Bacteroides nodosus causing ovine foot‐rot

et al. 1986

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“…The pathogenic mechanisms of B. nodosus are not clearly defined. Virulent strains are characterized by a distinctive papillate or beaded-colony morphology (B type), a high level of proteolytic elastase activity, and abundant surface pili (29,133). It is widely held that the pilus is an important protective immunogen and has served as the basis for much vaccine research (144,149,155).…”

Section: Pili Of Other Veterinary Bacterial Pathogensmentioning

confidence: 99%

Current and Future Applications of Monoclonal Antibodies against Bacteria in Veterinary Medicine

Sherman

Markham

1986

Monoclonal Antibodies Against Bacteria

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“…Bacteroides nodosus is the essential causative agent of ovine foot rot (1,9). Virulent isolates of this organism contain fimbrial structures (21) which have been implicated as playing a central role in both the infectivity of the bacteria and the immunological response of the host (12,26,27,29,39). The presence offimbriae on B. nodosus appears to be an unstable characteristic (26).…”

mentioning

confidence: 99%

“…Virulent isolates of this organism contain fimbrial structures (21) which have been implicated as playing a central role in both the infectivity of the bacteria and the immunological response of the host (12,26,27,29,39). The presence offimbriae on B. nodosus appears to be an unstable characteristic (26). Benign isolates show altered colony morphology with reduced fimbriae; similar variants may also be generated in vitro by serial culture of virulent isolates, especially in liquid media (26,36).…”

mentioning

confidence: 99%

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Isolation and characterization of Bacteroides nodosus fimbriae: structural subunit and basal protein antigens

Mattick¹,

Anderson²,

Mott³

et al. 1984

J Bacteriol

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We examined the isolation of fimbriae from Bacteroides nodosus. It was found that the best preparations were obtained from the supernatant of washed cells cultured on solid medium, from which fimbriae could be recovered in high yield and purity by a simple one-step procedure. Analysis of such preparations by sodium dodecyl sulfate gel electrophoresis showed that greater than 98% of the protein consisted of fimbrial structural subunits whose molecular weight was ca. 17,000. These preparations also usually exhibited minor contamination with a polypeptide of ca. 80,000 molecular weight, as well as trace amounts of lipopolysaccharide. Attempts to release additional fimbriae by the traditional means of subjecting the bacterial cells to physical stress, such as shearing or heating, resulted primarily in an increase in the level of contamination, without significant gain in the yield of fimbriae. Removal of the 80,000-dalton component could not be achieved by any of a variety of techniques normally used in fimbriae purification, including isoelectric precipitation, MgCl2 precipitation, and CsCl gradient ultracentrifugation, implying a direct physical association with the fimbrial strand. Electron micrographs of fractions containing this protein show cap-shaped structures attached to the ends of what appeared to be fimbrial stubs. These observations suggest that the 80,000-dalton polypeptide may actually constitute the basal attachment site which anchors the fimbria to the outer membrane, analogous to a similar protein recently described in enterotoxigenic strains of Escherichia coli. In B. nodosus, this 80,000-* Corresponding author. which is then followed by a lengthy series of purification steps (e.g., see reference 10). However, using these procedures, we noted substantial contaminants and, in particular, the presence and persistence of a protein of about 80,000 molecular weight. Our results indicate, first, that the current methods used for the production offimbriae from B. nodosus may be inappropriate, even counterproductive, and we present a simplified protocol for the isolation of fimbriae in high purity. Second, our evidence suggests that the 80,000dalton component may actually be physically associated with fimbriae in vivo and in vitro, and that it probably represents the basal protein which anchors the fimbrial strand to the outer surface of the cell.MATERIALS AND METHODS Bacterial culture. Bacteroides nodosus 198, originally obtained from the Commonwealth Scientific and Industrial Research Organization McMaster Laboratory (Sydney, Australia) culture collection, was used in these studies. This strain is the prototype assigned to serogroup A, subtype 1 and has been reclassified as VCS 1001 (3). Strain VCS 1006 (serogroup B, subtype 1) was also used in some experiments. Seeding cultures were purified from freeze-dried samples by two subcultures on 4% hoof-agar (HA) medium (35). For large-scale cultures, cells were grown on 2% HA medium and harvested by being scraped into 10 mM sodium phosphate buffer (pH 7.2) co...

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Differentiation of Bacteroides nodosus biotypes and colony variants in relation to their virulence and immunoprotective properties in sheep

Abstract: MATERIALS AND METHODS Cultural methods. Liquid and solidified Trypticase (BBL Microbiology Systems, Cockeysville, Md.) arginine-serine (TAS) media (25) were used for maintaining strains and as basal media for biochemical tests. TAS isolation medium was also used as originally

Cited by 42 publications

References 29 publications

The pathogenicity and cultural characteristics of virulent, intermediate and benign strains of Bacteroides nodosus causing ovine foot‐rot

The pathogenicity and cultural characteristics of virulent, intermediate and benign strains of Bacteroides nodosus causing ovine foot‐rot

Current and Future Applications of Monoclonal Antibodies against Bacteria in Veterinary Medicine

Isolation and characterization of Bacteroides nodosus fimbriae: structural subunit and basal protein antigens

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