Bio-imaging of the Bacillus anthracis spore coat

The deliberate release of pathogenic bacterial spores is recognised as presenting a major threat to civilian health, a major economic risk, and a significant hazard to military personnel. Addressing this threat will require a combination of fundamental scientific insight to the resilience, infectivity, and super-dormancy of Bacillus anthracis spores, coupled with the power of emerging portable sensor technology that will make it possible to detect the biohazard and to validate its decontamination with far greater effectiveness than is possible today. In the current project, the structure of the B. anthracis spore coat will be mapped with unprecedented resolution using the ellipsoid localisation microscopy technique developed recently by the PI and departmental colleague Dr Eric Rees. Conserved near-surface protein markers for identifying the presence of B. anthracis contamination will be characterised, and fine structural changes associated with decontamination will be quantified. Insight from these laboratory studies will elucidate mechanisms of decontamination, and potentially super-dormancy of spores. The second aspect of the work will entail the development of 'spray and visualise' chemical technology. Click-chemistry refers to a suite of modern detection reagents that can involve the creation of a fluorescent dye when a probe molecule encounters its target. By combining this method with nanobody-binding to conserved B. anthracis target proteins identified in the first part of the project, a 'spray and visualise' protocol for highlighting contaminated surfaces will be developed. This will first be demonstrated with laboratory analysis of swabs, but with a view to in-situ highlighting of contamination in the field