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Evaluation of Analytical Methods for Detection of Bacillus anthracis Surrogate Spores: Compatibility with Real-World Maritime Environmental Samples Collected from the USCG Assets and Facilities
Citation:
Shah, S., W. Calfee, K. Hofacre, S. Nelson, R. James, AND P. Keyes. Evaluation of Analytical Methods for Detection of Bacillus anthracis Surrogate Spores: Compatibility with Real-World Maritime Environmental Samples Collected from the USCG Assets and Facilities. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-21/226, 2021.
Impact/Purpose:
The U.S. Coast Guard (USCG) continues to enhance and expand their capabilities to respond to bioterrorism incidents such as Anthrax and effectively protect human health, and coastal zone assets and facilities (Maritime Environmental Response mission). Under the Department of Homeland Security (DHS) funded Analysis for Coastal Operational Resiliency (AnCOR) program, EPA scientists and engineers have been working with the USCG in their efforts to be better prepared to respond to bioterrorism incidents. EPA-HSRP has developed extensive protocols for sampling, analysis, and decontamination to respond to biological contamination incidents, however, responding to any contamination incident is specific to the affected site and surrounding environment. The USCG’s coastal zone facilities, and assets, including small and large boats and other vehicles, in diverse geographical areas and maritime environmental conditions can pose more complex and unique challenges for adapting existing methods or developing new ones for sampling, analysis, and decontamination to respond to biological contamination incidents. The purpose of this project was two folds: 1. To evaluate the Microbiological Plate-Culture and the Rapid Viability PCR (RV-PCR) analytical methods included in the EPA-ORD “Protocol for Detection of Bacillus anthracis from Environmental Samples During the Remediation Phase of an Anthrax Incident” for their compatibility with detection of Bacillus anthracis surrogate in real-world maritime environmental samples collected from the USCG coastal zone assets, and their close-proximity surroundings; and 2. To understand difficulties associated with processing and analyzing samples collected during the post-incident remediation of USCG bases and identify the capability gaps in this mission space. This report describes the performance of the analytical methods for the USCG base relevant environmental samples which will help the USCG to select the method during a real-world biothreat incident such as anthrax.
Description:
This product is a final report on the project on “Evaluation of Analytical Methods for Detection of Bacillus anthracis Surrogate Spores: Compatibility with Real-World Maritime Environmental Samples Collected from the U.S. Coast Guard (USCG) Assets and Facilities.” In this effort, two analytical methods; 1) Microbiological Plate-Culture; and 2) Rapid Viability PCR (RV-PCR) included in the EPA-ORD “Protocol for Detection of Bacillus anthracis from Environmental Samples During the Remediation Phase of an Anthrax Incident” were evaluated. Employing the commonly used surface sampling tools such as Sponge-Sticks and vacuum filter cassettes, samples containing background materials were collected from the response boat-small, response boat-medium, navigation computer screens on these boats, pier concrete, and wood surfaces at the USCG base in Portsmouth, VA. Also collected were grab samples (e.g., grass, gravel, boat wash-water). These samples were spiked in the laboratory with the spores of Bacillus thuringiensis which is commonly used as a surrogate for Bacillus anthracis, the bacteria that cause anthrax. The spiked samples were then processed to recover spores and analyzed by the two analytical methods. This report describes the performance of the two analytical methods to detect the surrogate spores from the USCG base relevant environmental samples. The results of this work will help the USCG to select the method during a real-world biothreat incident such as anthrax. The effort also identified R&D gaps in the sample processing procedures and limitation of the analytical methods.