United States Environmental Protection Agency	Office of Research and Development

National Exposure Research Laboratory
FY02 Research Abstract

Government Performance Results Act (GPRA) Goal 8.3.1
APM257

Significant Research Findings:

Develop Routine and Rapid Methods to Analyze Biological Agents

In the fall of 2001 terrorist attacks with Bacillus anthracis spores
overwhelmed the analytical laboratory capacity at the Centers for Disease
Control and Prevention (CDC), at various state health laboratories, and at the
U.S. Army Medical Research Institute of Infectious Diseases.

Research	On October 25, 2001, Administrator Whitman directed that the U.S.

Approach	Environmental Protection Agency's (EPA) biosafety level 3 microbiology

laboratory located in the Andrew W. Briedenbach Environmental Research
Center (AWBERC) in Cincinnati, Ohio, be upgraded in order to increase
laboratory capacity at this time of national need.

Results and	In response to the Administrator's directive, numerous physical

Implications improvements to the biosafety level 3 microbiology laboratory have

occurred. The cast-iron waste-water line connecting the laboratory to the
sanitary sewer was replaced with chemically resistant plastic pipe. Four
new biosafety cabinets, which are 100% vented, replaced recirculating
models. In order for these biosafety cabinets to work properly and be
certified, four new exhaust fans were installed on the roof of the AWBERC
to replace lower capacity models. Four incubators, four refrigerators, a
centrifuge, a microfuge, and a freezer were purchased to meet the
requirements for Bacillus anthracis analysis and spore production. Sensors
that digitally monitor and record the temperatures of all incubators,
refrigerators, and freezers were installed to alert the staff should the
respective instrument exceed set upper and lower temperature points.

Digital magnahelix devices were installed to monitor the pressure
differential in each laboratory module to ensure that the laboratories maintain
negative pressure with regard to the rest of the building. All walls, ceilings,
and floors were resealed with epoxy to preclude penetration of pathogens
into these porous surfaces. Eye wash stations were installed at each
laboratory sink to meet current safety requirements. In addition, foot
operated faucets were installed at each sink. Where there were insufficient
ultraviolet bacteriocidal ceiling lamps, new lamps were installed to
complete the coverage of the whole laboratory. Bulbs were replaced with
new bulbs in existing ultraviolet bacteriocidal lamps, so that the ultraviolet

Scientific
Problem and
Policy Issues

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light output would be as effective as possible. Laptop computers were
installed in various locations to facilitate digitally recording data to obviate
the use of laboratory notebooks, which potentially could become
contaminated. Furthermore, a digital camera was purchased to allow for the
photography of all samples as they arrive. A specially secured area on the
AWBERC local access network (LAN) was established. Only approved
investigators and analysts have access to this secured area of the LAN. Of
those with approved access, only two have the ability to edit the material.
Some of the investigators have been granted top secret security clearance.
All of the protocols required for analysis of anthrax spores were assembled,
adjusted to the needs of the laboratory, and stored electronically on the LAN.
In turn, these protocols were amended further with the addition of a quality
assurance plan and a comprehensive program plan. All investigators
scheduled to work with anthrax samples have been fit tested for respirators.
Several positive air pressure respirators have been purchased and are on
hand, if ultra-hazardous samples prone to aerosolization should need to be
analyzed. The apparatus producing reagent grade water has been upgraded
with a Milli-Q Synthesis, model A10.

For additional safety, a new sample receiving building was positioned
outside and to the side of the AWBERC loading dock. The intent is to
repackage any sample shipments and decontaminate the exterior of the sealed
repackaging in the chemical storage building before transporting the samples
through the AWBERC to the biosafety level 3 microbiology laboratory.

The laboratory was registered with the Laboratory Registration and Select
Agent Transfer Program. The Agency's Registration is effective from 14
November 2001 until 14 November 2004. The registration gives the Agency
access to reagents which are not commercially available but are prescribed
by the CDC. Besides Bacillus anthracis, the Agency is registered to analyze
samples for Burkholderia mallei, B. pseudomallei, and Clostridium
botulinum. Because the Agency is registered with the Laboratory Response
Network, investigators with potential exposure to B. anthracis are eligible to
receive anthrax vaccinations consisting of a series of six inoculations
administered over an eighteen-month period. Nine investigators have started
the vaccination series. A vaccination declination form for investigators with
potential exposure to anthrax, but refusing the vaccine, has been prepared and
reviewed by the Office of General Council.

Research	None.

Collaboration

and Publications

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Future Research To operate at the level of safety set by Agency officials, a number of tasks
must be completed before actual research and sample analysis can begin.
All of the investigators fitted for respirators must receive respirator training.
Bacillus anthracis spore background levels must be determined both inside
and outside the biosafety level 3 laboratory. The anthrax sample analytical
capacity must be determined for the federal in-house staff. To support this
capacity, an on-site support contract that would be used to assist in-house
staff in the event of a surge of samples must be established. Based on these
points, an acceptance criteria protocol must be prepared. Sterne strain B.
anthracis spores must be produced for the various research projects
awarded to AWBERC investigators as part of the Safe Building and Safe
Water Initiatives. The Agency needs to obtain and install the laboratory
information system and ancillary hardware currently being used by the CDC
as part of the Laboratory Response Network. A memorandum of
understanding between the U.S. Environmental Protection Agency and the
U.S. Food and Drug Administration to maximize the potential of their
respective biosafety level 3 laboratories and staffs during a time of national
crisis is planned.

Questions and inquiries can be directed to:

Dr. Frank W. Schaefer, III
US EPA, Office of Research and Development
National Exposure Research Laboratory
26 West M.L. King Drive
Cincinnati, Ohio 45268-1320

Phone: 513/569-7222
E-mail: schaefer.frank@epa.gov

Contacts for

Additional

Information

National Exposure Research Laboratory — November 2002


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