Optimization of a sample processing protocol for recovery of Bacillus anthracis spores from soil

Journal of Microbiological Methods
US Environmental Protection Agency; Pegasus Technical Services, Inc.
By: , and 



Following a release of Bacillus anthracis spores into the environment, there is a potential for lasting environmental contamination in soils. There is a need for detection protocols for B. anthracis in environmental matrices. However, identification of B. anthracis within a soil is a difficult task. Processing soil samples helps to remove debris, chemical components, and biological impurities that can interfere with microbiological detection. This study aimed to optimize a previously used indirect processing protocol, which included a series of washing and centrifugation steps. Optimization of the protocol included: identifying an ideal extraction diluent, variation in the number of wash steps, variation in the initial centrifugation speed, sonication and shaking mechanisms. The optimized protocol was demonstrated at two laboratories in order to evaluate the recovery of spores from loamy and sandy soils. The new protocol demonstrated an improved limit of detection for loamy and sandy soils over the non-optimized protocol with an approximate matrix limit of detection at 14 spores/g of soil. There were no significant differences overall between the two laboratories for either soil type, suggesting that the processing protocol will be robust enough to use at multiple laboratories while achieving comparable recoveries.

Publication type Article
Publication Subtype Journal Article
Title Optimization of a sample processing protocol for recovery of Bacillus anthracis spores from soil
Series title Journal of Microbiological Methods
DOI 10.1016/j.mimet.2016.08.013
Volume 130
Year Published 2016
Language English
Publisher Elsevier
Publisher location Amsterdam
Contributing office(s) St. Petersburg Coastal and Marine Science Center, Toxic Substances Hydrology Program
Description 8 p.
First page 6
Last page 13
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