Advances in decontamination technologies and strategies were tested to help improve response to biological incidents. The Biological Operational Test and Evaluation Project tested three decontamination methods - vaporized hydrogen peroxide, bleach, and chlorine dioxide fumigation. Chlorine dioxide was most effective at reducing spore levels, while bleach produced the most waste. Overall costs were dominated by waste management. The methyl bromide fumigation study demonstrated the feasibility of using this technology for decontamination of structures. Efforts are also underway to develop strategies to safely restore contaminated underground transportation systems like subways following an incident.

1. Advances and Lessons
Learned in Decontamination
U.S. EPA
Erica Canzler
For Official Use Only

2. Outline
• CMAD Background
• Advances in Decontamination since Capitol Hill
• BOTE
• Methyl Bromide
• Strategies and Initiatives Moving Forward
• Underground Transportation Restoration Project

3. US EPA – Special Team for CBRN
CBRN Consequence Management Advisory Team (CMAT)
Mission: provides scientific and technical expertise
for all phases of CBRN consequence management
and is available to support the On-Scene
Coordinators (OSC) 24/7
Focus: Operational preparedness for CBRN agents.
Maintain ASPECT and PHILIS
Buildings, infrastructure, indoor and outdoor environments, transportation sectors
Support: All phases of CBRN response,
including characterization,
decontamination, clearance and waste
management

4. 4
Renovation
Reoccupation
decision
Long-term
environmental
and public health
monitoring
Clearance
environmental
sampling
and analysis
Clearance
decision
Decontamination
strategy
Remediation Action Plan
Worker health and safety
Site preparation
Source reduction
Waste disposal
Decontamination of sites
or items
Decontamination
verification
characterization of
biological agent
Characterization of
affected site
Site containment
Continue risk
communication
Characterization
environmental sampling
and analysis
Initial risk assessment
Clearance goals
Initial threat assessment
HAZMAT and emergency
actions
Forensic investigation
Public health actions
Screening sampling
Determination of agent
type, concentration, and
viability
Risk communication
Receive
information on
biological
Incident
Identification of
suspect release
sites
Notification of
appropriate
agencies
ClearanceDecontaminationCharacterization
Restoration
(Reoccupancy)
Remediation/CleanupFirst ResponseNotification
(CONSEQUENCE MANAGEMENT)(CRISIS MANAGEMENT)
RESPONSE AND RECOVERY ACTIVITIES
IBRD SCOPE
Response and Recovery
Actions

5. Recent Examples Of Support to Bio Incidents
• Multiple Ricin Responses (Wisconsin, Oklahoma, Mississippi)
• Provided onsite technical support
• Developed responder training
• Ebola
• Developed operational guidance on decon and health & safety
• Trained over 200 responders in procedures
• DoD Anthrax Lab Contamination
• Provided technical support to CDC and State & Local Labs
• Coordinated issuance of expedited crisis exemptions
• Burkholderia pseudomallei
• Onsite technical support; led development of all sampling and decon strategies and
plans

6. Examples of Past Projects
• Biological Operational Test and Evaluation (BOTE)
• Phase 1 Report has been released
• Development of Ricin Training
• NYC CM Plan
• Non Traditional Threat Agents Efforts
• EPA/CDC Interim Clearance Guidance for Anthrax
• Post Fukushima Assistance in Japan
• WARRP/IBRD
For more info, CMAT Annual Reports can be found on our webpage at
http://www2.epa.gov/emergency-response/cmat
6

7. R&D Highlights
• Decon Efficacy Studies (bleach, vaporized hydrogen peroxide, fumigants)
• Material Compatibility (ex. ethylene oxide with electronics)
• Development of RV-PCR
• Robotic Sampling Devices
• Decon Selection Tool
• Waste Management Tool
• Personnel Decon Line Protocol

8. For Official Use Only – Do Not Distribute
Scenario for gap analysis based on National
Planning Scenario #2
Problem:
• Two surreptitious releases of Bacillus
anthracis spores in downtown Seattle and
at Fort Lewis (can be any urban area, but
Seattle is IBRD test city)
Initial conditions:
• BioWatch positives for Bacillus anthracis
• Confirmatory tests have been made
• Emergency response has been activated
and is underway
• Mass prophylaxis distribution has begun
• Hospitals are in surge mode and
overwhelmed with sick and worried well
• People who were contaminated live in other
communities as well
• Some contaminant has been tracked into
surrounding areas
Area for restoration is on the order of tens of square miles,
including hundreds of contaminated facilities.

9. For Official Use Only – Do Not Distribute
Time-frames and Costs of Fumigation
Site Cleanup
timeframe
Cost of
remediation
Comments
Capitol Hill 3 mos. $28M total;
$2M Daschle
Intense pressure from Hill
to re-open (24/7
response)
Brentwood 23 mos. >$200
million for
both
cleanups
Actual remediation: 9
mos.
Trenton 28 mos. Actual remediation: 6
mos.
AMI Building >39 mos. <$5 million Actual remediation: 8
mos.
DOJ mail room 4.5 mos. ~$500,000 Fumigation only for mail
machines in tent
GSA Bldg 410 >29 mos. $4 million Actual remediation: 9
mos.
DOS SA-32 26 mos. ~$9 million Actual remediation: 12
mos.

10. Objectives
• Decrease clean up time
• Decrease clean up cost
• Protect public health
• Focus on chokepoints – lab and decon capacity and capability
• How does intensive MCM campaign impact/alter decontamination and
clearance strategy?

11. Lab Capacity
 Establishment of Bio Laboratory Capability &Capacity
 RV-PCR capability
 EPA Microbiology Lab at Ft. Meade, MD
 The NEIC Lab in Denver, CO
 University of Cincinnati, Cincinnati, OH
 Lawrence Livermore National Laboratory, Livermore, CA
 2 Bio-safety Level 3 facilities owned and operated by EPA
 OEM has developed partnership with OCSPP, NEIC, and NHRSC to integrate
anthrax analysis into labs daily operations
 Equip and train the labs to analyze anthrax samples in order to increase capacity
utilizing the RV-PCR technique
 Field Exercises and Methods Evaluations 11

12. What is RV-PCR?
1
2
RV-PCR Protocol
Endpoint
response,
Ct (Tf) Time 0
response,
Ct (T0)

13. Why RV-PCR?
1
3
RV-PCR Method
 Rapid, sensitive, and high-throughput sample analyses
 Increased lab capacity to analyze large numbers of samples during a
wide area anthrax event
 Expedite remediation and recovery decisions
 Significantly less laboratory waste compared to traditional culture method –
A Green Solution!

14. RV-PCR vs. Culture
14
Culture Method
• 1 sample  11-12 culture plates + culture tube 
presumptive B. anthracis colonies  2-5 PCR analyses/sample
• Confirmed results in ~ 48 − 72 hr
Sample Dilutions, Plating, and Incubation + PCR
>1000 plates/96 samples
~ 3 incubators
Incubate for 18-24 hours
Filtration and platingSerial dilution – plating &
Enrichment culturing

15. RV-PCR vs. Culture…….continued
1
5
RV-PCR Method
• 1 sample  1 filter cup  2 PCR analyses
• Confirmed results in ~14-16 hr
88 samples + 8 controls/incubator
(22 samples/manifold;
4 manifolds/incubator rack)
Incubate for 9 hours

16. 16
RV-PCR Method Culture Method
Qualitative Analysis Semi-Quantitative
Method performs well even in
the presence of residual decon
agents and spores exposed to
decon agents.
Method is not tested/validated
in the presence of residual
decon agents and spores
exposed to decon agents.
Liquid growth medium is more
conducive for spore germination
and growth (Ref. 2001
Amerithrax).
Plating uses solid growth
medium. Some preps of spores
may not germinate and form
colonies (Ref. 2001 Amerithrax).
RV-PCR vs. Culture …….continued

17. Bio-response Operational
Testing and Evaluation
(BOTE) Project

18. Disclaimer
18
Disclaimer of Endorsement: Reference herein to any specific commercial products,
process, or service by trade name, trademark, manufacturer, or otherwise, does not
necessarily constitute or imply its endorsement, recommendation, or favoring by the
United States Government. The views and opinions of authors expressed herein do
not necessarily state or reflect those of the United States Government, and shall not
be used for advertising or product endorsement purposes.

19. Objectives
19
• Phase 1– Remediation Study (April – May 2011)
• Conduct and evaluate field-level facility remediation studies of three decontamination
technologies
• Assess potential risk of exposure to spores
• Evaluate effectiveness of waste/washwater collection, treatment, and disposal
procedures
• Determine total cost of applying selected decontamination technology or remediation
method/strategy (i.e., including waste management)
• Identify any damage to building or contents
• Phase 2 – Interagency Exercise (September 2011)
• Operationally test and evaluate biological incident response from health/law
enforcement response through environmental response (remediation).

20. Facility
20
PBF-632 at Idaho National Laboratory

21. Example Rooms
21
Mail Room Shop

22. Phase I: Remediation Study
22
• Three Separate Rounds - Conducted in April/May 2011
• A Round is defined as:
• Dissemination of Bacillus atropheus (subspecies globigii) spores in
facility
• First Floor – high contamination (~106 spores/ft2)
• Second Floor – low contamination (~102
spores/ft2)
• Pre-decontamination sampling
• Application of specified
decontamination procedure(s)
• Post-decontamination sampling
• Post-test analysis
(assessment of effectiveness)
• Reset facility for next round of
testing

23. Pre- and Post-Decon Sampling
23
• Pre-moistened Sponge-sticks
• Wetted swabs
• Vacuums socks
• Analysis via LRN
• Use of BROOM for sampling data management
• Aggressive air sampling (Post-decon)

24. Phase 1: Decontamination Methods
24
• Round 1: Fumigation with STERIS
Vaporized Hydrogen Peroxide (VHP®)
• Round 2: Treatment Process
incorporating pH-adjusted bleach
• Round 3: Fumigation with Sabre
chlorine dioxide (ClO2)

25. STERIS VHP®
25
• Full-facility fumigation with vaporized hydrogen peroxide
• Two generators
• Separate injection points on top and bottom floors
• Target conditions:
• 250 ppmv for minimum 90 min
• Temp>70 °F
• No tenting/sealing of facility
• No removal of materials
• Biological indictors (G. stearothermophilus) and chemical indicators placed
throughout facility
• 3 days (setup, fumigation, aeration)

26. pH-adjusted Bleach Process
26
• Procedure:
• Negative air machines to clean air ([re]aerosolized
spores)
• Removal of all porous materials in facility (PPE Level
C)
• Bagging and spraying with pH-adjusted bleach
• Spraying of all remaining surfaces in the facility with
pH-adjusted bleach solution (PPE Level B)
• Target minimum 10 min wetted
• Vacuum standing water
• Decontamination of HVAC return with pH-adjusted
bleach
• HVAC supply lines were capped and not
decontaminated
• 3 days for removal of porous material and
decontamination of facility
• 3 days for drying of facility

27. Sabre ClO2 Fumigation
27
• Fumigation of entire facility w/ ClO2
• Sealing of facility via tenting (under outer containment
and draw through NAM)
• Removal of some porous materials due to potential off-
gassing (longer aeration times)
• Target conditions:
• 3000 ppmv for min 3 hrs (9000 ppmv-hrs)
• >65% RH, >65 °F
• 6 Log Biological indictors (B. atropheus) on stainless steel
placed on each floor
• ClO2 sampling with impinger/titration & EPA with
prototype remote sensor
• 3 days (setup, fumigation, aeration) [plus 2 pre-staging
days]

28. Results (Positive Samples)
Description Floor 1 Floor 2
Pre-Decon VHP 151/153 125/133
Post VHP 44/153 7/134
Pre-Decon AB 146/147 109/124
Post AB 1/134 7/111
Pre-Decon ClO2 138/142 114/129
Post ClO2 3/138 3/127
28

29. 0
2000
4000
6000
8000
10000
12000
14000
VHP AB ClO2
QuantityofWaste(lb)
Decon & Other
Sampling
Phase I: Solid Waste
Generation
29

30. $-
$50,000
$100,000
$150,000
$200,000
$250,000
$300,000
$350,000
$400,000
VHP® pH-Adjusted
Bleach
Process
ClO2
Waste Management Costs
Cost of Decon Materials
Decon Contractor Fixed
Costs
Cost of Removal Teams
Entering
Cost of Decon Teams
Entering
Cost of Decon Line
Operations
30
Based on medium “difficulty” in disposal
30
Phase I: Cost Breakdown

31. Phase 1 Summary
31
• Conclusions from data assessment for the BOTE Project
scenario:
• Effectiveness:
• ClO2 fumigation ≈ pH-adj bleach process >> VHP® fumigation
• Waste generation:
• pH-adj bleach process >> VHP® fumigation > ClO2 fumigation
• Decontamination costs roughly equivalent for all three rounds;
Overall costs significantly dominated by waste management costs
• Spores were detected outside the facility (within the outer enclosure)
• migration outside facility (included dissemination phase)
• Reaerosolization of spores seemed evident during sampling
activities

32. MeBR Fumigation: Problem Statement
• BOTE = unique opportunity to test and evaluate new decon technologies
• The US lacks the capacity to clean up a large Bacillus anthracis (Ba) release in a
timely manner.
• It will take time to ramp up capacity, adding to the impact of an incident.
• Many technologies cause corrosion and collateral damage; creating large amounts
of waste.
• In the case of sensitive or historic infrastructure, corrosive remediation
technologies may not even be a viable option.
32

33. Me BR- When an Exercise became
Real Life
33
• Vendor did not bring enough MeBR to
achieve specified concentration from
RAP (300 mg/L at 24 hours)
• Issues heating building to specified
temperature;
• Vendor planned to vent building without
air filtration prior to clearance samling
• Ended up hitting concentrations of 80-
155 mg/L and temperature between 83-
97 degrees F

34. Why Methyl Bromide (MB)
 MB is efficacious against Ba
 There is an industry that is skilled in it’s application
 MB is less corrosive than alternatives
 MB can be captured after fumigation
34

35. Fl School of Fumigation MB Study

36. Key Steps include:
• Tenting
• Humidification preparation
• Temperature and Relative Humidity Monitoring
• Coupon Loading
• Coupon Placement

37. Wood and Glass
Coupons
37
Spores of Bacillus anthracis Sterne
34F2, the vaccine strain (strain
obtained from Colorado Serum Co.,
Denver, CO), were used as
surrogates for fully-virulent Bacillus
anthracis spores.

38. 38

39. Conditions
• 212 mg/l MB
concentration (5%)
• 80 degrees F
• 70 percent RH
• 48 hours
• Series coupon
extractions
• 16
• 24
• 32
• 40 hours
39

40. Activated Carbon
System
40

41. MB Operational Results
• Safely Completed
• Successfully met
parameters
• Monitored in &
outside structure
• All test coupons
were negative
• Successfully
scrubbed MB
41

42. Underground Transport Restoration (UTR)
CBRN Consequence Management Advisory Division
United States Environmental Protection Agency

43. Collaborative Effort
• Focus is on restoration of an underground transportation system
contaminated with Bacillus anthracis
• Collaborative effort between:
• US Department of Homeland Security (DHS)
• US Environmental Protection Agency (EPA)
• Sandia National Laboratory (SNL)
• Lawrence Livermore National Laboratory (LLNL)
• Argonne National Laboratory (ANL)
• MIT – Lincoln Laboratory (MITLL)

44. Subway recovery challenges
44
Problem:
Subway systems not prepared to quickly remediate and re-open after bioterrorism event
 Loss of billions of dollars to city and businesses (Super-storm Sandy - $125M – 3 days)
 Logistical nightmare for commuters (NYC > 5 mil riders per weekday)
 Above-ground release can also contaminate system
Pre-Event
Surveillance & Planning
Early ID
Real Time Detection
Rapid Response/CM
Situation Awareness
Recovery
“Urban Dispersion Program”
Tracer Concentration versus Distance
(Tracer Release: 1100-1130 EDT Aug. 8, 2005)Avg.TracerConc.
1200-1230EDT
Wind
Subway
Concentration
Above Ground Release
South-North Distance (Miles)
Watson / Heiser - BNL
Subway contamination presents challenges (e. g., bio-load, grime, metal
particulates, airflows, materials) unlike a ‘clean’, indoor environment

45. EPA knows how to ‘decon’ an office
building, NOT a subway
Project Objectives:
 Deliver first comprehensive
Federal Operational Guidance to
decrease time to return a
subway system to service
following a biological agent
event
 Field-test decontamination
technologies and isolation
techniques
 Reduce burden on laboratory
network performing sample
analysis
 Earlier start of decontamination
phase
Faster re-opening
45
Gap: Translate what we know
from a ‘clean’ building
to a ‘dirty’ complex
environment

46. EPA’s Primary Focus
• Bench-scale testing of decontamination options for challenging
environments (ex. fogging)
• Decontamination of rolling stock
• Demonstration of a field-scale decontamination of a section of a
subway platform/tunnel
• Operational Technology Demonstration (OTD)
• Exercise use of RV-PCR

47. Leveraging EPA Bio-Decon Tests to inform
subway system field tests and OTD
47
University of Florida Hurricane House, Davie, FL.
(Dec. 7-12, 2013)
Subway system field test with MeBr
(late FY14 at customer site)

48. Summary
Active field R&D program aimed at enhancing capabilities
and capacity for analysis and decontamination
Looking for partners for the next project – Outdoor decon!
EPA has responsibilities applicable to a wide range of
homeland security threats

49. Questions