APPENDIX B SAMPLE/SPECIMEN COLLECTION AND MANAGEMENT
Reading edition
APPENDIX B SAMPLE/SPECIMEN COLLECTION AND MANAGEMENT
Section I. INTRODUCTION
B-1. General
a. Critical elements for accuracy in analysis of NBC samples and physiological specimens are correct collecting, packaging, handling, and transporting techniques. The quality of any analytical evaluation is directly related to the quality of the sample/specimen and the degree of postcollection degradation that occurs prior to testing. Health service support personnel collect and submit specimens for suspect NBC hazards/agents involving humans and animals. Chemical corps and other nonmedical units collect and submit environmental (air, plant, and soil) samples for suspect NBC hazards/agents. Preventive medicine personnel collect and submit water and ice samples for suspect NBC hazards/agents. Veterinary personnel collect and submit food samples, such as fruits and vegetables, and specimens from animals for suspect NBC hazards/agents. Specimens collected from patients that are suspect of being exposed to a biological agent are forwarded to the supporting medical laboratory (such as the TAML, AML or US Navy Forward Deployed PVNTMED Unit) for analysis.
b. Essentially all military operations from war to stability operations and support operations may generate medical laboratory testing requirements. Each scenario, geographical region, population base, and suspect agent will impact on the type and amount of samples/specimens required and the collection process. During all operations, express permission is required before collecting specimens from civilians because of religious or sociological beliefs in many cultures. To obtain such specimens without permission could result in unnecessary mission complications.
NOTES
1. The term "sample" refers to nonhuman and nonanimal origin. The term "specimen" refers to human and animal origin. 2. Always consider that chemical agents may have been employed. Check for chemical agents before collecting a biological sample/specimen. Chemical agents can damage or destroy biological agents. Also, chemical agents not identified in the sample/specimen can pose a hazard to receiving laboratory personnel. Mark all samples that are potentially contaminated with chemical agents as such. 3. Precautions should be taken to protect the sample/specimen collector from potential BW agents; at a minimum, respiratory protection and rubber gloves must be worn. Additional care must be taken when collecting samples/specimens to prevent cross-contamination. Gloves must be changed or decontaminated between sample/specimen collections. 4. Samples will not be delivered to the clinical laboratory of an MTF for analysis. They must be delivered to the designated supporting medical laboratory for processing. This will prevent accidentally spreading a biological agent in the MTF.
c. Coordination for follow-on testing is absolutely critical to the sample/specimen collection process.
d. Coordination with the receiving laboratory should be made to establish sample requirements, preferred collection techniques, methods of preservation, and transportation conditions, when the tactical situation and/or mission permits.
e. The number of medical specimens that need to be collected varies with the type of analysis performed and the impact of the values determined. The number and types of "control" samples/specimens required to validate test information is determined by the supporting medical laboratory personnel. Random sampling, matched with control populations, or other techniques will be employed as the requirements are identified.
B-2. Sample/Specimen Background Information
a. A complete history of the circumstances about each sample's/specimen's acquisition must be provided to the agency conducting the analysis.
b. Critical information includes, but is not limited to—
Meteorological conditions. Describe what the meteorological conditions were at the time of the alleged attack and at the time of the sampling.
Attack to collection time. State the length of time after alleged attack when sample/specimen was taken.
Circumstances of acquisition. Describe how the sample/specimen was obtained and the source of the sample/specimen.
Physical description. Describe the physical state of the sample/specimen (solid, liquid, powder, apparent viscosity), color, approximate size, identity of the sample/specimen (that is, dirt, leaves, blood, tissue), and dose rate (if radiologically contaminated).
Circumstances of the agent deposition. Describe 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 the burst, and colors of smoke, flames, or mists that may be associated with the attack.
Agent effects on vegetation. Describe the general area (jungle, mountain, grassland) and changes in the vegetation after the agent deposition (that is, color change, wilting, drying, dead) in the main attack and fringe areas.
Agent effects on humans. 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 before, during, or after an attack; measures taken that alleviated or worsened the effects; and the approximate number of victims and survivors (include age and gender).
Agent effects on animals. Describe how they are affected.
Grid coordinates or other descriptive information on sample collection location.
B-3. Sample/Specimen Collection and Preservation
a. Ante mortem Specimens. Physiological specimens from living human or animal patients can include just about any conceivable body source or excreted by-product. It must be noted that specimen types are seldom interchangeable; the exact type and amount of specimen required for a specific assay must be known before a collection procedure is initiated (see Table B-1).
Patients seen in an MTF may be the first and in some cases the only source for sampling for suspect biological agent release. The primary medical care provider will determine the level of treatment for these patients and the specimens required for laboratory diagnosis. The MTF laboratory is not equipped to handle biological agents and, therefore, specimens generated will be forwarded to the supporting medical laboratory for analysis. Patient disposition will be based on evacuation policies, exposure, suspect agent, clinical symptoms, and required treatment/isolation.
Blood specimens represent the most common analytical sample. Certain techniques and special care must be exercised to ensure an acceptable specimen is collected and to minimize an adverse affect to the patient or specimen collector. In general, phlebotomy requires the use of a 20 to 22-gauge needle to minimize mechanical hemolysis during aspiration using a syringe or VacutainerTM tube collection system. Blood collected with a syringe and needle should be transferred to an appropriate VacutainerTM tube immediately after collection. The type of tube, type of anticoagulant or preservative, and amount of blood collected will vary with the specific assay requested. Unless some special sample preparation step is required, the blood is best left in the original rubber-stopper tube for transport.
Urine specimens are best collected using a clean-catch (midstream, if possible) technique in a sterile urine cup. The volume of sample required will vary depending on the specific assay requested; however, 25 to 50 ml is sufficient for most tests.
Tissue specimens can originate from any body source accessible by scraping, swabbing, or minor excision. Tissue specimens are collected only by medical trained personnel. Specific techniques for collecting these specimens are not provided in this appendix.
Sputum specimens are best collected using a sterile cup. The volume of specimen will normally be very small. However, a sufficient quantity must be collected to provide for in-theater testing and to provide for CONUS laboratory testing.
Nasal swabs should be collected using sterile cotton-tipped swabs. The swabs with specimen from each person should be placed in a separate sterile container to prevent cross-contamination.
NOTE
In cases where the supporting laboratory cannot be contacted, as a minimum the following specimens should be collected: Urine—25 to 50 ml in a sterile container. Blood—two 7 to 10 ml tubes without anticoagulant (red-stopper VacutainerTM); two 7 to 10 ml tubes with potassium or sodium ethylenediaminetetraacetate (EDTA) (lavender-stopper VacutainerTM).
All specimens (regardless of physiological source) must be labeled to positively identify the individual or animal from whom it was collected; at a minimum, the individual's full name, unique personal identification number (social security number, when possible), military unit and location, and date and time of collection should be written on the label of the specimen container.
All specimens are collected using aseptic techniques. All specimens are packaged, handled, and transported in a manner that ensures they arrive at the final destination laboratory in a testable condition. Personal protection guidelines must be adhered to when collecting or processing specimens; at a minimum, this includes gloves and a mask. In the laboratory, a gown or other protective items may also need to be used. In the field, under suspect NBC conditions, collectors should be in MOPP Level 4 or inside NBC-protected vehicles. Common sense and the clinical and/or tactical situation will determine the extent of personal protection necessary.
Preservation of specimens, either chemically or mechanically (cooling), will be necessary to minimize the amount of analyte degradation that occurs after removing the specimen from its physiological microenvironment. The optimal preservation technique will vary with different laboratory tests and must be confirmed for each requested assay. While freezing may preserve some serum constituents, freeze-thawing cycles may denature others. Freezing may also completely destroy certain microorganisms. This caution also applies to tissue specimens since "fixing" tissue with a standard 10 percent formalin solution will preserve tissue for special staining techniques; however, it renders the specimens completely useless for microbiological culture. Always verify specimen preservation requirements for storage and transport with the supporting medical laboratory before processing the specimen. Ideally, confirmation of the correct handling conditions should be coordinated before collection.
The importance of coordinating sample/specimen collection with the supporting laboratory facility cannot be overstated. Contact the receiving laboratory for instructions when doubt exists about the appropriate source, collection technique, storage and preservation conditions (such as, aerobic or anaerobic environment), and transportation requirements for samples/specimens. Extremely small volumes of samples/specimens, properly collected and handled, can yield a tremendous amount of information to assist in making medical, tactical, and strategic decisions. Conversely, very large quantities of poorly collected and insufficiently preserved samples/specimens are essentially worthless for most analytical techniques.
Analysis beyond intratheater capabilities will be coordinated by the supporting laboratory, when deployed, or through medical channels in the absence of an in-theater supporting laboratory.
b. Post mortem and Forensic Specimens. The analysis of specimens from deceased humans and animals can provide valuable information about the disease, organisms, injuries, or environmental conditions at the time of death. This information can greatly enhance the treatment of others affected by the same, or physiologically similar, process. Specimen collection for post mortem or forensic examination is very important; the techniques involved reflect a significant degree of training, experience, and skill. Most specimens will be of the same type and size as for ante mortem specimens, but types and amounts of specimens will be determined by the collector.
(1) The collection of specimens from remains should be conducted exclusively by a pathologist, or other personnel specifically trained in forensic collection techniques. An exception is when Special Operations Forces (SOF) personnel are operating under radio silence conditions; the most qualified medical person with the operation collects, preserves, and transports or coordinates transport of specimens for evaluation. The same chain of custody requirements applies to specimens collected by SOF personnel, as with all other specimens.
(2) A large amount of support information can be gained by analyzing the site of injury and subsequent death. This "site scene" investigation requires a tremendous attention to detail and a trained observer. If forensic personnel cannot be contacted, or will be unduly delayed in arriving at the scene, then photographs of the victim and the immediate surroundings should be made. The scope and extent of the photographs should be composed to reflect as much detail as possible to assist forensic personnel in reviewing the scene retrospectively. In the event that photography is not feasible, detailed sketches of the scene should be made to assist the forensic investigation.
(3) Techniques such as cardiac or bladder puncture, needle biopsy of organs, spinal tap, or exploratory laparotomy will not be performed by untrained personnel unless specifically requested and directed by forensic investigators.
Table B-1. Specimen Collection for Suspect Biological Warfare Agents
EARLY POSTEXPOSURECLINICALCONVALESCENT/TERMINAL/POSTMORTEM
ANTHRAX
0 TO 24 HOURS.24 TO 72 HOURS.3 TO 10 DAYS.
NASAL AND THROAT SWABS, AND INDUCED RESPIRATORY SECRETIONS FOR CULTURE, FA, AND PCR. SERUM (TT OR RT) FOR TOXIN ASSAYS. BLOOD (E, C, H) FOR PCR. BLOOD (BC OR C) FOR CULTURES. SERUM (TT OR RT) FOR TOXIN ASSAYS. BLOOD (BC OR C) FOR CULTURE. PATHOLOGY SPECIMENS.
PLAGUE
0 TO 24 HOURS.24 TO 72 HOURS.>6 DAYS.
NASAL SWABS, SPUTUM, AND INDUCED RESPIRATORY SECRETIONS FOR CULTURE, FA, AND PCR. BLOOD (BC AND C) FOR CULTURE AND BLOODY SPUTUM (C) FOR FA. SERUM (TT OR RT) FOR F-1 ANTIGEN ASSAYS. BLOOD (E, C, OR H) FOR PCR. SERUM (TT OR RT) FOR IgM, LATER FOR IgG. PATHOLOGY SPECIMENS.
TULAREMIA
0 TO 24 HOURS.24 TO 72 HOURS.>6 DAYS.
NASAL SWABS, SPUTUM, AND INDUCED RESPIRATORY SECRETIONS FOR CULTURE, FA, AND PCR. BLOOD (BC OR C) FOR CULTURE. BLOOD (E, C, OR H) FOR PCR. SPUTUM FOR FA AND PCR. SERUM (TT OR RT) FOR IgM AND LATER IgG, AGGLUTINATION TITERS. PATHOLOGY SPECIMENS.
MELIOIDOSIS/GLANDERS
0 TO 24 HOURS.24 TO 72 HOURS.>6 DAYS.
NASAL SWABS, SPUTUM, AND INDUCED RESPIRATORY SECRETIONS FOR CULTURE AND PCR. BLOOD (BC OR C) FOR CULTURE. BLOOD (E, C, OR H) FOR PCR. SPUTUM AND DRAINAGE FROM SKIN LESIONS FOR PCR AND CULTURE. BLOOD (BC OR C) AND TISSUE FOR CULTURE. SERUM (TT OR RT) FOR IMMUNOASSAYS. PATHOLOGY SPECIMENS.
BRUCELLOSIS
0 TO 24 HOURS.24 TO 72 HOURS.>6 DAYS.
NASAL SWABS, SPUTUM, AND INDUCED RESPIRATORY SECRETIONS FOR CULTURE AND PCR. BLOOD (BC OR C) FOR CULTURE. BLOOD (E, C, AND H) FOR PCR. BLOOD (BC OR C) AND TISSUE FOR CULTURE. SERUM (TT OR RT) FOR IMMUNOASSAYS. PATHOLOGY SPECIMENS.
Q FEVER
0 TO 24 HOURS.2 TO 5 DAYS.>6 DAYS.
NASAL SWABS, SPUTUM, AND INDUCED RESPIRATORY SECRETIONS FOR CULTURE AND PCR. BLOOD (BC OR C) FOR CULTURE IN EGGS OR MOUSE INOCULATION. BLOOD (E, C, AND H) FOR PCR. BLOOD (BC OR C) FOR CULTURE IN EGGS OR MOUSE INOCULATION. PATHOLOGY SPECIMENS.
BOTULISM
0 TO 24 HOURS.24 TO 72 HOURS.>6 DAYS.
NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR PCR (CONTAMINATING BACTERIAL DNA) AND TOXIN ASSAYS. SERUM (TT OR RT) FOR TOXIN ASSAYS. NASAL SWABS AND RESPIRATORY SECRETIONS FOR PCR (CONTAMINATING BACTERIAL DNA) AND TOXIN ASSAYS. USUALLY NO IgM OR IgG. PATHOLOGY SPECIMENS (LIVER AND SPLEEN FOR TOXIN DETECTION).
RICIN INTOXICATION
0 TO 24 HOURS.36 TO 48 HOURS.>6 DAYS.
NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR PCR (CONTAMINATING CASTOR BEAN DNA) AND TOXIN ASSAYS. SERUM (TT OR RT) FOR TOXIN ASSAYS. SERUM (TT OR RT) FOR TOXIN ASSAY. TISSUE FOR IMMUNOHISTOLOGICAL STAINING. PATHOLOGY SPECIMENS. SERUM (TT OR RT) FOR IgM AND IgG IN SURVIVORS.
STAPH ENTEROTOXICOSIS
0 TO 3 HOURS.2 TO 6 HOURS.>6 DAYS.
NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR PCR (CONTAMINATING BACTERIAL DNA) AND TOXIN ASSAYS. SERUM (TT OR RT) FOR TOXIN ASSAYS. URINE FOR IMMUNOASSAYS. NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR PCR (CONTAMINATING BACTERIAL DNA) AND TOXIN ASSAYS. SERUM (TT OR RT) FOR TOXIN ASSAYS. SERUM FOR IgM AND IgG.
T-2 TOXICOSIS
0 TO 24 HOURS POSTEXPOSURE1 TO 5 DAYS.>6 DAYS POSTEXPOSURE.
NASAL AND THROAT SWABS AND INDUCED RESPIRATORY SECRETIONS FOR IMMUNOASSAYS, HPLC/MASS SPECTROMETRY. SERUM (TT OR RT) AND TISSUE FOR TOXIN DETECTION. URINE FOR DETECTION OF TOXIN METABOLITES.
EQUINE ENCEPHALOMYELITIS
(VEE, EEE, AND WEE VIRUSES)24 TO 72 HOURS.>6 DAYS.
0 TO 24 HOURS. NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR RT-PCR AND VIRAL CULTURE. SERUM (TT OR RT) AND THROAT FOR CULTURE. SERUM (E, C, H, TT, OR RT) FOR RT-PCR. THROAT SWABS UP TO 5 DAYS FOR CULTURE THEN CSF. SERUM (TT OR RT) FOR ANTIGEN ELISA. SERUM (TT OR RT) FOR IgM. PATHOLOGY SPECIMENS PLUS BRAIN.
POX)
(SMALLPOX AND MONKEYPOX)2 TO 5 DAYS.>6 DAYS.
0 TO 24 HOURS. NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR PCR AND VIRAL CULTURE. SERUM (TT OR RT) FOR VIRAL CULTURE. SERUM (TT OR RT) FOR VIRAL CULTURE. DRAINAGE FROM SKIN LESIONS/ SCRAPINGS FOR MICROSCOPY, EM, VIRAL CULTURE, AND PCR. PATHOLOGY SPECIMENS.
EBOLA
0 TO 24 HOURS.2 TO 5 DAYS.>6 DAYS.
NASAL SWABS AND INDUCED RESPIRATORY SECRETIONS FOR RT-PCR AND VIRAL CULTURE. SERUM (TT OR RT) FOR VIRAL CULTURE. SERUM (TT OR RT) FOR VIRAL CULTURE. PATHOLOGY SPECIMENS PLUS ADRENAL GLAND.
LEGEND:
BCBlood culture
CCitrated blood
CSFcerebrospinal fluid
DNAdeoxyribonucleic acid
EEDTA
EEEeastern equine encephalitis
ELISAenzyme-linked immunosorbent assay
EMelectron microscopy
F-1fraction-1
FAfluorescent antibody
HHeparin
HPLChigh-pressure liquid chromatography
IgGimmunoglobulin class G
IgMimmunoglobulin class M
PCRpolymerase chain reaction
RTRed Top, if TT is not available
RT-PCRreverse transcriptase/ polymerase chain reaction
TTTiger top
VEEVenezuelan equine encephalitis
WEEwestern equine encephalitis
c. Water Sample Collection.
(1) Water samples for identification or verification of biological agent contamination are collected by PVNTMED personnel. The supporting laboratory should provide guidance on sampling procedures and collecting kits for use in collecting the samples. In the absence of guidance, a technique for use of the Sep-PakTM is described in FM 3-19.
(2) When sampling kits are not available, samples may be collected in other available sterile containers. The best containers for use are the 100-ml glass bottles used for collecting routine water samples. All water samples must be collected and placed in a cooler or refrigerator until the sample is transported to its destination. During transportation the samples must be maintained at a temperature between 1°C and 4°C.
d. Food Samples. Veterinary personnel must collect suspect biologically contaminated food samples for submission to the supporting laboratory for in-theater verification of contamination. All food samples must be collected and placed in sterile containers. Place the samples in a cooler or refrigerator until the sample is transported to its destination. During transportation the samples must be maintained at a temperature between 1°C and 4°C.
e. Animal Specimens. Veterinary personnel collect specimens from suspect biologically contaminated/diseased animals. The same types and amounts of specimens are prepared and shipped in the same manner as are human specimens.
f. Environmental Samples. Environmental samples are collected as directed in the operators' manual or other publications for operating collection systems. Example: The Biological Integrated Detection System (BIDS) collects an environmental sample using a single liquid sample collector. The collector is a high-volume aerosol sampling and collection device. On demand it samples ambient air through a two-stage virtual impactor that concentrates aerosol particles in the 2 to 10 micrometer diameter-size range. The concentrate particle stream is directed through a wet collector containing a buffer solution and, over a 45-minute period, a 40 to 50 ml sample is collected. On order or when test results indicate a suspected agent, the sample and associated documentation are packaged and transported IAW FM 3-101-4.
B-4. Chain of Custody
a. A strict chain of custody must be maintained for every sample/specimen collected. Use DD Form 1911 (Material Courier Receipt), or other document (such as DA Form 4137 [Evidence/Property Custody Document]) as directed for each sample/specimen collected. The chain of custody document must accompany the sample/specimen during transport from the point of collection to the final receiving laboratory. Each time the sample/specimen is transferred to another individual, the receiving person must sign the document to show that they received the sample/specimen and state what happened to the sample/specimen while in their custody. The document will provide the answer to the following questions:
When was the sample/specimen collected?
Who has maintained custody of the sample/specimen?
What has been done with the sample/specimen at each change of custody?
b. The samples/specimens must be appropriately packaged, labeled, and evacuated to the designated medical laboratory for confirmation of a biological attack. The standard chain of custody for the evacuation would be as follows:
Sampling unit.
Unit S2/security office or medical operations officer.
Technical escort unit or other command-designated escort personnel.
In-theater supporting medical laboratory.
Designated CONUS laboratory.
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
AMOUNTDESCRIPTIONSTOCK NUMBER
20 LABELS, PAPER, PRESSURE SENSITIVE7530-00-577-4376
2GLOVES, 8-8½, EDMONT WILSONTM8415-00-JO2-2902
2GLOVES, 9-9½, EDMONT WILSONTM8415-00-634-4639
1TAPE, ADHESIVE, PRESSURE SENSITIVE, 2 INCH7510-00-159-4450
1PLIERS, #47, 5 INCHES6520-00-543-5330
1SCREWDRIVER, FLAT TIP, ¼ INCH5120-00-596-865
1TONGS, TEFLONTM TIPSAF 15-202-5
2MICROSPATULA, WITH TEFLONTM ENDSAF 21-401-50A
1SCISSORS, UNIVERSAL TYPEAF 08-951-30
1SCOOP, POLYPROPYLENE, 5X2X2ASP S1021-5
2SPOON/SPATULA WITH TEFLONTMAF 14-356-10
1KNIFE, POCKET5110-00-526-8740
5BOTTLES, SAMPLE, POLYETHYLENE, 6 OUNCECP J-6103-50
1PIPET, JUMBO, TRANSFER TYPE (500/PKG)AF 13-711-7
10PIPET, GRADUAL, TRANSFER TYPE (500/PKG)AF 13-711-9A
10BAG, INSULATED, TYPE IAF 01-814-8
10BAG, INSULATED, TYPE 2[*]AF 01-814-10
1BAG, WHIRL/PAK, 6 OUNCE (500/PKG)AF 01-812-6B
1STRIP, pH TESTING, NONBLEEDING, PLASTICSW S-65271
1SEP-PAKTM C18W51910 (50/BOX)
2SYRINGE, HYPODERMIC, 50 OR 60 ml6515-00-168-6913
2STOPCOCK, THREE-WAYASP S8965-2
1TUBING, LABORATORY, R3602 CLEARAF 14-169-3B
1PEN, MARKING, WATERPROOFAF13-381 (12/PKG)
2TUBES, TENAXTMEC ST-023
1BLADE, SURGICAL, CS2L 150S6515-01-009-5297
2PACK, ICECP TR-6345-20
6PAD, NONADHESIVE, 3X4, 100s6510-00-111-0708
4PAD, COOLING, CHEMICAL, 4S6530-00-133-4299
2PIGLETTESSPECIAL ORDER
1TAPE, ANTISEIZE8030-00-889-3535
1AIR SAMPLER, PERSONALLSS G4980
1KIT, METRIC, POCKET BUBBLEGL4981
2METHANOL
1WATER, DISTILLED (5 BOTTLE/PKG)
1MATCHES, WATERPROOF
20RAZOR, SURGICAL PREP6515-00-926-2089
10WATCH, WRIST6645-00-066-4279
2PARAFILM WITH DISPENSER6640-01-185-3289
2FLOOR SWEEP (VERMICULITE)8720-01-026-9419
100SEALS, TAMPER-RESISTANT
1A GAS METER CAPABLE OF PROVIDING ON-STATION ANALYSIS/DETECTION CAPABILITY FOR MULTIPLE GASES TO INCLUDE INDUSTRIAL GASES.
1A COMBUSTIBLE GAS INDICATOR THAT INDICATES PERCENTAGE OF OXYGEN AND EXPLOSIVITY.
1A GAS METER THAT DETECTS VAPOR IN PARTS PER MILLION (PPM) AND INDICATES PRESENCE OF VAPOR AND ITS STRENGTH.
1SWABS, THROAT
2CAN, 6 POUND, METAL
10BAG, MYLAR
1CONTAINER, LEAD SHIELDING (FOR RADIATION SAMPLES)
1CONTAINER, SHIPPING, IATA
1CHEST, ICE
10BAG, 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 TYPEINFO VALUETO COLLECTTIME REQUIREDRELIABILITY
AIRGOODGOOD20 MINHIGH
WATERGOODGOOD5 MINHIGH
SOILFAIRFAIR5 MINMODERATE
VEGETATIONFAIRPOOR10 MINLOW
TISSUEEXCELLENTFAIR30 MINHIGH
BLOODGOODFAIR10 MINHIGH
URINEGOODFAIR10 MINHIGH
MUNITION FRAGMENTSFAIRFAIR10 MINFAIR
PACKING MATERIALSFAIRFAIR10 MINFAIR
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 TenaxTM tube for a total of three to four minutes when possible. Upon completion of sampling, place the TenaxTM tube in a 2¼-inch piglette. Seal the piglette around the cap with either pressure-sensitive or TeflonTM 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-PakTM technique or 100 ml in a sterile bottle when Sep-PakTM 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-PakTM 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 TeflonTM cup or glass jar.
d. Prior to collecting each sample, prime the Sep-PakTM system in the following manner:
Step 1. Attach Sep-PakTM directly to 60 cc syringe.
Step 2. Pour small amount of sterile water or methanol into container.
Step 3. Insert tip of Sep-PakTM into container.
Step 4. Withdraw at least 40 cc of solution.
Step 5. Detach Sep-PakTM from syringe and discard solution from syringe.
Step 6. Repeat steps 3 through 5 using the same syringe.
e. After priming the Sep-PakTM, assemble the components in the following configuration:
Attach the 3-way stopcock to a 60 cc syringe.
Attach the Sep-PakTM to the opposite end of stopcock.
Attach the plastic tubing to the open end of the Sep-PakTM.
f. Use the following procedures to collect samples with Sep-PakTM:
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-PakTM, and discard it as contaminated waste.
Step 8. Detach Sep-PakTM 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-PakTM 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 TeflonTM jar as the container when available. When these containers are not available, Mylar bags may be used. When using a glass bottle, jar, or TeflonTM jar, seal the cap with either pressure-sensitive or TeflonTM 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 VacutainerTM system. When using a VacutainerTM 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).
VacutainerTM 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 TeflonTM 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 TeflonTM 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 TeflonTM 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
TYPESIZENOTES
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 AGENT10 ML
WATER500 ML (MAXIMUM)
C18 SEP-PAKTM200 ML
VEGETATION(EQUIVALENT TO 3 LEAVES OR 3 HANDSFUL OF GRASS)DEPENDS ON AMOUNT OF CONTAMINATION. 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
LIQUID25 TO 50 MLDO NOT USE C18 SEP-PAKTM WITH MEDICAL SPECIMENS
VEGETATIONSIZE OF SOFT DRINK CANBEST SAMPLES DEPEND ON THE AMOUNT OF CONTAMINATION FOUND NEAR THE RELEASE POINT
MEDICAL SPECIMENS
URINE20 TO 50 MLMUST OBTAIN CONSENT TO COLLECT SPECIMENS FROM OTHER THAN US CASUALTIES
WHOLE BLOOD OR SERUM5 MLMUST OBTAIN CONSENT TO COLLECT SPECIMENS FROM OTHER THAN US CASUALTIES
CEREBRAL SPINAL FLUID2 MLMUST OBTAIN CONSENT TO COLLECT SPECIMENS FROM OTHER THAN US CASUALTIES
ORGAN TISSUE30 G (MINIMUM)
MEDIASTINAL LYMPH NODES2SHOULD BE REMOVED BY A SURGEON 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, TeflonTM 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 TeflonTM 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 NUMBERMESSAGE REFERENCE
TH-850102-001AG THRU TH-850102-005AGBANGKOK 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 NUMBERMESSAGE REFERENCE TH-8501
1AG THRU TH-850102-005AGBANGKOK DDTTTTZ JAN 0202-00
4. USDAO HAS STATED THAT THIS SHIPMENT IS PARTIAL FULFILLMENT OF CIR.
Figure B-4. Sample/specimen shipping report.