Partial characterization of a Moraxella bovis cytolysin

https://doi.org/10.1016/0378-1135(94)00084-AGet rights and content

Abstract

Moraxella bovis (M. bovis) is the etiologic agent infectious bovine keratoconjunctivitis and M. bovis hemolysin is believed to be an important virulence factor. Two strains of M. bovis were compared, Epp 63(300) (Epp), a known virulent and hemolytic strain, and IBH 63 (IBH), a known avirulent and nonhemolytic strain. Sterile 10-fold (10 ×) supernatant concentrates were obtained from cultures grown in TSB broth with 10 mM CaCl2. Supernatant hemolysin titers for Epp, were 1:1024 and 1:8192 for unconcentrated (1 ×) and 10 ×, respectively. Supernatant cytotoxin titers to bovine mononuclear cells were 1:32 and 1:128 for 1 × and 10 ×, respectively, for Epp. Cytolytic (hemolytic and cytotoxic) activities declined 10-fold but were still measurable for > 1 wk at 4°C. Both activities were inactivated by trypsin and by heating at 56°C for 20 min. A cytotoxic effect was observed on cultured bovine and ovine corneal epithelial cells with Epp. All cytolytic effects were neutralized with antiserum to 10 × Epp. No cytolytic activities were detected for 10 × IBH. SDS-PAGE electrophoresis and related immunoblots indicate a high molecular weight protein at 110 kDa for the 10 × Epp preparation when stained with silver or probed with monoclonal antibodies to the E. coli alpha hemolysin. No 110 kDa band is observed for 10 × IBH. These data suggest that hemolytic and cytotoxic activities are important in the pathogenesis of infectious bovine keratoconjunctivitis and identify the protein as a possible RTX related toxin of 110 kDa. Stability of the M. bovis cytolysin for > 1 week should allow further characterization and purification of the protein.

References (32)

  • P.J. Fedorka-Cray

    Actinobacillus (Haemophilus) pleuropneumoniae virulence factors: Partial characterization and efficacy studies in swine

  • P.J. Fedorka-Cray et al.

    Efficacy of a cell extract from Actinobacillus (Haemophilus) pleuropneumoniae serotype 1 against disease in swine

    Infect. Immun.

    (1990)
  • S.K. Frank et al.

    Hydrolytic enzymes of Moraxella bovis

    J. Clin. Microbiol.

    (1981)
  • J.D. Gerber

    Enhancement of Moraxella bovis-induced keratitis of mice by exposure of the eye to ultraviolet radiation and ragweed extract

    Am. J. Vet. Res.

    (1983)
  • P.S. Hoien-Dalen et al.

    Comparative characterization of the leukocidic and hemolytic activity of Moraxella bovis

    Am. J. Vet. Res.

    (1990)
  • D.E. Hughes et al.

    A five-year study of infectious bovine keratoconjunctivitis in a beef herd

    J. Am. Vet. Med. Assoc.

    (1970)
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