Chapter XVI: Section II: Sampling Techniques and Procedures

B-5. General

The collection of environmental, and background (control) samples/medical specimens is an integral part of investigating allegations pertaining to the first use of chemical or biological agents. The types of samples/specimens taken and the collection methods primarily depend upon the circumstances encountered by the collector. During all chemical and biological sampling operations, the commander establishes the required protective equipment to fit the situation. This appendix includes a recommended list of equipment for use during chemical and biological sampling operations (Table B-2).

_Table B-2. Example NBC Collection and Shipping Equipment List_

======================================================================
AMOUNT DESCRIPTION STOCK NUMBER
----------------------------------------------------------------------
20 LABELS, PAPER, PRESSURE SENSITIVE 7530-00-577-4376
2 GLOVES, 8-8½, EDMONT WILSON^{TM} 8415-00-JO2-2902
2 GLOVES, 9-9½, EDMONT WILSON^{TM} 8415-00-634-4639
1 TAPE, ADHESIVE, PRESSURE SENSITIVE, 2 INCH 7510-00-159-4450
1 PLIERS, #47, 5 INCHES 6520-00-543-5330
1 SCREWDRIVER, FLAT TIP, ¼ INCH 5120-00-596-865
1 TONGS, TEFLON^{TM} TIPS AF 15-202-5
2 MICROSPATULA, WITH TEFLON^{TM} ENDS AF 21-401-50A
1 SCISSORS, UNIVERSAL TYPE AF 08-951-30
1 SCOOP, POLYPROPYLENE, 5X2X2 ASP S1021-5
2 SPOON/SPATULA WITH TEFLON^{TM} AF 14-356-10
1 KNIFE, POCKET 5110-00-526-8740
5 BOTTLES, SAMPLE, POLYETHYLENE, 6 OUNCE CP J-6103-50
1 PIPET, JUMBO, TRANSFER TYPE (500/PKG) AF 13-711-7
10 PIPET, GRADUAL, TRANSFER TYPE (500/PKG) AF 13-711-9A
10 BAG, INSULATED, TYPE I AF 01-814-8
10 BAG, INSULATED, TYPE 2[*] AF 01-814-10
1 BAG, WHIRL/PAK, 6 OUNCE (500/PKG) AF 01-812-6B
1 STRIP, pH TESTING, NONBLEEDING, PLASTIC SW S-65271
1 SEP-PAK^{TM} C18 W51910 (50/BOX)
2 SYRINGE, HYPODERMIC, 50 OR 60 ml 6515-00-168-6913
2 STOPCOCK, THREE-WAY ASP S8965-2
1 TUBING, LABORATORY, R3602 CLEAR AF 14-169-3B
1 PEN, MARKING, WATERPROOF AF13-381 (12/PKG)
2 TUBES, TENAX^{TM} EC ST-023
1 BLADE, SURGICAL, CS2L 150S 6515-01-009-5297
2 PACK, ICE CP TR-6345-20
6 PAD, NONADHESIVE, 3X4, 100s 6510-00-111-0708
4 PAD, COOLING, CHEMICAL, 4S 6530-00-133-4299
2 PIGLETTES SPECIAL ORDER
1 TAPE, ANTISEIZE 8030-00-889-3535
1 AIR SAMPLER, PERSONAL LSS G4980
1 KIT, METRIC, POCKET BUBBLE GL4981
2 METHANOL
1 WATER, DISTILLED (5 BOTTLE/PKG)
1 MATCHES, WATERPROOF
20 RAZOR, SURGICAL PREP 6515-00-926-2089
10 WATCH, WRIST 6645-00-066-4279
2 PARAFILM WITH DISPENSER 6640-01-185-3289
2 FLOOR SWEEP (VERMICULITE) 8720-01-026-9419
100 SEALS, TAMPER-RESISTANT
1 A GAS METER CAPABLE OF PROVIDING
ON-STATION ANALYSIS/DETECTION CAPABILITY
FOR MULTIPLE GASES TO INCLUDE INDUSTRIAL
GASES.
1 A COMBUSTIBLE GAS INDICATOR THAT INDICATES
PERCENTAGE OF OXYGEN AND EXPLOSIVITY.
1 A GAS METER THAT DETECTS VAPOR IN PARTS
PER MILLION (PPM) AND INDICATES PRESENCE
OF VAPOR AND ITS STRENGTH.
1 SWABS, THROAT
2 CAN, 6 POUND, METAL
10 BAG, MYLAR
1 CONTAINER, LEAD SHIELDING (FOR RADIATION
SAMPLES)
1 CONTAINER, SHIPPING, IATA
1 CHEST, ICE
10 BAG, PLASTIC, RECLOSABLE

[*] WILL BE REPLACED BY MYLAR BAGS
======================================================================

B-6. Expended Material

The NBC recon units collect samples under various circumstances. For example, a recon unit may collect samples in an area free of hostile forces. The Special Forces NBC Reconnaissance team may collect samples within the enemy area of operations or deep into the enemy's rear area. Samples include toxic agent munitions, chemical products, air, water, soil, and vegetation. In addition, all expended material used to collect the samples should be turned in to the laboratory with the samples. This material includes items such as expended M256A1 kits, M8 and M9 paper. These items should be recovered, packaged, and shipped with the suspected samples for analysis. Different information may be derived from each type of sample; Table B-3 compares different types of samples.

B-7. Environmental Samples

Control or background samples that are collected from clean samples must be identical to the samples collected from the areas near the attack areas as baseline data. The contaminated samples must be compared to the baseline data (control samples). This is especially true if unknown or nonstandard chemical and/or suspected biological agents were employed. The analysis center uses the control samples to compare with a contaminated one. The recon unit collects control samples of soil, water, and vegetation from areas about 500 meters upwind of an alleged attack area. Control samples generically are the same as those collected in an alleged attack area. For example, if leaves from an apple tree in an attack area were collected as a suspected contaminated sample, the recon team should collect leaves (as a control sample) from an apple tree outside of the contaminated area. If water from a pond in the attack area is collected, the recon unit should collect control samples of water from a pond (not a moving stream) in a nearby clean area. The size of an environmental control sample should be about the same as the suspected contaminated sample collected from the attack area (see Table B-4, page B-20).

_Table B-3. Comparison of Sample Types_

======================================================================
SAMPLE STABILITY ANALYSIS
SAMPLE TYPE INFO VALUE TO COLLECT TIME REQUIRED RELIABILITY
----------------------------------------------------------------------
AIR GOOD GOOD 20 MIN HIGH
WATER GOOD GOOD 5 MIN HIGH
SOIL FAIR FAIR 5 MIN MODERATE
VEGETATION FAIR POOR 10 MIN LOW
TISSUE EXCELLENT FAIR 30 MIN HIGH
BLOOD GOOD FAIR 10 MIN HIGH
URINE GOOD FAIR 10 MIN HIGH
MUNITION FRAGMENTS FAIR FAIR 10 MIN FAIR
PACKING MATERIALS FAIR FAIR 10 MIN FAIR
======================================================================

B-8. Collection of Air and Vapor Samples

_a._ Air is a good sample matrix since it is a well-mixed medium. Air from a sample site contains a static concentration of contaminants. The concentration of contaminants depends upon the flow rate of the contaminant into the environment, the wind speed, and the physical state of the contaminant, the terrain contours, and temperature as a variable. The sample should be taken within 102 meters of a contaminated surface and at the downwind edge of a contaminated area. The method should consist of pumping a given volume of air, by hand or electric pump, through sample tubes.

_b._ To avoid contamination, persons conducting air sampling should not use cologne, perfume, insect repellent, medical creams, or strong soaps before taking a sample. The fragrances from these products are volatile organic compounds that may be absorbed on the filter and skew analytical results. Smoke also severely interferes with air sampling, therefore, avoid tobacco and vehicle exhaust smoke.

_c._ The primary method for collecting air samples is with the PAS 1000 automatic air sampler in conjunction with a Tenax^{TM} tube for a total of three to four minutes when possible. Upon completion of sampling, place the Tenax^{TM} tube in a 2¼-inch piglette. Seal the piglette around the cap with either pressure-sensitive or Teflon^{TM} tape. Once sealed, place the piglette into a Mylar or reclosable bag. Fold the bag around the piglette in a circular motion, then apply another bag and fold again. Once the bag is folded around the piglette, use any type tape to secure the bag around the piglette. Place the piglette into a refrigerator or cooler until the sample is transported to its destination.

_d._ When chemicals are permitted into the atmosphere from a facility, the best places to obtain samples are close to the emission source where the concentration of the chemical is not diluted. The further from the original point of release, the more diluted the sample becomes from mixing with air, water, or environmental pollutants.

_e._ Natural and man-made terrain features such as hills, valleys, and rows of buildings, sometimes aid the collector by channeling emissions. When these features are associated with a particular facility, their downwind side is a suitable place to collect a sample because the emission remains more concentrated further from the release point.

_f._ For collection in a possibly contaminated location, and if the situation permits, initially use a detection kit such as the M18A2/M256AI to determine if a possible vapor hazard exists from known chemical agents. Also, use the kit when personnel are required to examine possible toxic agent munitions. In any case, collect air samples with the white-band tubes and save for identification and analysis.

_g._ Small air samplers also enable the collector to obtain vapor samples from alleged toxic agent munitions at a safe distance while explosive ordnance disposal (EOD) operations are performed. If EOD personnel are not on the scene, the air sampler can be activated, and the collector can stand at a safe distance while the sampler is operating.

_h._ Perform sampling operations as soon as possible when directed by a higher headquarters or after suspected chemical or biological contamination is encountered.

B-9. Collection of Water Samples

_a._ Water sampling is a matter of collecting enough water to get acceptable information about the contaminants. The collector should provide the analysis center with one C18 and one silica cartridge when using the Sep-Pak^{TM} technique or 100 ml in a sterile bottle when Sep-Pak^{TM} is not available.

_b._ General guidelines: If it is believed that the threat has used standard chemical agents during an attack, use the M272 chemical agent water test kit for initial screening and sampling.

_c._ When collecting water samples using the Sep-Pak^{TM} C18 cartridge, the following items are required:

· One 60 cc syringe without needle.

· One 3-way sterile, plastic, stopcock with protective covers.

· One piece of plastic tubing (3/16" inner diameter × 6" long
minimum).

· Sterile water or methanol.

· One clean container, such as a Teflon^{TM} cup or glass jar.

_d._ Prior to collecting each sample, prime the Sep-Pak^{TM} system in the following manner:

· =Step 1.= Attach Sep-Pak^{TM} directly to 60 cc syringe.

· =Step 2.= Pour small amount of sterile water or methanol into
container.

· =Step 3.= Insert tip of Sep-Pak^{TM} into container.

· =Step 4.= Withdraw at least 40 cc of solution.

· =Step 5.= Detach Sep-Pak^{TM} from syringe and discard
solution from syringe.

· =Step 6.= Repeat steps 3 through 5 using the same syringe.

_e._ After priming the Sep-Pak^{TM}, assemble the components in the following configuration:

· Attach the 3-way stopcock to a 60 cc syringe.

· Attach the Sep-Pak^{TM} to the opposite end of stopcock.

· Attach the plastic tubing to the open end of the Sep-Pak^{TM}.

_f._ Use the following procedures to collect samples with Sep-Pak^{TM}:

· =Step 1.= Ensure that the lever on the stopcock is turned
sideways with the off arrow pointed toward the large outlet
port.

· =Step 2.= Place the open end of the plastic tubing into the
water near the bottom, without touching the bottom or sides of
the body of water.

· =Step 3.= Draw 60 cc of water into the syringe.

· =Step 4.= Turn the stopcock lever to the off position by
positioning the lever to point toward the stopcock.

· =Step 5.= Push the plunger all the way in, discharging the
water from the syringe through the outlet port.

· =Step 6.= Repeat steps 1 through 5.

· =Step 7.= Detach a plastic tubing from the Sep-Pak^{TM}, and
discard it as contaminated waste.

· =Step 8.= Detach Sep-Pak^{TM} from 3-way stopcock; place into
sample container; seal with pressure-sensitive tape; and mark
for Identification.

NOTE

You should take a minimum of four samples: three samples of the
suspected contamination and one control sample from a nearby
unaffected (none contaminated) area for reference.

· =Step 9.= Dispose of the syringe and stopcock as contaminated
waste.

· =Step 10.= Insert the sample container in a cooler or
refrigerator until the sample is transported to its destination.

_g._ For samples to be representative of the overall contaminated area, the collection point should be carefully selected. Collect samples from--

· Drains and slow-moving streams, since contamination and
dilution from other sources are minimized.

· Stagnant pools of water if the pools of water are part of
chemical waste areas, such as a landfill or chemical disposal
area. Chemicals may percolate into stagnant pools or sumps
close to the site.

NOTE

If an oil film, globules of organic materials, or an unnatural
appearing powder-like material is visible on the water's
surface, collect a surface sample of the material. If not,
collect the sample from near the bottom of the water source
(stream, lake, pond, water container). The upper layers of
water may have lesser amounts of contaminants, due to higher
temperatures that promote decomposition. Since most chemicals
of interest are more dense than water, contaminants usually
sink to the bottom of the water source.

_h._ Collect the sample without the Sep-Pak^{TM} by immersing a capped or stoppered container to the desired depth, removing the cap or stopper, letting the container fill, and then capping the container. An alternate method for deeper water is to use a plastic, pump-operated siphon to pump water from a specific depth.

_i._ The best time to collect a sample of water from a location is when intelligence or local reports indicate that a process of possible interest is ongoing. In the absence of reliable reporting, this may be indicated by increased activity, higher than normal amounts of security, or increased flow from facility chimneys or water discharge pipes. In field areas where a toxic agent has been sprayed or disseminated over a land area, the best time to collect water samples is just after the start of a rainstorm when runoff is beginning. Natural surface drainage will concentrate any remnants of toxic compounds in depressions, streams, or ditches.

B-10. Collection of Soil Samples

Soil is a suitable medium to collect as samples for toxic organic compounds. A critical point, however, is that the precise site of the agent deposition must be sampled for best results. Contamination may be recognized by discoloration or apparent deposition of material on the soil's surface. If discoloration or deposits of material are evident, only collect the discolored soil or deposited materials, if possible. Dead, malformed, and wilted foliage is an indicator of contamination. Soil samples should be collected from open areas, along the drip line tents, stationary equipment, bottom of ditches and terrain depressions.

_a._ Collect the soil samples by using a knife, spoon, spatula, or similar item to scrape a square of topsoil (2×5×1 centimeters) from areas that appear to have been contaminated in to a collection container. If chunks or clods of earth are collected, select those that are no larger than 10×5×1 centimeters (see Table B-4). Also, collect a control sample of soil of the same type and texture from a nearby uncontaminated area.

_b._ Use a glass bottle, jar, or Teflon^{TM} jar as the container when available. When these containers are not available, Mylar bags may be used. When using a glass bottle, jar, or Teflon^{TM} jar, seal the cap with either pressure-sensitive or Teflon^{TM} tape, and mark for identification. When using Mylar bags, place each sample in a separate bag, push excess air out, and seal by folding the open end over two to three times and wrapping the bag with tape. Insert the first bag into a second bag, seal, tape, and mark for identification. If possible, place the samples in a piglette.

+-----------------------------------------------------------+
| CAUTION |
| |
| Avoid direct contact with the sample to prevent exposing |
| yourself to the suspect agent (MOPP 4 is required). |
+-----------------------------------------------------------+

_c._ Collect samples as soon as possible when directed, upon detection of a suspect substance, or after the alleged incident.

B-11. Collection of Contaminated Vegetation

As with soil samples, vegetation is also a suitable medium to collect as samples for toxic organic compounds.

_a._ Collect samples of vegetation that appear to be different from normal. Select leaves that have wilted or appear to have been chemically burned. Collect samples of vegetation that appear to have liquid or solid substances deposited on their surfaces (this may be noticed as a shiny or moist area).

_b._ Collect samples of vegetation at several locations within the suspected contaminated area. Using a cutting tool or any sharp object, cut several affected leaves and/or a handful of grass whenever possible. Do not crush the sample. Place the sample into a Mylar or reclosable bag. Squeeze excess air out of the bag and seal it. Fold the open end of the Mylar bag over two to three times, and wrap it with tape. The minimum size for a sample is three leaves or three handsful of grass. One leaf is of little value, but is better than nothing. Bark is acceptable but not preferred. Mark the bag for identification. Take a control sample of similar material from an unaffected (uncontaminated) area. Fold, seal, tape, and mark the control sample in the same manner as the actual sample.

_c._ When it is possible to determine a probable center of attack in an area, collect vegetation samples near the center of the area, about 100 meters upwind of the area, and in several 100-meter increments downwind of the area. If the collector can discern a contamination pattern in the area, this should be reported.

B-12. Medical Specimens

_a._ Just as blood and urine specimens are taken from humans who were allegedly exposed in an attack, also collect specimens from individuals who claim not to be affected by a toxic agent and are from the same group as exposed personnel. The purpose is the same as collecting environmental control specimens; that is, to determine if a toxic substance is present in the individuals' natural environment or if it has been artificially introduced.

_b._ Selection of humans for control sampling is somewhat more complicated than selection of environmental control samples. This is because ethnic diets, racial differences, physiological makeup, and actual living conditions of persons who are outwardly similar may introduce potentially large deviations. Each of these factors must be accurately considered before selecting subjects as controls.

_c._ Consideration of ethnic diets is important because of unique foods or methods of food preparation that may exist. As an example, individuals in settled areas may purchase beer that has been carefully filtered and sterilized, while individuals in a nearby unsettled area may ferment their own beer by burying home crafted jugs in the ground and extracting the product little by little over several months.

_d._ Racial differences can account for differences in mortality and morbidity rates in specific populations. One example of this could be the high rate of hemophilia in a population versus the rarity of the disease in another.

_e._ Physiological makeup is critical because of the differences in hormone balance and tissue composition in males, females, adults, and juveniles. For this reason, medical control specimens should be drawn from individuals of the same gender and approximate age as specimens from exposed personnel, if possible.

_f._ Differences in the actual living conditions of people also require a close look. The point here is that conditions in remote, semicivilized camps are seldom the same as those in a well-established camp that has access to modern amenities.

_g._ The bottom line in selecting subjects for medical control sampling is that they be as similar in all aspects as possible.

B-13. Collection of Medical Specimens

_a._ Trained medical technicians or physicians should collect medical specimens (human or animal); however, Special Forces NBC Reconnaissance team personnel are trained to do this procedure. Remember, the collector must have express permission (authority) to collect medical specimens from the dead, because of religious beliefs in many cultures. To obtain such specimens without permission may result in unnecessary mission complications. Ensure all personnel handling or collecting medical specimens have received proper immunizations for their own protection. They must be inoculated IAW The Surgeon General's guidance.

_b._ Medical specimens collected during an investigation include blood, urine, sputum, nasal swabs, and tissue specimens from living victims and blood and urine specimens from unexposed persons (background control specimens).

_c._ Collect blood specimens using either a standard 10 cc disposable syringe with a 1- to 1½-inch needle (20 to 22 gauge), or by using a Vacutainer^{TM} system. When using a Vacutainer^{TM} system, ensure that multiple specimen needles and "red top" vacuum tubes are used. Ten cc of blood is sufficient for analytical testing. Do not take more than 5 cc from small, malnourished children. After blood is collected, it should be transferred to a polypropylene-type container and sealed with parafilm before transporting.

All body fluids should be collected without violating antiseptic techniques. Also, prior to transporting specimens, collectors need to check specimen containers for paper labels IAW guidelines for labeling medical specimens. Collect blood specimens using the following materials equipment:

· Gloves.

· 10 cc sterile, disposable syringe.

· 1- to 1.5-inch sterile needle (20 to 22 gauge).

· Vacutainer^{TM} device (adapter with needle).

· Constricting band.

· Disinfectant pads, Betadine, or alcohol.

· Sterile 2×2-inch gauze pads.

· Labels.

NOTE

Gloves should be worn whenever handling medical specimens. Do
not freeze liquid blood and urine specimens (ideal cooling
temperature is between 35° and 40°F [2° to 4°C].)

_d._ Collect urine specimens using either a standard urine cup or by a urine catheter and urine cup. When collecting the specimen directly into a urine cup, the person must urinate into the cup until sufficient fluid is collected (40 cc of urine is preferable, although 10 cc can support analytical testing). When the person is unable to urinate, the catheterization technique is preferable. The catheterization technique is best performed in a clinical environment. As with other body fluids collected, urine must be kept cold. Do not freeze.

NOTE

For correct procedures on catheterization refer to STP
8-91W15-SM-TG.

_e._ Collect tissue specimens using sterile scissors and forceps or as directed by the attending physician.

(1) When casualties have unidentified skin lesions, photographs
of the lesion(s) and overall photos of the extent of the
lesion(s) should be taken, using color film before biopsy. A
specimen of the lesion should be obtained. This is done by
surgically removing a portion of the skin with a sterile pair
of scissors and forceps.

(2) Place tissue specimens in a Teflon^{TM} container filled
¼ inch from the bottom with a preservative, (formalin 10%)
for preservation of the specimen until it reaches its proper
destination. Seal the container and lid with parafilm. As with
any other medical specimens, tissue specimens are refrigerated
prior to shipment; but do not freeze tissue specimens.

_f._ Collect nasal swabs by using a cotton-tipped swab. Place the swab with collected specimen in a Teflon^{TM} container filled ¼ inch from the bottom with a preservative for preservation of the specimen until it reaches its destination. Seal the container and lid with parafilm. Refrigerate the specimen for shipment, but do not freeze.

_g._ Collect sputum by having the patient discharge the sputum into a small, sterile screw-top jar or urine specimen cup. Seal the container and refrigerate the specimen for shipment, but do not freeze.

B-14. Post mortem Specimens

Post mortem specimens should be collected by individuals trained in forensics. When forensics-trained individuals are not available, the most qualified medical person should collect human specimens. Specimens from animals should be collected by veterinary personnel. In either case, the following specimens are collected:

· =Blood.= Use a 50 to 60 cc sterile syringe with an 18-gauge,
5-inch (large bore) needle to collect blood from the heart, and
urine directly from the bladder. Use a spinal needle to collect
cerebral spinal fluids. Three of each type of specimens must be
collected.

· =Lungs.= A biopsy needle is needed to properly collect lung
tissue specimens. After collecting specimens from the lungs,
place specimens in a plastic or Teflon^{TM} container filled
with 10% formalin (preservative) and seal the container for
transporting to its destination.

· =Liver.= If possible collect liver core specimens, using a
large-gauge needle (18-gauge, 5-inch long) via intercostal
or abdominal puncture. Or, if the family consents, perform
a minilaparotomy and obtain one or two 2×2×2 cm sections of
liver. Store and package the specimen as directed for tissue
specimens. For suspect biological agents, see Table B-1 for
specific types of specimens, amount, collection medium, and
from whom to collect.

NOTE

Before attempting any of the above procedures, collector must
be certified by a qualified person (medical doctor) on the
correct procedures to collect specimens from cadavers.

_Table B-4. Standard Sizes of CB Samples/Specimens to be Collected_

========================================================================
TYPE SIZE NOTES
------------------------------------------------------------------------
=CHEMICAL WARFARE SAMPLES=
SOIL (10 CM X 5 CM X 1 CM) CIGARETTE-PACK SIZE OR LARGER
AREA IS MORE USEFUL THAN
GREATER DEPTH
DILUTE AGENT 10 ML
WATER 500 ML (MAXIMUM)
C18 SEP-PAK^{TM} 200 ML
VEGETATION (EQUIVALENT TO 3 LEAVES DEPENDS ON AMOUNT OF CONTAMIN-
OR 3 HANDSFUL OF GRASS) ATION. BEST SAMPLES WILL BE
FOUND NEAR THE RELEASE POINT
------------------------------------------------------------------------
=BIOLOGICAL WARFARE SAMPLES=
SOIL (10 CM X 5 CM X 1 CM) CIGARETTE-PACK SIZE OR LARGER
AREA IS MORE USEFUL THAN
GREATER DEPTH
LIQUID 25 TO 50 ML DO NOT USE C18 SEP-PAK^{TM}
WITH MEDICAL SPECIMENS
VEGETATION SIZE OF SOFT DRINK CAN BEST SAMPLES DEPEND ON THE
AMOUNT OF CONTAMINATION
FOUND NEAR THE RELEASE POINT
------------------------------------------------------------------------
=MEDICAL SPECIMENS=
URINE 20 TO 50 ML MUST OBTAIN CONSENT TO COLLECT
SPECIMENS FROM OTHER THAN US
CASUALTIES
WHOLE BLOOD OR 5 ML MUST OBTAIN CONSENT TO COLLECT
SERUM SPECIMENS FROM OTHER THAN US
CASUALTIES
CEREBRAL SPINAL 2 ML MUST OBTAIN CONSENT TO COLLECT
FLUID SPECIMENS FROM OTHER THAN US
CASUALTIES
ORGAN TISSUE 30 G (MINIMUM)
MEDIASTINAL 2 SHOULD BE REMOVED BY A SURGEON
LYMPH NODES DURING AN AUTOPSY
========================================================================

B-15. Reporting, Packaging, and Shipment

Although a sample/specimen collected from an alleged attack area can be significant, it can become useless if proper steps are not taken to record critical information about its collection or if it is improperly packed and breaks during shipment to an analysis center. This section discusses the information needed when acquiring samples/specimens and the preferred methods for handling and packing samples/specimens for shipment.

_a._ A complete background information history of the circumstances about each sample's/specimen's acquisition must be provided to the agency analyzing the sample/specimen.

_b._ Critical background information includes--

· Circumstances of acquisition. How the sample/specimen was
obtained, where it was found, and how it was collected.

· Physical description. The physical state (solid, liquid,
powder, apparent viscosity), color, approximate size, identity
of the specimen (such as military nomenclature), dirt, leaves,
or so forth.

· Circumstances of agent deposition. The type of delivery
system, a description of how the weapon functioned, how the
agent acted on release, sounds heard during dissemination, a
description of any craters or shrapnel found associated with
a burst, and colors of smoke, flames, or mist that may be
associated with the attack.

_c._ Provide information on the agent effects on vegetation for soil or environmental samples. A description of the general area (jungle, mountain, grassland) and changes in the vegetation after agent deposition (such as color change, wilting, drying, dead) in the main attack and fringe areas.

_d._ Provide information on the agent effects on humans for medical specimens. Describe how the agent affected personnel in the main attack area versus fringe areas; the duration of agent effects; peculiar odors that may have been noticed in the area prior to, during, and/or after an attack; measures taken that alleviated or deteriorated the effects; and the approximate number of victims and survivors, to include their ages and genders.

_e._ Describe the agent effects on animals. Provide information on the types of animals that were or were not affected by an attack and of how they were affected.

B-16. Handling and Packaging Materials

Materials used for packaging samples/specimens primarily consist of Mylar collection bags, Teflon^{TM} specimen jars and tubes, pigs and piglettes, ice chests, sealing materials, and wrapping and cushioning supplies.

_a._ _Collection Bag._ Use the Mylar bag as the initial container for such samples as protective masks and filter canisters, individual antidote and decon kits, munition fragments, and other items too large to place in a specimen jar. Use it also to package sample/specimen containers to ensure a vapor barrier in case the container is broken in transit. The bag acts as an initial or secondary vapor barrier to prevent air from leaking inward and toxic material outward. Follow the procedures below when using the bag.

· If packaging a specimen container or nonenvironmental
sample/specimen, first, verify it has a sample/specimen number.
Carefully place the sample/specimen in a bottom corner of the
Mylar bag.

· Squeeze all the air out of the bag and seal it by removing
the adhesive's protective strip, and pressing the two sides
together.

· Place a piece of 2-inch-wide fiber or cloth tape across the
end of the bag that you just sealed to reseal the Mylar bag
on the outside. This serves as extra insurance in case the
internal seal is broken.

· With the bag lying in front of you and the seal at the top,
fold the bag across its width to as small a size as possible
without damaging the sample/specimen. At this point, use tape
to hold the fold. Next, fold the bag from the top down to the
bottom of the bag to as small a size as possible. The sealing
of the bag is the most critical step during the packaging
process.

· At this point, turn the bag over and use a marker or file
label to put the sample/specimen number on the outside of
the bag so that analysis center personnel can identify the
sample/specimen.

· Place the folded Mylar bag in a clear plastic reclosable
bag, if available. Following the same steps you used for
the Mylar bag, fold and seal the plastic bag. When this has
been completed, again mark the sample/specimen number on the
exterior of the bag.

_b._ _Glass Specimen Jars and Polypropylene Tubes._ Use glass containers to hold small environmental samples, water samples, and medical and post mortem specimens. Use polypropylene containers to hold medical specimens such as blood or urine. Polypropylene containers may be used for post mortem specimens if required; however, glass containers are preferred. The use of glass rather than plastic containers is preferred for environmental samples because toxic agents may leach chemicals from plastics into a sample, introducing contamination and confusing the analysis efforts.

· If the container has a screw-on lid, place Teflon^{TM}
plumber's tape (NSN 8030-00-889-3535; Tape, Antiseize) on the
threads of the container before putting on the lid. This helps
to limit the leakage of liquids and vapor from the container
and to assure the lid will not fall off while in transit. If
the lid has a cardboard liner, remove the liner and replace it
with one or two layers of parafilm (a laboratory sealant film).

· Once the lid is on, stretch parafilm around the outside of
the container at the junction of the lid and the glass. Two
wraps of the film are enough to provide a leakage barrier and
more assurance that the lid cannot fall off.

· At this point, ensure the sample/specimen number is on the
outside of the container. Use a diamond etching pencil or
an adhesive label to put the sample/specimen number on the
exterior of the container.

_c._ _Six-Pound Metal Can._ Use metal cans as the external container for packaging small items that have been sealed in Mylar bags, specimen jars, and polypropylene tubes containing medical specimens. The metal can helps absorb shock from rough handling during shipment and eliminates the spread of contamination if a specimen container is broken. The six-pound metal can is capable of holding more than one sample/specimen (depending upon size of samples/specimens).

· Before placing samples/specimens in the can for shipping,
ensure a sample/specimen number is assigned and is visible on
each item.

· Place about 1 to 2 inches of packing material in the bottom
of the can.

· Wrap jars and tubes in plastic bubble wrap or ⅛- to
¼-inch-thick foam rubber sheeting, secure the wrap with tape or
a rubber band, and place the wrapped item in the can.

· If bubble wrap or foam rubber is not available, use
newspaper. The guiding principle is that the sample/specimen
containers should fit snugly and not be able to move in the can.

_d._ _Ice Chest._ Standard polyethylene or metal ice chests are the most easily procured items used for transworld shipment of CB samples/specimens. The most easily used size is about 24 inches long by 18 inches high by 15 inches deep. This size permits the sender to ship two or three 6-pound metal cans in each chest with sufficient dry ice to maintain freezing temperatures for about four days. Also, each chest remains at a weight that one individual can handle.

_e._ _Transport Container._ When the samples/specimens must be transported on commercial aircraft, an IATA-approved sample transport container must be used for shipment/delivery to the CONUS laboratory.

_f._ _Coolants._ Samples/specimens submitted for laboratory analysis must be properly packaged, labeled, and shipped to ensure they arrive in an analytically acceptable condition. All samples should be maintained at a temperature of 1° to 4°C during transport. Ideally, samples/specimens should arrive at the in-theater laboratory within 6 hours of collection. The samples/specimens should be delivered to the CONUS laboratory within 24 to 48 hours. If the samples/specimens cannot be delivered to the CONUS laboratory within this time, then they should be flash frozen to -165°C, if capabilities are available. If available, dry ice should be used when flash freezing cannot be accomplished. If the samples/specimens cannot be delivered to the CONUS laboratory within 24 hours, the supporting laboratory should subculture the samples/specimens and send the subculture with the samples/specimens to the CONUS laboratory. The subculturing date should also be provided.

_g._ _Internal Insulation._ While a commercial ice chest provides good insulation of both the samples/specimens and the coolant, it is best to place extra insulation and cushioning around the metal cans inside the chest. Newspapers, plastic bubble wrap, and foam rubber may all be used with almost equally good results except newspapers and standard ice do not mix well.

B-17. Collection Reporting

_a._ The collector must provide a formatted message for transmission as soon as possible to report acquisition and shipment of samples/specimens. During special operations in a theater in which a Special Forces Group (SFG) is deployed, the message is transmitted by the fastest means through the fewest channels to the NBC control (NBCC) center. If a NBCC center has not been deployed to the area of operations, as in low-sample/specimen volume peacetime NBC sampling operation, the message is transmitted by the fastest means through the fewest channels to the message addressees below. In addition, a written report accompanies each sample/specimen or batch of samples/specimens. The collector ensures that the acquisition message has been properly classified.

_b._ The collection report includes at least the following addressees:

SECSTATE WASHDC
SECDEF WASHDCHOSD-ISA/OUS-DREH
JCS WASHDC//J-3/J-5H
CIA WASHDCHOSWR-STD-LSBNIC-NIO(STP)H
DIA WASHDC//DT-3B/DT-5A//
DIR AFMIC FT DETRICK MD//AFMIC-CR/AFMIC-SA//
DA WASHDC//DAMI-FIT/DAMO-SWC//
CMDT USACMLS FT LEONARD WOOD MO//ATSN-CM-CO//
CDR SBCCOM APG
MDHSMCCR-OPF/SMCTE-OPE-RA-ID2H
CDR FSTC CHARLOTTESVILLE VA//AIAST-RA-ID2H
CDR USAMRIID FT DETRICK MD (For suspect biological
samples/specimens only.)

_c._ A collection message contains the following information:

· The sample/specimen identification number is part of the
subject line if only a single sample/specimen is referred to in
the text. Otherwise, refer to the sample/specimen number within
the message body with its background information.

· The shipment date, mode of transportation, courier
identification, air bill of lading number, flight number
destination, and estimated time of arrival are included if
the sample/specimen is to be shipped immediately. Also, the
material courier receipt form (DD Form 1911) should be used to
maintain chain of custody.

· Background information on the sample/specimen. Questionable
circumstances surrounding acquisition of a sample/specimen.
The name of another country or agency that acquired a
sample/specimen from the same event or area and is not shown on
the message address.

· A recommended priority and rationale for analysis to guide
the analysis center on the assessment of the potential value of
the sample/specimen.

· All details relating to the collection of the
sample/specimen, regardless of how insignificant they may seem
to the collector.

_d._ Ship all samples/specimens by the fastest, safest means, preferably by a technical escort unit (TEU) to the theater Chemical-Biological Sampling Control Element (CBSCE) or to a location the CBSCE designates. If there is no CBSCE in the theater, send the samples/specimens IAW preplanned instructions from the Chemical-Biological Sampling Control Center (CBSCC) at CBDA, Aberdeen, Maryland. The CBSCC uses the following criteria to determine the final destination of each sample:

· Is the sample/specimen chemical or biological in content?

· Is the sample/specimen content completely unknown?

· Is the sample/specimen a possible biological material?

(1) In any case, the NBCC center must be notified in advance of
shipment of the sample so additional instructions or deviations
from standard instructions can be given. Figure B-l shows an
example of a shipping notification message. The NBCC center
will direct, in advance, that samples be sent to one or more
of the following locations, depending on the category of the
samples. Prior to shipment of samples/specimens, contact must
be made with--

Commander
Technical Escort Unit
ATTN: SMCTE-OPE
Aberdeen Proving Ground, MD 21010
DSN: 584-4381 (Duty hours) DSN: 584-2773 (After duty hours)

(2) This unit controls the transport of samples/specimens
to their final destination(s). Do not ship suspected toxic
samples/specimens or munition systems to CONUS technical
centers or intelligence agencies without coordination and prior
approval by the recipient.

NOTE

Suspect CB samples/specimens are first delivered to the
supporting medical laboratory in the AO for in-theater
analysis before they are transported out of the AO. The
supporting laboratory will withdraw an aliquot of selected
samples/specimens for analysis. The supporting medical
laboratory is responsible for providing the AO commander
confirmatory identification within the AO. The CONUS-based
reference laboratory is responsible for providing confirmatory
identification for President and Secretary of Defense purposes.

FM AMEMBASSY DDTTTTZ JAN 02
TO CDR TEU APG MD//SMCTE-OPE//
SECSTATE WASHDC
SECDEF WASHDC//OSD-ISA/OUS-DRE//
INFO CIA WASHDC//OSWR-STD-LSB/NIC-NIO(STP)//
JCS WASHDC//J-3/J-5//
DIA WASHDC//DT-3B/DT-5A//
DIR NSA FT MEADE MD
DIR AFMIC FT DETRICK MD//AFMIC-CR/AFMIC-SA//
DA WASHDC/DAMI-FIT/DAMO-SWC//
CDR FSTC CHARLOTTESVILLE VA//AIAST-RA-ID2//
CDR CBDA APG MD//SMCCR-OPF//
CDR USACMLS FT MCCLELLAN//ATZN-CM-CU//

CLASSIFICATION

SECSTATE FOR...
SECDEF FOR...
CIA FOR...
JCS FOR J-3/J-5 FOR...
DA FOR DAMO-SWC FOR...
AFMIC FOR...
CBDA FOR FIO...
FSTC FOR AMXST-FW...
USACMLS FOR THREAT MGR...

E.0.12356: DECL: OADR (Note: This is included if the message is
classified.)
TAGS: ...

Subject: Shipment of CB Samples/Specimens
REF(S): TEU MSG #, (DTG DDTTTT [time zone] JAN 02)

1. (W) SHIPPING INFORMATION:

A. DATE SHIPPED: JANUARY 11, 2002.
B. MODE OF TRANSPORTATION: AIR EXPRESS, AIR BILL NUMBER RPT
C. FLIGHT SCHEDULE: TO TYO BY JAL XXX, JANUARY 11, 2002. TO JFK
BY JAL YYY, JANUARY 12, 2002. TO IAD BY DEC ZZZ,
JANUARY 12, 2002.
D. DESTINATION: DULLES INTERNATIONAL AIRPORT.
E. ESTIMATED TIME OF ARRIVAL: 2010 HOURS, JANUARY 12, 2002.

2. SPECIAL HANDLING REQUIREMENTS: DRY ICE ENCLOSED AS COOLANT.

3. SHIPMENT CONSISTS OF TWO ICE CHESTS (1 FOR CRDEC AND 1 FOR AFMIC)
CONTAINING SIX SAMPLES/SPECIMENS. ALL LIQUID SAMPLES/SPECIMENS ARE
IN POLYPROPYLENE TUBES AND HAVE BEEN CAREFULLY PACKED TO AVOID
BREAKAGE. THE FOLLOWING SAMPLES ARE INCLUDED IN THE SHIPMENT:

SAMPLE/SPECIMEN NUMBER MESSAGE REFERENCE
TH-850102-001AG THRU TH-850102-005AG BANGKOK DDTTTTZ JAN 02

4. USDAO HAS STATED THAT THIS SHIPMENT IS PARTIAL FULFILLMENT OF CIR.

_Figure B-l. Sample shipping notification message._

B-18. Sample/Specimen Background Documents

The sample/specimen background document allows a collector to note the most relevant details associated with pre- and post-sample/specimen collection conditions. Do not consider the report to be all-inclusive. The information collected should include at least the items listed in Figure B-2. Interviews should be conducted with individuals exposed to the CB agent as well as individuals not exposed (see Figure B-3).

1. ID NUMBER___________

2. COLLECTION (DATE/TIME):______________________

3. COLLECTOR/UNIT:_______________________________

4. TYPE: ENVIRONMENTAL___ BIOMEDICAL___ SINGLE___ MULTIPLE___

5. PURPOSE: ATTACK___ CHEM/BIO ALARM___ CHEM DETECT___
RECON ILLNESS/DEATH___ OTHER___

6. POSTEXPOSURE: HOURS___ DAYS___ WEEKS___ UNKNOWN___

7. LOCATION: TOWN_______________ COORDINATES__________________________

A. TERRAIN: FLAT___ HILLS___ MOUNTAIN___ DESERT___ JUNGLE___
SPARSE TREES___ GRASS___ BODY OF WATER/TYPE___

B. WEATHER: CLEAR___ CLOUDY___ RAIN___ FOG___ SNOW___ DUST___

C. WIND: LIGHT___ HEAVY___ GUSTY___ NONE___

D. ODOR: SWEET___ FRUITY___ PEPPER___ FLOWER___ IRRITATING___
CHANGING___ NONE___ OTHER_____

E. TEMPERATURE AT TIME OF ATTACK:_____ TEMPERATURE AT TIME OF SAMPLE
COLLECTION:_____

8. COMMENTS:
______________________________________________________________________
______________________________________________________________________

9. ATTACK: DATE/TIME_______ METHOD: ARTILLERY___ ROCKET___ AIRCRAFT___
MORTAR___ RPG/GRENADE___ OTHER, DESCRIBE:__________________________

A. EXPLOSION: AIR_______ (HEIGHT)_______ GROUND_________ SIZE_______
DISTANCE_________ DESCRIBE:____________________

B. CONSISTENCY: SMOKE___ MIST___ DUST___ RAIN___ GEL___
INVISIBLE, DESCRIBE:_______

10. ENVIRONMENTAL SAMPLE: SOIL__ WATER__ VEGETATION__ AIR__ OTHER___

11. BIOMED SPECIMEN: ACUTE___ CONVALESCENT___ EXPOSED___ NOT ILL___
POSTMORTEM___ CONTROL, EXPLAIN:_________________ BLOOD___ LIVER___
LUNG___ SPLEEN___ BRAIN___ SKIN___ KIDNEY___ URINE___
OTHER, DESCRIBE:_________________________

12. COMMENTS:
__________________________________________________________________

13. CASUALTY: SSN_______________ UNIT_______________________ SEX______

14. SIGNS/SYMPTOMS: ONSET__ DURATION__

A. HEAD: FEVER___ CHILLS___ HEADACHE___ FLUSHED___ DIZZINESS___
UNCONSCIOUSNESS___ COMA___ HALLUCINATIONS___

B. EYES: SUNLIGHT SENSITIVE___ PAINFUL___ BURNING___
DROOPY EYELIDS___ DOUBLE VISION___ BLURRED VISION___
LARGE PUPILS___ PINPOINT PUPILS___

C. NOSE: RUNNY___ BLEEDING___

D. THROAT: SORE___ DRY___ SALIVATING___ BLOODY SPUTUM___
HOARSENESS___ DIFFICULTY SPEAKING___

E. RESPIRATION: DIFFICULTY BREATHING___ CHEST/PAIN DISCOMFORT___
WHEEZING (IN/OUT)___ COUGHING___ LABORED BREATHING___

F. HEART POUNDING OR RUNNING___ IRREGULAR HEARTBEAT___

G. GI: LOSS OF APPETITE___ NAUSEA___ FREQUENT VOMITING___
FREQUENT DIARRHEA___ VOMITING BLOOD___ DIARRHEA WITH BLOOD___

H. URINARY: BLOODY URINE___ UNABLE TO URINATE___

I. MUSCULOSKELETAL: NECK PAIN____ MUSCLE TENDERNESS___
MUSCLE TREMBLING/TWITCHING___ WEAKNESS___
PARALYSIS, DESCRIBE:_____________________ CONVULSIONS___
TREMORS___ MUSCLE ACHES___ BACK PAIN___ JOINT PAIN___

J. SKIN: RASH___ REDDENING___ ITCHING___ BLISTERS___ PAIN___
NUMBNESS___ PROFUSE PERSPIRATION___

15. COMMENTS:
______________________________________________________________________

16. ANIMALS AFFECTED: YES___ NO___ DESCRIBE:__________________________

17. RELATED SPECIMENS_________________________________________
ID NUMBER_____________________________________
DESCRIPTION__________________________________

18. COLLECTOR
SIGNATURE____________________________
NAME___________________________
PHONE NUMBER______________________
E-MAIL________________________

19. REVIEWER
SIGNATURE
NAME
PHONE NUMBER_________________
E-MAIL_________________

_Figure B-2. Sample/specimen background document._

CB INCIDENT INTERVIEW

DATE:_____________________ INTERVIEWER: ______________________________
SUBJECT'S NAME: ______________________________________________________
ALIAS #1_____________________________ #2______________________________
AGE:____________ SEX: ____ M ____F YEAR OF BIRTH:__________
NATIONALITY:__________________________________________________________
SUBJECT'S ADDRESS:____________________________________________________
IDENTITY CARD #:______________________________________________________
DELIVERY METHODS:
TYPE: ____UNKNOWN _____GROUND ____AIR _____ARTILLERY/ROCKET ____MINE
OTHER, DESCRIBE:______________________________________________________
HEIGHT: ______(M)
SIZE: _____________ (AFFECTED AREA IN METERS)
DISTANCE: __________(M)
AGENT CHARACTERISTICS
ODOR: _____NONE _____SWEET _____FRUITY _____IRRITATING _____PEPPER
_____FLOWER _____CHANGING _____OTHER, DESCRIBE:_______________________
COMMENTS:_____________________________________________________________
______________________________________________________________________
CONSISTENCY:
______SMOKE ______MIST _____DUST _____RAIN ______GEL ______DRY
____VISIBLE ____INVISIBLE ____OTHER, DESCRIBE:________________________
COLOR:_____________ DESCRIBE DEVELOPMENT OF COLOR:____________________
AREA COVERAGE:________________________________________________________
PHYSICAL DISSEMINATION/COVERAGE (i.e., DROPLET SIZE AND DISTRIBUTION):
WRITE OR DRAW_________________________________________________________
SYMPTOMS:_____________________________________________________________
______________________________________________________________________
INDIVIDUAL'S ACTIONS:
DURING ATTACK:________________________________________________________
______________________________________________________________________
AFTER ATTACK:_________________________________________________________
______________________________________________________________________
PROTECTIVE MEASURES:__________________________________________________
TREATMENT RECEIVED:___________________________________________________
______________________________________________________________________
ENVIRONMENTAL EFFECTS: VEGETATION CHANGE? ___YES ___NO
DESCRIBE:_____________________________________________________________
______________________________________________________________________
ANIMALS AFFECTED? ___YES ___NO
DESCRIBE:_____________________________________________________________

OTHERS AFFECTED:

NAME AGE SYMPTOMS RESOLUTION
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________

_Figure B-3. Chemical/biological incident interview._

FM AMEMBASSY DDTTTTZ JAN 02

1.00
SHIPPING

TO CDR TEU APG MD/SMCTE-OPEI/
SECSTATE WASHDC
SECDEF WASHDC//OSD-ISA/OUS-DRE//
INFO CIA WASHDC//OSWR-STD-LSB/NIC-NIO(STP)//
JCS WASHDC//J-3/J-5//
DIA WASHDC//DT-3B/DT-5A//
DIR NSA FT MEADE MD
DIR AFMIC FT DETRICK MD//AFMIC- CR/AFMIC-SA//DA WASHDC//
DAMI-FIT/DAMO-SWC//
CDR FSTC CHARLOTTESVILLE VA//AIAST-RA-ID2//
CDR CRDEC APG MD//SMCCR-OPE//
CDR USACMLS FT LEONARD WOOD MO//ATSN-CM-CO//
CDR USAMRIID FT DETRICK MD// (FOR SUSPECT BIOLOGICAL
SAMPLES/SPECIMENS ONLY)

CLASSIFICATION

SECSTATE FOR...
SECDEF FOR
CIA FOR
JCS FOR J-3/J-5 FOR
DA FOR DAMO-SWC FOR
AFMIC FOR
CRDEC FOR FIO
FSTC FOR AMXST-FM
USACMLS FOR THREAT MGR
E.O. 12356: DECL: OADR (NOTE: This is included if the message is
classified.) TAGS:

=SUBJECT: SHIPMENT OF CB SAMPLES=
REF(S): TEU MSG, #______. (DTG DDTTTT [time zone] Jan 02)

1. INFORMATION:
A. DATE SHIPPED: JANUARY 11.2002.
B. MODE OF TRANSPORTATION: AIR EXPRESS. AIR BILL NUMBER RPT
C. FLIGHT SCHEDULE: TO TYO BY JAL XXX JANUARY 11, 2002. TO JFK BY
JAL YYY, JANUARY 13, 2002. TO IAD BY DEC ZZZ, JANUARY 12, 2002.
D. DESTINATION: DULLES INTERNATIONAL AIRPORT
E. ESTIMATED TIME OF ARRIVAL; 2010 HOURS. JANUARY 12, 2002.

2. SPECIAL HANDLING REQUIREMENTS: DRY ICE ENCLOSED AS COOLANT.

3. SHIPMENT CONSISTS OF TWO ICE CHESTS (1 FOR CRDEC AND 1 FOR AFMICO)
CONTAINING SIX SAMPLES/SPECIMENS. ALL LIQUID SAMPLES/SPECIMENS ARE
IN POLYPROPYLENE TUBES AND HAVE BEEN CAREFULLY PACKED TO AVOID
BREAKAGE. THE FOLLOWING SAMPLES/SPECIMENS ARE INCLUDED IN THE
SHIPMENT:

SAMPLE/SPECIMEN NUMBER MESSAGE REFERENCE TH-8501
1AG THRU TH-850102-005AG BANGKOK DDTTTTZ JAN 0202-00

4. USDAO HAS STATED THAT THIS SHIPMENT IS PARTIAL FULFILLMENT OF CIR.

_Figure B-4. Sample/specimen shipping report._