Isolation and characterization of toxic fractions from Brucella abortus

Abstract

Two types of toxic fractions, protein-rich and carbohydrate-rich, were isolated from attenuated (strain 19) and virulent (strain 2308) Brucella abortus organisms. Polyacrylamide gel electrophoresis of the protein-rich fraction, in the presence and absence of sodium dodecyl sulfate, revealed qualitative and quantitative differences in the protein bands derived from the attenuated and virulent strains. Sodium dodecyl sulfate-gel electrophoresis indicated that the major differences between these protein fractions were in the molecular weight range from 14,000 to 40,000. Immunoelectrophoresis of these fractions from the attenuated and virulent strains revealed differences in the antigenic spectrum. Polypeptides in the carbohydrate-rich fraction could be visualized on polyacrylamide gels only when reacted with fluorescamine before electrophoresis. Immune sera did not precipitate the components of the carbohydrate-rich fraction. Intradermal injecttion of the protein and carbohydrate-rich fractions resulted in different types of skin lesions in guinea pigs, i.e., edematous/erythematous and necrotic lesions, respectively. Fractions derived from attenuated and virulent strains of B. abortus were equally toxic in the guinea pig skin test. The toxic activity of both types of fractions was susceptible to pronase and heat treatment.