The objective of this research was to study the rate of structural damage and survival of Bacillus sporothermodurans spores following treatment at high temperatures by determining the amount of Dipicolinic acid (DPA) and soluble protein leakage over time. A reference strain of B. sporothermodurans (DSM 10599) and a South African strain (UP20A) isolated from UHT milk were used. To determine the survival of spores at 130 degrees C, spores were heated for 4, 8 and 12min. To check the viability of spores plate counts were determined, while structural damage was determined using the Transmission Electron Microscopy. The filtrate of the heated spore suspension was analysed for the amount of DPA and soluble protein release due to heating. The amount of DPA released was quantified by HPLC analysis while the amount of soluble protein released from heated spores was quantified using the Bradford method. The log values of spore counts, released DPA and soluble proteins from triplicate experiments were analysed. The results of this study indicate that the inactivation of B. sporothermodurans spores during wet heat treatment is due to the penetration of hot moisture into the spore which then moistens the spore components, and inactivates enzymes, and because of the high water pressure, vital spore components such as proteins and DPA in solution leak out of the spore. Interestingly a vast majority of heated spores were inactivated before a significant amount of DPA was released. This research is the first to determine the effect of high temperature wet heat treatment on the structure of B. sporothermodurans spores and has given an insight regarding the mechanisms of destruction of B. sporothermodurans spores by wet heat.
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