Linking Geospatial and Laboratory Sciences to Define Mechanisms behind Landscape Level Drivers of Anthrax Outbreaks.

Background: A seasonal predictor of anthrax outbreaks is rainfall, which may be approximated by NDVI using remote sensing. How rainfall or vegetative green-up influences bacterial physiology or microecology to drive anthrax outbreaks is not known. Methods: Rainfall and NDVI dependency of anthrax epizootics was demonstrated with global and local phenological analysis. Growth analysis of B. anthracis in response to pH and calcium gradients was carried out. The influence of pH and calcium levels on expression of toxin and sporulation related proteins in broth culture models was characterized using engineered B. anthracis luminescent reporter strains. Results: Short-term bacterial growth and longer-term bacterial survival were altered by pH and calcium. These conditions also played a major role in pagA and sspB promoter-driven luminescent expression in B. anthracis. Conclusions: Rainfall induced cycling of pH and calcium in soils plays a plausible role in amplifying spore load and persistence in endemic anthrax zones. Observed evidence of B. anthracis favoring soil alkalinity and high soil calcium levels in the environment were linked to physiological conditions that promote bacterial growth, survival, toxin secretion and spore formation; illustrating the utility of bringing laboratory-based (controlled) microbiology experiments into the fold of zoonotic disease ecology.