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(i) Higher KCl concentrations increased the amount of MRS101 spore germination. With ��2?mmol?l?1 KCl, >80% of spores germinated http://www.selleckchem.com/products/AZD6244.html in 90?min, although the average Tlag values and their variation increased as the KCl concentration decreased. KCl concentrations http://www.selleckchem.com/products/SP600125.html (Fig.?4c,d; bold arrows). (v) Average Ilag values also decreased somewhat as average Tlag values increased, perhaps because there was more slow CaDPA release from spores with long Tlag periods. Similar results were obtained when multiple spores�� germination at various KCl concentrations were monitored by Method C (Fig.?4e). Again there was a small amount (c.?10%) of germination of the MRS101 spores in 2?h in Na-Hepes buffer alone, but the time of spores�� germination in the buffer alone was spread out over the 2?h measurement period and was actually greatest in the last 20?min. In contrast, with 100?mmol?l?1 KCl, c.?93% of the spores germinated in 2?h, and almost all within 20?min. With lower KCl concentrations, germination http://www.selleck.cn/products/Imatinib(STI571).html was less complete in 2?h and the germination times of individual spores in the population were shifted to slightly longer times, consistent with the longer Tlag times of MRS101 spores germinating at lower KCl concentrations as measured by Method B. Another parameter that alters the germination heterogeneity of spores of Bacillus species is the degree of spore heat activation, as optimal heat activation decreases Tlag values and their variability as well as increasing the percentage of spores that germinate (Zhang et?al. 2010a). Therefore, we examined the effects of heat activation conditions on Cl.?perfringens MRS101 spore germination and its heterogeneity (Fig.?5; Table?2). This work led to a number of notable conclusions as follows. (i) With no heat activation, only c.?2% of these spores germinated with 100?mmol?l?1 KCl in 90?min (Fig?5a). (ii) Heat activation at higher temperatures allowed more spores to germinate with KCl, but heat activation at 80��C for 10 or 30?min had similar effects, suggesting that 10?min at 80��C gives optimal heat activation for KCl germination. (iii) ��80% of the spores heat activated at 80��C for 10 or 30?min that germinated in 90?min germinated in the first c.?15?min, in the first c.?30?min after heat activation at 80��C for 5?min or 70��C 10?min, and in the first c.