GUIDELINES FOR ACCEPTABLE TECHNIQUES (BY SPECIES) FOR SURVIVAL BLOOD WITHDRAWAL

The following guidelines cover recommended techniques for the collection of blood in laboratory animals. For all of these techniques, repetitive sampling is assumed as is venous access. Withdrawal volumes must conform to the “Maximum Blood Withdrawal Volume” Guidelines (see IACUC website for this and other guidelines).

Specialized training, use of a two-person team for collection, and/or demonstration of proficiency may be required for any of these techniques. Investigators are encouraged to consult the veterinarians for information or training.

Regardless of the technique used, adequate hemostasis is required to limit blood loss or bruising within tissues, minimize animal pain and distress, and avoid blood smearing or droplets in housing units. Commercially available products such as HemaBlock™ (microporous polysaccharide beads), QR™ (hydrophilic polymer & potassium salt), or styptic powder may be applied to control bleeding (except for retroorbital blood collection), or pressure can be applied to the phlebotomy site with clean gauze.

METHODS FOR BLOOD COLLETION IN ANIMAL EXPECTED TO SURVIVE (NON-TERMINAL)

SPECIES CONSCIOUS1 SEDATED / ANESTHETIZED

Cardiac puncture6 FROG

Submental / submandibular vein (Angle of jaw) 9 Retro-orbital3 MOUSE Lateral saphenous vein5 Tail tip snip 2 Tail nick7

Tail vein Vena cava Tail nick7 or tip puncture Retro-orbital3 RAT Tail snip4 Sublingual vein Lateral saphenous vein5 Jugular vein

Marginal ear vein RABBIT Lateral saphenous vein Jugular vein

SWINE Anterior vena cava8 Umbilical vein Jugular vein8 Lateral saphenous vein Ear vein Cephalic vein

Updated 1-17-18 1

Footnotes: 1. May also be performed with or require the animal be under sedative or anesthesia. 2. Guidelines for Blood Withdrawal Tail Snip Procedures in the Mouse: • The tail should be wiped clean with alcohol before snipping. • Use sterile sharp or to perform tail snips. Wipe the instrument with alcohol between animals. • The length to excise should not be longer than 1-2 mm. • Hemostasis is required. • Because repeated blood collection is often required for research purposes, removal of the scab from the site is recommended for additional blood collections. If repeated excising is necessary, the length of the tail excised should not be longer than 1 additional mm repeated at most 2 additional times. 3. Retro-orbital. Retro-orbital method is used only when other options are not viable. Animals are always anesthetized unless scientifically justified. A single collection from each eye is allowed. The procedure can be repeated in a week utilizing the opposite eye if scientifically justified. Use of sterile pipette is required. Ophthalmic-specific ointment will be applied to the eye if necessary, by observation of dryness. 4. Guidelines for Repetitive Sampling of Blood from Rats using Tail Tip Snip: • The tail should be wiped clean with alcohol before snipping. • Use sterile sharp scissors or scalpel to perform tail snip. Wipe the instrument with alcohol between animals. • Only the tip of the tail should be removed, approximately 1-2 mm. • Subsequent blood collection may be taken by removal of the tail scab. • Hemostasis must be applied 5. Guidelines for Repetitive Sampling of Blood from Rodents using the Saphenous Vein: • The saphenous vein can be punctured using a 20 – 25 gauge needle. • The scab or clot is removed for repetitive samples. 6. Wright, K. 1995. A Simple Method for Obtaining Blood Samples from Mature Frogs. Laboratory Animal Science 1983:386-387. 7. Guidelines for Repetitive Sampling of Blood from Rats and Mice using Tail Nick: • The tail is nicked preferably using the tip of a for a controlled depth and length, but use of a lancet is also allowed. If there are concerns with the depth or length of the nick, monitor the tail for signs of necrosis • The scab or clot is removed for repetitive samples or the tail is slightly twisted and the tail is stroked gently from the base of the tail towards the tip to facilitate blood flow. 8. Must draw blood from the right side only 9. Angle of jaw is not recommended for multiple, closely spaced samples but is fine for multiple samples, when used no more than once weekly, alternating sides.

References: Fox, JG, Anderson, LC, Loew, FM, Quimby, FW 2002 Laboratory Animal Medicine, 2nd edition. Academic Press, ACLAM series. Parasuraman, S, Raveendran, R, and Kesavan, R 2010 Blood sample collection in small laboratory animals. J Pharmacology and Pharmacotherapeutics. 1(2): 87-93. Quesenberry, K.E. and Carpenter, J.W. Ferrets, Rabbits, and Rodents 3rd edition. 2012 St. Louis, MO: Saunders. Suckow, MA, Stevens, KA, and Wilson, RP. 2012 The Laboratory Rabbit, Guinea Pig, Hamster, and Other Rodents. Academic Press, ACLAM series.

Updated 1-17-18 2