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Related Experiment Video

Updated: Jun 30, 2026

Physical Isolation of Endospores from Environmental Samples by Targeted Lysis of Vegetative Cells
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Exploring improved sample recovery and processing methods targeting Bacillus anthracis spores from vegetation

Michael Pirhalla1, Denise Aslett2, Worth Calfee3

  • 1U.S. Environmental Protection Agency (EPA) Office of Air and Radiation/Office Radiation and Indoor Air, Radiation Protection Division (formerly: Office of Research and Development (ORD)/Center for Environmental Solutions and Emergency Response (CESER)/Homeland Security and Materials Management Division (HSMMD) at the time this manuscript was developed), Research Triangle Park, NC, USA.

Journal of Microbiological Methods
|December 18, 2025
PubMed
Summary
This summary is machine-generated.

Optimizing Bacillus anthracis (Ba) spore recovery from vegetation involves reducing sample agitation time and testing various treatments. While some methods improve enumeration in ground litter and leaves, grass samples remain challenging.

Keywords:
AnthraxBacillus anthracisEmergency responseEnvironmental samplingMatrix interferenceRV-PCRSample processingSample recovery optimization

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Area of Science:

  • Environmental Science
  • Microbiology
  • Analytical Chemistry

Background:

  • Environmental contamination by persistent biological agents like Bacillus anthracis (Ba) necessitates effective remediation strategies.
  • Vegetation matrices (grass, litter, leaves) can harbor background microorganisms that interfere with target analyte recovery during contamination assessments.
  • Standardized sample collection and processing are crucial for accurate environmental monitoring.

Purpose of the Study:

  • To optimize the recovery of surrogate Ba spores from vegetation samples.
  • To investigate methods for separating target spores from non-target compounds.
  • To explore techniques for reducing background microorganism interference in vegetation samples.

Main Methods:

  • Reduced sample agitation time in Phosphate-Buffered Saline with Tween 20® (PBST) from 2 min to 30 s.
  • Evaluated a combination of heat, chemical, antifungal, and enzyme treatments for background mitigation.
  • Assessed traditional culture plating and Rapid Viability-Polymerase Chain Reaction (RV-PCR) for spore enumeration.

Main Results:

  • Sample agitation time with PBST was significantly reduced to 30 s.
  • No single treatment protocol completely eliminated background interference in grass samples.
  • Successful enumeration of Ba surrogate spores was achieved in ground litter and leaf samples using specific treatments (e.g., H₂O₂ and heat).

Conclusions:

  • Optimized sample processing can improve Ba surrogate spore recovery from certain vegetation matrices.
  • Grass samples present unique challenges for accurate spore enumeration due to persistent background microorganisms.
  • Further development of standardized methods is needed for reliable environmental monitoring of biological agents in diverse vegetation types.