Decontamination Related Products

Bio-Response Operational Testing and Evaluation (BOTE) Project – Phase 1: Decontamination Assessment:  Technical Brief and Report

The Bio-Response Operational Testing and Evaluation (BOTE) Project was a multi-agency effort designed to test and evaluate a complete response to a biological incident – from the initial public health and law enforcement reaction through environmental remediation. The BOTE Project was conducted in two distinct phases. Phase 1 was a field-level decontamination assessment. Phase 2 was an operational exercise involving key federal agencies that are responsible for the forensic investigation, public health assessment, and remediation following a biological incident. This technical brief and report are focused on Phase 1 of the project. Phase 1 was designed to assess three approaches to site remediation after the release of Bg spores within a building. The assessment incorporated recent advances in biological sampling and decontamination that had previously been tested in small-scale applications.

Inactivation of Bacillus Spores in Decontamination Wash Down Wastewater using Bleach Solution:  Journal Article

  • Bacillus anthracis spores are highly resistant to disinfection.
  • The effectiveness of bleach solution for the inactivation of Bacillus atrophaeus subspecies globigii spores (a surrogate for Bacillus anthracis) in wash down water was evaluated by bench-scale and pilot-scale experiments
  • Approximately 5% (v/v) bleach solution was adequate for a ≥7 log10 inactivation of Bacillus spores in wastewater after 10 min of contact time at room temperature.
  • This bleach dose is less than the U.S. National Response Team-recommended formulation of 10% (v/v) bleach solution with 10% (v/v) vinegar
  • For the wash down waters evaluated, the addition of vinegar was not required
  • Inactivation was faster when a detergent was added to the wash down waters.


At the request of Japan’s Ministry of the Environment, the U.S. Department of State, through its Embassy Science Fellow (ESF) Program, provided experts in radiological decontamination, one from the U.S. Environmental Protection Agency (EPA) and two from the U.S. Department of Energy (DOE) national laboratories, to provide recommendations on the continued GOJ’s off-site remediation activities.

 Sampling and Analysis Related Products

 Evaluation of Sampling Methods for Bacillus Spore-Contaminated HVAC Filters.:  Journal Article

  • Spore recovery from neat and dust-loaded HVAC filters was determined.
  • Vacuum- and extraction-based methods were compared.
  • Neither the filter type nor the presence of dust significantly affected recovery.
  • Recoveries by vacuum methods were typically 30% of those by extraction.
  • Results may guide selection of sampling methods following a biological incident.

Evaluation of swabs and transport media for the recovery of Yersinia pestis:  Journal Article

  • We evaluate four swab types, four processing methods, and six transport media.
  • We investigate storage time and temperature for recovery of Yersinia pestis.
  • Macrofoam and rayon swabs performed better than cotton or polyester swabs.
  • The optimum conditions differ by swab type and Y. pestis strains tested.
  • The best conditions provided for ≥ 83% recovery of viable cells from the swabs.

A Rapid and Repeatable Method to Deposit Bioaerosols on Material Surfaces:  Journal Article

  • A reliable and simple method for inoculating material surfaces with aerosolized spores is presented.
  • Repeatability (%CV) was similar to that of liquid-based inoculation methods.
  • A variety of surface types can be inoculated with this method.
  • This method offers significant utility to studies of agent detection, sampling, and decontamination.

Evaluation of Vacuum-based Sampling Devices for Collection of Bacillus spores from Environmental Surfaces:  Report and journal article

  • Four sampling devices (vacuum socks, mixed cellulose ester (MCE) filter cassettes, umpolytetrafluoroethylene (PTFE) filter cassettes, and 3M™forensic filters) were evaluated for their ability to recover spores from concrete, carpet, and upholstery
  • Results indicated that the MCE method exhibited higher relative recoveries than the other methods when used for sampling concrete or upholstery.
  • Vacuum sock resulted in the highest relative recoveries on carpet; however, the difference was not statistically significant.

These results may be used to guide selection of sampling approaches following biological contamination incidents.

High Throughput Determination of Ricinine, Abrine, and Alpha-Amanitin in Drinking Water by Solid Phase Extraction and High Performance Liquid Chromatography Tandem Mass Spectrometry (HPLC/MS/MS):  Analytical Method

This document provides the standard operating procedure for determination of ricinine (RIC), abrine (ABR), and α-amanitin (AMAN) in drinking water by isotope dilution liquid chromatography tandem mass spectrometry (LC/MS/MS). This method is designed to support site-specific cleanup goals of environmental remediation activities following a homeland security incident involving one or a combination of these analytes.

 Water Security Related Products

Inactivation of Bacillus Spores in Decontamination Wash Down Wastewater using Chlorine Bleach Solution:  Journal Article

The results of bench- and pilot-scale bleach inactivation tests conducted in this study using three different wash down wastewaters suggest that the currently published formulation of 10% bleach solution with 10% vinegar at 60 minutes of contact time for the inactivation of Bacillus spores in wash down wastewater is more rigorous than necessary.

Protecting Water Supply Critical Infrastructure: An Overview:  Book chapter

This book chapter discusses the general principles and characteristics of water and wastewater system security. The chapter also summarizes current research as it relates to system security, focusing on intentional threats to water systems.

Water Security Toolkit User Manual: Version 1.1:  User manual: Data Product/Software

The Water Security Toolkit (WST) assists in the evaluation of multiple response actions in order to select the most beneficial consequence management strategy. It includes hydraulic and water quality modeling software and optimization methodologies to identify: (1) sensor locations to detect contamination, (2) locations in the network in which the contamination was introduced, (3) hydrants to remove contaminated water from the distribution system, (4) locations in the network to inject decontamination agents to inactivate, remove, or destroy contaminants, (5) locations in the network to take grab samples to confirm contamination or cleanup, and (6) valves to close in order to isolate contaminated areas of the network. This user manual describes the different components of WST and how to use the software.

CANARY Training Tutorials:  Report

The CANARY event detection software was developed to enhance the detection of contaminants in drinking water. Many drinking water utilities collect real-time data from sensors located throughout the water distribution network. The sensors measure water quality parameters such as pH, residual chlorine, total organic carbon, and specific conductance. CANARY analyzes these data rapidly to identify anomalous or abnormal periods of water quality that might indicate contamination incidents. CANARY has been used as a research platform for testing and developing new capabilities for event detection system (EDS) algorithms and sensor performance. CANARY Training Tutorials provide practical examples of the full range of CANARY’s capabilities. Some basic functions are used each time the software is used, and others are used mainly in specific situations. Each example is documented with a step-by-step approach using text and screen-capture figures illustrating the changes made to the configuration file parameters and discussing the impact of those changes on the CANARY output.

A complete collection of NHSRC products can be found at www.epa.gov/nhsrc


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