deletions | additions       diff --git a/section_Methods_subsection_Bacterial_Strains__.tex b/section_Methods_subsection_Bacterial_Strains__.tex  index bf593d9..b35e699 100644  --- a/section_Methods_subsection_Bacterial_Strains__.tex  +++ b/section_Methods_subsection_Bacterial_Strains__.tex 
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\textit{B. dubia} roaches were inoculated with 10^6 \textit{F. tularensis} LVS cells as described previously. Cockroaches were incubated at 37°C with access to dry dog food and carrot slices (to prevent dehydration). Hemolymph was extracted from roaches at each designated time point of 6, 12, 24, 50, 72, and 96 hours post initial infection. Two groups were used for each extraction time point. One group of roaches received a 16μg dose of gentamicin 2 hours prior to hemolymph extraction, and a second group received an equal amount of sterile PBS in parallel. The roaches were cleansed with 70\% isopropyl alcohol and decapitated using sterile surgical scissors. Hemolymph was immediately drained into a 1.5mL Eppendorf tube containing 10μl chilled PBS with anticoagulant (0.05\% N-Phenylthiourea, Sigma). Tubes were weighed before and after addition of hemolymph in order to estimate the volume of hemolyph collected from each roach. In order to quantify the number of \textit{F. tularensis} LVS CFU present in the hemolymph, the harvested sample was serial diluted 1:10 in PBS and aliquots of each dilution were plated on Chocolate II agar plates supplemented with ampicillin and trimethoprim. The total number of \textit{F. tularensis} LVS per mL of hemolymph was determined based on the number of CFU observed from PBS-treated cockroaches and the number of intracellular \textit{F. tularensis} LVS per mL of hemolymph was determined based on the number of CFU observed from gentamicin-treated cockroaches.   \subsection{Statistial analysis}  The R programming environment (version 3.2.1; \cite{R}), accessed via the RStudio interactive development environment \cite{RStudio}, was used for all comparisons. Lethal doses 50\% and corresponding 95\% confidence intervals were estimated by non-linear regression using the 'drc' package \cite{Ritz}. Differences in survival outcome between groups that received equivalent doses of \textit{F. tularensis} with or without antibiotics were determined by log-rank (Mantel-Cox) analysis using the 'survival' package \cite{survival-book, survival-package}. For analysis of \textit{in vivo} growth patterns of \textit{F. tularensis} LVS, the mean, median, and interquartile range of each group was calculated using plot.ly \cite{plotly}.