During development of BGas a simulant for B. anthracis, strains were selected that exhibited the most desirable characteristics, those being rapid growth, high spore yield, and experimental reproducibility. Without being aware of the nature of the genetic alterations in their ''optimized'' strains, BW workers at Camp Detrick selected a mutant that provided dramatically higher total and relative spore yields, and generated consistent experimental results [6] . These strains were adopted into the inventories of numerous biodefense laboratories and have been used for many Figure 7 . The spo0F(H101R) and spo0F(A98P) alleles are associated with hypersporulation. Phase-contrast microscopy of BG strains after two days of growth on SBA. Vegetative cells appear as phase-dark rods, while spores appear as round, phase-bright globules. The mean percentage sporulation of each strain in a representative experiment is given 6SEM. The experiment was repeated on three consecutive days; representative results of a single experiment are shown. Statistical significance was determined by mixed ANOVA (Tukey's method, p,0.05). doi:10.1371/journal.pone.0017836.g007 Figure 6 . Omnilog phenotypic arrays of B. atrophaeus subsp. globigii strains. Six strains were each inoculated into twenty 96-well Omnilog plates and grown at 37uC. Reduction of tetrazolium dye by respiring cells was measured every 15 minutes by optical density. Dye reduction relative to the 1942 strain is shown; the red ratio values indicate less respiration while the green ratio values indicate more respiration as compared to the 1942 strain. Individual arrays or strains are displayed in each of the six major columns labeled Detrick 1, Detrick 2, Detrick 3, 1013-1, 1013-2, and Dugway. A) Heat map of all conditions for each strain. Each of the twenty plates for each strain is represented by the notation PM01-PM20 (left-toright for each strain) along the x-axis. The rows represent the well position, and are denoted as A i to H i (i = 1 to 12) from the bottom to the top of the plot in each array along the y-axis. Each cell ratio value represents the average of two biological replicates for each strain. Plates PM01-PM10 contains single wells for each growth condition, while plates PM11-PM20 contain quadruplicate wells for each growth condition. Solid circle indicates wells containing sodium lactate; dotted circle indicates well containing L-serine at pH 4.5. The details of the 1920 growth conditions can be found in the first worksheet labeled ''All strain AUC data'' in Table S4 . B) Most significant phenotypes for each of the six test strains as compared to the 1942 strain. The phenotypes with statistically significant increases and/or the decreases in ratio values for each of the six strains are presented. For the 1013 isolates only the conditions giving the five largest changes are presented. The number in each color block indicates the ratio for the test strain relative to the parent strain for the phenotype specified. The details of all significant phenotypes for each test strain can be obtained in Table S4 . Bold Italic font indicates p,0.05. doi:10.1371/journal.pone.0017836.g006 decades in simulations of decontamination and dispersal [12] . By applying a combination of genomic and biochemical profiling techniques, our data demonstrate that the BG isolates were ''enhanced'' by researchers at Camp Detrick during the development of the organism as a simulant.