Dose–response analysis of Bacillus thuringiensis HD‐1 cry‐ spore reduction on surfaces using formaldehyde with pre‐germination

AIM: To establish a basis for rapid remediation of large areas contaminated with Bacillus anthracis spores. METHODS AND RESULTS: Representative surfaces of wood, steel and cement were coated by nebulization with B. thuringiensis HD‐1 cry‐ (a simulant for B. anthracis) at 5.9 ± 0.2, 6.3 ± 0.2 and 5.8 ± 0.2 log10 CFU per cm(2), respectively. These were sprayed with formaldehyde, either with or without pre‐germination. Low volume (equivalent to ≤2500 L ha(−1)) applications of formaldehyde at 30 g l(−1) to steel or cement surfaces resulted in ≥4 or ≤2 log10 CFU per cm(2) reductions respectively, after 2 h exposure. Pre‐germinating spores (500 mmol l(−1) l‐alanine and 25 mmol l(−1) inosine, pH 7) followed by formaldehyde application showed higher levels of spore inactivation than formaldehyde alone with gains of up to 3.4 log10 CFU per cm(2) for a given dose. No loss in B. thuringiensis cry‐ viability was measured after the 2 h germination period, however, a pre‐heat shock log10 reduction was seen for B. anthracis strains: LSU149 (1.7 log10), Vollum and LSU465 (both 0.9 log10), LSU442 (0.2 log10), Sterne (0.8 log10) and Ames (0.6 log10). CONCLUSIONS: A methodology was developed to produce representative spore contamination of surfaces along with a laboratory‐based technique to measure the efficacy of decontamination. Dose–response analysis was used to optimize decontamination. Pre‐germinating spores was found to increase effectiveness of decontamination but requires careful consideration of total volume used (germinant and decontaminant) by surface type. SIGNIFICANCE AND IMPACT OF THE STUDY: To be practically achievable, decontamination of a wide area contaminated with B. anthracis spores must be effective, timely and minimize the amount of materials required. This study uses systematic dose–response methodology to demonstrate that such an approach is feasible.