Effect of inoculation method on the determination of decontamination efficacy against Bacillus spores.

Decontamination studies investigating the effectiveness of products and processes for the inactivation of Bacillus species spores have traditionally utilized metering viable spores in a liquid suspension onto test materials (coupons). The current study addresses the representativeness of studies using this type of inoculation method compared to when coupons are dosed with a metered amount of aerosolized spores. The understanding of this comparability is important in order to assess the representativeness of such laboratory-based testing when deciding upon decontamination options for use against Bacillus anthracis spores. Temporal inactivation of B. anthracis surrogate (B. subtilis) spores on representative materials using fumigation with chlorine dioxide, spraying of a pH-adjusted bleach solution, or immersion in the solution was investigated as a function of inoculation method (liquid suspension or aerosol dosing). Results indicated that effectiveness, measured as log reduction, was statistically significantly lower when liquid inoculation was used for some material and decontaminant combinations. Differences were mostly noted for the materials observed to be more difficult to decontaminate (i.e., wood and carpet). Significant differences in measured effectiveness were also noted to be a function of the pH-adjusted bleach application method used in the testing (spray or immersion). Based upon this work and the cited literature, it is clear that inoculation method, decontaminant application method, and handling of non-detects (i.e., or detection limits) can have an impact on the sporicidal efficacy measurements.

Evaluation of sampling methods for Bacillus spore-contaminated HVAC filters.

The objective of this study was to compare an extraction-based sampling method to two vacuum-based sampling methods (vacuum sock and 37mm cassette filter) with regards to their ability to recover Bacillus atrophaeus spores (surrogate for Bacillus anthracis) from pleated heating, ventilation, and air conditioning (HVAC) filters that are typically found in commercial and residential buildings. Electrostatic and mechanical HVAC filters were tested, both without and after loading with dust to 50% of their total holding capacity. The results were analyzed by one-way ANOVA across material types, presence or absence of dust, and sampling device. The extraction method gave higher relative recoveries than the two vacuum methods evaluated (p≤0.001). On average, recoveries obtained by the vacuum methods were about 30% of those achieved by the extraction method. Relative recoveries between the two vacuum methods were not significantly different (p>0.05). Although extraction methods yielded higher recoveries than vacuum methods, either HVAC filter sampling approach may provide a rapid and inexpensive mechanism for understanding the extent of contamination following a wide-area biological release incident.

Comparative evaluation of vacuum-based surface sampling methods for collection of Bacillus spores.

In this study, four commonly-used sampling devices (vacuum socks, 37 mm 0.8 µm mixed cellulose ester (MCE) filter cassettes, 37 mm 0.3 µm polytetrafluoroethylene (PTFE) filter cassettes, and 3M™ forensic filters) were comparatively evaluated for their ability to recover surface-associated spores. Aerosolized spores (~10(5)CFUcm(-2)) of a Bacillus anthracis surrogate were allowed to settle onto three material types (concrete, carpet, and upholstery). Ten replicate samples were collected using each vacuum method, from each material type. Stainless steel surfaces, inoculated simultaneously with test materials, were sampled with pre-moistened wipes. Wipe recoveries were utilized to normalize vacuum-based recoveries across trials. Recovery (CFUcm(-2)) and relative recovery (vacuum recovery/wipe recovery) were determined for each method and material type. Recoveries and relative recoveries ranged from 3.8 × 10(3) to 7.4 × 10(4)CFUcm(-2) and 0.035 to 1.242, respectively. ANOVA results indicated that the 37 mm MCE method exhibited higher relative recoveries than the other methods when used for sampling concrete or upholstery. While the vacuum sock resulted in the highest relative recoveries on carpet, no statistically significant difference was detected. The results of this study may be used to guide selection of sampling approaches following biological contamination incidents.

Evaluation of surface sampling for Bacillus spores using commercially available cleaning robots.

Five commercially available domestic cleaning robots were evaluated on their effectiveness for sampling aerosol-deposited Bacillus atrophaeus spores on different indoor material surfaces. The five robots tested include three vacuum types (R1, R2, and R3), one wet wipe (R4), and one wet vacuum (R5). Tests were conducted on two different surface types (carpet and laminate) with 10(6) colony forming units of B. atrophaeus spores deposited per coupon (35.5 cm × 35.5 cm). Spores were deposited on the center surface (30.5 × 30.5 cm) of the coupon's total surface area (71.5 × 71.5 cm), and the surfaces were sampled with an individual robot in an isolation chamber. Chamber air was sampled using a biofilter sampler to determine the potential for resuspension of spores during sampling. Robot test results were compared to currently used surface sampling methods (vacuum sock for carpet and sponge wipe for laminate). The test results showed that the average sampling efficacies for R1, R2, and R3 on carpet were 26, 162, and 92% of vacuum sock sampling efficacy, respectively. On laminate, R1, R2, R3, R4, and R5 average sampling efficacies were 8, 11, 2, 62, and 32% of sponge wipe sampling efficacy, respectively. We conclude that some robotic cleaners were as efficacious as the currently used surface sampling methods for B. atrophaeus spores on these surfaces.

Experimental study on the effect of SO2 on PCDD/F emissions: determination of the importance of gas-phase versus solid-phase reactions in PCDD/F formation.

Cofiring coal in municipal solid waste incinerators (MSWls) has previously been reported to reduce polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/Fs) emissions due to increasing the flue gas SO2 concentration. The present study was focused on understanding the primary mechanism responsible for the suppressant effect of SO2 on total PCDD/F formation and toxic equivalent (TEQ) emissions. The addition of SO2, simulating the effect of coal addition on the flue gas composition, resulted in significant reductions in the TEQ emissions due to reactions involving SO2 in the postcombustion zone. However, emissions of total PCDDs/Fs, unlike the TEQ value, were dependent upon the Cl2 and SO2 injection temperatures due to increases in non-TEQ correlated isomers. The conversion of metal chlorides in the fly ash to sulfates, thus reducing the sites responsible for chlorination/oxidation reactions, was concluded to be the main suppressant mechanism; proposed reactions for copper and iron are presented. This mechanism was found to be independent of combustion conditions and could have prolonged effects on PCDD/F emissions from deposits formed with high flue gas S/Cl ratios.