Field Manual of Wildlife Diseases
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Chapter 3 - Specimen Shipment

Author:  J. Christian Franson

Procedures for shipping specimens vary with different disease diagnostic laboratories. Therefore, it is important to contact the receiving laboratory and obtain specific shipping instructions. This will facilitate processing of specimens when they reach the laboratory and assure that the quality of specimens is not compromised. Time spent on field investigation, specimen collection, and obtaining an adequate history will be of little value if specimens become contaminated, decomposed, or otherwise spoiled during shipping to the diagnostic laboratory.

There are five important considerations for proper specimen shipment: (l) prevent cross-contamination from specimen to specimen, (2) prevent decomposition of the specimen, (3) prevent leakage of fluids, (4) preserve individual specimen identity, and (5) properly label the package. Basic supplies needed for specimen shipment are shown in Fig. 3.l.

Figure 3.1 Basic specimen shipment supplies
Specimen identification	Primary containers	Secondary containers	Miscellaneous

Isolate individual specimens from one another by enclosing them in separate packages such as plastic bags. Protect specimens from direct contact with any coolant used (e.g., wet ice or dry ice), and contain all materials within the package so that leakage to the outside of the shipment container is prevented if breakage occurs (e.g., blood tubes) or materials thaw (wet ice and frozen carcasses) due to transit delays.

Plastic bags should be strong enough to resist being punctured by materials contained within them and from contact with other containers within the package.

Styrofoam® coolers, shipped in cardboard boxes, are useful for their insulating and shock absorbing qualities. Styrofoam® at least 1–inch thick is preferred. When possible, select Styrofoam® coolers that have straight sides. Coolers that are wider at the top than at the bottom are more likely to break during transit than those with straight sides. Fill the space between the outside of the Styrofoam® cooler and the cardboard box with newspaper or other packing material to avoid cooler breakage (Fig. 3.2). If coolers are not available, cut sheets of Styrofoam® insulation to fit the inside of cardboard boxes. 

The cardboard box protects the Styrofoam® cooler from being crushed during transit and serves as containment for the entire package (Fig. 3.3). The strength of the box should be consistent with the weight of the package. Cardboard boxes are not needed when hard plastic or metal insulated chests are used for specimen shipment, but boxes can be used to protect those containers from damage and to provide a surface for attaching labels and addresses to the shipment.

Figure 3.2 Proper packaging to prevent Styrofoam® coolers from becoming crushed during transit.

Place the sealed Styrofoam® cooler in a sturdy cardboard box. Use crumpled newspaper or other packaging material to fill all space between the cooler and the box.

Figure 3.3 Proper packaging to prevent Styrofoam® coolers from becoming crushed during transit.

This Styrofoam® cooler was not packaged in a cardboard box for shipping.

Chemical ice packs (Fig. 3.4) are preferable to wet ice because their packaging prevents them from leaking when they thaw. Ice cubes or block ice may be used if leakage can be prevented. This can be accomplished most easily by filling plastic jugs such as milk, juice, and soda containers with water and freezing them. The lids of these containers should be taped closed to prevent them from being jarred open during transit.

Use dry ice to keep materials frozen, but do not use it to ship specimens that should remain chilled because it will freeze them. Also, the carbon dioxide given off by dry ice can destroy some disease agents; this is of concern when tissues, rather than whole carcasses, are being shipped. Shipment of dry ice, formalin, and alcohol is regulated and should be cleared with the carrier before shipping.

Figure 3.4 Chemical coolants.

Chemical coolants are available in (A) soft plastic, (B) hard plastic, and (C) metal containers.

Other disease diagnostic laboratories may require minor variations in shipping procedures.

1. Call the NWHC (608-270-2400) to determine the optimal type and number of specimens for diagnostic procedures, how these specimens are best preserved during transit (whether they should be chilled or frozen), and when they should be shipped. In most cases, the NWHC requests that specimens be shipped the same day or within 24 hours.
2. Double-bag carcasses (Fig. 3.5) and place them in a Styrofoam® cooler lined with a plastic bag. When both frozen and fresh whole carcasses are shipped in the same container, the frozen carcasses can be used as a refrigerant to keep the fresh carcasses chilled. This can be accomplished by interspersing individually bagged frozen carcasses among the individually bagged fresh carcasses or by placing the fresh carcasses between two layers of frozen carcasses (Fig. 3.6). Blood tubes and other breakable containers of uniform size can be protected by packing them in a common plastic bag that is sealed within a metal can or a hard plastic container with a lid (Fig. 3.7). Pack any space around the specimen containers within the can (side and top) with paper or some other absorbent material to prevent jarring that could cause breakage and to collect fluids if tubes do break. Seal the can within a plastic bag before placing it in the Styrofoam® cooler.
3. When using chemical ice packs, intersperse them among specimens; place within the Styrofoam® container other types of coolants in locations that will provide maximum cooling for all contents or, if dry ice is used, will keep everything frozen (Fig. 3.8). Fill all empty space within the Styrofoam® cooler with newspaper to prevent materials from moving during transit. The insulating properties of newspaper will also help maintain cool temperatures within the package, and its absorbent qualities will help prevent fluid leakage outside of the box or container.
4. Close the plastic bag lining the cooler and seal the lid with strapping tape (Fig. 3.9). Tape the specimen data sheet and history, contained in an envelope within a waterproof plastic bag, to the top of the cooler (Fig. 3.10A).
5. Enclose the Styrofoam® cooler in a cardboard box and secure the contents with strapping tape (Fig. 3.10B).

Figure 3.5 Individual carcasses

Individual carcasses should be double-bagged to prevent leakage of fluids and cross-contamination of specimens.

Figure 3.6 Frozen carcasses used to keep fresh specimens chilled during short transit times.

Frozen carcasses (white bags) can be used to keep fresh specimens (dark bags) chilled during short transit times of 24 hours or less. Fill the space between the carcasses and the top of the container with newspaper to provide added insulation to maintain the cold temperature.

Figure 3.7 Packaging sequence for blood tubes.

(A) Pack blood tubes within Whirl-Pak® or other plastic bag; (B) place bag in metal can or hard plastic container and pack with crumpled newspaper or other absorbent, soft, space-filling material; and (C) enclose the can in a plastic bag, then seal the bag.

Figure 3.8 Packing specimens for shipment using different coolants.
(A) Packing specimens for shipment when ice packs are used as coolants. Note that the shipping container is always lined with a large plastic bag.	(B) Packing specimens for shipment when wet ice is used as a coolant. Note that the shipping container is always lined with a large plastic bag.	(C) Packing specimens for shipment when dry ice is used as a coolant. Note that the shipping container is always lined with a large plastic bag.

Figure 3.9 Closing a specimen container.

(A) Secure the large plastic bag containing the specimens by tying the top; (B) close the container lid; and (C) secure the container with several bands of strapping tape.

Figure 3.10 Completing the packaging process.

(A) Tape specimen data sheet and history, contained in an envelope within a waterproof plastic bag, to top of cooler. (B) Place cooler in cardboard box, secure box with several bands of strapping tape, and secure another copy of the specimen data sheet to the outside of the box. If the specimens were placed inside a styrofoam cooler, then use crumpled newspaper of other packing material to fill all spaces between the cooler and the box.

Your packaging and labeling of specimens must conform to the following regulations.

The Code of Federal Regulations (CFR) states under 50 CFR Part 14 of Fish and Wildlife Regulations that containers with wildlife specimens must bear the name and address of the shipper and consignee, and a list of the species and numbers of each species must be conspicuously marked on the outside of the container. You may instead conspicuously mark the outside of each package or container with the word "wildlife" or the common names of the species contained within the package. Secure an invoice or packing list that includes the name and address of the consignee and shipper and that accurately states the number of each species contained in the shipment to the outside of one container in the shipment.

In addition to Fish and Wildlife Service regulations, the interstate shipment of diagnostic specimens is subject to applicable packaging, labeling, and shipping requirements for disease-causing etiologic agents (42 CFR Part 72). These regulations do not require you to identify diagnostic specimens as etiologic agents when the disease agent is not known or is only suspected. However, all specimen packages sent to the NWHC should be prominently labeled with the words "DIAGNOSTIC SPECIMENS (WILDLIFE)." You can meet packaging requirements under 42 CFR Part 72 by following recommendations 2 through 5 above for enclosing specimens within two containers before enclosing them within the package.

Hazardous Materials Regulations of the Department of Transportation apply whenever dry ice is contained within the shipping container (49 CFR Part 172, 173, 175). Always call the carrier ahead of time for the current shipping and package labeling requirements. At the time of this writing, the following must be clearly visible on containers with dry ice: DRY ICE 9, UN1845, weight of dry ice (kilograms), a hazardous materials miscellaneous 9 sticker, and the complete addresses of the shipper and recipient. The dry ice labeling should go on the side of the container, so it is visible if something is stacked on top of it. Always include the words "DIAGNOSTIC SPECIMENS (WILDLIFE)" on the container. A properly labeled container is illustrated in Fig. 3.11. Label containers with permanent markers, if possible.

Figure 3.11 Proper package labeling.

Proper package labeling.

Specimens should be shipped by carriers that can guarantee 24-hour delivery to the location of the diagnostic laboratory. For many locations, commercial delivery services will pick up packages at the point of origin. When shipping arrangements have been made, contact the NWHC again and provide the airbill number and estimated time of arrival. This information is needed to allow prompt tracing of shipments that may not arrive on schedule and to schedule work at the laboratory.

Supplementary Reading

• Code of Federal Regulations. Title 42; Part 72.
• Code of Federal Regulations. Title 49; Parts 172, 173, 175.
• Code of Federal Regulations. Title 50; Part 14.