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\subsection{Effect of incubation temperature on \textit{F. tularensis} LVS virulence}  Temperature Dependent Pathogenesis of F. tularensis  The virulent activity of F. tularensis LVS at different temperatures was also determined. B. dubia roaches were injected with F. tularensis LVS as described previously, and groups of roaches were incubated at different temperatures (40°C, 37°C, 30°C, and 22°C). An increase in temperature correlated to a decrease in LD50 value. The 40C LD50 of F. tularensis LVS was calculated to be 1.24E+04, while the 22°C group had an LD50 value of 2.00+06. Survival of B. dubia roaches also reflected the decrease in virulence, with prolonged survival in the groups of roaches incubated at 22°C and 30°C compared to those incubated at 37°C and 40°C.  Table 2. Lethal dose 50\% values of F. tularensis LVS at variable temperatures.  Values calculated by the survival of roaches seven days post-infection of F. tularensis LVS using the Reed-Muench method. Increasing LD50 values reflect decreasing incubation temperatures.  Incubation Temperature   LD50 Value  40°C 1.24 x 104 CFU  37°C 2.86 x 104 CFU  30°C 1.93 x 105 CFU  22°C 2.00 x 106 CFU  Figure 4. Temperature dependent virulence of F. tularensis LVS.  The above graph shows survival of B.dubia roaches after >3x106 CFU inoculation of F. tularensis LVS via systemic injection. 0\% survival was observed in groups incubated at 40°C and 37°C after 5 days post infection. Prolonged survival was observed in groups of roaches incubated at 30°C and 22°C, with 20\% and 30\% survival after seven days, respectively.