Composite Sampling Approaches for Bacillus anthracis Surrogate Extracted from Soil

PLoS One. 2015 Dec 29;10(12):e0145799. doi: 10.1371/journal.pone.0145799. eCollection 2015.

Abstract

Any release of anthrax spores in the U.S. would require action to decontaminate the site and restore its use and operations as rapidly as possible. The remediation activity would require environmental sampling, both initially to determine the extent of contamination (hazard mapping) and post-decon to determine that the site is free of contamination (clearance sampling). Whether the spore contamination is within a building or outdoors, collecting and analyzing what could be thousands of samples can become the factor that limits the pace of restoring operations. To address this sampling and analysis bottleneck and decrease the time needed to recover from an anthrax contamination event, this study investigates the use of composite sampling. Pooling or compositing of samples is an established technique to reduce the number of analyses required, and its use for anthrax spore sampling has recently been investigated. However, use of composite sampling in an anthrax spore remediation event will require well-documented and accepted methods. In particular, previous composite sampling studies have focused on sampling from hard surfaces; data on soil sampling are required to extend the procedure to outdoor use. Further, we must consider whether combining liquid samples, thus increasing the volume, lowers the sensitivity of detection and produces false negatives. In this study, methods to composite bacterial spore samples from soil are demonstrated. B. subtilis spore suspensions were used as a surrogate for anthrax spores. Two soils (Arizona Test Dust and sterilized potting soil) were contaminated and spore recovery with composites was shown to match individual sample performance. Results show that dilution can be overcome by concentrating bacterial spores using standard filtration methods. This study shows that composite sampling can be a viable method of pooling samples to reduce the number of analysis that must be performed during anthrax spore remediation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anthrax / microbiology
  • Bacillus anthracis / isolation & purification*
  • Dust
  • Soil Microbiology*
  • Specimen Handling / methods*
  • Spores, Bacterial / isolation & purification

Substances

  • Dust

Grants and funding

The funder provided support in the form of salaries for all the authors [BF, WB, EC, TS]. The specific roles of these authors are articulated in the 'author contributions' section. This work was also supported by the Defense Threat Reduction Agency (DTRA) under contract HDTRA1-12-C-0071. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.