post decontamination sampling and analysis … · response and remediation efforts that have...

POST DECONTAMINATION SAMPLING AND ANALYSIS PLAN

DEPARTMENT OF STATE ANNEX 32

Prepared byEcology & Environment, Inc.

Technical Directive Document No.: SW3-02-07-0010Ecology & Environment, Inc. Project No.: 001262.0272.01TG

U.S. EPA Contract Number 68-S3-00-01Region III West START

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Table of ContentsPage

1.0 SITE BACKGROUND . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

2.0 DATA USE OBJECTIVES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

3.0 PROJECT OVERVIEW . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33.1 Sampling Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33.2 Project Scope . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43.3 Sampling Strategy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

3.3.1 Focused, Biased, and Random Sampling Strategies . . . . . . . . . . . . . . . . . . . . . . . . . . 53.3.2 Sampling Frequency Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

4.0 SAMPLING METHODOLOGIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64.1 Sample Types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64.2 Surface Sampling Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64.3 Surface Sampling Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74.4 Surface Sampling SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84.5 Air Sampling Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84.6 Air Sampling Preparations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94.7 Air Sampling Durations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104.8 Air Sampling SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

5.0 SAMPLE QUANTITIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135.1 Surface Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135.2 Air Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

6.0 SAMPLE DOCUMENTATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146.1 Sample Labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146.2 Field Data Sheets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146.3 Chain of Custody and Custody Seals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156.4 Sample Handling, Packaging, and Shipping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

7.0 LABORATORY SELECTION AND ANALYTICAL PROCEDURES . . . . . . . . . . . . . . . . . . . . . . . . . . . 167.1 Laboratory Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167.2 Sample Extraction Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167.3 Analytical Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

8.0 QUALITY ASSURANCE REQUIREMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

9.0 DELIVERABLES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179.1 Analytical Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189.2 Sample Location Maps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189.3 Final Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

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LIST OF ATTACHMENTS

Attachment ASite Location Map . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

Attachment BSummary of Historical Positive Sample Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Attachment CCDC Nov 8 & 20, 2001 Sample Location Map . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Attachment DFumigation Zone Layout Map . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Attachment ESurface Sampling SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Attachment FAir Sampling SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Attachment GField Data Sheet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

Attachment HChain of Custody and Custody Seal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Attachment ISummary of Sample Quantities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Attachment JZone (1-7) Specific Packages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

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1.0 SITE BACKGROUND

The Department of State’s (DoS) Annex - 32 Diplomatic Pouch and Mail Facility (SA-32) is located inSterling, Loudoun County, Virginia, at 44132 Mercure Circle in the Mercure Business Park. Thefacility grounds cover a three-acre plot and is a secured fenced facility that is monitored 24 hours a dayby armed guards and remote camera surveillance system. The site is surrounded to the south, southeast,east, and northeast by commercial businesses. There is a residential community approximately 500 feetto the northwest. The remainder of the surrounding area primarily consists of undeveloped land, publichighways, and Dulles International Airport lightly-developed property. See Attachment A for SiteLocation Map.

The single-story building has a floor space of approximately 70,000 square feet with 20 foot ceilings, foran estimated total volume of 1,400,000 cubic feet. The interior of the building was subdivided intooffice space (approximately 10%), various rooms (such as Electrical, Sprinkler Maintenance, and PouchStorage), Shipping and Receiving docks, and the central workroom floor where mail and parcels werehandled by large, automated sorting machines. On October 19, 2001, the United States Postal Service(USPS) stopped its practice of using compressed air to remove dust from their sorting machines andalerted other mail facilities of their change in practice. Prior to October 19, 2001, this DoS facilityfollowed the same practice of using compressed air to remove dust from the sorting machines.

On October 22, 2001, a mail handling worker developed symptoms of inhalation anthrax. The workerwas hospitalized and received treatment for inhalation anthrax. In response to this incident, the facilityceased operations on October 24, 2001, and has remained closed. The DoS assumed oversight of theresponse and remediation efforts that have followed under the authority of the National ContingencyPlan (NCP) and Executive Order 12580.

The initial investigation of anthrax contamination at Annex 32 consisted of four sampling events byDoS contractors and Center for Disease Control (CDC)/National Institute of Occupational Safety andHealth (NIOSH) personnel. Approximately 237 samples representing mail routes within the building,mail handling equipment, mail sorting bins, and personnel work stations were collected from lateOctober 2001 through late November 2001. Of the 237 samples, 28 locations were confirmed positivefor bacillus anthracis (BA) with two additional positives from co-located samples. The positivedetections were found in six distinct areas in the east side of the building, where mail receiving andsorting occurred, and also where the anthrax victim worked. See Attachment B for a summary ofprevious positive sample results, and Attachment C for a historic sample location map.

Following the initial investigative activities, the DoS conducted para-formaldehyde decontamination ofdiplomatic mail pouches within an enclosure built inside the site building.

Under the current phase of the SA-32 remediation process, DoS contractors are involved in thedismantling, sizing, decontamination, and packaging of all materials (i.e., paper, carpet, furniture, andmail sorting equipment) within the building for off-site treatment and/or disposal. The fumigationoperation is expected to occur during the summer of 2003 and will be conducted using vaporizedhydrogen peroxide (VHP). The current plan for the building condition, prior to fumigation, is that thebuilding will be “gutted” leaving only the walls, ceiling, concrete floor, some utilities, and minimal


equipment (i.e., forklifts, scissors lift, ladders, etc.). The existing HVAC system (14 rooftop units) hasbeen removed and is not included as part of this sampling. In addition, no duct work is present thatwould necessitate sampling; however, sections of the diffusers may be present and will be sampled aspart of the surface sampling phase of this plan.

DoS requested the U.S. Environmental Protection Agency (EPA) provide technical assistance during thedecontamination of SA-32 under an Inter-Agency Agreement (IAG). EPA Region III On-SceneCoordinator (OSC) Michael Terrain, directed Ecology and Environment, Inc. (E&E), under theSuperfund Technical Assessment and Response Team (START) West, contract, to develop a PostDecontamination Sampling and Analysis Plan (SAP) for the site.

2.0 DATA USE OBJECTIVES

The overall objective of this SA-32 SAP is to determine if the comprehensive SA-32 decontaminationactivities have been effective (i.e. zero growth for BA spores in the facility within the limitations ofsampling methodology and approach). The decontamination procedures for SA-32 began with theremoval of known contaminated objects and continued with physically cleaning the floor and ceilingusing various methods such as wet wiping large surfaces with soapy water, High Efficiency ParticulateAir (HEPA) vacuuming, and utilizing a 0.5 percent bleach application for those areas that previouslytested positive for solution for BA. These activities are preparatory for the final SA-32 decontaminationstep, in which the SA-32 interior will undergo complete fumigation using vaporized hydrogen peroxide(VHP). Post decontamination sampling will be conducted after this fumigation operation.

Presently, there no validated sampling and analytical methods specifically for BA in environmentalsamples. However, the SA-32 remediation project applies a subcommittee, Technical Working Group(TWG), and Environmental Clearance Committee (ECC) project approval structure similar to thatfollowed at prior anthrax cleanup sites. These various groups consist of agency and industry personnelof various applicable backgrounds assembled to form advisory and review boards regarding theparameters to be followed for a successful cleanup project.

The TWG is a technical and scientific oversight body that serves to review and approve all plannedmajor activities under the SA-32 cleanup. The TWG reviews the findings and recommendations of thevarious subcommittees. The ECC is an objective peer review group of professionals with expertise inthe multiple disciplines relevant to the remediation of sites. The ECC remains involved for the durationof the project and reviews individual activities but is tasked with performing a final evaluation of thecleanup procedure and cleanup-derived data. The ECC remains an independent, objective body thatperforms a review of the final project data, provides an objective finding on the adequacy of thecleanup, and ultimately issues a final recommendation to DoS regarding site clearance for re-occupancy.

The evaluation of fumigation efficacy by the ECC will involve collective consideration of all theparameters of the decontamination efforts including:

• Systematic removal and cleaning of material from the building • Extensive cleaning of the facility with HEPA vacuums and wet methods• Use of a proven fumigant (VHP) following established crisis exemption techniques and


protocols• Monitoring of chemical and physical parameters of the fumigation process to ensure a thorough

fumigation effort• Placement and results of spore strips and chemical indicators to ensure complete fumigation

coverage and penetration• Surface sampling to maximize the likelihood of finding BA• Aggressive air sampling of each fumigated area• Additional surface and/or air sampling after enclosure removal

3.0 PROJECT OVERVIEW

3.1 Sampling Approach

The post decontamination sampling project approach at SA-32 consists of applying multiplesampling methods and multiple collection media types to maximize the strengths of the respectivemethods, and media. The overall objective of this SAP is to determine if the comprehensive SA-32decontamination activities have been effective (i.e., zero growth for BA spores in the facility withinthe limitations of sampling methodology and approach).

This SAP follows standard, government agency and industry-accepted sampling methodologies aswell as newer methodologies or recommendations developed by groups such as U.S. Environmental Protection Agency (EPA), Center for Disease Control (CDC) / Agency for ToxicSubstances and Disease Registry (ATSDR), CDC/National Institute for Occupational Safety andHealth (NIOSH), Occupational Safety and Health Administration (OSHA), and US Army Centerfor Health Promotion and Preventive Medicine (USACHPPM) in response to the anthrax mailattacks of 2001.

The health risk from residual anthrax exposure is currently unknown, which makes a statistically-based sampling strategy unrealistic. Therefore, the rationale for post decontamination clearancesampling at SA-32 is based on an empirical approach (observation, good practice), combined withappropriate statistical considerations (i.e., random assignment of sample locations, estimates ofsampling sensitivity, etc.). Sample numbers and surface sample sizes cannot be calculated based onknown risk, so they will be determined based on practical experience, observation, and standardindustrial hygiene practices.

Due to the fact there are no statistical “targets” for comparison between or among samples, and thetargeted criteria for environmental samples are “0”, very large surface areas/composite areas can besampled. This allows for a reduction in the overall numbers of samples for analysis while coveringlarge targeted areas and maintaining a high level of confidence that BA, if present, will be detected. Similarly, for the evaluation for the presence of airborne BA, a properly designed and implementedaggressive air sampling scheme (to maximize the mobilization and thus enhance the capturepotential of any spores) is an appropriate means of detecting this contaminant in the atmosphere. This SAP describes the multi-media, multi-technique sampling approach to be used at SA-32 todetect any remnant BA spores as a means of determining decontamination efficiency.


The SA-32 sampling subcommittee, which includes members from DoS, EPA, CDC/ATSDR,CDC/NIOSH, OSHA, and USACHPPM was formed to determine the technical needs andapplication methods of implementing a post decontamination sampling plan. This SAP describesthe SA-32 post decontamination sampling plan as determined by that subcommittee.

3.2 Project Scope

This subsection outlines the general sampling program scope. Section 4 describes the specificsampling design, analytical methods to be applied, and the amounts and general locations of thesamples to be collected.

As described in Section 1, the removal of all materials inside SA-32 will occur prior to fumigation. Per the SA-32 Remedial Action Plan (RAP), the empty building will be subdivided into sevenpartitioned Zones to facilitate the fumigation process, each with a floor space of approximately 10,000 ft2 and a volume of approximately 200,000 ft3. The planned Zone configuration is shownin Attachment D. Each Zone will be fumigated individually, one at a time. The postdecontamination sampling activities will be conducted following the fumigation of all Zones, withthe Zone partitions to be left in place until the post-decontamination sampling results have beenreceived and reviewed. Therefore, the post-decontamination sampling process will adopt thefumigation’s Zone configuration scheme and treat each Zone as a distinct area for samplingpurposes.

Post decontamination sampling will take place in three phases: • Phase 1 - Surface Sampling• Phase 2 - Air Sampling• Phase 3 - Zone Partition Surface Sampling

The post decontamination sampling process will begin during the fumigation process. The initialstep for the sampling process is for a sampling supervisor to enter each Zone and determine samplegrid layouts, and then within each layout, specific sampling points. A specific sampling design willresult for each Zone, which sampling teams can then implement. The sampling grid system, Zone-specific sampling designs, and sampling point selection is discussed further in Section 4. OnceZone-specific sampling designs are ready, the zones will be sampled in order beginning with Zone1.

After the floor grids are designated and marked, surface sampling (Phase 1) will occur. Sampleswill be submitted to the designated laboratory as sampling occurs. After receiving final analyticalfresuts from all surface samples, air sampling can be initiated. This pattern of sampling will befollowed throughout all seven zones. The partitioned condition of SA-32 will aid in the progressionof sampling. Zone partition surface sampling (Phase 3 ) will then occur once non-detect results areobtained for the air samples and zone partitions are removed. The zone partition surface samplingwill take place as each individual partition is removed.

3.3 Sampling Strategy


As a result of actions at prior anthrax contamination sites, general strategies for sampling forresidual BA contamination have been developed to facilitate the building clearing (re-occupancyapproval) process. The strategies are based on a delineation approach and are derivative of theinitial detection or characterization data but expand further by accounting for a variety of potentialinfluences, including the distribution or migration of contaminants resulting from mechanicalactions (air flow, foot-traffic, mail movements during processing), site decontamination methodsand activities, and the amount and type of objects or materials remaining after decontamination. This data is combined with the most effective and appropriate means of contamination discovery(sampling techniques), contamination identification (analytical methods), and target area coverage(sample placement and/or distribution patterns) to produce an overall sampling approach.

3.3.1 Focused, Biased, and Random Sampling Strategies

This SAP applies focused, biased, and random sampling strategy criteria similar to that used fordesigning sampling investigations at other anthrax contamination sites. Attachment B showsthe historical list of BA positive samples at SA-32 and the post decontamination samplingclassification as either a focused or biased sample.

A Focused sampling approach consists of thoroughly sampling objects and/or areas wherepositive results for anthrax were previously found. The sample intensity of the objects and/orareas meeting these criteria is extreme, both in terms of the type, and number of samples to betaken. The logic behind Focused sampling is that finding the areas of previous contamination tobe clean likely validates the remedial measures and increases the likelihood of the rest of thebuilding (where no anthrax was detected) to be clean. As a result of the extensive removal ofmail equipment “objects” in SA-32 where the majority of positive BA results were observedduring the initial investigations, the number of potential post decontamination focused samplelocations has been reduced significantly, and only one location is characterized as focused.

Biased sampling is used to preferentially target objects and/or areas that are adjacent to knowncontaminated locations, high traffic areas, and objects and/or areas that are likely to experiencehigh contact by personnel eventually re-occupying the facility. Although it is not possible toexhaustively sample each area or object, meeting the Biased sampling definition, the sampledensity is increased to accommodate those objects and areas near contaminated sites or onesthat are preferentially used. Those locations in SA-32 where anthrax was previously detectedon objects (now removed), or surfaces (now cleaned), are considered biased sampling locations. The “objects” and their respective locations will be considered areas of interest and receivebiased sampling.

Random sampling is used to provide a certain amount of minimal coverage over areas notnecessarily targeted by Biased or Focused sampling. Under Random sampling, and whereapplicable under Focused and Biased sampling, samples are often allocated to multiplelocations at multiple samples for ground, middle, and at a height as necessary to meet thevertical and horizontal distribution need of a given area. The majority of post decontaminationsamples to be collected at SA-32 are considered random.


3.3.2 Sampling Frequency Parameters

Although the initial anthrax sampling detections occurred in the eastern portion of SA-32, thepotential exists for spores to have been transported to other parts of SA-32 through variousmechanisms. As a result, the SA-32 decontamination efforts will involve the entire buildinginterior, per the RAP. To verify the efficacy of the entire decontamination effort, postdecontamination sampling will also occur throughout SA-32.

The post decontamination sampling subcommittee has determined a surface and air samplingfrequency rate for the random type samples. This rate is intended to represent the entire SA-32interior. That rate is based on the goal of collecting surface samples from a minimum of 10% ofall original surfaces (pre-existing floors, walls, and ceiling.) Zones containing focused orbiased sampling locations (previous anthrax detection sites) will receive the random 10%sampling frequency, as well as the additional focused or biased sampling. Air sampling for thebuilding will be accomplished through a horizontal and vertical distribution method. The airsampling rate is intended to meet or exceed the surface area representation criteria. For bothsurface and air sampling, the zone’s floor surface area will be divided into 1000 ft2 grids. Thenumber of samples will be based on the number of these grids within each zone. Thedistribution of surface and air samples within this grid system is as follows:

• The number of random surface sample locations are based on the number of 1000 ft2 floorsurface area grids. A 100 ft2 surface area sample will be collected within each 1000 ft2 grid. This provides for the 10% overall surface coverage.

• The number of floor-based air sampling locations will be matched to the number of 1,000 ft2

grids falling on a Zone’s floor, with the number of air sampling locations to sequentiallydecline over the two elevated-height air sampling levels (mid-Zone height and ceiling level).

4.0 SAMPLING METHODOLOGIES

This section explains the sample types, methods, and standard operating procedures (SOPs) to beutilized for the post decontamination sampling of SA-32.

4.1 Sample Types

Two general types of samples will be collected under this SAP: surface samples and aggressive airsamples. Surface samples will consist of residue (dust, etc.) to be collected from horizontal andvertical surfaces inside SA-32. Multiple surface sample collection methods and media will beemployed. The concept of “aggressive air sampling”, which involves the use of particulatedisturbance and air circulation techniques to help mobilize particulates that might not otherwisebecome airborne and thus detectable under air sampling, will be applied to the collection of SA-32air samples. Air samples will consist of airborne particulate matter found in ambient SA-32 interiorair, to be collected by varying collection methods and media.

4.2 Surface Sampling Methods


As a particulate matter, capable of being airborne, BA spores could potentially reside on anysurface reachable via air currents or other physical distribution methods. A variety of surfacesampling methods exist, and their application varies according to the type of material beingsampled and the ability to access the target surface. The interior surface areas of SA-32 consistmainly of smooth concrete floors, semi-smooth cement and concrete block walls, and a ceilingcomprised of metal trusses and sheet metal panels. During post decontamination sampling, cracksor small holes can be expected in the floor and walls, and there will be various-shaped metalsurfaces (painted and un-painted), including electrical conduit, electrical panel boxes, ceiling I-beams and cross member beams, and plumbing pipes remaining inside SA-32. All other objects inthe building, including porous materials, will be removed and disposed of prior to the fumigationactivities and, therefore, are not expected to be present during this sampling event.

A study conducted at United States Postal Service (USPS) facilities, just after the October 2001anthrax mail attacks, indicated that HEPA Vacuum Dust Collection (HEPA Vac) and wet wipesamples were the favored surface sampling methods for their ability to collect anthrax spores from a majority of surface types. These two sample types can also be used to sample large surface areas,while wet swab samples, the other common surface sampling method, are limited by the smallamount of surface they can adequately cover. Wet swabs are, therefore, less desirable for samplinglarge areas but are very effective for collecting surface samples from localized or hard to reachareas.

The SA-32 Sampling Sub-Committee has chosen HEPA Vac sampling as the primary technique tocollect surface samples, based upon the positive results in similar sampling programs and thepredominantly expansive, flat, and non-porous surfaces in SA-32 that are to be sampled. Wet wipeand wet swab sampling equipment will be available and the technique used if the HEPA Vactechnique does not appear to be adequately removing dust from the specific surface location, or iflimited space restrictions dictate or favor the use of one of these alternate methods. Decisions toapply the alternate methods will be made during the sampling operation by the sampling supervisor. With the pre-cleaning scheduled for the building surfaces, the sub-committee’s consensus is that the100 ft2 sample area will not result in overloading of the HEPA Vac samples.

Zone 2's specific package information found in Attachment J, Zone Specific Packages is anexample of an interior enclosure with anticipated dimensions and corresponding total number ofsamples with an approximate breakdown of each sample type.

4.3 Surface Sampling Preparation

Prior to surface sampling, all original floor, wall (pre-existing walls - not fabricated Zonepartitions) and ceiling surfaces will be divided into approximate 1000 ft2 grids. The grids will bephysically numbered, using chalk or grease markers, sequentially from the number 1, starting withthe south end of a Zone. The Acceptance Dock and Dispatch Dock rooms are oriented lengthwisewest to east. For these two rooms the grid numbering will continue sequentially from west to east.

In addition to the zone grid designation, the floor, east wall, west wall, and ceiling will be furtherdesignated by letters. In the case of interior rooms, the rooms will be identified by a roomabbreviation (see Section 6.0 Sample Documentation). Grid placement will result in the generation


of a Zone Specific Surface Sampling Map, which will show the grid identification for referencingpurposes. Surface sample identification numbers, to be logged onto a Field Data Sheet at the timeof collection, will include references to the Zone number and additional sample locationidentification. See Section 6.0 Sample Documentation for further details.

4.4 Surface Sampling SOPs

Surface sampling SOPs for anthrax have been developed and modified from existing environmentalsampling SOPs. CDC/NIOSH, EPA, and other governmental agencies have participated in thedevelopment and modifications to the current versions of these SOPs. See Attachment E forSurface Sampling SOPs. These SOPs will be utilized and followed as closely as possible; however,actual site conditions and equipment limitations may cause the procedures to vary slightly. In allinstances, the actual procedures employed will be documented and presented in the final reportrequired by this SAP (see Section 9.0 Deliverables).

Three surface sampling methods, HEPA Vac, wet wipe, and wet swab, are to be utilized at the site. Each method calls for specific equipment during the collection of the samples. In all methods thesamples will be handled in accordance with Section 6.4 Sample Handling, Packaging, andShipping. All sampling methods require the use of the following equipment/supplies: samplelocation tags/labels, clean examination gloves, field data sheets, writing pens, permanent markers,self sealing plastic bags, garbage bags, sample transport containers, spray bottles of 0.05% sodiumhypochlorite bleach solution, camera, and film/disks. The sampling equipment needed for eachSOP is listed below:

HEPA Vac Samples• HEPA Vacuum (i.e., Omega Vac Supreme - or equivalent)• Dust collection sock (P161 Dust Collection Sock® 1.1 micron - Midwest Filtration) or

equivalent• Disposable sampling sock holder• 50 ml sterile conical vial container

Wipe Samples• 3" x 3" sterile rayon polyester gauze pad (individually packaged)• Sterile water, sterile saline, or sterile phosphate-buffered solution• 50 ml sterile conical vial container

Swab Samples• Rayon polyester swab (individually packaged) • Sterile water, sterile saline, or sterile phosphate-buffered solution• 15 ml sterile conical vial container

4.5 Air Sampling Methods

The selection of an air sampling method involves consideration of the target analyte, the dynamicsof the targeted environment, and the intended data use. There are a variety of standardized,government-approved airborne particulate matter air sampling methods commonly utilized for


industrial hygiene, process or point-source characterization, and ambient air characterizationapplications. The goal of air sampling under the SA-32 post decontamination sampling program isthe capture and detection of residual airborne anthrax (particulate) spores. Sampling devicessensitive to the capture of BA (such as impactors) are desired, as are high-flow instruments for thepurpose of sampling representative volumes of ambient air inside SA-32 (such as DFUs). Toachieve this goal, the following air sample types have been selected for use at SA-32: Dry FilterUnits (DFUs) with polyester filters, Mattson-Garvin Slit-to-Agar (M-G) impaction samplers usingSBA agar plates, low-flow air sampling pumps and high-flow air sample pumps connected toTeflon® air filter cassettes.

4.6 Air Sampling Preparations

Independent of the 1,000 ft2 floor grid system, each Zone’s main floor and interior rooms will befurther divided lengthwise into three equal rows, respectively. This process will form a smaller,tighter grid pattern for use in the specific distribution and location of air sampling devices. Gridsizes may vary from one room to another, but are only applicable for location referencing purposesrather than for designating a representative sampled area. Air sampling grids will be identified byan alphanumeric code of the format #XY, where the # relates to the floor surface sampling grid oforigin, the first letter, X, is a code indicating the sampling height level (i.e., floor, middle, ceiling),and the last letter, Y, indicates the A, B, or C-lettered row the sampler was placed in. The gridswill be labeled as reference points and recorded on the Field Data Sheet. See Section 6.0 SampleDocumentation for further details.

The basis for the Aggressive Air Sampling guideline is the EPA’s Asbestos Air Sampling SOPs. The current aggressive air sampling guidelines have been developed and modified for use duringthe post decontamination sampling efforts at anthrax contaminated sites since November 2001. Theguideline to be followed at SA-32 is based on several techniques used at Capitol Hill, Wallingford,CT and Brentwood, DC, USPS Facilities, and GSA 410 Mail Facility.

The aggressive air sampling process begins with the placement of air sampling devices at theirdesignated locations. The sampling devices will be loaded with their respective pre-labeled(numbered) sample collection media. The sampling equipment will be set up securely to withstandoccasional gusts of air created during the air disturbance of the Zone. Once all sampling equipmentis deployed, twenty-inch diameter household fans will be turned on and set to run on slow speed forthe duration of the air sampling period. The fans will be positioned to direct the air flow toward theceiling for circulation. The number of fans will be based on one fan per each 10,000 ft3 of thezone’s air volume. As the volume of each Zone is approximately 200,000 ft3, this equates to 20fans per Zone. The fans will be stationed to allow slight mixing of air from all spaces within theZone. Sampling personnel will activate the sampling devices and then perform air disturbanceusing leaf blowers.

The air disturbance effort will be conducted using a minimum of two hand-held leaf blowers(minimum 1 horsepower) for aggressive mixing of the zone’s air and any particulate matter. Theleaf blowers will be operated throughout the Zone by the sampling personnel for a minimum of 30minutes with the intent that all surfaces will be disturbed twice during this period. One person willconcentrate on blowing surfaces from the floor to 12 feet heights and one person on a man-lift will


concentrate on surfaces from 12 feet to the ceiling. Begin the air blowing in the northwest (NW)corner of the zone, closest to the entry/exit point for the majority of the zones. Blow the width ofthe area moving to the northeast (NE) corner of the zone. Work from the entry/exit end of the zonetoward the opposite end of the zone. At the far end of the zone, the workers will turn around andrepeat this procedure back to the entry/exit point of the zone.

Interior rooms within a Zone, especially ones with walls that extend to the ceiling, present achallenge to the attempt to thoroughly mix all the air within a zone and keep it mixed during thesampling period. This challenge will be met by setting an additional box fan (also at the slowsetting) at the doorway to the room positioned to circulate air from the smaller space into the largerspace. In addition, these interior spaces will be sampled using all three types of sampling devicesdistributed both horizontally and vertically to ensure these spaces are representatively sampled.

All instruments will be started at the commencement of the air disturbance/leaf blowing aggressiveair procedure with the exception of all the M/G samplers. The M/G samplers will be turned on in astaggered approach. Air sampling will occur in each Zone at three designated height levels: floor(2-4 feet above the floor), mid-height (10-14 feet above the floor), and ceiling (17-20 feet above thefloor). According to the objective of matching the number of floor-based air sampling instrumentsto the number of floor grids, there will be between 10 and 12 floor-based air sampling instrumentsin each Zone. The number of air sampling instruments to be deployed at the elevated heights (mid-height and ceiling levels) decreases by approximately 1/3 as the height levels increased (forexample, Zone 4 will have 11 floor-based samplers, 7 mid-height samplers, and 4 ceiling samplers). The rational for decreased sampling as height increases is based on particulate settling actionpotential. The types and amounts of each type of sampling instrument is indicated per Zone and perheight level in the Zone Specific Design Table.

4.7 Air Sampling Durations

Air sampling within each Zone will occur over a 24-hour period. Varying numbers of all three ofthe air sampling equipment types will be operated in a Zone, some simultaneously, for time periodsspecified by the sampling method or objective.

The information below represents instrument operational periods under ideal conditions. Difficulties such as sample overloading or power interruptions could alter the intended samplingperiods. All collection media is subject to overloading, and therefore, has to be monitoredthroughout the sampling period. If the media is determined to approach an overloading conditionby the professional judgement of the samplers, the sampling period for that sample will be stoppedand the sample will be collected and considered a discrete sample from that location. If anyinterruptions or deviations occur, attempts will be made to adhere to the planned sample collectionprogram as much as possible as determined appropriate by the sampling supervisor. For example,another piece of sample media may be placed into the sampling device as a second sample from thesame location if the initial sample is considered compromised or inadequate. Sampling periodinformation and related data will be entered onto the Field Data Sheet to be maintained for eachsample.


The planned sampling period for a DFU is 24 hours, with the intention that the instrument operatesuninterrupted over that period with the same filter media installed. The M/G samples will becollected over the first eight hours of the air sampling. The starting of the M-G instruments will be staggered and each location will collect two 1-hour samples during the initial eight hour period.

The planned sampling period for the high-flow Teflon® Air Cassette fixed-location pumps to beplaced at pre-determined locations is eight hours. Additionally, one pump will be operated in thedecontamination station located at the zone entry/exit point, with the intention of determiningwhether BA is migrating outside the zone. At the entry/exit point to each zone is a two stagedecontamination chamber. The decontamination chamber is constructed with two chambersseparated by heavy duty polyethylene sheeting. The sheeting has an industrial zipper installed toprovide a physical barrier between the chambers. The entry/exits sides of the decontamination chamber are constructed of triple layered Z-flaps. One high-flow Teflon® Air Cassette sample willbe placed in the decontamination chamber outside of the zone. The air cassette sample will bestarted before the leaf blowers are turned on and will sample until the sampling team exits the zonefor the final time (after the collection of the DFU samples). The decontamination station areasample may be operated for periods greater than eight hours. Additional sealed filter cassettes willbe available within the Zone to change filter cassettes as needed.

Two low-flow Teflon® Air Cassette pumps will be set up as personal sampling pumps to be carriedon sampling team members while they perform sample collection work in a Zone. Those personnelwill wear the pumps when in a zone, and upon exiting a zone, will switch the power source for thepumps from battery to AC and leave these sampling devices running at opposite ends of the zone’sMain Floor area between existing operating instruments. These samplers will be run for 24 hours.

4.8 Air Sampling SOPs

Air sampling SOPs for anthrax have been developed and modified from existing environmentalsampling SOPs. CDC/NIOSH, EPA, and other governmental agencies have participated in thedevelopment and modifications to the current versions of these SOPs. See Attachment F for AirSampling SOPs. These SOPs will be utilized and followed as closely as possible; however, actualsite conditions and equipment limitations may cause the procedures to vary slightly. In allinstances, the actual procedures employed will be documented and presented in the final reportrequired by this SAP (See Section 9.0 Deliverables).

Three air sampling methods/media, DFU/polyester filter, Impactor/agar plates, and Low-flow/Hi-flow Teflon® air filter cassettes, are to be utilized at the site.

DFUs will be employed due to their ability to intake relatively large portions of a zone’s airspaceover an extended sampling period (designated as 24 hours under the SA-32 project). DFUs are runat approximately 350 liters per minute (Lpm) and contain two polyester felt filters as the capturingmedia, placed in-series within the sampling inlet. For purposes of estimating sample numbers, it isexpected that both filters can be extracted and analyzed as one sample. DFUs are a relatively newair sampling device and were developed as a bio-particulate sampling device by the U.S. ArmedForces’ Joint Program Office for Biological Defense. The instrument is portable and designed forcontinuous operation.


The M-G sampling device is an active impaction sampler that uses a pump to draw in sample airwhile the capturing media, a 150 mm, a 5% Sheeps Blood Agar (SBA) plate, rotates in place for upto a 60 minute sampling period. The M-G impactors provide a very sensitive method fordetermining the presence of airborne BA spores via direct sample impaction onto the agar plate. This plate contains a culturing media that eliminates the media extraction procedure in a laboratory,which is a step the other air sampling methods must use. As the agar plate is rotated on theinstrument’s base, a slit on the instrument’s cover plate selectively exposes a portion of the agarplate at a time to the incoming sampled air. Over the course of the sampling period, the entire plateis exposed. This allows for the even distribution of a sample onto the plate and avoids potentiallyoverloading any one part of the plate to sample material, which can be detrimental to the BAdetection process.

Even longer sampling periods are accomplished by replacing an exposed SBA plate with a newplate. The sampling period is controlled by selecting the correct motor for the desired samplingtime (60 minutes maximum). Compared to other impactor types, the larger M-G plate and samplerpump design allow for a longer-than-usual agar plate sampling period without drying out the platemedia. Additionally, the M-G collects one sample (entailing one analysis) over a 60 minute timeframe, whereas shorter-term impaction sampling devices typically sample over a 15-minute period,thereby using four individual samples and four individual analyses to cover the same 60 minutesampling period. The M-G is calibrated at the factory for a flow rate of 60 ft3/hr, which is indicatedby a marker on the device’s rotometer.

Low and high-flow, air sampling pumps will be utilized for the collection of particulates ontopolytetrafluoroethylene (PTFE) Teflon® filters. The pumps can be run at flow rates ranging fromless than 3 Lpm to 15 Lpm. These pumps sample a smaller volume of air compared to a DFUsampler but provide a different capture medium and are easily deployed and operated, making themeffective for attaining wide spatial coverage.

Each method calls for specific equipment during the collection of the samples. In all methods, thesamples will be handled in accordance with Section 6.4 Sample Handling, Packaging, andShipping. All sampling methods require the use of the following equipment/supplies: samplelocation tags/labels, clean examination gloves, field data sheets, writing pens, permanent markers,self sealing plastic bags, garbage bags, sample transport containers, spray bottles of 0.05% sodiumhypochlorite bleach solution, camera, and film/disks. The sampling equipment needed for eachmethod is listed below:

Dry Filter Units• DFU pump• Polyester felt filter 1 micron mesh, 47 mm diameter

Mattson-Garvin Slit-to-Agar Impactor• Mattson-Garvin Slit-to-Agar (M 220) Air Sampler• 150 mm 5% SBA plate filled according to M 220 impactor specifications


Teflon® Air Filter CassettesC 1 -5 Lpm Low flow personal air sampling pumpC 10-15 Lpm High flow asbestos-type air pumpC 37mm diameter cassette with polytetrafluoroethylene PTFE (Teflon®) filter 0.8 pore sizeC Tygon® tubingC Tripod stand where needed

5.0 SAMPLE QUANTITIES

Within each zone, both surface and air sampling will be conducted. Due to internal structures, eachzone is shaped differently and divided based on available dividing points and support structures. Thenumber of samples is based on guidance provided by the sampling sub-committee

Air sampling involves homogenizing the air through fan disturbance in order to blow potential sporesfrom their current location back into the air for collection on collection media. Air sampling considers athree dimensional space in which three levels have been selected for sample collection. The number ofair floor samples is based on one sample per 1000 ft2 floor grid. As you increase to the next height levelthe total overall number of samples decrease and it likewise decreases again for the ceiling samples.

At a minimum, there will be an additional 10% of field blanks and 5% media blanks collected forQuality Assurance/Quality Control (QA/QC).

The number of samples (including QA/QC) anticipated for the SA-32 site is 575 with an additional 70optional samples for a maximum total of 645. The total number of samples for each zone have beensummarized in Attachment I Summary of Sample Quantities.

5.1 Surface Samples

The interior surface areas of Annex 32 consist mainly of concrete block walls, smooth concretefloors, and a ceiling comprised of metal trusses and sheet metal. These types of materials are easilysampled by using surface techniques discussed in this plan. The surface area of each of the zoneshave been measured based on preliminary construction drawing in order to calculate a total surfacearea for each zone. From these measurements, the number of surface samples have been estimatedbased on a square footage area. The sampling sub-committee has provided guidance on arecommended number of samples per 1000 ft2 of floor, ceiling, and wall area. The sub-committeehas recommended one surface sample be representative of 1000 ft2 of which 100 ft2 (or 10%) wouldphysically be sampled. The anticipated number of surface samples to be collected is 224 with anadditional 70 optional samples for a total of 294.

5.2 Air Samples

The total number of air samples are more difficult to determine due to many variables that maycause extra samples to be taken. Air samples will be collected from three height levels within eachzone. As a basis, the number of floor air sampling locations is based on one location per 1000 ft2

floor surface area. The number of devices at the middle and ceiling heights will vary based on adistribution of sampling devices. As a general guideline, the number of sample locations decrease


for the middle height and decreases further for the ceiling height locations. With the exception ofoverloading, each Area Air Cassette and DFU location will produce a single sample. Theanticipated total number of air samples is 259.

6.0 SAMPLE DOCUMENTATION

All sample documents will be completed legibly in black ink. Any corrections or revisions will be madeby lining through the incorrect entry and initialing the strikeout entry.

Sample data within the hot zone is to be collected on field data sheets. Due to the decontaminationprocedures and the potential contamination of the data sheets, the original field data sheets will not beremoved from the hot zone and archived; however, written transcriptions or electronic copies (scans orfaxes) will be made, verified by the sample team, and signed. The signed copies will be archived in thesite file.

6.1 Sample Labels

Sample labels will be securely affixed to the sample container or written directly on the samplecontainer with permanent ink. Sample labels will clearly identify the sample number. At aminimum, the sample label will identify the sample number. The sample number will be in theformat of X-0001 :• “X” signifies the Zone the sample is collected from (i.e., 1-7)• “0001” signifies a series of sequential numbers for that zone

6.2 Field Data Sheets

Field data sheets supplement the sample label and ensure that all pertinent data is captured for theChain of Custody (COC), as well as additional sample locations and sampling information such asstart and stop times for the air samples. See Attachment G for the contents of the Field Data Sheet.

For this site, the definitions of a composite and grab sample vary slightly from the standardenvironmental definitions. A composite sample is defined as a sample collected from more thanone discrete location or a large area (i.e., greater than 10 ft2) representing an even larger samplingarea (i.e., 1000 ft2). A grab sample is defined as a sample collected from one discrete location suchas an object or specific part of an object.

Additional location identifiers will be documented on the field data sheets. The followingabbreviations will be utilized as necessary:• the room designations for the samples are: MF = Main Floor, OA = Office Area, AD =

Acceptance Dock, ER = Electrical Room, DD = Dispatch Dock, and SR = Storage Room, andSS = small Side Storage

• the descriptions of surface samples are: F = Floor, C = Ceiling, N = North Wall, S = SouthWall, E = East Wall, W = West Wall

• The height level designation for air samples are: F = Floor (2-4 ft), M = Middle (10-14 ft), andC = Ceiling (17-20 ft)


• the grid designations for surface samples are: 01, 02, 03, ...and grid designations for air samplelocations are: A1, A2, A3, B1, B2, ...

• the type of sample is designated by: HE - HEPA Vac Dust Collection filter, WP-wipe, SW-swab, PC-Personnel Teflon® Air Cassette, AC- Area Teflon® Air Cassette, MG-Mattson-GarvinSheep’s Blood Agar plate, and DF-Dry Filter Unit Polyester Filter

6.3 Chain of Custody and Custody Seals

Proper chain of custody will be maintained from the time the sample is collected to its finaldeposition. Every transfer of custody must be noted and signed for. When samples are not underdirect control of the individual responsible for them, they must be stored in a locked containersealed with a custody seal. EPA Region III COC record forms will be utilized. See Attachment Hfor an example COC form.

Custody seals demonstrate that a sample container has not been tampered with nor opened. Theindividual in possession of the sample(s) will sign and date the seal, affixing it in such a mannerthat the container cannot be opened without breaking the seal. EPA Region III Custody Seals willbe utilized. See Attachment H for an example EPA Region III Custody Seal.

6.4 Sample Handling, Packaging, and Shipping

The samples collected in the hot zone will be sealed inside a primary sample container (i.e., plasticconical tube) and placed inside a self sealing plastic bag. This plastic bag cannot be opened in thedecontamination reduction or support zones. The inside of this bag has been exposed in the hotzone and therefore is considered contaminated. The outside of this bag will be decontaminated witha 0.5% sodium hypochlorite solution, before placement inside a transport container (i.e., 5-gallonplastic pail). The transport container will be prepared to receive samples by spraying the inside ofthe container with the decontamination solution and allowed to remain moist. The sample transportcontainer will be transported around the zone with the sample teams until the samples are taken tothe decontamination reduction zone for processing. As the samples pass from the decontaminationzone to the support zone, the decontaminated bagged samples are placed in a clean second self-sealing plastic bags and wiped dry, if necessary by the samplers. After the samples are transferredfrom the decontamination zone to the support zone, the samples will be taken to the samplepreparation area by the respective sampling personnel for further labeling and packaging. Thesamplers will maintain physical custody of the samples throughout this entire process. If necessary,a sample custodian or transporter will be designated and take custody of the samples until they aredelivered to the sample processing team or the laboratory.

Since August 2002, the remediation contractor has performed air sampling with Teflon® aircassettes, HEPA wipe and wet wipe samples from the outsides of building materials that have eitherbeen disposed of or sent for additional sterilization techniques, and wet wipe samples frompersonnel respirators. Over 830 air samples, 100 HEPA/wipe samples from the outsides of buildingitems, and 50 wet wipes from personnel respirators have been collected with no positive results. Inaddition, over 700 Biological Indicator (BI) spore strips used to demonstrate the efficacy of theVHP fumigation will have been collected and analyzed with all negative results. Based on thisinformation, the post decontamination samples will be classified as Non-Hazardous, and therefore,


exempt from Department of Transportation’s (DOT’s) Hazardous Materials Regulations (HMR; 49CFR Parts 171-180) requirements. The sample containers will be placed in a plastic film bag that istreated with a disinfectant after it is sealed and then placed in a clean second plastic bag as thesample is processed through the decontamination area. The plastic bag is then placed in a strongouter packaging (i.e., cooler, paint can). A plastic liner may be used inside the outer packaging inorder to facilitate its reuse. A label with the words “Non-regulated Biological Samples” will beplaced on the outside of the container along with the proper consignee and consignor addressinformation, if needed.

7.0 LABORATORY SELECTION AND ANALYTICAL PROCEDURES

7.1 Laboratory Identification

Laboratory selection was based on multiple criteria, including sample media extraction protocols,laboratory analytical protocols (ability to analyze the entire sample), experience in processing andidentifying positive anthrax samples, the laboratory’s processing capacity, and turn-around time forcertified sample results. Consideration was also given to the location of the laboratory (affectsshipping and/or delivery time), the means of sample delivery (courier vs. mailing), and quality ofdata deliverables.

The laboratory selected for the project, pending CDC’s evaluation of the lab’s extraction andanalysis protocols, is the Navy Medical Research Center (NMRC) located in Silver Spring, MD atWalter Reed Army Institute of Research Facility.

7.2 Sample Extraction Protocols

Each type of sampling media projected to be utilized under this sampling program has a distinctextraction protocol. One exception is the agar plate media which is placed directly into anincubator for culturing and is not discussed here. These extraction protocols can vary betweenlaboratories. The extraction protocols for NMRC are explained below for each media type.

It is important to remember that, in some cases, it may be necessary to adjust the protocol due tocharacteristics unique to some samples. In some cases, the sample will be heat shocked at 65oC for15 minutes. This will reduce the background from other organisms, as well as synchronize thegrowth of the spores if they are present in the sample. The protocols listed below are for thepurpose of recovering BA spores from different sample types, and thus should not be used forextraction and culturing if other organisms are suspected. The buffer solution used for all theextractions will be reagent grade water unless specified in the protocols.

The High Efficiency Particulate Air (HEPA) sock should be received in a 50 ml tube. Open thesock under the laminar flow hood. Place 2 ml of reagent grade water into the HEPA sock. Using a5 ml pipet move the contents of the HEPA sock up and down in the pipet, simultaneously mixingthe contents of the bag. Using the same pipet, remove all of the aqueous solution from the tube andculture this solution on blood agar plates. Plate 250 ml of solution onto each plate until the entiresolution is used.


The wipe will be saturated with reagent grade water until there is 1-1.5 ml of fluid in the tube that isnot absorbed by the wipe. Using 5 ml pipet, vigorously agitate the filter so as to release as muchmaterial as possible from the filter. Using the same pipet, remove all of the aqueous solution fromthe tube and culture this solution on blood agar plates. Plate 250 ml of solution onto each plateuntil the entire solution is used.

The swab should be received in a 15 ml conical tube. Place 1 ml of reagent grade water into thetube containing the swab. Vortex the sample on high for 30 seconds in order to release materialsfrom the swab. Remove all of the aqueous solution from the tube and culture on blood agar plates. Plate 250 ml of solution onto each plate until the entire solution is used.

The polyester dry filter will be placed in 4 ml of reagent grade water. Using a 5 ml pipet,vigorously agitate the filter to release as much material as possible from the filter. Using the samepipet, remove all of the aqueous solution from the tube and culture this solution on blood agarplates. Plate 250 ml of solution onto each plate until the entire solution is used.

7.3 Analytical Procedures

Following extraction, all samples collected under this program will undergo the same analyticalprocedure for the detection of BA. Examine plates for the presence of colonies with morphologiesthat are consistent with BA following a 24-hour culture at 37oC. Any suspicious growth whichcannot be identified by the morphologies, will undergo polymerase chain reaction (PCR) analysisfor confirmation.

8.0 QUALITY ASSURANCE REQUIREMENTS

Field blanks are collected to determine the contribution of sampling handling, shipping and storagebefore analysis. Field blanks will be prepared and handled the same as samples. Field blanks areprepared in a clean area (i.e., support zone). Field blanks should consist of 10% of each sample mediatype.

Media blanks are collected to determine the contribution of contaminant(s), in this case to the overallconcentration of BA in the sample results. Based on the experience of receiving all negative BA mediablank sample results during the sampling work at other, recent anthrax contaminated sites, media blanksshould consist of 5% of each sample media.

9.0 DELIVERABLES

Prior to sampling, zone specific sampling instruction packages will be completed for sub-committeereview as the zone enclosures are constructed on site. Each zone specific package will consist of:written instructions to the samplers, a table of types, and quantities of samples and their generallocation, and a location diagram of air sampling devices. The following deliverables will be providedunder this project:

9.1 Analytical Report


A tabulated analytical summary report will be prepared after receiving the final analytical resultsreport for each zone. The analytical summaries report will be included as an attachment in the finalreport.

9.2 Sample Location Maps

Sample location maps will be generated after all phases of each zone is sampled. These samplelocation maps will be included as an attachment in the final report.

9.3 Final Report

A final report will be prepared within 30 days of receiving the final sampling results for the project. The final report will correlate available background information with data generated under thissampling event. The report will be a summary of the sampling events as well as identifying anyvariations of the sampling SOPs. Quality Assurance/Quality Control (QA/QC) results, chain ofcustody documentation, and laboratory correspondence will be provided within this deliverable, oras required by the EPA OSC.


Attachment ASite Location Map


Site Location MapDepartment of State Annex 32 Site

44132 Mercure CircleSterling, Loudoun County, Virginia

Department ofState Annex 32

Site

post decontamination sampling and analysis … · response and remediation efforts that have...

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