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Host-Parasite Relationships in Experimental Airborne Tuberculosis II. Reproducible Infection by Means of an Inoculum Preserved at –70 C

Department of Medical Microbiology, University of Wisconsin, Madison, Wisconsin 53706

ABSTRACT

The use of an inoculum preserved at low temperature for the infection of guinea pigs by the respiratory route was evaluated. In a preliminary study with Mycobacterium bovis (BCG), some of the conditions required for maximal recovery of viable cells stored at low temperature were examined. Survival of BCG was decreased by rapid freezing to –70 C and by storage at –20 C, but there was no decrease when BCG was frozen slowly and stored at –70 C or –196 C. In a subsequent study, the effect of storage at –70 C on viability and infectivity of M. tuberculosis (H37Rv) was considered. There was no loss of viability of H37Rv cells suspended in Dubos broth and stored 1 year at –70 C. This suspension showed no loss of infectivity as assessed by the number of primary pulmonary lesions initiated in guinea pigs. Constant viability and infectivity of a suspension stored at low temperature assures the reproducibility of the amount of infection and facilitates comparisons between experiments. This advantage, as well as others, of storage at low temperature are discussed.

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