EPA/600/R-20/412 | February 2021
www.epa.gov/homeland-security-research
United States
Environmental Protection
Agency
oEPA
Literature Search and Review
for Sampling, Analysis, and
Decontamination of Biological
Warfare Agent —
Contaminated Maritime Vessels
Office of Research and Development
Homeland Security Research Program

-------
EPA/600/R-20/412
February 2021
LITERATURE SEARCH AND REVIEW FOR SAMPLING,
ANALYSIS, AND DECONTAMINATION OF BIOLOGICAL
WARFARE AGENT— CONTAMINATED MARITIME VESSELS
Lukas Oudejans, U.S. Environmental Protection Agency
and
David See, Battelle
Homeland Security and Materials Management Division (HSMMD)
Center for Environmental Solutions and Emergency Response (CESER)
Office of Research and Development
U.S. Environmental Protection Agency
Research Triangle Park, NC 27711

-------
DISCLAIMER
The U.S. Environmental Protection Agency (EPA) through its Office of Research and
Development funded and managed the research described herein under Contract Number EP-C-
15-002, Task Orders 68HE0C18F0807 and 68HERC20F0074, with Battelle. This study was
funded through the Vessel Detection US Coast Guard Project by the U.S. Department of
Homeland Security Science and Technology Directorate under EPA-DHS interagency agreement
No. 7095923201 (DHS IAANo: HSHQM-17-X-00245). This report has been subjected to the
Agency's review and has been approved for publication. Note that approval does not signify that
the contents necessarily reflect the views of the Agency. Any mention of trade names, products,
or services does not imply an endorsement by the U.S. Government or EPA. The EPA does not
endorse any commercial products, services, or enterprises. The contractor role did not include
establishing Agency policy.
Questions concerning this document, or its application should be addressed to:
Lukas Oudejans, Ph.D.
U.S. Environmental Protection Agency
Center for Environmental Solutions and Emergency Response
Homeland Security and Materials Management Division
109 T.W. Alexander Drive
Research Triangle Park, NC 27711
Phone: 919-541-2973
Fax:919-541-0496
E-mail: Oudeians.Lukas@epa.gov
li

-------
FOREWORD
The U.S. Environmental Protection Agency (EPA) is charged by Congress with
protecting the Nation's land, air, and water resources. Under a mandate of national environmental
laws, the Agency strives to formulate and implement actions leading to a compatible balance
between human activities and the ability of natural systems to support and nurture life. To meet
this mandate, EPA's research program is providing data and technical support for solving
environmental problems today and building a science knowledge base necessary to manage our
ecological resources wisely, understand how pollutants affect our health, and prevent or reduce
environmental risks in the future.
The Center for Environmental Solutions and Emergency Response (CESER) within the
Office of Research and Development (ORD) conducts applied, stakeholder-driven research and
provides responsive technical support to help solve the Nation's environmental challenges. The
Center's research focuses on innovative approaches to address environmental challenges
associated with the built environment. We develop technologies and decision-support tools to
help safeguard public water systems and groundwater, guide sustainable materials management,
remediate sites from traditional contamination sources and emerging environmental stressors,
and address potential threats from terrorism and natural disasters. CESER collaborates with both
public and private sector partners to foster technologies that improve the effectiveness and
reduce the cost of compliance, while anticipating emerging problems. We provide technical
support to EPA regions and programs, states, tribal nations, and federal partners, and serve as the
interagency liaison for EPA in homeland security research and technology. The Center is a
leader in providing scientific solutions to protect human health and the environment.
This report summarizes the current scientific literature on remediation-related activities
associated with US Coast Guard vessels and other assets. It also provides descriptions of
research gaps and needs for consideration by US Coast Guard and other federal agencies to
improve on sampling and decontamination of vessels following a biological warfare agent
release scenario.
Gregory Sales, Director
Center for Environmental Solutions and Emergency Response
111

-------
ACKNOWLEDGMENTS
This effort was directed by the principal investigator (PI) from the Office of Research and
Development's (ORD's) Homeland Security and Materials Management Division (HSMMD)
within the Center for Environmental Solutions and Emergency Response (CESER). The
contributions of the following individuals have been a valued asset throughout this effort.
EPA Project Team
Lukas Oudejans, ORD/CESER/HSMMD (PI)
Shannon Serre, OLEM/OEM/CMAD
Worth Calfee, ORD/CESER/HSMMD
Sandip Chattopadhyay, OCSPP/OPPT
Battelle Memorial Institute
David See
Ryan James
U.S. EPA Technical Reviewers of Report
Katherine Ratliff, ORD/CESER/HSMMD
Mace Barron ORD/CESER/HSMMD
U.S. EPA Quality Assurance
Ramona Sherman ORD/CESER/HSMMD
U.S. EPA Editorial Review
Joan Bursey ORD/CESER/HSMMD
iv

-------
TABLE OF CONTENTS
DISCLAIMER	II
FOREWORD	Ill
ACKNOWLEDGMENTS	IV
LIST OF ACRONYMS	VII
1.	INTRODUCTION	1
1.1.	Background	1
1.2.	Scope and Purpose	1
1.3.	Objectives	2
1.4.	Use of Secondary Data	4
1.5.	Organization of Report	4
2.	LITERATURE SEARCH	6
2.1.	Literature Search Approach	6
2.1.1.	USCG Vessels	6
2.1.2.	Target BWAs	8
2.2.	Literature Search Resources	8
2.3.	Literature Search Criteria and Strategy	9
2.4.	Literature Source Quality Assessment	11
2.4.1.	Qualitative Quality Assessment	11
2.4.2.	Quantitative Quality Assessment	12
2.5.	Literature Search Results	13
2.6.	Source Compilation Document	13
3.	KNOWLEDGE REVIEW	14
3.1.	BWA Fate and Transport	14
3.2.	BWA Contamination Management and Response	15
3.3.	BWA Decontamination Efficacy	18
3.3.1.	BWA Decontamination State of the Science	18
3.3.2.	Vaporous/Volumetric Decontamination Technologies	19
3.3.3.	Liquid-Based/Applied Decontamination Technologies	24
3.3.4.	UV Irradiation and Photodegradative Decontamination Technologies	27
3.3.5.	Physical Removal-Based Decontamination Approaches	28
3.3.6.	Thermal Decontamination	29
3.3.7.	Additional Approaches, Information, and Data	29
3.4.	BWA Decontaminant Material Compatibility	31
3.5.	BWA Sampling and Analysis	34
3.6.	Comprehensive Summary Tables	37
v

-------
4.	KNOWLEDGE/CAPABILITY GAP ASSESSMENT AND RESULTS
4.1.	Knowledge/Capability Gaps	
4.2.	Additional Information, Discussion, and Notes	
4.3.	Gap Table	
5.	REFERENCES	
ATTACHMENTS
Attachment A - Literature Assessment Factor Rating
Attachment B - Source Quality Evaluations
LIST OF TABLES
Table 1.	Focus Materials	
Table 2.	Source Document Types	
Table 3.	Research Focus Areas and Sources Collected	
Table 4.	BWA Decontamination and Sampling Summary (USCG Materials)	
Table 5.	BWA Decontamination and Sampling Summary (Additional Materials)
Table 6.	Gap Table	
LIST OF FIGURES
Figure 1.	Project Overview and Progression	
Figure 2.	1st Search Run	
Figure 3.	2nd Search Run	
Figure 4.	3rd Search Run	

-------
LIST OF ACRONYMS
ฐc
Degree(s) Celsius
ฐF
Degree(s) Fahrenheit
AHP
Accelerated hydrogen peroxide
APC
Aircraft Performance Coating
BI
Biological indicator
BiSKit
Biological Sampling Kit
BSA
Bovine serum albumin
BSL
Biosafety level
BWA
Biological warfare agent
CARC
Chemical agent resistant coating
CASCAD
Canadian Aqueous System for Chemical/Biological Agent Decontamination
CD
Compact disk
CESER
Center for Environmental Solutions and Emergency Response
CDC
Centers for Disease Control and Prevention
CFU
Colony forming unit(s)
ClO"
Hypochlorite
C102
Chlorine dioxide
cm2
Square centimeter(s)
CMAD
Consequence Management Advisory Division
COTS
Commercial off-the-shelf
CPU
Central processing unit
CT
Contact time
CWA
Chemical warfare agent
DHS
U.S. Department of Homeland Security
DTIC
Defense Technical Information Center
DVD
Digital video disk
ECL
Electrochemiluminescence
eC102
Electrochemically generated chlorine dioxide
ELISA
Enzyme-linked immunosorbent assay
EMS
Emergency Medical Service
EPA
U.S. Environmental Protection Agency
EtO
Ethylene oxide
EtOH
Ethanol
FMDV
Foot-and-mouth disease virus
FOUO
For Official Use Only
EVD
Ebola virus disease
g
Gram(s)
GPU
Graphics processing unit
H2O2
Hydrogen peroxide
HDPE
High density polyethylene
HEPA
High efficiency particulate air
HINS-light EDS
High-intensity narrow-spectrum light environmental decontamination system
HOC1
Hypochlorous acid
HSMMD
Homeland Security and Materials Management Division
HVAC
Heating, ventilation, and air conditioning
vii

-------
in2
Square inch(es)
kGy
Kilogray(s)
L
Liter(s)
LDPE
Low density polyethylene
LOD
Limit of detection
LR
Log reduction
3
m
Cubic meter(s)
MCE
Mixed cellulose ester
MDR
Multidrug resistant
MeBr
Methyl bromide
Mel
Methyl iodide
mg
Milligram(s)
MgF2
Magnesium fluoride
mL
Milliliter(s)
MRSA
Methicillin-resistant Staphylococcus aureus
NaOCl
Sodium hypochlorite
NFB
Non-freezing bleach
NHSRC
National Homeland Security Research Center
NIOSH
National Institute for Occupational Safety and Health
nm
Nanometer(s)
NTC
Navy (ship) top-coat
03
Ozone
OCSPP
Office of Chemical Safety and Pollution Prevention (U.S. EPA)
OEM
Office of Emergency Management (U.S. EPA)
OLEM
Office of Land and Emergency Management (U.S. EPA)
OPPT
Office of Pollution Prevention and Toxics (U.S. EPA)
ORD
Office of Research and Development (U.S. EPA)
pAB
pH-adjusted bleach
PC
Personal computer
PCR
Polymerase chain reaction
PDA
Personal digital assistant
PI
Principal Investigator
PPE
Personal protective equipment
ppm
Part(s) per million
ppmv
Part(s) per million by volume
PTFE
Polytetrafluoroethylene
PVC
Polyvinyl chloride
PX-UV
Pulsed xenon ultraviolet
RB-M
Response Boat - Medium
RB-S
Response Boat - Small
RH
Relative humidity
RTU
Ready to use
RV-PCR
Rapid Viability PCR
SDF
Surface decontamination foam
USCG
United States Coast Guard
USPS
U. S. Postal Service
viii

-------
uv
Ultraviolet
UV-A
UV light within the "A" band
UV-B
UV light within the "B" band
UV-C
UV light within the "C" band
VHP
Vaporous hydrogen peroxide
VRE
V ancomy cin-resi stant Enter ococcus

-------
1. INTRODUCTION
1.1.	Background
The U.S. Environmental Protection Agency (EPA) is responsible for preparing for, responding
to, and recovering from threats to public health, welfare, or the environment caused by actual or
potential hazardous materials incidents. Hazardous materials include chemical, biological, and
radiological substances, whether accidentally or intentionally released.
In 2002, Congress passed the Public Health Security and Bioterrorism Preparedness and
Response Act (Bioterrorism Act). The Office of the President issued a series of Homeland
Security Presidential Directives to specify the responsibilities of federal agencies as related to the
Bioterrorism Act. EPA's roles and responsibilities include protecting human health and the
environment from bioterrorism. Included within the scope of these responsibilities are the
personnel and assets of the U.S. Coast Guard (USCG), the principal federal agency responsible
for maritime safety, security, and environmental stewardship in U.S. ports and waterways.
The USCG protects and defends more than 100,000 miles of U.S. coastline and inland
waterways. To this end, the USCG may be responsible for countering and responding to
incidents involving weapons of mass destruction, including biological warfare agents (BWAs)
and chemical warfare agents (CWAs). To carry out their mission, the USCG maintains a fleet of
small boats, larger cutters and aircraft, as well as a network of fixed infrastructure. Such assets
are likely to be utilized in the event of a USCG response to an incident involving BWAs and/or
CWAs and would likely become contaminated as a result.
Following a contamination incident, decontamination is necessary so that assets may be returned
to service and USCG capability can be maintained. Efficacious decontamination strategies are
thus necessary. Further, effective sampling is necessary to determine the extent and magnitude of
contamination, inform responders on selection of decontamination strategies, determine the
success of decontamination strategies, and determine the presence/absence of residual
contaminants to clear assets for return to service. USCG vessel usage scenarios, operating
environments, and materials of construction present unique challenges to BWA and CWA
decontamination and sampling that have not been previously addressed.
1.2.	Scope and Purpose
The purpose of this project was to: (1) conduct a review of existing BWA and CWA
contamination response and management, decontamination, and sampling strategies; (2) assess
their applicability to USCG vessels and the associated materials of construction; and (3) identify
knowledge/capability gaps associated with decontamination and sampling of USCG vessel
materials. The review of existing BWA and CWA contamination response and management,
decontamination, and sampling strategies was accomplished through completion of a systematic
search of the open literature, focused primarily on representative USCG vessels and contaminant
groups (refer to Sections 2.1.1 and 2.1.2, respectively). The results of the literature search and
1

-------
discussion of knowledge/capability gaps related to methodologies, procedures, and technologies
for decontamination and sampling of CWA contamination on USCG assets is provided in a
separate report.
1.3. Objectives
Specific objectives to support the purpose of this project included the following:
•	Conduct a search of the open literature (including existing guidance documentation,
information, scientific literature, secondary data, etc.) for BWA and CWA contamination
response and management, decontamination, and sampling strategies that are potentially
applicable to USCG vessels and associated materials of construction.
•	Develop summaries of the guidance documentation, information sources, scientific
literature, secondary data sources, etc., that were identified and collected during the
search and develop a source compilation document to collate the findings from the
literature search according to primary research focus.
•	Identify knowledge/capability gaps associated with decontamination and sampling of
USCG vessel materials due to their unique vessel usage scenarios, operating
environments, and materials of construction.
•	Develop reports to summarize and discuss:
o The approach and resources used to conduct the literature search and the results of
the search.
o The approaches, procedures, and methodologies for BWA and CWA
contamination response and management, decontamination, and sampling and
analysis that were identified during the search and that may have relevance to
USCG vessels and associated materials of construction.
o The knowledge/capability gaps identified that relate to the unique challenges
presented by USCG vessel operations and materials that must be overcome for
development of effective decontamination and sampling strategies for BWA-
and/or CWA-contaminated USCG vessels that allow for prompt and safe return to
service.
Figure 1 provides an overview of the steps taken during the project to accomplish the above
project objectives to conduct the literature search, summarize the search results and collate the
summaries into the source compilation document, identify knowledge/capability gaps based on
the search results, and develop reports to present the search results, information and data
collected, and gaps identified.
2

-------
Definition of Project
Scope and Objectives
* Defined fey EPA in
consultation •with project
stakeholders
P] e-Search Activities
•Selection of focus
USCG vessels materials
* Selection of focus
BWAs CWAs
•Development of the
search strategy
(including selection of
search tetins and
resources'
Literature Search
~Application of the
search terms to the
search re s ources to
collect information and
secondary' data
•Refinement of the search
terms and re application
to the search resources
to elicit identification
Mid collection of
additional data and
increase data relevancy
to the project objectives
Summary ofSouices
ami Development of
Source C ompiladon
Document
* Summary of
information s eeondaxy
data sources using a
standardized template
•Assessment of source
quaity
•Colation of sourer
summaries into the
Identification and
Discussioii of
Knowledge C apnbility
Gaps
•	Presentation of the
information and
sec ondaty data c olle cte d
during the searchto
project stakeholders
•	Identification of
knowledge capability'
saps, based on the
search re suits
Reporting
•	Presentation of the
project objectives and
search strategy
•	Pre s entation of the
information and
secondary data collected
•Presentation of
knowledge capability
gaps identified
•	Separate reports for
BWAs and CWAs
document based on
pnmary research focus
Figure 1. Project Overview and Progression
3

-------
1.4.	Use of Secondary Data
Secondary data are defined as existing data, also termed nondirect measurements, that were not
developed originally through the project to which they are being applied 1. For this project,
secondary data that were gathered consisted of information and data related to BWA and CWA
contamination response and management, decontamination, and sampling strategies that are
potentially applicable to USCG vessels and associated materials of construction. These data were
collected from various sources, including government reports and publications in the open
literature.
1.5.	Organization of Report
This report is organized by section to: (1) describe how the literature search was conducted to
review existing BWA contamination response and management, decontamination, and sampling
strategies, and how the quality of the secondary data and information that were collected were
assessed; (2) present and review the secondary data and information collected; and (3) describe
the approach for identification of knowledge/capability gaps associated with decontamination
and sampling of USCG vessel materials and present the outcomes of the gap discussions. The
three primary sections of the report are outlined and described as follows:
•	Literature Search
The approach for conducting the literature search is described (Section 2.1). To focus the
literature search efforts, specific USCG vessels and their associated materials of
construction and specific BWAs were selected as focus materials/contaminants (Sections
2.1.1 and 2.1.2). Search criteria including lists of strategic keywords (i.e., search terms)
and the arrangements of the keywords with Boolean operators to execute the searches
were developed (Section 2.3), and the criteria were applied to a variety of
repositories/resources (Section 2.2) to identify and collect information and secondary
data. Quality of the information and secondary data sources that were collected was
assessed qualitatively and quantitatively (Section 2.4). Following completion of the
search but prior to development of this report, the articles, reports, guidance documents,
and other information and secondary data sources of sufficient quality that were collected
during the literature search were summarized using a standardized summary structure,
categorized according to content and primary research focus, and the summaries were
collated into a source compilation document (Section 2.6).
•	Knowledge Review
The information and secondary data collected during the literature search are presented,
categorized according to content and primary research focus, and bibliographic citations
are provided for the literature sources from which the information and secondary data
were collected. The research focus areas include: (1) BWA fate and transport, (2) BWA
contamination management and response, (3) BWA decontamination efficacy studies, (4)
4

-------
BWA decontaminant material compatibility studies, and (5) BWA sampling and analysis
methodologies (Sections 3.1 through 3.5).
• Knowledge/Capability Gap Assessment
Based on the results of the literature search, gaps in information/secondary data related to
methodologies, procedures, and technologies for decontamination and sampling of BWA
contamination on USCG assets were identified. Project stakeholders met to discuss the
literature search and the identified gaps. The knowledge/capability gaps identified and
other information, discussions, and notes from the meeting are presented in Section 4.
As discussed in Section 1.2, the scope of this project included: (1) review of existing
contamination response and management, decontamination, and sampling strategies, (2)
assessment of the applicability of the strategies to USCG vessels and their associated materials of
construction, and (3) identification of knowledge/capability gaps associated with
decontamination and sampling of USCG vessel materials for both BWA and CWA
contaminants. The approach for conducting the literature search to include both BWAs and
CWAs is included in Section 2 of this report, but the results of the literature search and
discussions of knowledge/capability gaps provided in Sections 3 and 4, respectively, are focused
only on BWAs in this report. Literature search results and discussion of knowledge/capability
gaps related to methodologies, procedures, and technologies for decontamination and sampling
of CWA contamination on USCG assets are provided in a separate report.
5

-------
2. LITERATURE SEARCH
2.1. Literature Search Approach
The literature search was conducted using the resources described in Section 2.2 and the search
criteria and strategy described in Section 2.3. The secondary data gathered during this effort
include information on BWA and CWA contamination response and management,
decontamination, and sampling strategies that are potentially applicable to USCG vessels and
associated materials of construction. For the purpose of this search, BWA and CWA
contamination response and management, decontamination, and sampling were defined as:
•	Contamination response and management - Initial action(s) taken in response to an
incident involving creation or spread of contamination by BWA, CWA, or similar agents,
as well as ongoing actions taken to assess the initial response and to direct and modify
subsequent response steps. Such actions may include: initial steps to stop the spread of
contamination and contain existing contamination (in the case of USCG vessels, such
actions may include storing/staging of contaminated vessels that cannot be
decontaminated immediately); procedures and guidance for development of response
plans and for continuous assessment and modification of plans, as necessary; guidance
and considerations regarding response safety and personal protective equipment (PPE)
requirements; and information, considerations, and guidance related to disposition and
disposal of contaminated and decontaminated wastes.
•	Decontamination - General guidance and procedures for decontamination/neutralization
of contamination by BWA, CWA, or similar agents. Information and secondary data
related to use and efficacy of specific methodologies and technologies for
decontamination of BWA, CWA, or similar agents.
•	Sampling and analysis - General guidance and procedures for qualitative and/or
quantitative detection of BWA and/or CWA (or similar agent) contamination, either prior
to or following decontamination. Information and secondary data related to use,
resolution, precision, and accuracy of specific methodologies and technologies for
decontamination of BWA, CWA, or similar contamination.
To focus search efforts, information and data related to specific USCG vessels and associated
materials of construction and specific BWAs were sought primarily.
2.1.1. USCG Vessels
2.1.1.1. Vessels and Vessel Missions
Four (4) USCG vessels were selected as focus assets for directing the literature search efforts.
The vessels and their primary missions are provided and summarized below:
•	Response Boat - Medium (RB-M) - The RB-M is a 45-foot multimission capable, all-
aluminum utility boat. The RB-M includes wireless crew communication systems and is
powered by twin diesel engines and water jet propulsion. RB-M missions include search
6

-------
and rescue, living marine resources, recreational boating safety, enforcement of laws and
treaties, and port, waterway, and coastal security 2
•	Response Boat - Small (RB-S) - The RB-S (also referred to as the Defender-class boat)
is a 25-foot boat introduced by the USCG in 2003 to replace shore-based nonstandard
boats. RB-S assets serve a variety of law enforcement, security, and vessel safety
missions 2
•	RB-S II - The RB-S II, a 29-foot boat designed as an upgrade and replacement to the 25-
foot RB-S, is a high-speed deployable asset designed to operate year-round in shallow
waters along coastal borders. The RB-S II supports search and rescue, recreational
boating safety, law and treaty enforcement, marine environmental protection, defense,
and port, waterways, and coastal security missions 2
•	Marine Protector-class Patrol Boat - The 87-foot Marine Protector-class patrol boat is a
multimission vessel capable of supporting search and rescue, law enforcement, fishery
patrol, drug interdiction, illegal immigrant interdiction, and homeland security missions
up to 200 miles offshore. The vessel includes improved seakeeping abilities and
enhanced habitability compared to other vessels, capability to interface with surface
search radars used by U.S. warships, and is designed to maintain compliance with current
and projected environmental protection laws 3.
2.1.1.2. Vessel Materials of Construction
Specific materials used in construction of the vessels described in Section 2.1.1.1 were selected
as focus materials for the purpose of further directing search efforts. The focus materials
selected, categorized by vessel, are provided in Table 1.
Table 1. Focus Materials

45-foot RB-M
Vessel Type
25-foot RB-S 29-foot RB-S II
87-foot Patrol Boat
Hull material
• Aluminum
• Aluminum
• Aluminum
• Coated steel
Decking material
on hull material
• Nonskid coatings
• Nonskid coatings
• Nonskid coatings
• Nonskid coatings
Sensitive
equipment/
components
•	Propulsion (air and
seawater intakes)
•	Other electronic
systems
•	Propulsion (air and
seawater intakes)
•	Other electronic
systems
•	Propulsion (air and
seawater intakes)
•	Other electronic
systems
•	Ventilation
•	Propulsion (air and
seawater intakes)
•	Other electronic and
internal systems
Additional
relevant
materials
•	Foam
•	Glass
•	Glazing materials
•	Insulation and other
bulkhead coverings
•	Foam
•	Glass
•	Glazing materials
•	Insulation and other
bulkhead coverings
•	Foam
•	Glass
•	Glazing materials
•	Insulation and other
bulkhead coverings
•	Glass
•	Glazing materials
•	Insulation and other
bulkhead coverings
7

-------
2.1.2. Target BWAs
Two (2) BWAs were selected in consultation with stakeholders as the focus contaminants for the
purpose of directing the literature search efforts. The two BWAs include:
•	Bacillus anthracis Ames - Virulent strain of gram-positive spore-forming bacterium that
is the causative agent of anthrax disease 4
•	Ebola virus - Refers to one of six known viral species within the genus Ebolavirus. It is
the single member of the species Zaire ebolavirus, which is the type species for the genus
Ebolavirus, family Filoviridae, order Mononegavirales. Causes Ebola virus disease
(EVD), a severe and often fatal hemorrhagic fever 5.
In addition, Bacillus anthracis A Sterne was included in the literature search as a simulant.
Bacillus anthracis A Sterne is an avirulent Bacillus anthracis strain. This strain is attenuated
through loss of the pXOl (toxin synthesis) and pX02 (capsule synthesis) plasmids 6.
2.2. Literature Search Resources
The following resources were utilized to identify information and secondary data related to
BWA and CWA contamination response and management, decontamination, and sampling
strategies that are potentially applicable to USCG vessels and associated materials of
construction:
•	SciTech Premium (also or formerly known as ProQuest Science & Technology)
Multidisciplinary content collection of scholarly material in the natural sciences,
technology, engineering and related disciplines. Includes numerous databases, including a
military database that indexes over 700 scholarly journal articles, trade and industry
journals, magazines, technical reports, conference proceedings, government publications,
etc. Included as part of the military database is the National Technical Information
Service, which provides summaries of U.S. government research, development, and
engineering, plus analyses prepared by federal agencies or their contractors.
•	Scopus
An abstract and citation database of peer-reviewed literature, with bibliometric tools to
track, analyze and visualize research. Scopus contains over 22,000 titles from more than
5,000 publishers around the world, covering the fields of science, technology, medicine,
social sciences, and others. Scopus has 55 million records dating back to 1823, and 84%
of these contain references dating from 1996.
•	Battelle Library
The Battelle library holds over 20,000 volumes and subscribes to over 10,000 print and e-
journal titles in a range of scientific and technical disciplines, both foreign and domestic.
In addition, the library manages access to more than 150 foreign and domestic databases,
including eBrary™, Hooversฎ, Applied Science & Technology, EBSCOhostฎ, and
National Technical Reports Library.
8

-------
• Internet
Internet searches (e.g., - EPA website, Google, Google Scholar™, etc.) were used to
identify available information on studies conducted using BWAs and CWAs focused on
vessel response, decontamination, and sampling.
2.3. Literature Search Criteria and Strategy
Prior to initiation of the literature search, the criteria used to perform the search were developed.
The criteria included lists of strategic keywords anticipated to elicit identification of relevant
secondary data and information, and the arrangement of the keywords with Boolean operators to
execute the searches. Following each iteration of the search and subsequent review of the results,
the arrangement of keywords and Boolean operators was revised to further focus the search and
attempt to identify additional relevant secondary data and information.
To ensure identification of a wide breadth of response and management, decontamination, and
sampling strategies potentially applicable to BWA- and CWA- contamination on USCG vessels,
initial search criteria were comprehensive, including provisions for identification of procedures,
methodologies, techniques, and technologies for contamination response and management,
decontamination, and sampling for any type of contaminant (toxic chemicals, biological
contaminants, radionuclides, etc., beyond the focus BWAs discussed in Section 2.1.2) from any
maritime vessel or environment-related material (beyond the focus materials provided in Table 1
in Section 2.1.1.2). Information regarding fate and transport of persistent BWAs and CWAs on
and across various materials was sought as well. Furthermore, the search criteria were developed
to elicit collection of information related to all relevant aspects of BWA and CWA
contamination on USCG vessel materials and impacts of use of the contamination response and
management, decontamination, and sampling strategies identified.
Boolean searches were performed using strategically selected keywords with the operators AND
and OR. After each search run (with a run defined as application of a particular arrangement of
the keywords with the operators to the sources provided in Section 2.2), the resulting identified
literature was reviewed to determine the effectiveness of the search and the relevancy of the
results. Based on the search run results, the Boolean search strategy was revised, and another run
was performed. Three runs were performed in this manner using the keywords and operators in
different arrangements to refine and focus the searches to maximize the potential of identifying
meaningful and relevant results. Figures 2, 3, and 4, below, provide the search strategies used.
All search runs were conducted simultaneously for CWA and BWA, and the results from the third
search run were used to compile the literature/references for this review.
9

-------
Related to response	Related to	Related to	.	Related to materials,
Related to contaminants
and management	decontamination	sampling	systems, and environments

Respon*


Decontaminat*


Surface


Chemical agent


Marine
OR
Manag*

OR
Detoxif*

OR
Sampl*

OR
Chemical warfare agent

OR
Maritime
OR
Mitigat*

OR
Disinfect*

OR
Analy*

OR
CWA

OR
Vessel
OR
Hazard*

OR
Clean*

OR
Wip*

OR
Distilled Mustard

OR
Boat
OR
Incident*

OR
Remov*

OR
Headspace

OR
HD

OR
Seawater
OR
Remediat*

OR
Attenuat*

OR
Vapor

OR
Sarin

OR
Foul*

AND
OR
Steriliz*
AND
OR
Qualitative

OR
GB

OR
Hull

OR
Neutraliz*
OR
Quantitative

OR
VX

OR
Bulkhead


OR
Oxidiz*

OR
Detect*

OR
Biological agent

OR
Sensitive equipment


OR
Hydroly*

OR
Inhibit*

OR
Biological warfare agent

OR
Propulsion


OR
Degrad*

OR
Interfer*

OR
BWA
AND
OR
Electronic


OR
Efficac*

OR
Sorbent

OR
Bacillus anthrads

OR
Ventilation


OR
Damag*

OR
Fate

OR
Anthrax

OR
Intake


OR
Compatib*

OR
Transport
AND
OR
Ebola

OR
Deck*






OR
Radiological agent

OR
Aluminum






OR
Radionuclide

OR
Non-skid






OR
Radioactive

OR
Glazing






OR
Radioisotope

OR
Steel






OR
Cesium*

OR
Insulation






OR
Cs-137

OR
Foam






OR
Radiological dispersal device

OR
Glass






OR
Improvised nuclear device








OR
Fallout








OR
Nuclear








OR
Chlorinated biphenyl








OR
PCB








OR
Contamina*


Figure 2, 1st Search Run
Related to response	Related to	Related to	„ , ,	,	Related to materials,
,	.	. . . ..	..	Related to contaminants	,	. .
ana management	decontamination	sampling	systems, and environments

Mitigat*


Decontaminat*


Surface


Chemical agent


Marine
OR
Hazard*

OR
Detoxif*

OR
Sampl*

OR
Chemical warfare agent

OR
Maritime
OR
Incident*

OR
Disinfect*

OR
Analy*

OR
CWA

OR
Vessel
OR
Remedial*

OR
Clean*

OR
Wip*

OR
Distilled Mustard

OR
Boat


OR
Remov*

OR
Headspace

OR
HD

OR
Seawater


OR
Attenuat*

OR
Vapor

OR
Sarin

OR
Foul*


OR
Steriliz*
AND
OR
Qualitative
AND
OR
GB

OR
Hull

AND
OR
Neutraliz*
OR
Quantitative
OR
VX

OR
Bulkhead


OR
Oxidiz*

OR
Detect*

OR
Biological agent

OR
Sensitive equipment


OR
Hydroly*

OR
Inhibit*

OR
Biological warfare agent

OR
Propulsion


OR
Degrad*

OR
Interfer*

OR
BWA
AND
OR
Electronic


OR
Efficac*

OR
Sorbent

OR
Bacillus anthrads

OR
Ventilation


OR
Damag*

OR
Fate

OR
Anthrax

OR
Intake


OR
Compatib*

OR
Transport

OR
Ebola

OR
Deck*


OR
Inanivat*





OR
Aluminum








OR
Non-skid








OR
Glazing








OR
Steel








OR
Insulation








OR
Foam








OR
Glass
Figure 3. 2"'' Search Run
10

-------
Related to response	Related to	Related to	_ . . ,. ...	Related to materials, systems,
Related to contaminants
and management	decontamination	sampling	and environments

Mitigat*
AND

Decontarninat*
AND

Sam pi*
AND

Chemical warfare agent
AND

Marine
OR
Hazard*
OR
Detoxif*
OR
Analy*
OR
Distilled Mustard
OR
Maritime
OR
Incident*
OR
Disinfect*
OR
Wip*
OR
HD
OR
Boat
OR
Remediat*
OR
Remov*
OR
Headspace
OR
Sarin
OR
Seawater
OR
Emergency
OR
Steriiiz*
OR
Vapor
OR
GB
OR
Foul*
OR
Attack
OR
Neutraliz*
OR
Detect*
OR
VX
OR
Hull
OR
Rescue
OR
Oxidiz*
OR
1 nterfer*
OR
Biological warfare agent
OR
Bulkhead
OR
Contain*
OR
Hydroly*
OR
Sorbent
OR
BWA
OR
Sensitive equipment
OR
Risk
OR
Degrad*
OR
Fate
OR
Bacillus anthraris
OR
Ventilation
OR
Cleanup
OR

OR
Transport
OR
Anthrax
OR
Deck-

OR
Ebola
OR
Aluminum
OR
Vesicant
OR
Non-skid
OR
Nerve agent
OR
Steel


OR
Foam

OR
Glass
OR
Sea
OR
Military
OR
Coast Guard [coast + guard)
Figure 4. J'd Search Run
2.4. Literature Source Quality Assessment
2.4.1. Qualitative Quality Assessment
During the literature search, information and secondary data sources were qualitatively assessed
according to the source document type. Table 2 provides the source document type list used
during the literature search (not all source document types were accumulated during the search).
Knowledge of the document type provided an indication of trustworthiness of the
information/secondary data contained therein, based on general professional judgment of each
document type.
11

-------
Table 2. Source Document Types
Designation
Description
A
Technical Report, U.S. Government
B
Technical Report, Contractor for U.S. Government
C
Translated Foreign-Language Document
D
Translated Foreign-Language Abstract
E
Untranslated Foreign-Language Document
F
Untranslated Foreign-Language Abstract
G
Peer-Reviewed English Language Literature, post-1975
H
Peer-Reviewed English Language Literature, 1925-1975
I
Peer-Reviewed English Language Literature, pre-1925
J
Government Website, with citations
K
Government Website, without citations
L
Non-Government Website, with citations
M
Non-Government Website, without citations
N
Book Chapter or Book, with peer-review and/or editorial oversight
0
Book Chapter or Book, no peer-review nor editorial oversight
P
Book Chapter or Book, peer review and editorial oversight unknown
Q
Patent (United States)
R
Patent (International)
S
Thesis/Dissertation
T
News Article
U
Manufacturer-Supplied Literature
V
Other
W
Analysis Pending
Most of the sources collected were of type A (Technical Report, U.S. Government), type B
(Technical Report, Contractor for U.S. Government), or type G (Peer-Reviewed English
Language Literature, post-1975).
2.4.2. Quantitative Quality Assessment
Each source of information and/or secondary data was evaluated according to the following
categories: focus, verity, integrity, rigor, utility, clarity, soundness, uncertainty and variability,
and evaluation and review. A description of each attribute is provided in the Literature
Assessment Factor Rating (Attachment A). Information sources were evaluated against the
Literature Assessment Factor Rating and assigned an overall rating to accomplish a
semiquantitative assessment of the quality of the source. For the quality of a source to be deemed
adequate, the source was required to receive an overall Literature Assessment Factor Rating
score of 15 or greater.
Source quality evaluations (document type designations and Literature Assessment Factor Rating
scores) for all sources of sufficient quality that were collected during the literature search,
included in the source compilation document (refer to Section 2.6), and discussed in Section 3
are included as Attachment B.
12

-------
2.5. Literature Search Results
During the literature search, 83 sources of information and secondary data related to BWA
contamination response and management, decontamination, and sampling strategies that are
potentially applicable to USCG vessels and the associated materials of construction (the focus
materials provided in Table 1 in Section 2.1.1.2) were collected. All articles, reports, guidance
documents, and other pertinent information sources of adequate quality that were collected
during the literature search were summarized using a standardized summary structure and
categorized according to content and primary research focus. Table 3 provides a list of the
research focus areas and the number of information/secondary data sources collected for each
category during the literature search.
Table 3. Research Focus Areas and Sources Collected
Research Focus Area
Number of
Sources Collected
BWA Fate and Transport
3
BWA Contamination Management and Response
8
BWA Decontamination Efficacy Studies
49
BWA Decontaminant Material Compatibility Studies
12
BWA Sampling and Analysis Methodologies
11
Total	83
2.6. Source Compilation Document
Following the literature search and summary of the information/secondary data sources that were
collected, the summaries were collated into a source compilation document according to the
primary research foci listed in Table 3. The source compilation document also provided
descriptions of the literature search approach, strategy, criteria, and resources (as described in
Sections 2.1, 2.2, and 2.3) and provisions for assessment of the quality of information/secondary
data sources that were collected during the search (as described in Section 2.4).
13

-------
3. KNOWLEDGE REVIEW
3.1. BWA Fate and Transport
The information and secondary data related to BWA fate and transport described below were
collected during the search and should be assessed, as applicable, alongside the information and
data on decontamination strategies (described in Section 3.3) and sampling strategies (described
in Section 3.5) that were collected when decisions are made and strategies are developed for
response to incidents that involve BWA contamination of USCG vessels. Fate and transport of
BWA contamination on USCG vessel construction materials will impact not only the efficacy of
decontamination and sampling methodologies but also the required extent of decontamination
and sampling efforts/operations and thus will drive contamination response and management
decisions.
Prime examples of the transportability of BWAs are the anthrax attacks that occurred over the
course of several weeks in 2001 (also known as "Amerithrax"), during which letters containing
anthrax spores were mailed to news media and federal government offices. Widespread
contamination of the affected U. S. Postal Service (USPS) facilities and exposure of multiple
personnel to Bacillus anthracis spores resulted from the letters being routed through normal mail
processing procedures 7 Aerosolized bacterial spores, including those of B. anthracis, can
remain aloft for hours and are capable of wide dispersion 8. B. anthracis spores are also very
resistant to inactivation by biocides and may survive on surfaces for centuries if not remediated
8. "Amerithrax" remediation efforts were of an unprecedented scale, with costs reaching
approximately $320 million 8. Following the attacks, several studies (including the following)
were conducted to evaluate and characterize the threat presented by spore-contaminated letters,
potential mitigation strategies, and the propensity for spread of spores by normal handling and
processing of contaminated letters:
• A study of potential mitigation procedures intended to deal with letters contaminated with
B. anthracis spores using a B. anthracis simulant spore release scenario within an actual
office building was conducted 9 Spore aerosols were created by opening letters
containing 0.1 gram (g) of dry powdered Bacillus atrophaeus spores. The movement of
B. atrophaeus spores throughout an office building was evaluated based on various
mitigation strategies including moving away from the letter opening area, closing doors,
turning off heating, ventilation, and air conditioning (HVAC) systems, water spray
mitigation, and letter opener clothing removal and showering. Potential total inhalational
hazard for the letter opener ranged from 4.1 x 105 to 1 .6 x 106 colony forming units
(CFU) compared to 3.9 x 105 CFU for controls. Surface contamination of the letter
opener was highest on the right hip (4.8 x 104 to 1.0 x 105 CFU/square centimeter [cm2])
and lowest on the right or left side of the head (2.2 x 102 to 3.7 x 103 CFU/cm2).
Mitigation procedures tested in this study generally did not reduce aerosol hazard or
surface contamination.
14

-------
• Letters contaminated with fluorescent tracer powder were opened using various
techniques in an office setting 10. Spread of contamination was qualitatively assessed
using an ultraviolet (UV) light source. Results clearly demonstrated that when letters
containing powdery contaminants are opened, the contaminant can be dispersed both on
the immediate and surrounding areas, on the person, on objects nearby, and into the
HVAC system. Potentially contaminated persons are not limited to those in direct contact
with the envelope and/or its contents.
The information and secondary data presented in these studies highlight the high potential and
high risk for spread of BWA contamination. Given the operational setting of USCG vessels,
spread of BWA contamination into water/marine environments must also be considered, in
addition to the spread of contamination throughout an affected vessel itself. The fate of viruses
and mechanisms controlling virus inactivation in coastal waters have been evaluated and
discussed u. Inactivation rates in estuarine and marine waters in laboratory studies range from 1
to 43 days when expressed as the time required for a tenfold reduction in concentration.
3.2. BWA Contamination Management and Response
USCG vessel materials and operational settings present unique challenges to the development of
effective, efficient, and safe strategies for management of and response to BWA contamination
incidents. Despite these challenges, the objectives of USCG-vessel BWA contamination incident
management, response, and remediation operations must remain unchanged from the objectives
of any other BWA contamination incident: (1) Personnel safety must be ensured; (2) BWA
contamination must be identified, contained, and adequately decontaminated; and (3) deleterious
environmental, equipment/infrastructure, and financial impacts must be avoided or minimized to
the greatest extent possible.
BWAs are highly pathogenic, and exposure (as a direct result of a BWA attack, during incident
response and remediation actions, after an incident due to residual contact hazard, etc.) can often
be lethal. Numerous studies, literature assessments, and data analyses are available
12,13,14,15,16,17,18 couate available information and summarize and describe: (1) BWA
types/classes, (2) pathogenic characteristics and mechanisms of pathogenesis, (3)
delivery/exposure routes, (4) exposure symptoms and medical diagnosis guidance, and (4)
prevention and prophylaxis, treatment options, and intervention, etc. These data and
considerations highlight the lethality of BWAs and the need for effective, comprehensive, and
rapidly mobilized contamination management, response, and remediation strategies following
incidents involving USCG vessels/assets and BWA.
Fitch, Raber, and Imbro discuss considerations related to response to a biological terrorist attack
and identify and discuss numerous technologies for detection, sampling, and decontamination of
BWAs 19 The primary focus of the review is on field systems, and emerging laboratory
technologies and a general strategy for characterization of and response to a BWA contamination
incident are provided and discussed. The overall response to an incident involving BWAs is
15

-------
broken down into four main phases: (1) monitoring and notification (i.e., contamination
detection), (2) first response, (3) characterization (i.e., sampling to discern contamination extent
and severity), and (4) restoration (i.e., decontamination). Several technologies for BWA
sampling and decontamination are identified and discussed, including liquid and vaporous
technologies (multiple liquid-applied decontamination technologies as well as chlorine dioxide
[CIO2] gas and vaporized hydrogen peroxide [H2O2, VHP] are highlighted).
Existing plans governing operations and procedures for sampling and detection of BWAs,
contamination management, and equipment and infrastructure for other organizations, sites,
facilities, installations, etc., can be used as templates for development of effective similar plans
and guidance documents tailored to specific USCG sites, vessels, and/or assets. Existing plans
may often also provide insight into strategies for selection of adequate PPE and development of
emergency response and exposure control/response plans. Similarly, assessment of the
effectiveness of response and management strategies used during previous BWA outbreak and
contamination incidents (whether accidental or terrorism-related), the associated outcomes, and
the successes and failures that were experienced can provide invaluable insight during post-
incident refinement and improvement of procedures and development of new and/or modified
procedures and/or capabilities based on any knowledge/capability gaps that were exposed.
Examples from the literature include the following:
•	Specialized training for ambulance staff, decontamination logistics considerations,
procedures for infection control (PPE and engineering and administrative controls), and
ambulances with special technical features (e.g., controlled ventilation, high efficiency
particulate air [HEPA] filtration, intercom systems, separation of drivers from patients,
etc.) for Emergency Medical Service (EMS) transport of patients with confirmed or
suspected EVD are discussed 20.
•	Procedures and equipment for containment and treatment of EVD patients are discussed
21. Use of Trexler isolator tents is discussed as a means of providing critical care to EVD
patients while minimizing risk to care providers.
•	Following the anthrax attacks that occurred in October 2001, the USPS developed plans
to install HEPA filtration systems at mail processing facilities to capture and
prevent/minimize airborne/aerosolized spores (most notably aerosolized B. anthracis
spores) in the event of a release. A report from the U.S. General Accounting Office
describes the results of government review of the proposed designs of the HEPA
filtration system 22 Results include consideration of necessary air sampling and detection
equipment to monitor/confirm effectiveness of the filtration systems and ensuring
existing infrastructure can accommodate the needs of the system (e.g., logistics, power,
etc.).
•	Describes dispatch and operation of a mobile Biosafety Level (BSL)-3 laboratory and
well-trained diagnostic team to Sierra Leone to assist in EVD diagnosis when the largest
16

-------
outbreak of EVD to date emerged in West Africa in 2014 23. The setup allowed for the
diagnosis of suspected EVD cases in less than four (4) hours following receipt of
samples. The mobile laboratory was composed of three (3) container vehicles and was
equipped with an advanced ventilation system, communication system, and electricity
and gas supply systems.
While, as discussed, review, assessment, and consideration of the approaches, procedures, and
technologies used by other organizations for BWA contamination management and response
operations, as well as review of past incidents and the associated response actions utilized (both
successful and unsuccessful), can be valuable for directing the development of new and
improvement of existing strategies for management and response during USCG vessel-related
BWA incidents, as mentioned previously. The environmental settings in which the USCG
operates and the assets requiring decontamination are likely to be very different from those
involved during incidents in civilian/urban/etc., settings. Nonetheless, various basic principles
for contamination management and response strategies may still be translatable.
Quantitative risk analysis systems, computer simulations, and mathematical models have also
been developed as valuable tools used to inform decision makers and assist with BWA
contamination management and response operation planning, such as in the following examples.
•	A mathematical model was utilized to simulate a release of anthrax in lower Manhattan to
compare a HEPA air cleaner/vacuum cleaner remediation plan with vaccinations to a
CIO2 fumigation remediation plan 24. Cost, recovery time, and number of inhalation
anthrax cases among reoccupants were the metrics of interest. The study suggested that a
HEP A/vaccine approach is viable for most buildings after a large-scale anthrax attack.
•	Available information on five BWAs (including B. anthracis, Yersiniapestis, Francisella
tularensis, Variola major, and Lassa fever) was collected (including fate and transport
and sampling data) and assessed to develop quantitative guidelines for the relationship
between environmental pathogen concentrations and human health risk in an indoor
environment 25. An integrated model of environmental BWA transport and exposure was
constructed that: (1) included effects of environmental attenuation, (2) considered
different pathogens instead of just B. anthracis, (3) considered contact exposure (e.g.,
ingestion or dermal risk) as well as inhalational exposure, and (4) included an uncertainty
analysis and identified key input uncertainties (which may inform the direction of future
research). Findings provide a framework for developing the standards required for
making risk-informed response decisions.
•	An integrated mathematical model was developed that included environmental transport,
exposure, and health risk following a release of B. anthracis spores 26. The model linked
environmental concentrations of B. anthracis to health risk so that once a target level of
health risk was specified, environmental concentration standards corresponding to the
17

-------
risk level could be computed. Intended use of the model was to support prospective risk
analysis (i.e., determining future risk, which can/will inform remediation activities).
3.3. BWA Decontamination Efficacy
Remediation of BWA contamination can be accomplished using a wide variety of strategies,
approaches, and technologies including use of volumetric decontaminants (i.e., vaporous or
gaseous technologies, fumigants, fogs, etc.), liquid-applied technologies (via spray, foam, or
simply through use of a mop or sponge), photodegradative/photolytic approaches and ionizing
radiation, cold atmospheric plasma, physical removal approaches (e.g., washing, vacuuming,
etc.), thermal decontamination approaches (e.g., hot air, autoclaving, etc.), and others 8.
Furthermore, the activities of reactive BWA decontaminants are based on a variety of
chemistries including hypochlorous acid (HOCl)/hypochlorite (CIO"), H2O2, peracetic acid,
CIO2, aldehydes, and others 8.
As discussed by Wood and Adrion 8, the specific material contaminated by BWA can often be a
critical factor affecting the efficacy of decontamination approaches/technologies. Nonporous,
hard, and inorganic materials (e.g., glass and stainless steel) are typically more easily
decontaminated than porous, permeable, or organic materials (e.g., wood, concrete, soil, etc.).
Soil and other/similar organic materials may impart a deleterious effect on the efficacy of
decontaminants that rely on oxidative degradation/inactivation of contaminants. Spores may
orient within microlocations throughout porous materials and be shielded from contact with
decontaminants. Regarding the target USCG vessel-related materials provided in Table 1 in
Section 2.1.1.2, high/sufficient decontamination efficacy may be easier to achieve from hard
nonporous materials such as aluminum and glass but more difficult to achieve from porous
materials such as foam and insulation. Efficacy of decontaminants on coated steel and nonskid
coatings will likely be dependent on the nature/characteristics of the coating (e.g., permeability).
3.3.1. BWA Decontamination State of the Science
Numerous sources are available that provide reviews of the open literature to summarize the state
of the science with respect to BWA decontamination. Such sources collected during the literature
search performed for this effort include the following:
• Wood and Adrion provide an extensive review of a wide variety of approaches and
technologies for decontamination of B. anthracis spores 8. Reviewed/discussed
decontamination methodologies are primarily categorized as liquid-based sporicides
(including HOC1 and CIO", peroxide, and aldehyde-based technologies), gaseous
decontaminants (including CIO2 gas, VHP, methyl bromide [MeBr], methyl iodide [Mel],
metam sodium, formaldehyde gas, and ozone [O3]), and physical-based decontaminants
(including thermal decontamination approaches and UV radiation). Focus was placed
primarily on technologies that inactivate spores (i.e., technologies that simply remove
spore contamination are not considered), that are commercially available, and that are
potentially applicable to use during large-scale BWA decontamination efforts.
18

-------
•	A comprehensive report commissioned by the EPA National Homeland Security
Research Center (NHSRC) summarized data and information related to multiple
characteristics of a wide variety of decontamination technologies, including general
decontaminant applicability and principles of operation, available decontamination
efficacy and material compatibility data, technical maturity, possible user concerns, and
cost considerations 27 Decontaminants were broadly categorized as: (1) liquid-based, (2)
foams and gels, and (3) gas and vapor decontaminants, and included specific technologies
such as hypochlorite (bleach, dilute bleach, pH-adjusted bleach [pAB]), aqueous CIO2,
aqueous H2O2, TechXtractฎ, SandiaFoam, Decon Green™, Canadian Aqueous System
for Chemical/Biological Agent Decontamination (CASCAD™) surface decontamination
foam (SDF), L-Gel, CIO2 gas, H2O2 vapor, paraformaldehyde, and MeBr.
•	An EPA technical brief discussed several BWA decontamination methodologies that
could be considered for use during outdoor and/or wide-area B. anthracis contamination
remediation operations 28. Methodologies included liquids, foams, fumigants, gels, and
wipes based on a variety of active species that have demonstrated efficacy against B.
anthracis spores on contaminated surfaces. Application procedures/conditions, available
efficacy data, and other considerations were provided in tabular form for EPA-registered
liquid B. anthracis decontaminants, liquid EhCh/peracetic acid decontaminants, H2O2
foams, HOC1 liquids and foams, liquids and fumigants for soil decontamination, and
commercial off-the-shelf (COTS) sodium hypochlorite (NaOCl) wipes. Effective surface
decontamination options according to surface type were provided, with multiple
decontaminant options provided for nonporous materials, porous materials, and soil.
Considerations related to use of decontamination approaches in outdoor settings were
discussed, with the primary consideration being the potential presence of grime on
surfaces to be decontaminated (liquid sporicides are generally less effective on heavily
grimed surfaces). Sodium persulfate may be the best liquid sporicidal option for
decontamination of soil. MeBr has decontaminated B. anthracis on outdoor building
materials while CIO2 may be effective in decontaminating soil and surfaces covered with
dirt or grime. Metam sodium is the most widely used soil fumigant in the United States.
Metam sodium achieved a > 6 log reduction (LR) of B. anthracis on topsoil. In the same
study, MeBr [180 milligrams per liter (mg/L) for a 36-hour exposure] also achieved > 6
LR of B. anthracis on topsoil at 25 degrees Celsius [ฐC],
3.3.2. Vaporous/Volumetric Decontamination Technologies
Vaporous/volumetric technologies provide decontamination approaches that can be used on
accessible surfaces, across larger/wide areas, and in confined or "hard to reach" areas. Hazardous
residues left following application and use of volumetric decontaminants and fumigants may still
need to be mitigated/remediated, as applicable, and compatibility of the decontaminated
items/surfaces/structures with fumigants must also be considered (e.g., corrosion and/or
condensation in electronic equipment). Secondary information and data from studies collected
19

-------
during the literature search that are focused on evaluation of vaporous/volumetric
decontamination methodologies include the following:
•	Gaseous O3 at 3 mg/L (1,500 parts per million [ppm]) produced approximately a 3 LR of
Bacillus subtilis spores (a simulant for B. anthracis) within 4 hours of exposure at 90%
relative humidity (RH) on glass surfaces 29. Inactivation rates on vinyl floor tile and
office paper were nearly the same as on glass. Slower inactivation was measured from
carpet (approximately 2 LR after roughly 4 hours) and hardwood (approximately 1.5 to 2
LR after approximately 4 hours).
•	Treatment with Mel for a duration of 12 hours at generally ambient laboratory condition
yielded a 6 LR in B. anthracis spores on stainless steel strips 30. Efficacies greater than 6
LR reduction were achieved at 55ฐC after an hour.
•	Complete inactivation of biological indicators (Bis) was achieved using H2O2
(22%)/peracetic acid (4.5%) fog, and the decontaminant also reduced aerosol-deposited
B. anthracis spores to less than 1 log CFU (with numerous samples having no detectable
spores)31. A 4 LR of viable spores was achieved on wood and stainless steel. Results for
concrete were generally not significantly different from zero.
•	Carpet, Mylarฎ coating, aluminum, rubber, upholstery, fiberglass siding, air filter
materials, and unpainted concrete were inoculated with B. anthracis Ames or B.
atrophaeus (approximately 1x10s CFU per coupon). Peracetic acid and H2O2
decontaminant fogs achieved > 6 LR of B. anthracis Ames from rubber, upholstery,
aluminum, and Mylar 32 Efficacy on unpainted concrete was generally lower.
•	Geobacillus stearothermophilus biological indicators (Bis) and stainless steel and cotton
carriers containing greater than 4 logio viable multidrug-resistant (MDR) methicillin-
resistant Staphylococcus aureus (MRS A), vancomycin-resistant Enterococcus (VRE), or
MDR Acinetobacter baumannii were treated with H2O2 vapor 33. G. stearothermophilus
spore Bis were inactivated (representing > 6 LR) and no MRS A, VRE, or MDR A.
baumannii were recovered from the steel and cotton carriers.
•	Exposure to 35% H2O2 vapor for a period of 40 minutes achieved full inactivation of
foot-and-mouth disease virus (FMDV) on Bis on a repeated basis 34. The results
demonstrate a higher decontamination efficacy for 35% H2O2 vapor compared to
formaldehyde (considered to be the primary decontamination agent for FMDV), which
requires 10 hours of contact time.
•	The efficacy of H2O2 vapor (concentration in excess of 100 ppm) and aerosolized H2O2
(less than 50 ppm) was evaluated against 4- and 6-log G. stearothermophilus Bis and test
disks containing approximately 106 spores of MRS A, Clostridium difficile or A.
baumannii35. H2O2 vapor generally achieved a 6 LR, whereas aerosolized H2O2
generally achieved less than a 4 LR.
20

-------
•	Carpet, pine wood, painted concrete, glass, Formica laminate, galvanized metal, and
painted drywall were inoculated with lxlO8 spores of B. anthracis Ames, B. subtilis, or
G. stearothermophilus. Contaminated coupons were exposed to > 1,000 ppm H2O2 gas
for 20 minutes. Mean LR values of B. anthracis Ames spores ranged from 3.0 (carpet) to
7.9 (glass and laminate). B. subtilis LR values ranged from 1.6 to 7.7. G.
stearothermophilus LR values ranged from 0.81 to 6.0. All mean LRs were statistically
significantly different from zero, except G. stearothermophilus reduction on carpet36.
•	Stainless steel coupons contaminated with Bacillus spores and Bis preloaded with greater
than 106 spores of B. atrophaeus or G. stearothermophilus were exposed to H2O2 vapor at
500 ppm to 750 ppm for 20 minutes to 60 minutes at 35ฐC or to CIO2 gas at 396 ppm for
60 minutes at 25ฐC 37. H2O2 vapor achieved a 6 LR of B. atrophaeus within 6 minutes, a
5 LR of G. stearothermophilus within 20 minutes, and a 6 LR of Bacillus thuringiensis
within 20 minutes. CIO2 gas achieved a 5 LR of G. stearothermophilus in 60 minutes, a 5
LR of B. atrophaeus after 60 minutes, and B. thuringiensis was not significantly reduced
after 60 minutes of treatment with CIO2 gas.
•	H2O2 vapor at 250 and 50 ppm by volume (ppmv) was used to decontaminate B. subtilis
on galvanized metal and fiberglass HVAC duct liner (using both coupons and actual lined
HVAC ducts) 38. Decontaminant contact durations of 90 or 240 minutes were used. The
lined duct exhibited significant H2O2 vapor desorption during the post-decontamination
aeration phase, contributing approximately 75% of the total H2O2 exposure. High
efficacy (> 7.3 LR) was achieved using 250 ppmv for both the 90-minute and 240-minute
decontaminant contact durations. Fumigation at 50 ppmv resulted in lower efficacy (4.7
LR during fumigation for 90 minutes and no measurable reduction during the desorption
phase).
•	A condensing H2O2 vapor system, in which H2O2 is injected into the air until saturation
and H2O2 begins to condense on surfaces, was tested against five viruses inoculated on
stainless steel disks 39 H2O2 exposure periods of 2 to 3 hours were utilized. Viruses were
inactivated completely after H2O2 vapor exposure in 25-milliliter (mL), 27-mL, and 33-
mL cycles.
•	EPA testing has demonstrated B. anthracis reductions by H2O2 vapor of 6.9 LR or better
from nonporous surfaces and 3.0 LR or better from porous surfaces. Other studies have
shown that 500 ppm H2O2 vapor with 30 ppm ammonia can achieve a 6 LR of B.
anthracis spores within 5 minutes on operationally relevant materials. Decontamination
with 300 ppm H2O2 vapor for 2.5 hours decontaminated G. stearothermophilus Bis 40.
•	Carpet, ceiling tile, unpainted cinder block, painted steel, painted wallboard, and
unpainted wood were inoculated with 106, 107, or 108 spores of B. anthracis NNR1A1.
Coupons were fumigated with either CIO2 gas or H2O2 vapor. In general, mean spore
21

-------
recovery from the different material surfaces ranged between 24% and 78% of the
inoculated spores. LR was observed to be a strong function of material type 41.
•	An EPA technical brief presented the results of multiple studies evaluating the efficacy of
volumetric decontamination technologies against BWAs on subway-related materials 42:
o The efficacies of sporicidal liquid fogs (specifically peracetic acid and H2O2 fogs)
against B. anthracis were evaluated on carpet, aluminum, upholstery, rubber,
Mylarฎ, fiberglass siding, air filters, and unpainted concrete. Efficacy of > 6 LR
was achieved on every material except unpainted concrete and carpet. Lower
efficacy was measured at lower temperature (10ฐC). Similar results were achieved
using 35% H2O2 and peracetic acid fogs. Comparatively lower efficacy was
measured using a 22% H2O2 fog.
o MeBr fumigation was performed on coupons contaminated with B. anthracis
surrogate spores. MeBr at a concentration of 212 mg/L (with no chloropicrin) and
conditions of 24ฐC and RH greater than 75% was applied for a 36-hour exposure
to contaminated coupons of carpet, fiberglass, aluminum, rubber, Mylarฎ, and
vinyl. No viable spores were recovered from fiberglass and aluminum coupons.
Viable spores were detected on only a limited number of carpet, rubber, Mylarฎ,
and vinyl coupons.
o CIO2 fumigation of grimed subway materials (concrete, painted steel, and ceramic
tile) was evaluated. A 6 LR in viable spores was achieved at 24ฐC and > 75% RH
using 230 ppmv CIO2 for a 12-hour exposure duration or 3500 ppmv CIO2 for a 4-
hour duration. The impact of dirt and grime on decontamination efficacy was less
noticeable than the impact of temperature and was dependent on the material.
o MeBr fumigation of ceramic tile, painted steel, concrete, and granite was
assessed. Materials were tested with and without simulated subway grime.
Fumigation with MeBr was evaluated at a concentration of 212 mg/L at
conditions of 4.5ฐC or 10ฐC and 50% or 75% RH and using exposure times
ranging from 2 to 9 days. Fumigant conditions were found to affect the efficacy of
MeBr. Grime increased the time required to achieve a 6 LR.
o A mock subway system was contaminated with a B. anthracis surrogate.
Decontamination via either bleach fogging or spray-application of pAB was
performed. Eleven (11) of 132 post-decontamination samples were positive
following bleach fogging. Five (5) of 138 post-decontamination samples were
positive following pAB spraying.
•	An EPA technical brief summarized the results of multiple studies and discusses the
effectiveness of various volumetric decontamination technologies as a function of the
operational conditions under which they are applied 43:
22

-------
o MeBr is efficacious against B. anthracis Ames spores at concentrations ranging
from 212 to 300 mg/mL, RH of 75%, temperatures ranging from 22ฐC to 32ฐC,
and exposure durations ranging from 18 to 36 hours.
o CIO2 fumigation at high concentrations (e.g., 1,000 to 3,000 ppm) has
demonstrated efficacy against B. anthracis Ames spores, but such high
concentrations are likely to create issues related to material compatibility and/or
generation capacity. Additional testing has thus been performed to evaluate the
efficacy of lower concentration CIO2 fumigation (e.g., 100 to 300 ppm) for longer
exposure durations (e.g., 3 to 12+ hours).
o Formaldehyde fumigation for 10 hours at a concentration of 1,100 ppm at
conditions of 16ฐC to 32ฐC and 50% to 90% RH can effectively inactivate B.
anthracis Ames spores on several surfaces, including industrial carpet, bare pine
wood, painted concrete, glass, decorative laminate, and galvanized metal
ductwork.
o EPA has tested VHP generators and identified the H2O2 concentration of 400 ppm
with minimum exposure duration of 6 hours (cumulative exposure of 2,400 ppm-
hours at a temperature of 18ฐC or higher) to be efficacious against B. anthracis
Ames spores.
o Ethylene oxide (EtO) has been found to be an effective decontaminant against B.
anthracis Ames under optimal conditions of concentration, contact time,
temperature, and RH. On glass and carbon steel, efficacious EtO application
conditions range from > 600 mg/L EtO, 50% RH, 50ฐC, and a contact duration >
180 minutes, to > 300 mg/L EtO, 75% RH, 37ฐC, and at least a 90-minute contact
duration.
o EPA has measured the value of 6 LR for B. anthracis Ames from glass, ceiling
tile, carpet, painted wallboard paper, wood, and unpainted concrete using 200
mg/L Mel at conditions of 25ฐC and greater than 70% RH for a 12-hour contact
duration.
o Effective peracetic acid fogging against B. anthracis requires at least 10 mL of
4.5%) peracetic acid per 1 cubic meter (m3) volume with a contact duration of
three (3) or more hours at 75% to 80% RH and a temperature of 21 to 27ฐC.
o Efficacious O3 parameters against B. anthracis Ames spores on building materials
were identified as a concentration of 12,000 ppm, a 9- to 12-hour exposure time,
85%) RH, and temperature of 21 to 27ฐC.
• Vaporous decontaminants including CIO2 vapor, formaldehyde vapor, and H2O2 vapor
are discussed 44. Benefits, drawbacks, and the intended-use BWAs are identified and
discussed for each.
23

-------
o Adequate formaldehyde efficacy requires RH to be maintained at or above 70%.
Formaldehyde is toxic, an irritant, and is classified as a human carcinogen.
o Gaseous CIO2 is not stable and cannot be stored, thus it must be generated on site.
CIO2 decontamination during the 2001 anthrax attacks involved treatment with
CIO2 at 750 ppm for 12 hours while maintaining temperature above 75 degrees
Fahrenheit (ฐF) and RH above 75%. These CIO2 conditions have achieved a 6 LR
of B. anthracis spores and are consistent with laboratory data. CIO2 gas can
penetrate some materials (e.g., porous materials, plastic, rubber). However, large
volumes of liquid waste are generated, and CIO2 concentrations above 10% pose a
risk for explosion.
o H2O2 vapor is typically produced by heating a 30 to 35% solution of H2O2 in
water, and numerous commercial H2O2 vapor generators are available. Scalability
of VHP treatment up to 5,660 m3 has been demonstrated.
•	Decontamination using VHP can lead to absorption of H2O2 into treated porous or
permeable materials (e.g., polymethyl methacrylate [Plexiglasฎ]). Subsequent outgassing
can then allow H2O2 vapor concentrations to reaccumulate to levels capable of
inactivating B. anthracis spores 45.
•	Automated H2O2 vapor room disinfection was found to reduce the incidence of C.
difficile infection in a hospital setting 46.
3.3.3. Liquid-Based/Applied Decontamination Technologies
Several secondary information and data sources were collected during the literature search from
studies focused on evaluation of decontaminants based on a variety of active species applied as
liquids, including hypochlorite-based decontaminants (e.g., NaOCl [household bleach], dilute
bleach, CASCAD™ SDF, pAB, etc.), H2O2 and peracetic acid-based decontaminants (including
Peridoxฎ Ready to Use [RTU], Spor-Klenzฎ RTU, etc.), and other decontamination
technologies. BWA decontamination efficacy data collected on liquid-applied technologies
include the following:
•	CASCAD™ SDF achieved B. atrophaeus spore LR values of 9.1, 9.2, and 9.0 from steel,
brick, and lumber, respectively, outperforming Peridoxฎ RTU which achieved LR values
of 4.7, 9.3, and 8.0, respectively, and pAB (4.8, 8.6, and 4.9, respectively) 47
•	Aqueous CIO2 achieved an 8 LR of viable B. anthracis Sterne spores in suspension in
only three (3) minutes. Spraying or spreading liquid CIO2 onto surfaces (type 304
stainless steel and polystyrene) resulted in only a 1 LR because CIO2 gas was rapidly
vaporized from the solutions 48. Full potency of the sprayed aqueous CIO2 solution was
restored by preparing the CIO2 solution in 5% bleach (0.3% NaOCl).
•	B. anthracis spores, Burkholderia thailandensis, Vibrio cholerae, Salmonella en/erica,
aflatoxin, and brevetoxin were decontaminated from seven (7) types of pipe materials
24

-------
including high density polyethylene (HDPE), polyvinyl chloride (PVC), aged black iron
pipe, and epoxy-coated steel pipe using NaOCl, Pipe-Kleanฎ, and Simple Greenฎ 49.
NaOCl was the most effective for reducing the adherence of bacteria and levels of B.
anthracis spores but not as effective against aflatoxin as the other treatments.
•	Burkholderiapseudomallei was more easily decontaminated from nonporous materials
(e.g., glass and aluminum) than porous materials (e.g., wood, concrete, and carpet) using
pAB, citric acid (1%), ethanol (EtOH; 70%), quaternary ammonium, and PineSolฎ 50.
Citric acid demonstrated poor efficacy. pAB, 70% EtOH, quaternary ammonium, and
PineSolฎ demonstrated > 6 LR on glass and aluminum at both 20ฐC and 12ฐC, but
achieved varying results for decontamination from wood, carpet, and concrete.
•	Coupons of aluminum, wood, glass, concrete, and carpet were inoculated with
approximately 1x10s CFU per coupon of one of Y. pestis, F. tularensis, Burkholderia
mallei, or V. cholerae, then decontaminated with pAB (pH approximately 6.8,
approximately 6,200 ppm chlorine), 1% citric acid, quaternary ammonia, 70% ethanol, or
Pine-Solฎ 51. Decontaminants were applied via spray and utilized 15-minute (nonporous
materials) or 30-minute (porous materials) decontaminant contact periods. Complete
inactivation was achieved more often with Pine-Solฎ and pAB than with other
decontaminants (particularly from nonporous materials). Roughly 7 to 8 LR was achieved
for both Pine-Solฎ and pAB for all four organisms from aluminum and glass. Citric acid
(1%) demonstrated the lowest efficacy for all four organisms on all materials.
•	Bleach (5,250 ppm NaOCl), SDF, and Virkon (2%) were used to decontaminate spores
(G. stearothermophilus, 4.6 x 106 CFU) on stainless steel carrier disks with and without
light or heavy organic load at -20ฐC, 4ฐC, 10ฐC, or 23ฐC for predefined time periods up
to 24 hours 52 At -20ฐC, less than 2.0 LR was achieved. With light organic load after two
(2) hours at 4ฐC and 10ฐC, bleach achieved 4.4 and 4.7 spore LR values, respectively.
After 24 hours with light organic load at 4ฐC and 10ฐC, both bleach and SDF LR were
more than 5.0. Virkon was less efficacious at all temperatures with light organic load.
With heavy organic load, all three decontaminants produced less than 2 LR within two
(2) hours at either 4ฐC or 10ฐC. Efficacy of SDF was 4.5 LR at both temperatures after 24
hours. With both organic loads at 23ฐC, SDF and Virkon achieved 5.5 LR in 24 hours.
Bleach was comparable with the light organic load but not with the heavy organic load.
•	Bleach at 2%, 5%, and 10% (by volume in water, without adjusting pH), Virkon (5%),
Spor-Klenzฎ RTU, Rescue Sporicidal Liquid (4.5% H2O2, accelerated hydrogen
peroxide [AHP]) and Allen Vanguard SDF were used to decontaminate 106 CFU of B.
anthracis Sterne spores on stainless steel 53. Bleach (10%) consistently achieved > 6 LR
of spores after 5 minutes at room temperature, whereas it took >10 minutes of contact
time for AHP and at least 20 minutes for 5% bleach and Spor-Klenzฎ.
25

-------
•	Bleach, Spor-Klenzฎ RTU (peracetic acid and H2O2), and Metricide 14-Day (2.6%
glutaraldehyde) were used to decontaminated, subtilis on glass 54 Three efficacy levels
per decontaminant were evaluated, and LR values increased with increasing efficacy
level.
•	Bleach (5,000 ppm available chlorine), 70,000 ppm accelerated H2O2, 1,000 ppm CIO2,
and 3,000 ppm peracetic acid were used to decontaminate various species of Bacillus
spores on stainless steel disks 55. CIO2 achieved a LR of > 6 of B. anthracis Sterne spores
at all the contact times tested. B. licheniformis exhibited the highest resistance to
inactivation by CIO2. Peracetic acid showed a faster level of sporicidal activity (higher
levels of spore kill at 5 minutes than the other three decontamination formulations tested).
Peracetic acid activities at 10 and 20 minutes were roughly comparable to diluted bleach
and accelerated H2O2.
•	Tyvek, butyl rubber, stainless steel, and polycarbonate were inoculated with B. anthracis
A Sterne or ricin protein toxin and disinfected with pAB or dilute Peridoxฎ RTU 56. No
viable Bacillus spores were recovered from any surface after 15 minutes of treatment
with pAB or dilute Peridoxฎ RTU (representative of > 6 LR). Ricin protein toxin was not
detected on any surface after 15 minutes of treatment with pAB. Ricin toxin was detected
on all surfaces after 30 minutes of treatment with dilute Peridoxฎ RTU.
•	Ebola virus surrogates (bacteriophages MS2, M13, Phi6, and PR772) were inoculated
onto stainless steel disks and decontaminated using 0.1% and 0.5% dilute bleach 57
Contact periods for inactivation ranged from 1 to 10 minutes. Dilute bleach (0.5%)
achieved 3.4 and 3.5 LR values against MS2 and M13, respectively, after a 10-minute
treatment time. PR772 achieved a 4.8 LR using 0.5% dilute bleach after only 1 minute.
Phi6 achieved a 4.1 LR using 0.5% dilute bleach after 5 minutes and was not detected
after 10 minutes.
•	An EPA technical brief discussed the test procedures and results of studies performed to
evaluate strategies and technologies for decontamination of BWAs in outdoor
environments and challenging settings 58:
o Evaluation of B. atrophaeus (B. anthracis surrogate) and bacteriophage MS2
decontamination efficacy of pAB, Spor-Klenzฎ RTU, and citric acid (2%)
applied using a handheld sprayer, backpack sprayer, and chemical sprayer on
concrete and treated plywood was performed. pAB was more effective than Spor-
Klenzฎ RTU against spores on concrete (7.3 LR using the backpack sprayer).
Spor-Klenzฎ RTU was more effective than pAB against spores on plywood (7.4
LR using backpack sprayer).
o Eight (8) nonfreezing bleach (NFB)-based formulations were evaluated for
efficacy against B. atrophaeus on concrete and glass when applied at temperatures
ranging from -25ฐC to 25ฐC. pAB was included during tests conducted above
26

-------
freezing temperatures (> 0ฐC) as a reference decontaminant. None of the NFB
formulations were as effective as pAB, and as test temperatures were lowered,
decontamination efficacies also tended to decrease.
•	B. anthracis A Sterne and B. anthracis Ames spores were inoculated onto stainless steel
coupons and decontaminated using electrochemically generated CIO2 (eClCh)59 The
eC102 decontaminant achieved a 7.0 ฑ 0.5 LR of spores following a contact period of 1
minute.
•	Coupons of aircraft performance coating (APC)-painted aluminum, stainless steel, Navy
ship top-coat (NTC)-coated stainless steel, chemical agent resistant coating (CARC)-W-
coated stainless steel, magnesium fluoride (MgF2)-coated glass, low density polyethylene
(LDPE), and Lexanฎ were inoculated with > 7 logs of spores of B. anthracis Ames, B.
anthracis A Sterne or B. thuringiensis A1 Hakam. Contaminated coupons were
decontaminated using PES-Solid at room temperature using a 15-minute decontaminant
contact period. Either no spore survival or less than 1 log of viable spores was recovered
from 56 of 63 possible test combinations (strain, decontaminant formulation, and test
material surface) after treatment with PES-Solid. Less than 2.7 log CFU survived in the
remaining test combinations 60.
3.3.4. UV Irradiation and Photodegradative Decontamination Technologies
Secondary information and data collected during the literature search from studies focused on
evaluation of UV irradiation and other photodegradative decontamination technologies include
the following:
•	UV irradiation within the "C" band (UV-C; specifically, 253.7 nanometers [nm] in this
study) was evaluated for inactivation of eight varieties of Bacillus spores (including B.
sub/ilis, B. anthracis Sterne, and others) 61. Percent kill on agar plates increased from an
average of 21% for a 30-second exposure to an average of 71% for a 120-second
exposure (a high value of 81 % was achieved at 120 seconds for a strain of B. subtilis).
•	Pulsed xenon UV (PX-UV) disinfection demonstrated a > 4 LR for canine parvovirus (an
Ebola virus surrogate) on glass carriers, face shield material, and gown material initially
inoculated at 5.98 log per carrier 62
•	The effect of simulated sunlight (UV irradiation in the A [380 to 320 nm] and B [320 to
290 nm] ranges, i.e., UV-A and UV-B) on the inactivation kinetics of virulent B.
anthracis Ames spores and B. subtilis spores was investigated 63. Bacillus spores were
dried on porous (including wood, unpainted concrete, and topsoil) and nonporous
materials (glass) at 1x10s CFU per coupon. Contaminated coupons were exposed to both
UV-A and UV-B radiation using elapsed times of 2, 14, 28, and 56 days. Data showed
that viable spore recovery is diminished when contaminated coupons are exposed to
simulated sunlight on all materials except topsoil. UV-A/B exposure resulted in roughly 1
27

-------
to 2 LR on glass, wood, and concrete. As high as 6 LR on glass at 56 days was observed.
Without exposure to UV, only approximately 1 LR on glass, concrete, and wood was
observed for both Bacillus species. Minimal reduction in/on topsoil was observed
regardless of spore species/UV exposure condition.
•	Suspensions of Candida albicans, Aspergillus niger, B. subtilis, Clostridium perfringens,
and Mycobacterium fortuitum were exposed to a UV light source at a distance of eight
feet for a duration of 30 minutes 64 An LR value of 4 was achieved for Bacillus subtilis
and C. perfringens. An LR of 3 was achieved for C. albicans and M fortuitum. LR for A.
niger was less than 3.
•	A ceiling-mounted 405-nm high-intensity narrow-spectrum light environmental
decontamination system (HINS-light EDS) achieved a decrease between 22% and 86% in
the mean number of surface bacteria (hospital room-related surfaces)65. When use of the
HINS-light EDS was discontinued, surface bacteria increased by 78% to 309%.
3.3.5. Physical Removal-Based Decontamination Approaches
As discussed earlier, physical removal approaches include methods such as washing, vacuuming,
wipe-removal of contaminants, etc. Such approaches decontaminate surfaces by removing
BWAs, but do not always inactivate spores/viruses/etc. Thus, rinsate, vacuuming waste, and used
wipes must be further decontaminated using other reactive approaches/technologies prior to
disposal. Addition of reactive components to wipe technologies (e.g., saturation of wipes with
reactive liquid decontaminants) can impart BWA inactivating/degradative qualities. Secondary
information and data collected during the literature search from studies focused on evaluation of
sporicidal and disinfecting wipe technologies include the following:
•	Four (4) hypochlorite-based sporicidal decontamination wipes (Cloroxฎ Healthcare™
Bleach Germicidal Wipe, Sani-Clothฎ Bleach Germicidal Disposable Wipe, Dispatchฎ
Hospital Cleaner Disinfectant Towel with Bleach, and Hype-Wipeฎ Disinfecting Towel
with Bleach), a H202/peracetic acid-based sporicidal wipe (Steriplexฎ SD Wipe), two
commercially available disinfecting wipes (Lysolฎ Disinfecting Wipe and Cloroxฎ
Disinfecting Wipe), and a pAB-wetted wipe were evaluated for efficacy in inactivation of
B. atrophaeus spores (B. anthracis surrogate) on glass Petri dishes, stainless steel,
composite epoxy, LDPE, and painted drywall 66. All four hypochlorite-based sporicidal
wipes achieved a LR of at least 7 of Bacillus spores, with the exception of the Dispatchฎ
wipe on painted drywall (5.71 LR).
•	Four sporicidal wipes (Cloroxฎ Healthcare™ Bleach Germicidal Wipe, Sani-Clothฎ
Bleach Germicidal Disposable Wipe, Dispatchฎ Hospital Cleaner Disinfectant Towel
with Bleach, and Hype-Wipeฎ Disinfecting Towel with Bleach) and three disinfecting
wipes (Steriplexฎ SD Wipe, Lysolฎ Disinfecting Wipe, and Cloroxฎ Disinfecting
Wipe) were used to inactivate B. atrophaeus on 12-inch by 12-inch coupons of stainless
steel, glass, composite epoxy, painted drywall, and LDPE 67. The sporicidal wipes
28

-------
achieved >6.1 LR on all materials with only a single exception (the Dispatchฎ wipe on
painted drywall, which achieved a 5.7 LR). Two sporicidal wipes were then used to
inactivate B. atrophaeus on larger glass and painted drywall surfaces with dimensions of
42-inches by 42-inches. The highest efficacy obtained when spores were evenly
distributed across the surfaces was 4.5 LR from glass by the Hype-Wipeฎ.
3.3.6.	Thermal Decontamination
Thermal decontamination can provide an efficacious and, depending on the surface, material-
compatible BWA inactivation approach. Secondary information and data collected during the
literature search from studies focused on evaluation of thermal decontamination include the
following:
•	Thermal decontamination (using hot, humid air) was capable of complete
decontamination of B. anthracis A Sterne or B. thuringiensis A1 Hakam spores (> 7 logio)
from aluminum, anti-skid material, and wiring insulation but not nylon webbing 68.
Nineteen (19) combinations of temperature (55ฐC, 65ฐC, and 75ฐC), RH (70%, 80%, and
90%>), and duration (1, 2, or 3 days) were evaluated for efficacy. Porous materials and
organic debris delay decontamination kinetics for hot, humid air.
•	APC-coated aluminum, anti-skid-coated aluminum, InsulFab insulation, wiring
insulation, nylon webbing, and polypropylene coupons were inoculated with > 7 logio of
B. anthracis A Sterne and B. thuringiensis A1 Hakam spores. Hot humid air
decontamination (upper limit of temperature and RH of 77ฐC and 90%>) was less effective
at inactivating spores on nylon. For wet spore controls and spores dried onto wiring
insulation, most of the test runs showed complete spore inactivation 69.
3.3.7.	Additional Approaches, Information, and Data
Plasma is an emerging technology that shows potential as a simultaneously efficacious and
material-compatible surface decontamination approach. A prototype nonequilibrium corona
plasma surface decontamination technology achieved B. subtilis destruction efficiencies of 98%>
on plastic, 99.9% on aluminum, and 99.4%> on cotton surfaces with a 60-second treatment
duration 70. Treatments of 5 minutes resulted in > 3 LR without harm to material surfaces. Plastic
and CARC-painted aluminum inoculated with 2.5 x 106 spores of B. anthracis were treated, with
up to 99.7%o of spores destroyed with a 60-second exposure.
Secondary information and data from other studies collected during the literature search that are
focused on decontamination of BWAs by a variety of technologies using multiple application
approaches include the following:
•	Acoustic ceiling tile, carpet, fabric, painted wallboard, concrete, and CARC-painted
metal were contaminated with spores of Bacillus globigii (B. anthracis simulant) via
aerosol spray (107 to 108 CFU per 4 square inches [in2]). University of Michigan
Nanotech (novel, broad-spectrum antimicrobial nanoemulsion), SNL aqueous foam, and
29

-------
L-Gel performed better than gaseous O3, activated hypochlorite, GD-5 (mixture of
aminoalcholates and surfactant), and metal oxide nanoparticles at inactivating spore
contamination 71.
•	Tile, fabric, wood, activated carbon-based PPE, glass, paper, plastic, and metal coupons
were contaminated with B. atrophaeus spores. NaOCl (5%, 0.5%, and 0.05%) at pH 7
and pH 12 (30 min contact time) resulted in no bacterial growth on any sample, except
for paper (0.5% and 0.05% NaOCl at pH 12 as well as 0.05% NaOCl at pH 7) and both
tile and metal (0.05% NaOCl at pH 12). Spores were detected on tile after sterilization
with EtO, but no spores were detected after autoclaving, 2% glutaraldehyde sterilization
for 30 minutes, boiling water treatment for 30 minutes, or treatment with 3% H2O2. UV
irradiation (24 h) completely removed contamination from fabric, wood, PPE, and paper.
Free chlorine solutions (1,000 mg/L and 10,000 mg/L) provided full disinfection of all
materials 72.
•	UV light-based systems have been shown to achieve up to a 4 LR of vegetative bacteria
on carriers in (up to) 20 minutes, and up to a 4 LR of C. difficile in (up to) 100 minutes.
H2O2 systems have been demonstrated to achieve complete inactivation (> 6 LR) of G.
stearothermophilus spores and all surface contamination of MRS A, VRE, M
tuberculosis, and other various spores, viruses, and multidrug-resistant gram-negative
bacilli 73.
•	Data from the open literature on UV light and H2O2 room decontamination systems were
reviewed 74 Efficacy of UV light and H2O2 systems against microbes experimentally
plated on carrier materials as well as MDR pathogens in hospital settings has previously
been demonstrated. Data presented for UV irradiation demonstrate MDR organism LR
values as high as 4.71 on carriers. H2O2 systems (aerosolized H2O2 and VHP) have
demonstrated MDR organism percent reduction values between 86% and 100% in
contaminated hospital rooms.
Decontamination approaches selected for remediation of BWA contamination on USCG vessels
must be simultaneously efficacious and material-compatible to ensure BWA surface hazards and
the potential for, e.g., spore re-aerosolization are sufficiently remediated/mitigated while not
compromising the integrity and function of vessel materials, construction, and mechanical and
electrical systems, allowing for unlimited return of vessels to service. Other characteristics of
decontamination technologies may also be particularly relevant to consideration of their use
during USCG vessel-related decontamination operations. Examples of other potentially relevant
technology characteristics given the operational settings of USCG vessels (beyond
decontamination efficacy and material compatibility) include: (1) any logistical burden related to
application and/or use of technologies (i.e., high utility requirements, bulky equipment, high raw
material or reagent needs, etc.), (2) production of large amounts of wastes and/or hazardous
wastes, (3) demonstration of the technology/approach at full-scale, (4) cost and availability, (5)
potential health and environmental impacts, and others 8.
30

-------
3.4. BWA Decontaminant Material Compatibility
Reactive decontaminants that have either demonstrated efficacy against BWAs or have been
evaluated for efficacy against BWAs are based on a variety of chemistries (e.g., hypochlorite-
based and oxidative technologies). However, not all BWA decontamination technologies are
appropriate for use in certain circumstances given the corrosive nature of some reaction
chemistries toward the surface(s) to be decontaminated, especially critical for sensitive
equipment and related surface materials, as such equipment and materials are often associated
with high procurement costs and long lead times for procurement. Sensitive equipment and
surfaces are incorporated extensively into the construction of USCG vessels. Consideration of
the compatibility of decontamination systems with the materials incorporated into the
construction of USCG vessels, including sensitive equipment and related materials, is critical to
ensuring a prompt and unlimited return to service of the assets following decontamination.
Secondary information and data collected during the literature search from studies focused on
evaluation of the compatibility of decontamination technologies with a variety of materials
(including sensitive equipment and sensitive equipment-related materials) include the following:
•	The compatibilities of unpainted concrete cinder block, standard stud lumber (fir), latex-
painted gypsum wallboard, ceiling suspension tile, painted structural steel, carpet, and
electrical circuit breakers with VHP (application conditions of < 30% RH and > 30ฐC)
were evaluated 15. Materials were exposed to either 250 ppm VHP for four (4) hours for a
total contact time (CT) of 1000 ppm-hours, or 125 ppm VHP for eight (8) hours also for a
total CT of 1000 ppm-hours. Generally, VHP-exposed building materials showed no
change in appearance or integrity compared to nonexposed samples.
•	The effects of MeBr fumigation and its interactions with 24 different test materials,
including HVAC duct and liner and painted metal, were evaluated 76. Materials were
exposed to MeBr at 1,000 ppm for 16 hours. Generally, MeBr did not ad/absorb
appreciably onto/into the materials tested, though some diffusion into porous materials
occurred.
•	Compatibilities of MeBr (300 mg/L with 2% chloropicrin for nine (9) hours at 37ฐC and
75% RH), CIO2 gas (CTs ranging from 900 ppmv-hours to 9,000 ppmv-hours), VHP, and
EtO gas with sensitive equipment (including functioning personal computers [PCs]) were
evaluated 77 CIO2 treatment caused material degradation, but PCs remained functional.
No changes in visual appearance or functionality due to exposure to VHP were observed.
Corrosion was observed on low carbon steel and steel outlet/switch boxes following
MeBr fumigation (no other materials were affected). Power supplies in all MeBr-
fumigated PCs failed (but this was attributed to the chloropicrin component of the MeBr
fumigant). Little to no impact to any of the materials following fumigation with EtO was
recorded.
31

-------
•	Fumigation technologies that have been used to decontaminate sensitive equipment
materials were reviewed 78. Fumigation with EtO generally demonstrates the greatest
degree of material compatibility. However, EtO is highly toxic and flammable, so ex-situ
decontamination (off-site or at a separate location on-site) is recommended. VHP
treatment is effective and material-compatible, but the application process must be
closely monitored and controlled to prevent damage (due to condensation). MeBr
demonstrates material compatibility but is toxic to humans. CIO2 gas is generally a more
efficacious decontaminant than VHP but demonstrates decreased material compatibility
in comparison to VHP.
•	Compatibility of CIO2 fumigation with sensitive electronic components and materials was
evaluated using multiple conditions including: (1) 3,000 ppmv CIO2 with 75% RH, (2) 75
ppmv CIO2 with 75% RH, (3) 75 ppmv CIO2 with 40% RH, and (4) 3,000 ppmv CIO2
with 90% RH 79. No visual or functional changes for stainless steel, laser-printed paper,
or gaskets were observed. Circuit breaker screws and inkjet-printed paper were affected
under every condition (including tests using only high RH [i.e., no CIO2]). RH at 75%
severely affected low carbon steel, copper, photographs, and drywall nails and screws.
CIO2 fumigation at high temperature and RH led to intermittent light switch failures. No
impacts to personal digital assistant (PDA) devices under any fumigation condition were
observed. Mild discoloration and fading of cell phone screens were noted under certain
conditions. CIO2 and condensing humidity caused severe corrosion of fax machine printer
bars, compact disks (CDs), and digital video disks (DVDs). At lower RH, these impacts
were not observed. Power state of PCs had an effect on material/decontaminant
compatibility. CIO2 and RH at least 75% resulted in corrosion of computer components.
CD/DVD drives were damaged by 3,000 ppmv CIO2 and RH greater than 75%.
•	Compatibility of sensitive electronic components and materials with MeBr was evaluated
at a MeBr concentration of 300 mg/L (with 2% chloropicrin), 75% RH, and 37ฐC for nine
(9) hours 80. Compatibility with CIO2 was also evaluated at 3,000 ppmv CIO2, 75% RH,
and 24ฐC for 3 hours (CT 9,000 ppmv-hours). MeBr fumigation with chloropicrin caused
some surface corrosion of low carbon steel and rusting around the edges of steel
outlet/switch boxes, but otherwise no effects were observed. CDs, DVDs, cell phones,
PDAs, and a fax machine all retained visual and functional integrity. PC parts affected by
the MeBr (with chloropicrin) and CIO2 fumigants included external and internal stamped
metal grids, external metal slot covers, and internal cut metal edges. All PCs exposed to
MeBr fumigation exhibited power supply failures, but the failures were traced to the
chloropicrin component of the MeBr fumigant. PC central processing units (CPUs) and
CPU and graphics processing unit (GPU) heat sinks were not impacted by either
fumigant.
•	Unpainted concrete cinder block, standard stud lumber (2-inch by 4-inch fir), latex-
painted 0.5-inch gypsum wallboard, ceiling suspension tile, painted structural steel,
32

-------
carpet, and electrical (circuit) breakers were exposed to CIO2 vapor 81. Fumigation
conditions were 2,000 ppm CIO2 for six (6) hours for a total CT of 12,000 ppm-hours, or
1,000 ppm CIO2 for twelve (12) hours also for a total CT of 12,000 ppm-hours. RH target
was 75%, and temperature target was 75 ฐF. No visual differences were observed for any
of the materials following CIO2 exposure. Tensile strength of standard stud lumber
furring strips was reduced by exposure to high concentrations of CIO2 for short durations.
Under a 30-amp load, C102-exposed circuit breakers tripped more slowly than the control
units. Otherwise, no functional impacts were observed.
•	MeBr (without chloropicrin) and Mel fumigation were performed under conditions of 26
to 30ฐC and 75 to 85% RH for 48 hours at target MeBr or Mel concentrations of 200 to
250 mg/L 82 Desktop computers were used as surrogates for high-value
sensitive/electronic equipment. Coupons of metals used frequently in electronics (copper,
aluminum, tin) were included to evaluate corrosion. Only copper coupons showed a
noticeable difference in appearance after fumigation (green and brown discoloration).
The most substantial effect observed on PC functionality was damage to displays
fumigated with Mel, particularly those that were powered on during fumigation
(demonstrated reduced brightness and a bluish tint).
•	Compatibility of gamma irradiation at 30 and 50 kilograys (kGy) with historical oil
paintings, archival documents, books, photographs, historical pastel paintings,
wood/furniture, porcelain/bisque, fabrics, metal and alloys, and leather was evaluated 83.
All materials showed some visual changes.
•	The material demand (mass flux) of selected materials for CIO2 under select fumigation
conditions (specifically, CIO2 concentrations of 1,000 ppmv or 2,000 ppmv to achieve a
total CT of 12,000 ppmv-hours) was determined 84 The materials included concrete,
painted steel, wood, gypsum wallboard, ceiling tile, and carpet. Required feed
concentration and the time required to reach the target CIO2 fumigant concentration were
found to be functions of building material. Rank of CIO2 demand for the building
materials over the 0 to 12,000 CT range was (from highest demand to lowest) ceiling tile
> wood > gypsum wallboard > carpet > concrete = steel = baseline for the 1,000 ppmv
tests, and ceiling tile > gypsum wallboard > carpet > wood > concrete = steel = baseline
for the 2,000 ppmv tests.
•	Compatibility of H2O2 (3% liquid), bleach (0.58% hypochlorite), and Oxoneฎ (1%) was
evaluated on metals (copper, brass, silver, tin, titanium, iron, and gold), inks, cellulose
from new and aged paper and cotton fabrics, collagen, keratin, and fibroin (silk)85.
Results demonstrated that the decontaminants damaged the materials, but the degree of
damage varied with the specific decontaminant and the material. Damage can be
minimized with the appropriate choice of decontaminant. H2O2 was generally the least
aggressive on metals. Oxoneฎ was the most aggressive on organic materials. Bleach
decontamination affected a higher percentage of inks.
33

-------
• The material demands of unpainted concrete cinder block, standard stud lumber, latex
painted gypsum wallboard, acoustical ceiling suspension tile, primer-painted structural
steel, and carpet for VHP was evaluated 86. The concrete cinder block coupon had the
greatest impact on maintaining the VHP concentration, while carpet and steel had a low
impact on the VHP concentration.
3.5. BWA Sampling and Analysis
To identify specific BWA contaminants, accurately and precisely determine the extent of BWA
contamination, and assess the effectiveness of BWA decontamination efforts, effective sampling
procedures, methodologies, and technologies for qualitative and/or quantitative measurement of
BWA concentrations/amounts in a variety of environmental matrices are necessary (including
measurement of surface concentration levels, concentrations in liquid matrices, and
aerosol/aerosolized concentrations). As with approaches for decontamination, the unique
operational settings and construction materials associated with USCG vessels can create
challenging sampling scenarios. Effective surface sampling can be challenged by the materials
themselves (which, at times, incorporate complex coating systems) and/or by various
contaminants and foulants introduced by USCG vessel operational settings (e.g., seawater,
grime, etc.).
Silvestri et al. summarized and discussed key challenges faced in collecting, analyzing, and
interpreting microbial field data from a contaminated site (consideration is given primarily to B.
anthracis contamination) 87 The implications and limitations of using field data for determining
environmental BWA concentrations both before and after decontamination were explored.
Considerations, challenges, and limitations associated with collection of field samples for BWA
contamination characterization and/or assessing remediation effectiveness as well as estimation
of environmental concentrations from interpretation of the data were also presented and
discussed.
In addition to the efficacy of BWA sampling methods for recovery of target contaminants from
intended matrices, other method characteristics must also be considered when decisions are
made regarding which methods to use during BWA contamination response and remediation
operations. One such important consideration is the potential for cross-contamination from
application of the sampling method. Fluorescing tracer powder was used to evaluate cross-
contamination potential during sample collection and packaging operations 88. B. atrophaeus
was used as a surrogate for B. anthracis. Sampling was performed according to Centers for
Disease Control (CDC), National Institute for Occupational Safety and Health (NIOSH) and
EPA methods using 3M Sponge-Sticks, and recommendations were provided (based on the
results) to minimize/eliminate the observed transfer of contamination that occurred during
application of the sampling approaches.
Once appropriate/effective BWA sampling methods have been selected and implemented,
collected samples must be analyzed to qualitatively/quantitatively determine BWAs. BWA
34

-------
analysis and detection technologies include culture-based assays, polymerase chain reaction
(PCR)-based technologies (including real-time PCR and Rapid Viability PCR [RV-PCR])
biosensors, microarrays, immunoassays, electrochemiluminescence (ECL), enzyme-linked
immunosorbent assay (ELISA), and others. A literature review by Herzog et al. (2009) includes
discussion of studies on detection of BWAs in soil and air, detection on fomites, and detection in
water 89 Methods for surface sampling include collection of BWA samples from stainless steel,
plastic, wood, glass, painted wallboard, carpet, and concrete using swabs, wipes, vacuum socks,
and a biological sampling kit [BiSKit; designed to sample surfaces for viruses, bacteria, and
toxins]). The data collected suggest that pre-moistened swabs perform better than dry swabs,
although the BiSKit outperforms swab sampling. The predominant methods used for detection of
BWAs are cultivation/culture-based assays and PCR-based methods (including RV-PCR).
Considering the median instrument limit of detection, real time PCR and PCR are the most
sensitive methods, with median instrument limits of detection (LODs) of 430 and 440 cells/mL,
respectively.
Common BWA surface sampling approaches include vacuum-based methods and wipe/sponge-
based methods. Secondary information and data collected during the literature search from
studies focused on evaluation of vacuum-based BWA sampling methodologies, including the
following.
•	Recovery of (aerosol-deposited) B. atrophaeus spores from pleated HVAC filters by an
extraction method (phosphate-buffered saline) and two vacuum methods (vacuum sock
and cassette filter) were compared 90. The HVAC filters were tested both with and
without dust loading to approximately 50% of their holding capacity. Recovery of spores
from the filters via extraction was higher than recovery by either of the vacuum methods.
Vacuum recovery was approximately 30% of the recovery achieved by the extraction
method. Recoveries between the two vacuum methods that were evaluated were not
significantly different. Although the extraction methods provided higher recovery, the
vacuum methods may provide a more rapid and inexpensive approach for confirming
BWA contamination. Dust loading did not affect recovery by the vacuum methods.
•	Commercially-available autonomous (robotic) floor cleaners were evaluated for efficacy
in sampling B. atrophaeus spores (B. anthracis surrogate) from the surface of various
indoor flooring materials (both porous [e.g., carpet] and nonporous [e.g., laminate and
tile]) and from concrete surfaces 91'92. Three vacuum-based robots, one wet vacuum-
based robot, and one wipe-based robot were evaluated (on appropriate surface types, i.e.,
the wet-vacuum and wipe technologies were not evaluated on carpet). Recoveries by the
robot technologies were compared to sponge wipe and vacuum sock methods to calculate
a comparative recovery value for each robot. Generally, the wipe and wet-vacuum-based
robots performed better than the dry vacuum robots on hard nonporous material surfaces.
The dry vacuum-based robots performed as well as or better than a vacuum sock method
35

-------
for spore recovery from carpet. A small but detectable amount of spore reaerosolization
due to operation of the robots was detected.
•	Vacuum socks, mixed cellulose ester (MCE) filter cassettes, polytetrafluoroethylene
(PTFE) filter cassettes, and 3M™ forensic filters were comparatively evaluated for
efficacy in recovery of B. atrophaeus (B. anthracis surrogate) spores from the surface of
concrete, carpet, and upholstery 93. Stainless steel surfaces were also inoculated with
spores and sampled using pre-moistened wipes to act as a control. The MCE filter
cassettes exhibited higher recoveries than the other vacuum-based sampling methods
when sampling spores from concrete and upholstery. Vacuum socks demonstrated the
highest relative recoveries of spores from carpet, but no statistically significant difference
between the methods was determined.
Secondary information and data collected during the literature search from studies focused on
evaluation of wipe and/or sponge-based BWA sampling methodologies (including composite
sampling methodologies) include the following.
•	Stainless steel, vinyl tile, and drywall were contaminated with B. atrophaeus spores.
Cellulose sponges were used to collect wipe samples using one of three composite
sampling approaches. A multiple medium/multiple pass composite sampling method
resulted in the highest recovery of Bacillus spores 94.
•	Two composite-based collection approaches using cellulose sponge samplers were
evaluated for efficacy in recovery of B. atrophaeus (B. anthracis surrogate) spores from
the surface of stainless steel (a CDC-defined method and a modified method) 95. Results
indicated that the composite sampling methods evaluated during the study can increase
the number of surface area samples without increasing laboratory processing time, labor,
or consumable materials. The results also suggest that the CDC method can be used with
fewer passes over a single sampling location without compromising the efficiency of the
method.
•	Recovery of bovine serum albumin (BSA; used as a BWA surrogate) deposited via
aerosol onto various materials (including glass, foliage, and sand) by wipe sampling
(glass) or extraction (foliage and sand) was evaluated 96. Results indicated that, for
retrospective verification of BWA following a contamination incident, cleaner matrices
and horizontal orientation of sampling surfaces provide optimal recoveries.
•	As part of an effort to develop and evaluate a fluorescent viability assay (developed as an
alternative to plate count methods to determine BWA), a swab and syringe/filter assay
sampling approach was developed 97 B. globigii spores were recovered via the
swab/syringe/filter approach from glass with efficiencies between 80% and 90%.
36

-------
3.6. Comprehensive Summary Tables
Table 4 provides a summary of techniques, technologies, and methodologies for decontamination
and sampling of BWAs (including the target BWAs [or simulants of the target BWAs] indicated
in Section 2.1.2) from the USCG vessel-relevant materials listed in Table 1 in Section 2.1.1.2,
based on the information and secondary data collected during the literature search.
Table 4. BWA Decontamination and Sampling Summary (USCG Materials)
Vessels
Material
Information/
Data Area
Bacillus anthracis Ames
Ebola
virus
Other BWAs A
25-foot RB-S, 29-foot RB-S II,
45-foot RB-M, 87-foot Patrol Boat
Aluminum
(hull)
Decontamination
•	MeBr fumigation 42
•	Peracetic acid fog 32>42
•	VHP 32>42
•	Thermal decontamination o8'09
•	Plasma 70
NA
•	pAB 50
•	Citric acid 50
•	Sanihol ST 50
•	CONFLIKT 50
•	PineSol50
•	Thermal decontaminationo8'09
•	Plasma 70
Sampling
NA
NA

Coated steel
(hull)
Decontamination
•	MeBr fumigation 42
•	Gaseous CIO2 42
•	Bleach, diluted bleach 49
•	Peracetic acid, PES-Solid 00
•	Simple Green 49
•	Pipe-Klean 49
NA
•VHP41
•	Gaseous CIO2 41
•	Bleach, diluted bleach 49
•	Simple Green 49
•	Pipe-Klean 49
•	Peracetic acid, PES-Solid60
Sampling
NA
NA
NA
Foam
Decontamination
NA
NA
NA
Sampling
NA
NA
NA
Non-skid coatings
(decking)
Decontamination
• Thermal decontamination o8<09
NA
• Thermal decontamination o8<09
Sampling
NA
NA
NA
Glass
Decontamination
•	Bleach, diluted bleach 72-H58
•	pAB 58
•VHP36
•	Mel fumigation 43
•	Peracetic acid, PES-Solid 00
•	Liquid CIO259
•	Liquid H2O272
•	Ozone 29
•	Wipes 00fil
•	Dichloroisocyanurate 72
•	Autoclave 72
•	EtO 72
•UV72
•	Boiling water 72
•	Spor-Klenz RTU 54
•	Aldehydes 72<54
•	Liquid CIO259
•	Ozone 29
•	Wipes 00>ฐ7
•UV62
•	Peracetic acid, PES-Solid00
•	pAB 50
•	Citric acid 50
•	Sanihol ST 50
•	CONFLIKT 50
•	PineSol50
Sampling
•	Sponge/swab 89>97
•	Wipe sampling %'89
•	Vacuum 89
•	BiSKit89
NA
NA
Insulation, other
bulkhead coverings
Decontamination
• Thermal decontamination o8<09
NA
• Thermal decontamination o8<09
37

-------
Vessels
Material
Information/
Data Area
Bacillus anthracis Ames
Ebola
virus
Other BWAs A

Insulation, other
bulkhead coverings
Sampling
NA
NA
NA

Glazing materials
Decontamination
NA
NA
NA

Glazing materials
Sampling
NA
NA
NA

Sensitive equipment
and components
Decontamination
• Thermal decontamination o8<09
NA
• Thermal decontamination o8<09

Sensitive equipment
Sampling
NA
NA
NA

and components
Sampling
NA
NA
NA
NA - Not applicable; no related information or secondary data were collected during the literature search
A Non-target BWAs (according to Section 2.1.2), e.g., Burkholderiapseudomallei. Refer to reference provided.
Table 5 provides a summary of techniques, technologies, and methodologies identified during
the search for decontamination and sampling of BWAs (including the target BWAs [or simulants
of the target BWAs] indicated in Section 2.1.2) from various other materials (apart from the
USCG vessel-relevant materials identified in Table 1 of Section 2.1.1.2).
Table 5. BWA Decontamination and Sampling Summary (AdditionalMaterials)
Material

Decontamination

Sampling
Noncoated metals
(excl. aluminum; e.g.,
stainless steel,
galvanized metal,
etc.)
•	Mel fumigation30
•	Peracetic acid fog 31
•	VHP 33,45,35,36,37,39
•	Bleach, diluted bleach 72,48,49,52,55,57
•	Peracetic acid, PES-Solid 55>60
•	pAB 47'5ฐ
•	Gaseous CIO237
•	Aldehydes 72
•	Liquid CIO2 48"
•	Liquid H2O2 72>53>55
•	Wipes 00fil
•	Peridox 47'5ฐ
•	CASCAD SDF 47>52>53
•	Dichloroisocyanurate 72
•	Autoclave 72
•	EtO 72
•UV72
•	Boiling water 72
•	Simple Green 49
•	Pipe-Klean 49
•	Virkon52'53
•	Spor-Klenz RTU 53
•	Sponge/swab
94,89,95
•	Wipe sampling
89,93
•	Vacuum 90'89
•	Extraction 90
•	BiSKit89
Tile (e.g., glazed
ceramic, vinyl,
acoustic ceiling, etc.)
•	Mel fumigation 43
•	MeBr fumigation 42
•VHP41
•	Bleach, diluted bleach 72
•	Gaseous CIO2 41'42
•	Liquid H2O272
•	Aldehydes 72
•	Reactive nanoparticles 71
•	L-Gel71
•	UM Nanotech 71
•	Sandia foam 71
•	Ca(C10)2 with surfactant
71
•	GD-5 71
•	Ozone 71<29
•	Dichloroisocyanurate
72
•	Autoclave 72
•	EtO 72
•UV72
•	Boiling water 72
•	Sponge/swab 94
•	Robots 91>92
Plastics (e.g., LDPE,
HDPE, acrylic,
laminate, etc.)
•	MeBr fumigation 42
•	Peracetic acid fog 32>42
•VHP36
•	Bleach, diluted bleach 72>48>49
•	Peracetic acid, PES-Solid 00
•	pAB 50
•	Liquid CIO2 48
•	Thermal
decontamination o8<09
•	Liquid H2O272
•	Wipes 00>ฐ7
•	Peridox 50
•	Dichloroisocyanurate 72
•	Autoclave 72
•	Aldehydes 72
•	EtO 72
•UV72
•	Boiling water 72
•	Simple Green 49
•	Pipe-Klean 49
•	Plasma 70
•	Sponge/swab 89
•	Wipe sampling 89
•	Vacuum 89
•	Robots 91
•	BiSKit89
Coated porous
materials (e.g.,
drywall, concrete,
wood, etc.)
•	Mel fumigation 43
•	Peracetic acid fog 31
•	VHP 36>41
•	Gaseous CIO2 41
•	Reactive nanoparticles 71
•	L-Gel71
•	UM Nanotech 71
•	Sandia foam 71
•	Ca(C10)2 with
surfactant71
•	GD-5 71
•	Ozone 71
•	Wipes 00>ฐ7
• Sponge/swab 94
Coated nonporous
(e.g., coated metals
excl. steel, CARC-
coated aluminum,
etc.)
• Peracetic acid, PES-Solid 00
• Plasma 70

NA
38

-------
Material

Decontamination

Sampling
Noncoated porous
materials (e.g.,
drywall, concrete,
wood, etc.)
•	Mel fumigation 43
•	MeBr fumigation 42
•	Peracetic acid fog 31<32
•	VHP 36>41
•	Bleach, diluted bleach 72<58
•	pAB 47>50>58
•	Gaseous CIO2 41>42
•	Liquid H2O272
•	Reactive nanoparticles 71
•	L-Gel71
•	UM Nanotech 71
•	Sandia foam 71
•	Ca(C10)2 with surfactant
71
•	GD-5 71
•	Ozone 7U9
•	Peridox 47
•	CASCAD SDF 47
•	Dichloroisocyanurate 72
•	Autoclave 72
•	EtO 72
•UV72
•	Boiling water 72
•	Citric acid 50
•	Sanihol ST 50
•	CONFLIKT 50
•	PineSol50
•	Spor-Klenz RTU 58
•	Sponge/swab 89
•	Wipe sampling 89
•	Vacuum 89>93
•	Extraction 90
•	Robots 92
•	BiSKit89
Fabric
•	Peracetic acid fog 32<42
•VHP33
•	Bleach, diluted bleach 72
•	Liquid H2O272
•	Reactive Nanoparticles 71
•	L-Gel 71
•	UM Nanotech 71
•	Sandia foam 71
•	Ca(C10)2 with surfactant
71
•	GD-5 71
•	Ozone 71
•	Dichloroisocyanurate 72
•	Autoclave 72
•	EtO 72
•UV72
•	Boiling water 72
•	Plasma 70
NA
Rubber
• MeBr fumigation 42
• Peracetic acid fog 32<42

NA
Carpet
•	Mel fumigation 43
•	MeBr fumigation 42
•	Peracetic acid fog 31>32>42
•	VHP 36>41
•	pAB 50
•	Gaseous CIO2 41
•	Reactive nanoparticles 71
•	L-Gel71
•	UM Nanotech 71
•	Sandia foam 71
•	Ca(C10)2 with surfactant
71
•	GD-5 71
•	Ozone 7U9
•	Citric acid 50
•	Sanihol ST 50
•	CONFLIKT 50
•	PineSol50
•	Robots 91
•	Vacuum 93
Developmental,
reactor studies, data
review, or similar
•Mel8
•	MeBr 8>43
•	Peracetic acid fog 43
•	VHP 74,73,38,46,8,43
•	Bleach, diluted bleach 8
•	pAB 43
•	Gaseous CIO28'43
•	Liquid CIO28
•	Thermal
decontamination8
•	Liquid H2O28
•	Ozone 8'43
•	EtO 43
•	UV 74<73<8
•	Aldehydes 8>43
NA

• Bleach, diluted bleach 72
• Peridox50
• EtO 72

PPE materials
•	pAB 50
•	Liquid H2O272
•	Dichloroisocyanurate 72
•	Autoclave 72
. UV 72<62
• Boiling water 72
NA
Soil
• Sodium persulfate 28
•UV63
• MeBr28
• Metam Sodium 28
• Extraction 89
Sand, foliage, or
similar
NA
• Wipe sampling 90
Paper
•	Bleach, diluted bleach 72
•	Liquid H2O272
•	Ozone 29
•	Dichloroisocyanurate 72
•	Autoclave 72
•	EtO 72
•UV72
• Boiling water 72
NA
Coatings
• MeBr fumigation 42
•VHP42
• Peracetic acid fog 32<42
• Wipes <",<ฐ7
NA
Biological indicators,
agar plates
• VHP 34>35>40
• UV 61>64>65

NA
NA - Not applicable; no related information or secondary data were collected during the literature search
39

-------
4. KNOWLEDGE/CAPABILITY GAP ASSESSMENT AND RESULTS
As described in Section 2.6, information/secondary data source summaries were collated into the
source compilation document according to the content and primary research focus of the sources.
This arrangement of the literature summaries in the source compilation document served to
illustrate the distribution of the information and secondary data that were collected during the
search across the research focus areas and support identification of gaps in
information/secondary data related to methodologies, procedures, and technologies for
decontamination and sampling of BWA contamination on USCG assets.
On 27 April 2020, project stakeholders from the USCG and the U.S. Department of Homeland
Security (DHS) joined with the EPA and Battelle in a meeting to review and discuss the
information and secondary data collected during the literature search and to identify
data/information/capability gaps related to decontamination and sampling capabilities for BWA
contamination on USCG assets. Prior to the discussion, EPA and Battelle provided expected
attendees with the complete results of the literature search for review (specifically, the source
compilation document was provided, which, as described in Section 2.6, included summaries of
all secondary data and information sources collected during the literature search categorized by
research focus). Meeting discussions focused on identification of knowledge/capability gaps
related to the unique challenges imposed on BWA contamination response and management,
decontamination, and sampling and analysis strategies due to USCG vessel operations and
operational environments and/or vessel construction materials (based on the
information/secondary data collected during the literature search).
The knowledge/capability gaps that were identified and discussed, as well as additional
discussion topics, are provided in Sections 4. 1 and 4.2, respectively.
4.1. Knowledge/Capability Gaps
•	Although outside the scope of the current work, data on efficacy of decontamination and
sampling technologies and methodologies in/on other matrices (not just the select
material surfaces) would be valuable.
o This knowledge may be especially true for air and liquid matrices (i.e., seawater,
bilge water, oil, lubricants, fluids, other vessel areas that may require liquid
sampling, HVAC, and exhaust systems, etc.).
o Liquid matrices can become contaminated during decontamination and/or
sampling efforts, spread contamination or recontaminate surfaces, and/or cause
further penetration into other contaminated materials, areas, and/or surfaces.
•	While some data were collected, collection of additional data (and thorough evaluation of
collected data) related to sampling porous materials would be valuable.
•	The generally smaller amount of data related to decontamination and sampling of BWAs
on coated steel that were collected is notable.
40

-------
o Some coated-steel-related BWA decontamination and material compatibility data
source summaries were presented, but none were focused on evaluation/efficacy
of sampling technologies or methodologies.
o Likely more data have been generated, but such data may be For Official Use
Only [FOUO] and/or unpublished.
•	No data that appear to be related to BWA decontamination on bumper foam were
collected.
o The term "bumper foam" may have been too restrictive. A use of "elastomeric
foam" or "thermoplastic polyurethane coating" may have been a better approach
for the literature search.
o Decontamination and/or sampling data related to bumper foam may have been
collected, though the report/article/data source/etc. does not refer to the material
as "bumper foam".
o Need to consider what the bumper foam material is comprised of to determine if
data related to BWA/CWA decontamination and/or sampling from the specific
composition material(s) have already been collected.
ฆ Determined that the bumper foam material is comprised of a polyurethane
fabric wrapped around a (rounded) interior foam.
o Data related to decontamination and/or sampling of BWAs on other types of
plastic and/or rubber may be applicable/translatable to bumper foam.
•	Additional data on and consideration of the impacts to decontamination and/or sampling
by fouling of/on a surface (i.e., salt/seawater, grime, etc.) would be valuable.
o Need data related to realistic surfaces (most of the collected studies used clean
[i.e., unused, pristine] materials/surfaces).
o A few search terms were included to attempt to collect some data related to
fouling/foulants, and some data related to BWA decontamination and sampling in
the presence of heavy/light organic load were collected 52, but more emphasis
should be placed on this topic/focus.
o Sea/saltwater was discussed as an "interferent" and/or "foulant" impacting
decontamination and sampling efficacy. Other specific foulants of interest should
be identified.
•	Additional data on and consideration of the impacts to decontamination and/or sampling
of environmental conditions (temperature, relative humidity) would be valuable
o Outdoor weather conditions cannot be controlled and may impact
decontamination processes.
•	In addition to vessels, an area of focus to consider would be areas/locations within USCG
bases or stations (and the specific/associated construction materials) that are more
susceptible to frequent contamination and/or recontamination/cross-contamination.
41

-------
o Analogous to subway trains/stations,
o Surfaces/items that are frequently touched/interacted with.
•	Additional data/information related to compatibility of hot/humid air decontamination
with the materials of interest would be valuable.
o Reports/articles on studies performed by other government agencies on efficacy
of hot/humid air decontamination of CWAs may also include data/information on
compatibility of the tested materials with the decontamination process.
o It was discussed that studies have been conducted/data have been generated
related to the operability of CI30 aircraft after hot/humid air decontamination.
•	Other characteristics of BWA decontamination and sampling technologies and
approaches must be considered also, beyond just efficacy of the technologies/approaches.
Such considerations include "scale-up" requirements (for larger areas and/or multiple
assets/vessels, etc.), supply-chain, availability limitations, surge capacities, etc.
•	With regard to sampling, clearly defined direction and/or a guiding framework for
utilization of sampling results is necessary (e.g., sampling results inform and drive
decisions for phase-based contamination response, management, and decontamination
strategies). Alongside this is the need for tools, strategies, and/or guidance for the
development of sampling and analysis plans based on the circumstances, characteristics,
and demands specific to an incident (e.g., location, setting, politics, response phase,
unknowns, etc.).
•	Although outside of scope for this effort, data related to decontamination and sampling of
BWAs on soil and vegetation (foliage, grass, etc.) would be valuable.
4.2. Additional Information, Discussion, and Notes
•	Scientific articles, reports, guidance documents, etc. that are publicly available
from/through the Defense Technical Information Center (DTIC) were collected and
summarized during the literature search and included in the source compilation
document. Anything not publicly available from/through DTIC (i.e., classified,
controlled, limited distribution, etc.) was not collected or included.
•	Quality of the scientific articles, reports, guidance documents, etc. collected during the
literature search was assessed both qualitatively through use of source document type
designations (refer to Section 2.4.1) and quantitatively through use of the Literature
Assessment Factor Rating (refer to Section 2.4.2).
•	The absence of data in the open literature related to decontamination and/or sampling of a
specific contaminant on the surface of a particular material does not necessarily indicate
that such knowledge/data/capabilities do not exist. Similarly, the presence/existence of
data related to decontamination and sampling of specific contaminants on specific
materials in the literature does not indicate that the decontamination and/or sampling
technologies/methodologies are efficacious.
42

-------
•	Although there do appear to be gaps in the data/information that were collected and/or
that exist related to decontamination and sampling of BWAs on the specified USCG
materials, sampling plans and future research can be directed at filling these gaps.
•	In addition to specific technologies for decontamination and sampling of BWAs, the
methods/procedures for use/application of technologies must be considered (i.e.,
decontamination and sampling tactics/strategies in addition to technologies).
4.3. Gap Table
The knowledge/capability gaps described in Section 4.1 and the additional information,
discussion topics, and notes described in Section 4.2 are summarized in Table 6.
Table 6. Gap Table
Knowledge/Capability Gaps
Data on efficacy of decontamination approaches in/on other
matrices (besides the target USCG vessel-related materials).
Limited data were collected related to decontamination of
BWAs on coated steel.
No data were collected related to decontamination of BWAs
on bumper foam.
Additional data on the impacts of foulants and (outdoor)
environmental conditions on the efficacy of
decontamination technologies/approaches would be
valuable.
In addition to vessels, consider areas/locations within USCG
bases or stations that are susceptible to contamination.
Additional data on the efficacy of hot, humid air
decontamination would be valuable.
Consider other characteristics of BWA decontamination
technologies/approaches (e.g., "scale-up" requirements,
supply-chain, availability, surge capacities, etc.).
Data related to decontamination of BWAs on soil and/or
vegetation would be valuable.
Additional Information, Discussion, Notes
Literature and data publicly available through DTIC
were collected. Anything not publicly available through
DTIC was not collected.
Literature and data source quality was assessed both
qualitatively and quantitatively (refer to Section 2.4).
The absence of data in the open literature related to
decontamination of BWA on a particular material does
not necessarily indicate that such knowledge/capability
does not exist. Similarly, the presence/existence of such
data does not indicate that the described
decontamination technology/method is efficacious.
Future research can be directed at filling the
knowledge/capability gaps identified during this effort.
In addition to specific technologies for decontamination
of BWA, methods/procedures for use/application of
decontaminants must be considered.
43

-------
Knowledge/Capability Gaps
Additional Information, Discussion, Notes
•	Data on efficacy of sampling approaches in/on other
matrices (besides the target USCG vessel-related materials).
•	Need additional data related to sampling from porous
materials.
•	Limited data were collected related to sampling of BWAs
on coated steel.
•	No data were collected related to sampling of BWAs on
bumper foam.
•	Additional data on the impacts of foulants on the efficacy of
sampling technologies/approaches would be valuable.
•	In addition to vessels, consider areas/locations within USCG
bases or stations that are susceptible to contamination.
•	Consider other characteristics of BWA sampling
technologies/approaches (e.g., "scale-up" requirements,
supply-chain, availability, surge capacities, etc.).
•	Clearly defined direction and/or a guiding framework for
utilization of sampling results is necessary.
•	Data related to sampling of BWAs on soil and/or vegetation
would be valuable.
•	Literature and data publicly available through DTIC
were collected. Anything not publicly available through
DTIC was not collected.
•	Literature and data source quality was assessed both
qualitatively and quantitatively (refer to Section 2.4).
•	The absence of data in the open literature related to
sampling of BWA on a particular material does not
necessarily indicate that such knowledge/capability does
not exist. Similarly, the presence/existence of such data
does not indicate that the described sampling
technology/method is efficacious.
•	Future research can be directed at filling the
knowledge/capability gaps identified during this effort.
•	In addition to specific technologies for BWA sampling,
methods/procedures for use/application of sampling
technologies must be considered.
44

-------
5. REFERENCES
1	U.S. Environmental Protection Agency. 2001. EPA Requirements for Quality Assurance
Project Plans, Office of Environmental Information: Washington, DC. EPA/240/B-01/003.
2	Coast Guard Assets, United States Coast Guard, U.S. Department of Homeland Security.
(2019, April 26). Retrieved from https://www.uscg.mil/Assets/?Page=2. accessed 11
November 2020.
3	Coast Guard Assets, United States Coast Guard, U.S. Department of Homeland Security.
(2019, April 24). Retrieved from https://www.uscg.mil/Assets/?Page=3. accessed 11
November 2020.
4	Read, T.D.; Peterson, S.N.; Tourasse, N.; Baillie, L.W.; Paulsen, I.T.; Nelson, K.E.; Tettelin,
H.; Fouts, D.E.; Eisen, J.A.; Gill, S.R.; Holtzapple, E.K.; Okstad, O.A.; Helgason, E.;
Rilstone, J.; Wu, M.; Kolonay, J.F.; Beanan, M.J.; Dodson, R.J.; Brinkac, R.M.; Gwinn, M.;
DeBoy, R.T.; Madpu, R.; Daugherty, S.C.; Durkin, A.S.; Haft, D.H.; Nelson, W.C.; Peterson,
J.D.; Pop, M.; Kouri, H.M.; Radune, D.; Benton, J.L.; Mahamoud, Y.; Jiang, L.; Hance, J.R.;
Weidman, J.F.; Berry, K.J.; Plaut, R.D.; Wolf, A.m.; Watkins, K.L.; Nierman, W.C.; Hazen,
A.; Cline, R.; Redmond, C.; Thwaite, J.E.; White, O.; Salzberg, S.L.; Thomason, B.;
Friedlander, A.M.; Koehler, T.M.; Hanna, P.C.; Kolst0, A.-B.; Fraser, C.M. The genome
sequence of Bacillus anthracis Ames and comparison to closely related bacteria. Nature.
2003. 423: 81-86, doi:10.1038/nature01586.
5	Ebola virus disease, World Health Organization. (2020, February 10). Retrieved from
https://www.who.int/news-room/fact-sheets/detail/ebola-virus-disease. accessed 11
November 2020.
6	Farchaus, J.W.; Ribot, W.J.; Downs, M.B.; Ezzell, J.W. Purification and characterization of the
major surface array protein from the avirulent Bacillus anthracis delta Sterne-1. Journal of
Bacteriology. 1995. 177(9): 2481-2489.
7	Dewan, P.K.; Fry, A.M.; Laserson, K.; Tierney, B.C.; Quinn, C.P.; Hayslett, J.A., Broyles,
L.N.; Shane, A.; Winthrop, K.L.; Walks, I.; Siegel, L.; Hales, T.; Semenova, V.A.; Romero-
Steiner, S.; Elie, C.; Khabbaz, R.; Khan, A.S.; Hajjeh, R.A., Schuchat, A.; and members of
the Washington, D.C., Anthrax Response Team. Inhalational anthrax outbreak among postal
workers, Washington, D.C., 2001. Emerging Infectious Diseases. 2002. 8(10): 1066-1072.
8	Wood, J.P.; Adrion, A.C. Review of decontamination techniques for the inactivation of
Bacillus anthracis and other spore-forming bacteria associated with building or outdoor
materials. Environmental Science & Technology. 2019. 53: 4045-4062.
9	Kournikakis, B.; Ho, J.; Duncan, S. Anthrax letters: personal exposure, building contamination,
and effectiveness of immediate mitigation measures. Journal of Occupational and
Environmental Hygiene. 2010. 7: 71-79.
10	Best, M.; McLennan, S.; Callaghan, M.; Raymond, D. Investigation on the dispersal patterns
of contaminants in letters. AppliedBiosafety. 2003, 8(2): 50-52.
11	Kapuscinski, R.B.; Mitchell, R. Processes controlling virus inactivation in coastal waters.
Water Research. 1980, 14: 363-371.
45

-------
12	Szinicz, L. History of chemical and biological warfare agents. Toxicology. 2005, 214: 167-
181.
13	Artenstein, A.W. Chapter 79 - Biological Attack in Ciottone, G.; Biddinger, P.D.; Darling,
R.G.; Fares, S.; Keim, M.E.; Molloy, M.S.; Suner, S. Ciottone's Disaster Medicine (Second
Edition) (480-488), 2016, Elsevier.
14	Pile, J.C.; Malone, J.D.; Eitzen, E.M.; Friedlander, A.M. Anthrax as a potential biological
warfare agent. Archives of Internal Medicine. 1998. 158; 429-434.
15	Hendricks, K.A.; Wright, M.E.; Shadomy, S.V.; Bradely, J.S.; Morrow, M.G.; Pavia, A.T.;
Rubenstein, E.; Holty, J.C.; Messonnier, N.E.; Smith, T.L.; Pesik, N.; Treadwell, T.A.;
Bower, W. A. Centers for Disease Control and Prevention Expert Panel Meetings on
Prevention and Treatment of Anthrax in Adults. Emerging Infectious Diseases. 2014. 20(2):
el30687. doi:10.3201/eid2002.130687.
16	Chugh, T. Bioterrorism: Clinical and public health aspects of anthrax. Current Medicine
Research and Practice. 2019. 9:110-111.
17	Abramova, F.A.; Grinberg, L.M.; Yampolskaya, O.V.; Walker, D.H. Pathology of inhalational
anthrax in 42 cases from the Sverdlovsk outbreak of 1979. Proceedings of the National
Academy of Sciences of the United States of America. 1993. 90: 2291-2294.
18	Savransky, V.; Ionin, B.; Reece, J. Current status and trends in prophylaxis and management
of anthrax disease. Pathogens. 2020. 9(5): 370. doi: 10.3390/pathogens9050370.
19	Fitch, J. P.; Raber, E.; Imbro, D.R. Technology challenges in responding to biological or
chemical attacks in the civilian sector. Science. 2003. 302: 1350-1354.
20	Fusco, F.M.; Puro, V.; Ippolito, G. Considerations regarding safe transport and transfer for
Ebola patients in western countries. Intensive Care Medicine. 2015. 41: 1175-1176.
21	Martin, D.; Howard, J.; Agarwal, B.; Rajalingam, Y.; Athan, B.; Bhagani, S.; Cropley, I.;
Hopkins, S.; Mepham, S.; Rodger, A.; Warren, S.; Jacobs, M. Ebola virus disease: the UK
critical care perspective. British Journal of Anaesthesia. 2016. 116(5): 590-596.
22	Diffuse Security Threats - USPS Air Filtration Systems Need More Testing and Cost Benefit
Analysis before Implementation, GAO-02-838; United States General Accounting Office;
Washington, D.C., August 2002.
23	Zhang, Y.; Gong, Y.; Wang, C.; Liu, W.; Wang, Z.; Xia, Z.; Bu, Z.; Lu, H.; Sun, Y.; Zhang,
X.; Cao, Y.; Yang, F.; Su, H.; Hu, Y.; Deng, Y.; Zhou, B.; Zhao, Z.; Fu, Y.; Kargbo, D.;
Dafae, F.; Kargbo, B.; Kanu, A.; Liu, L.; Qian, J.; Guo, Z. Rapid deployment of a mobile
biosafety level-3 laboratory in Sierra Leone during the 2014 Ebola virus epidemic. PLoS
Neglected Tropical Diseases. 2017. 11(5): 1-15.
24	Wein, L. M.; Liu, Y.; Leighton, T. J. HEP A/vaccine plan for indoor anthrax remediation.
Emerging Infectious Diseases. 2005. 11(1): 69-76.
25	Hong, T.; Gurian, P.L.; Huang, Y.; Haas, C.N. Prioritizing risks and uncertainties from
intentional release of selected category A pathogens. PLoS ONE. 2012. 7(3): 1-19.
26	Hong, T.; Gurian, P.L.; Dudley Ward, N.F. Setting risk-informed environmental standards for
Bacillus anthracis spores. Risk Analysis. 2010. 30(10): 1602-1622.
46

-------
27	Compilation of Available Data on Building Decontamination Alternatives; U.S. Environmental
Protection Agency, Office of Research and Development, National Homeland Security
Research Center; Washington, D.C., March 2005. EPA/600/R-05/036.
28	EPA Technical Brief- Surface Decontamination Methodologies for a Wide-Area Bacillus
anthracis Incident, U.S. Environmental Protection Agency, Office of Research and
Development, National Homeland Security Research Center; Research Triangle Park, NC,
2015. EPA/600/S-15/172.
29	Aydogan, A.; Gurol, M.D. Application for gaseous ozone for inactivation of Bacillus subtilis
spores. Journal of the Air & Waste Management Association. 2006. 56: 179-185.
30	Sutton, M.; Kane, S.R.; Wollard, J.R. Methyl Iodide Fumigation of Bacillus anthracis Spores.
LLNL-JRNL-648514. Lawrence Livermore National Laboratory. January 2014.
31	Wood, J.P.; Calfee, M.W.; Clayton, M.; Griffin-Gatchalian, N.; Touati, A.; Egler, K.
Evaluation of peracetic acid fog for the inactivation of Bacillus anthracis spore surrogates in
a large decontamination chamber. Journal of Hazardous Materials. 2013. 250-251: 61-67.
32	Richter, W.R.; Wood, J.P.; Wendling, M.Q.S.; Rogers, J.V. Inactivation of Bacillus anthracis
spores to decontaminate subway railcar and related materials via the fogging of peracetic
acid and hydrogen peroxide sporicidal liquids. Journal of Environmental Management. 2018.
206: 800-806.
33	Lemmen, S.; Scheithauer, S.; Hafner, H.; Yezli, S.; Mohr, M.; Otter, J.A. Evaluation of
hydrogen peroxide vapor for the inactivation of nosocomial pathogens on porous and
nonporous surfaces. American Journal of Infection Control. 2015. 43(1): 82-85.
34	Petit, B.M.; Almeida, F.C.; Uchiyama, T.R.; Lopes, F.O.C.; Tino, K.H.; Chewins, J.
Evaluating the efficacy of hydrogen peroxide vapour against foot-and-mouth disease virus
within aBSL4 biosafety facility. Letters in Applied Microbiology. 2017. 65: 281-284.
35	Fu, T.Y.; Gent, P.; Kumar, V. Efficacy, efficiency, and safety aspects of hydrogen peroxide
vapour and aerosolized hydrogen peroxide room disinfection systems. Journal of Hospital
Infection. 2012. 80: 199-205.
36	Rogers, J.V.; Sabourin, C.L.K.; Choi, Y.W.; Richter, W.R.; Rudnicki, D.C.; Riggs, K.B.;
Taylor, M.L.; Chang, J. Decontamination assessment of Bacillus anthracis, Bacillus subtilis,
and Geobacillus stearothermophilus spores on indoor surfaces using a hydrogen peroxide gas
generator. Journal of Applied Microbiology. 2005. 99: 739-748.
37	Pottage, T.; Macken, S.; Giri, K.; Walker, J.T.; Bennett, A.M. Low-temperature
decontamination with hydrogen peroxide or chlorine dioxide for space applications. Applied
and Environmental Microbiology. 2012. 78(12): 4169-4174.
38	Meyer, K.M.; Calfee, M.W.; Wood, J.P.; Mickelsen, L.; Attwood, B.; Clayton, M.; Touati, A.;
Delafield, R. Fumigation of a laboratory-scale HVAC system with hydrogen peroxide for
decontamination following a biological contamination incident. Journal of Applied
Microbiology. 2013. 116: 533-541.
39	Goyal, S.M.; Chander, Y.; Yezli, S.; Otter, J. A. Evaluating the virucidal efficacy of hydrogen
peroxide vapour. Journal of Hospital Infection. 2014. 86: 255-259.
47

-------
40	McAnoy, A.M.; Sait, M.; Pantelidis, S. Establishment of a Vaporous Hydrogen Peroxide Bio-
Decontamination Capability. DSTO-TR-1994. Human Protection Performance Division,
DSTO, Fishermans Bend, Victoria 3207, Australia, 2007.
41	Rastogi, V.K.; Wallace, L.; Smith, L.S.; Ryan, S.P.; Martin, B. Quantitative method to
determine sporicidal decontamination of building surfaces by gaseous fumigants, and issues
related to laboratory-scale studies. Applied and Environmental Microbiology. 2009. 75(11):
3688-3694.
42	EPA Technical Brief- Decontamination Options for Restoring Subway Systems following a
Bacillus anthracis Contamination Incident; U.S. Environmental Protection Agency, Office of
Research and Development, National Homeland Security Research Center; Research
Triangle Park, NC, 2018. EPA/600/S-18/286.
43	EPA Technical Brief- Summary of the Effectiveness of Volumetric Decontamination Methods
as a Function of Operational Conditions. U.S. Environmental Protection Agency, Office of
Research and Development, National Homeland Security Research Center; Research
Triangle Park, NC, 2015. EPA/600/S-15/190;
44	McAnoy, A.M. Vaporous Decontamination Methods: Potential Uses and Research Priorities
for Chemical and Biological Contamination Control. DSTO-GD-0465, Human Protection
and Performance Division, DSTO Defence Science and Technology Organisation, 2006.
45	Baron, P.A.; Estill, C. F.; Beard, J.K.; Hein, M.J.; Larsen, L. Bacterial endospore inactivation
caused by outgassing of vapourous hydrogen peroxide from polymethyl methacrylate
(Plexiglasฎ). The Society for Applied Microbiology, Letters in Applied Microbiology. 2007.
45: 485-490.
46	McCord, J.; Prewitt, M.; Dyakova, E.; Mookerjee, S.; Otter, J.A. Reduction in Clostridium
difficile infection associated with the introduction of hydrogen peroxide vapour automated
room disinfection. Journal of Hospital Infection. 2016. 94: 158-187.
47	Edmonds, J.M.; Rastogi, V.K. Decontamination Efficacy of Three Commercial Off-The-Shelf
Sporicidal Agents on Medium-Sized Panels Contaminated with Surrogates of Bacillus
Anthracis. ECBC-TR-939. Edgewood Chemical Biological Center, U.S. Army Research,
Development, and Engineering Command, Aberdeen Proving Ground, MD. April 2012.
48	Chatuev, B.M.; Peterson, J.W. Analysis of the sporicidal activity of chlorine dioxide
disinfectant against Bacillus anthracis (Sterne strain). Journal of Hospital Infection. 2010.
74: 178-183.
49	Chattopadhyay, S. Adhesion and decontamination of biological contaminants in drinking
water distribution systems. Exposure and Health. 2016. 8: 199-210.
50	Calfee, M.W.; Wendling, M. Inactivation of Burkholderiapseudomallei on environmental
surfaces using spray-applied common liquid disinfectant. Applied Microbiology. 2015. 61:
418-422.
51	Calfee, M.W.; Wendling, M. Inactivation of vegetative bacteria threat agents on environmental
surfaces. Science of the Total Environment. 2013. 443: 387-396.
48

-------
52	Guan, J.; Chan, M.; Brooks, B.W.; Rohonczy, L. Influence of temperature and organic load on
chemical disinfection of Geobacillus stearothermophilus spores, a surrogate for Bacillus
anthracis. The Canadian Journal of Veterinary Research. 2013. 77: 100-104.
53	Amoako, K.K.; Santiago-Mateo, K.; Shields, M. J.; Rohonczy, E. Bacillus anthracis spore
decontamination in food grease. Journal of Food Protection. 2013. 76(4): 669-701.
54	Tomasino, S. F.; Pines, R. M.; Cottrill, M. P.; Hamilton, M.A. Determining the efficacy of
liquid sporicides against spores of Bacillus subtilis on a hard nonporous surface using the
quantitative Three Step Method: collaborative study. Journal of AO AC International. 2008.
91(4): 833-852.
55	Majcher, M.R.; Bernard, K.A.; Sattar, S.A. Identification by quantitative carrier test of
surrogate spore-forming bacteria to assess sporicidal chemicals for use against Bacillus
anthracis. Applied and Environmental Microbiology. 2008. 74(3): 676-681.
56	Rastogi, V.K.; Smith, L.S.; Wallace, L.; Shah, S.S.; Sabol, J. Laboratory-Scale Study in
Determining the Decontamination Standards for Personnel Protective Equipment Used by
Homeland Defense Personnel: Evaluation of Commercial Off-the-Shelf Technologies for
Decontamination of Personnel Protective Equipment-Relevant Surfaces. ECBC-TR-631.
Edgewood Chemical Biological Center, U.S. Army Research, Development, and Engineering
Command, Aberdeen Proving Ground, MD. June 2008.
57	Gallandat, K.; Lantagne, D. Selection of a biosafety level 1 (BSL-1) surrogate to evaluate
surface disinfection efficacy in Ebola outbreaks: Comparison of four bacteriophages. PLoS
ONE. 2017. 12(5): 1-10.
58	EPA Technical Brief- Effectiveness of Outdoor Environment Decontamination for Biological
Agents; U.S. Environmental Protection Agency, 2018. Office of Research and Development,
National Homeland Security Research Center; Research Triangle Park, NC. EPA/600/S-
18/223.
59	Buhr, T.L.; Young, A.A.; Minter, Z.A.; Wells, C.M.; Shegogue, D.A. Decontamination of a
hard surface contaminated with Bacillus anthracis A Sterne and B. anthracis Ames spores
using electrochemically generated liquid-phase chlorine dioxide (eClCh). Journal of Applied
Microbiology. 2011. Ill: 1057-1064.
60	Buhr, T.L.; Wells, C.M.; Young, A.A.; Minter, Z.A.; Johnson, C.A.; Payne, A.N.; McPherson,
D.C. Decontamination of materials contaminated with Bacillus anthracis and Bacillus
thuringiensis A1 Hakam spores using PES-Solid, a solid source of peracetic acid. Journal of
Applied Microbiology. 2013. 115: 398-408.
61	Menetrez, M.Y.; Foarde, K.K.; Webber, T.D.; Dean, T.R.; Betancourt, D.A. Efficacy of UV
irradiation on eight species of Bacillus. Journal of Environmental Engineering and Science.
2006. 5: 329-334.
62	Jinadatha, C.; Simmons, S.; Dale, C.; Ganachari-Mallappa, N.; Villamaria, F.C.; Goulding, N.;
Tanner, B.; Stachowiak, J.; Stibich, M. Disinfecting personal protective equipment with
pulsed xenon ultraviolet as a risk mitigation strategy for health care workers. American
Journal of Infection Control. 2015. 43: 412-414.
49

-------
63	Wood, J.P.; Meyer, K.M.; Kelly, T.J.; Choi, Y.W.; Rogers, J.V.; Riggs, K.B.; Willenberg, Z.J.
Environmental persistence of Bacillus anthracis and Bacillus subtilis spores. PLoS One.
2015. 10(9): 1-17.
64	Katara, G.; Hemvani N.; Chitnis, S.; Chitnis, V.; Chitnis, DS. Surface disinfection by exposure
to germicidal UV light. Indian Journal of Medical Microbiology. 2008. 26(3): 241-242.
65	Bache, S.E.; Maclean, M.; Gettinby, G.; Anderson, J.G.; Macgregor, S.J.; Taggart, I. Universal
decontamination of hospital surfaces in an occupied inpatient room with a continuous 405
nm light source. Journal of Hospital Infection. 2018. 98: 67-73.
66	Meyer, K.M.; Tufts, J.A.; Calfee, M.W.; Oudejans, L. Efficacy of sporicidal wipes for
inactivation of a Bacillus anthracis surrogate. Journal of Applied Microbiology. 2014. 117:
1634-1644.
67	EPA Technical Brief- Effectiveness of Disinfecting and Sporicidal Wipes against Bacillus
atrophaeus, a Bacillus anthracis Surrogate; U.S. Environmental Protection Agency, 2017.
Office of Research and Development, National Homeland Security Research Center;
Research Triangle Park, NC. EPA/600/S-17/182.
68	Buhr, T.L.; Young, A.A.; Barnette, H.K.; Minter, Z.A.; Kennihan, N.L.; Johnson, C.A.;
Bohmke, M.D.; DePaola, M.; Cora-Lao, M.; Page, M.A. Test methods and response surface
models for hot, humid air decontamination of materials contaminated with dirty spores of
Bacillus anthracis A Sterne and Bacillus thuringiensis A1 Hakam. Journal of Applied
Microbiology. 2015. 119: 1263-1277.
69	Prokop, E.J.; Crigler, J.R.; Wells, C.M.; Young, A. A.; Buhr, T.L. Response surface modeling
for hot, humid air decontamination of materials contaminated with Bacillus anthracis A
Sterne and Bacillus thuringiensis A1 Hakam spores. AMB Express. 2014. 4(21): 1-15.
70	Moeller, T.M.; Luna, M.L.; Irving, P.M. Portable Nonequilibrium Plasma Surface
Decontamination System. IEEE conference record, 2001. doi: 10.1109/PPPS.2001.961003.
71	O'Connor, L.E.; Harper, B.; Larsen, L. A Comparison of Decontamination Technologies for
Biological Agents on Selected Commercial Surface Materials. U.S. Army Soldier and
Biological Chemical Command, Aberdeen Proving Ground, MD 21010. April 2001.
72	Kenar, L.; Ortatatli, M.; Yaren, H.; Karayilanoglu, T.; Aydogant, H. Comparative Sporicidal
Effects of Disinfectants after Release of a Biological Agent. Military Medicine. 2007. 172,
pp 616-621.
73	Rutala, W.A.; Weber, D.J. Disinfectants used for environmental disinfection and new room
decontamination technology. American Journal of Infection Control. 2013. 41: S36-S41.
74	Weber, D.; Rutala, W.; Anderson, D.; Chen, L.; Sickbert-Bennett, E.; Boyce, J. Effectiveness
of ultraviolet devices and hydrogen peroxide systems for terminal room decontamination:
Focus on clinical trials. American Journal of Infection Control. 2016. 44: e77-e84.
75	Brickhouse, M.D.; Lalain, T.; Bartram, P.W.; Hall, M.; Hess, Z.; Reiff, L.; Mantooth, B.;
Zander, Z.; Stark, D.; Humphreys, P.; Williams, B.; Ryan, S.; Martin, B. Effects of Vaporized
Decontamination Systems on Selected Building Interior Materials: Vaporized Hydrogen
Peroxide; ECBC-TR-661; Edgewood Chemical Biological Center, U.S. Army Research,
Development, and Engineering Command, Aberdeen Proving Ground, MD, January 2009.
50

-------
76	Corsi, R.L.; Walker, M.B.; Liljestrand, H.M.; Hubbard, H.F.; Poppendieck, D.G. Methyl
bromide as a building disinfectant: Interaction with indoor materials and resulting byproduct
formation. Journal of the Air & Waste Management Association. 2007. 57(5): 576-585.
77	EPA Technical Brief- Assessment of the Impact of Decontamination Fumigants on Electronic
Equipment; U.S. Environmental Protection Agency, 2014. , Office of Research and
Development, National Homeland Security Research Center; Research Triangle Park, NC.
EPA/600/R-14/316.
78	EPA Technical Brief- Decontamination Options for Sensitive Equipment in Critical
Infrastructure following a Bacillus anthracis Incident, U.S. Environmental Protection
Agency, 2017. Office of Research and Development, National Homeland Security Research
Center; Research Triangle Park, NC. EPA/600/S-17/166.
79	Compatibility of Material and Electronic Equipment with Chlorine Dioxide Fumigation; U.S.
Environmental Protection Agency, August 2010. Office of Research and Development,
National Homeland Security Research Center; Washington, D.C. EPA/600/R-10/037.
80	Compatibility of Material and Electronic Equipment with Methyl Bromide and Chlorine
Dioxide Fumigation; U.S. Environmental Protection Agency, October 2012. Office of
Research and Development, National Homeland Security Research Center; Research
Triangle Park, NC. EPA/600/R-12/664.
81	Effects of Vaporized Decontamination Systems of Selected Building Interior Materials:
Chlorine Dioxide; U.S. Environmental Protection Agency, April 2008. Office of Research
and Development, National Homeland Security Research Center; Research Triangle Park,
NC. EPA/600/R-08/054.
82	Adrion, A.C.; Scheffrahn, R.H.; Serre, S.; Lee, S.D. Impact of sporicidal fumigation with
methyl bromide or methyl iodide on electronic equipment. Journal of Environmental
Management. 2019. 231: 1021-1027.
83	Material Compatibility for Historic Items Decontaminated with Gamma Irradiation; U. S.
Environmental Protection Agency, February 2017. , Office of Research and Development,
National Homeland Security Research Center; Washington, D.C. EPA/600/R-16/264.
84	Material Demand Studies: Interaction of Chlorine Dioxide Gas With Building Materials, U.S.
Environmental Protection Agency, September 2008. Office of Research and Development,
National Homeland Security Research Center; Research Triangle Park, NC. EPA/600/R-
08/091.
85	Solazzo, C.; Erhardt, D.; Marte, F.; Von Endt, D.; Tumosa, C. Effects of chemical and
biological warfare remediation agents on the materials of museum objects. Applied Physics
A. 2004. 79: 247-252.
86	Procell, L.R.; Hess, Z.A.; Gehring, D.G.; Lyann, J.T.; Bartram, P.W.; Lalain, T.; Ryan, S.;
Attwood, B.; Martin, B.; Brickhouse, M.D. Material Demand Studies: Materials Sorption of
Vaporized Hydrogen Peroxide. ECBC-TR-778. Edgewood Chemical Biological Center, U.S.
Army Research, Development, and Engineering Command, Aberdeen Proving Ground, MD.
June 2010.
87	Silvestri, E.E.; Yund, C.; Taft, S.; Bowling, C.Y.; Chappie, D.; Garrahan, K.; Brady-Roberts,
E.; Stone, H.; Nichols, T.L. Considerations for estimating microbial environmental data
51

-------
concentrations collected from a field setting. Journal of Exposure Science and Environmental
Epidemiology. 2017, 27: 141-151.
88	Calfee, M.W.; Tufts, J.; Meyer, K.; McConkey, K.; Mickelsen, L.; Rose, L.; Dowell, C.;
Delaney, L.; Weber, A.; Morse, S.; Chaitram, J.; Gray, M. Evaluation of standardized sample
collection, packaging, and decontamination procedures to assess cross-contamination
potential during Bacillus anthracis incident response operations. Journal of Occupational
and Environmental Hygiene. 2016. 13(12): 980-992.
89	Herzog, A.B.; McLennan, S.D.; Pandey, A.K.; Gerba, C.P.; Hass, C.N.; Rose, J.B.; Hashsham,
S.A. Implications of limits of detection of various methods for Bacillus anthracis in
computing risks to human health. Applied and Environmental Microbiology. 2009. 75(1):
6331-6339.
90	Calfee, M.W.; Rose, L.J.; Tufts, J.; Morse, S.; Clayton, M.; Abderrahmane, T.; Griffin-
Gatchalian, N.; Slone, C.; McSweeny, N. Evaluation of sampling methods for Bacillus spore-
contaminated HVAC filters. Journal of Microbiological Methods. 2014. 96: 1-5.
91	Evaluation of Surface Sampling for Bacillus Spores Using Commercially Available Cleaning
Robots; U.S. Environmental Protection Agency, Office of Research and Development,
National Homeland Security Research Center; Research Triangle Park, NC, June 2013. EPA
600/R-13/100;
92	EPA Technical Brief - Sampling Procedures Using Commercially Available Robotic Floor
Cleaners for Bacillus anthracis Spores; U.S. Environmental Protection Agency, National
Homeland Security Research Center; Research Triangle Park 2017. EPA/600/S-17/261.
93	Calfee, M.W.; Rose, L.; Morse, S.; Mattorano, D.; Clayton, M.; Touati, A.; Griffin-Gatchalian,
N.; Slone, C.; McSweeney, N. Comparative evaluation of vacuum-based surface sampling
devices for collection of Bacillus spores. Journal of Microbiological Methods. 2013. 95(3):
389-396.
94	Hess, B.M.; Amidan, B.G.; Anderson, K.K.; Hutchison, J.R. Evaluating composite sampling
methods of Bacillus spores at low concentrations. PLoS One. 2016. 11(10): 1-22.
95	Tufts, J. A.M.; Meyer, K.M.; Calfee, M.W.; Lee, S.D. Composite sampling of a Bacillus
anthracis surrogate with cellulose sponge surface samplers. PLoS One. 2014. 9(12): 1-16.
96	Rajoria, S.; Kumar, R.B.; Gupta, P.; Alam, S.I. Postexposure recovery and analysis of
biological agent in a simulated biothreat scenario using tandem mass spectrometry.
Analytical Chemistry. 2017. 89: 4062-4070.
97	Tassell, R.L.V. Monitoring the decontamination of bacterial spores using fluorescent viability
assays. 45512.1-CH. U.S. Army Research Office. Durham, NC. September 12, 2002.
52

-------
vvEPA
United States
Environmental Protection
Agency
PRESORTED STANDARD
POSTAGE & FEES PAID
EPA
PERMIT NO. G-35
Office of Research and Development (8101R)
Washington, DC 20460
Official Business
Penalty for Private Use
$300

-------