Citation:

Nelson, S., K. Hofacre, Michael Calfee, S. Serre, E. Benard, C. Graham, L. Oudejans, Ronald Mickelsen, J. Tang, D. Bansleben, S. Taft, R. James, AND S. Shah. Evaluation of two methods for detection of viable Bacillus anthracis simulant spores in maritime environmental samples. ENVIRONMENTAL MONITORING AND ASSESSMENT. Springer, New York, NY, 195:257, (2023). https://doi.org/10.1007/s10661-022-10772-8

Impact/Purpose:

The United States has coastlines on multiple oceans, gulfs, and great lakes, and therefore, has thirty coastal states. Coastal cities in those states are ports of entry, shipping ports, and popular tourist destinations, and hence, have a significant portion of the U.S. population. Such cities are vulnerable to biohazard contamination incidents, including bioterrorism incidents involving anthrax. The coastal zone facilities and assets, including U.S. Coast Guard (USCG) boats, and maritime environmental conditions can pose complex and unique challenges for adapting existing methods or developing new methods for sample analysis to respond to such contamination incidents. This manuscript presents the data on the evaluation of performance of the traditional culture (TC) and the rapid viability-polymerase chain reaction (RV-PCR) methods for compatibility with detection of spores of Bacillus thuringiensis subspecies kurstaki (Btk), a surrogate for the anthrax causing Bacillus anthracis, in maritime environmental surface and grab samples. The representative samples collected from a USCG installation included surfaces such as aluminum on boats, nonskid tread on decks of watercraft, computer touchscreens, and concrete piers, and grab samples of boat washdown water, soil, vegetation, and gravel from surrounding areas. The results of this study will guide the laboratories for analyzing maritime environmental samples during a response to a biohazard incident, especially, one involving anthrax, and help the USCG in their efforts to be better prepared to respond to such incidents. The results will also provide a basis to scientists to further test the application of the analytical methods evaluated in this effort for maritime environmental samples. Additionally, the results will guide the researchers to develop processing methods for complex/difficult maritime sample types and matrices for effective analysis. The EPA Office of Land and Emergency Management's Environmental Response Laboratory Network (ERLN), public health laboratories, and the biodefense laboratories, in general, at the national and international level are likely to use the data in their efforts on response to potential anthrax incidents.

Description:

Analytical methods exist to detect biothreat agents in environmental samples during a response to biological contamination incidents. However, the coastal zone facilities and assets of the United States Coast Guard (USCG), including response boats in diverse geographical areas and maritime environmental conditions, can pose complex and unique challenges for adapting existing analytical detection methods. The traditional culture (TC) and the rapid viability-polymerase chain reaction (RV-PCR) methods were evaluated for their compatibility with detection of spores of Bacillus thuringiensis subspecies kurstaki (Btk), a surrogate for Bacillus anthracis, in maritime environmental surface and grab samples. The representative samples collected from a USCG installation included surfaces such as aluminum on boats, nonskid tread on decks of watercraft, computer touchscreens, and concrete piers, and grab samples of boat washdown water, soil, vegetation, and gravel from surrounding areas. Replicate samples were spiked with Btk spores at two to three 10-fold increasing levels and analyzed. Out of a total of 150 samples collected and analyzed, the TC method gave 10 false-positive and 19 false-negative results, while the RV-PCR method-based analysis resulted in 0 false-positive and 26 false-negative results. An abundance of microbial background and particulates in some samples interfered with true results, while both methods gave similar results for relatively clean samples. The TC method was more time-consuming in providing results compared to the RV-PCR method, with the latter generating relatively less laboratory waste. Improved and high-throughput sample processing methods are needed for analysis of complex environmental samples.

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