KINETICS OF DRY RUPTURE OF BACTERIAL SPORES IN THE PRESENCE OF SALT.

Sacks, L. E. (U.S. Department of Agriculture, Albany, Calif.), Peter B. Percell, Richard S. Thomas, and Glen F. Bailey. Kinetics of dry rupture of bacterial spores in the presence of salt. J. Bacteriol. 87:952-960. 1964.-The kinetics of breaking spores in the dry state by use of an excess of sodium chloride and a steel ball in a shaking device were investigated. Under most conditions, disruption is a first-order process. The disruption-rate constant varies directly with the weight of the ball and inversely with the weight of the capsule contents (spores plus salt). Different spore batches differ somewhat in susceptibility to dry rupture. The dry-rupture process is highly reproducible and it is relatively simple to obtain preparations in which exactly 50%, or 90%, of the spores are broken. The procedure is uniquely suited to the disruption of small (5 to 20 mg) samples, but 150 mg of spores have been handled with conventional equipment. Apparently, the chief function of the salt is to separate the spores from one another with a relatively hard, energy-nonabsorbing matrix, preventing aggregation and consequent cushioning of the ball's impact. However, under certain conditions (small ball, high salt, large crystals) appreciable breakage results from collisions of spores with the salt crystals. The minimal salt-spore ratio for efficient breakage depends on the spore batch, but is usually greater than 3:1. Fine glass beads or inorganic salts other than sodium chloride will also serve as the matrix. Electron micrographs of the spores in various stages of disruption are shown, as are electron micrographs of the spore coats of Bacillus macerans, B. megaterium, B. cereus, B. coagulans, and Clostridium bifermentans. Prolonged agitation disintegrates spore coats. The spore coats of B. macerans exhibit a characteristic ribbed structure, previously detected only by carbon replicas of intact spores. Possible application to other biological materials is considered.