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Effects of Thermoradiation Treatments on the DNA of Bacillus Subtilis Endospores

Jacobs, Jennifer A.; Turman, Bobby N.

Endospores of the bacterium, Bacillus subfilis, have been shown to exhibit a synergistic rate of cell death when treated with particular levels of heat and ionizing radiation in combination. This synergism has been documented for a number of different organisms at various temperatures and radiation doses (Sivinski, H.D., D.M. Garst, M.C. Reynolds, C.A. Trauth, Jr., R.E. Trujillo, and W.J. Whitfield, ''The Synergistic Inactivation of Biological Systems by Thermoradiation,'' Industrial Sterilization, International Symposium, Amsterdam, 1972, Duke University Press, Durham, NC, pp. 305-335). However, the mechanism of the synergistic action is unknown. This study attempted to determine whether the mechanism of synergism was specifically connected to the DNA strand breakage--either single strand breakage or double strand breakage. Some work was also done to examine the effect of free radicals and ions created in the spore body by the radiation treatments, as well as to determine the functionality of repair enzymes following heat, radiation, and thermoradiation treatments. Bacillus subtilis spores were treated at combinations of 33 kr/hr, 15 kr/hr, 105 C, 85 C, 63 C, and 50 C. Some synergistic correlation was found with the number of double strand breaks, and a strong correlation was found with the number of single strand breaks. In cases displaying synergism of spore killing, single strand breakage while the DNA was in a denatured state is suspected as a likely mechanism. DNA was damaged more by irradiation in the naked state than when encased within the spore, indicating that the spore encasement provides an overall protective effect from radiation damage in spite of free radicals and ions which may be created from molecules other than the DNA molecule within the spore body. Repair enzymes were found to be functional following treatments by radiation only, heat only, and thermoradiation.

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