Extension of Bacillus endospore gas dynamic heating studies to multiple species and test conditions.

AIMS: Shock wave-induced damage to a variety of Bacillus endospore species is studied for a wide range of postshock temperatures and test times in oxidative and non-oxidative gas environments. METHODS AND RESULTS: Bacillus atrophaeus and Bacillus subtilis endospores are nebulized into an aqueous aerosol, loaded into the Stanford aerosol shock tube (SAST) and subjected to shock waves of controlled strength. Endospores experience uniform test temperatures between 500 and 1000 K and pressures ranging from 2 to 7 atm, for either a short test time (∼2·5 ms) or a relatively long test time (∼45 ms). During this process, the bioaerosol is observed using in situ laser absorption and scattering diagnostics. Additionally, shock-treated samples are extracted for ex situ analysis including viability plating and flow cytometry. For short test times, results are consistent with previous studies; all endospore species begin to lose the ability to form colonies when shock-heated to temperatures above 500 K, while significant breakdown in morphology is observed for postshock temperatures above 700 K. Oxidative bath gases did not affect viability losses or morphological breakdown rates. Experiments with extended postshock test time showed increased viability loss with minimal morphological damage for shocks between 600 and 700 K. CONCLUSIONS: Genetic differences between B. subtilis and B. atrophaeus endospores do not confer noticeable gains in resistance to shock heating. Oxidative environments do not exacerbate shock-induced damage to endospores. Extended test time experiments reinforce our hypothesis that a temperature/time-dependent inactivation mechanism that does not involve morphological breakdown exists at low-to-moderate postshock temperatures. SIGNIFICANCE AND IMPACT OF THE STUDY: The methodology and experiments described in this paper extend the study of the interactions of endospores with shock/blast waves to new species and environmental conditions.

Bacillus endospore resistance to gas dynamic heating.

AIM: To develop a novel laboratory procedure for the study of shock wave-induced damage to Bacillus endospores. METHODS AND RESULTS: Bacillus atrophaeus endospores are nebulized into an aerosol, loaded into the stanford aerosol shock tube and subjected to shock waves of controlled strength. Endospores experience uniform test temperatures between 500 and 1000K and pressures ranging from 2 to 7atm, for a relatively short time (2-3ms). During this process, the bioaerosol is observed using in situ laser absorption and scattering diagnostics. Additionally, shock-treated samples are extracted for ex situ analysis including viability plating, flow cytometry and SEM imaging. Measurements indicate that endospores lose the ability to form colonies when heated to test temperatures above 500K while significant breakdown in morphology is observed at test temperatures above 750K. CONCLUSION: These results demonstrate the disruption of essential biochemical pathways or biomolecules prior to the onset of significant endospore morphological deterioration. SIGNIFICANCE AND IMPACT OF THE STUDY: This novel laboratory approach to study the interaction of endospores with shock waves provides an experimental means to investigate the mechanisms of endospore resistance to rapid heating. In addition, this methodology allows for the direct simulation of a blast wave-bioaerosol interaction in an atmospheric environment.

Vaccination with a recombinant vaccinia vaccine containing the B7-1 co-stimulatory molecule causes no significant toxicity and enhances T cell-mediated cytotoxicity.

B7-1 is a co-stimulatory molecule that provides a second signal for T-cell activation. Several studies have demonstrated that vaccination with a vector containing genes encoding B7-1 and an antigen appears to be efficacious at promoting immune responsiveness to the antigen. To evaluate the safety of such a protocol and determine the effect of the B7-1 vector on immune responsiveness, female C57BL/6 mice were administered Wyeth wild-type vaccinia virus (V-WT) or V-WT containing the gene for B7-1 (rV-B7-1) as a single s. c. injection or 3 monthly s.c. injections. Immunologic parameters were evaluated in half of the mice and general toxicity in the other half. Immunologic end points included determination of splenic lymphocyte phenotypes, mitogen-induced T- and B-cell proliferation, T-cell proliferation in response to alloantigens, cell-mediated cytotoxicity (CMC), natural killer cell activity and serum anti-nuclear antibody (ANA) titers. No significant signs of general toxicity were noted. The primary immunologic effect was an increase in the ability of spleen cells to lyse allogeneic targets and to proliferate in response to allogeneic stimulation. Numbers of splenic CD8(+) cells were also increased. These effects were more pronounced after 3 vaccinations than after a single vaccination. Minimal differences in ANA were observed between mice immunized with V-WT and rV-B7-1. In addition, no serum antibodies against B7-1 were detected in any mice. The data suggest that vaccination with rV-B7-1 augments CMC with minimal toxicity.