Date: October 1998
Reference:  Administrative Panel on Laboratory Animal Care

DIRECTIONS: Review following material. Keep copies of guidelines with applicable protocols. You may find it helpful to post a copy of these guidelines in your laboratory. Questions should be forwarded to the A-PLAC office, 723-4550.

TRAINING: Training in these techniques and the humane treatment of laboratory animals during the procedures is taught by the Veterinary Service Center (VSC) Staff. All new personnel who will be performing these techniques should contact VSC staff for training (723-6735).

1. Cardiac bleeding requires general anesthesia unless interference of anesthetic drugs with experimental objectives is adequately documented and specifically approved by the A-PLAC. This technique should generally be limited to terminal collections due to the danger of cardiac tamponade or pulmonary hemorrhage and pneumothorax. Safer alternative techniques for obtaining repeated large volumes of blood are available. Anesthesia is also required for periorbital bleeding.
   
   
2. Use of topical irritants such as xylene to dilate vessels of the ear pinna is discouraged. If used, the irritant must be immediately removed after bleeding (alcohol followed by soap and water) and a soothing lotion such as Alpha-Keri applied. Vessels can generally be adequately dilated by increasing body temperature by use of a heat lamp or by increasing room temperature. A tranquilizer such as Innovar-Vet also works well.
   
   
3. Use of suction devices applied to the ear is generally unnecessary and may excessively damage tissues. Approval of VSC is required prior to use of such suction devices.
   
   
4. Use of commercially available rabbit restraint devices is acceptable provided adequate care is taken to prevent injury. Use of a tranquilizer such as Innovar-Vet may eliminate or reduce the need for restraint devices.
   
   
5. The volume of blood removed at any one survival bleeding should not exceed 1.0% of body weight, assuming 1 ml blood weighs 1 gm (40 ml for a 4 kg rabbit). In healthy rabbits this procedure may be repeated every other week without serious hematological consequences. If more frequent bleeding is needed, this volume should be proportionately reduced (e.g. only 50% of this volume taken if weekly bleeds are to be performed). More severe or more frequent bleeding requires monitoring of hematocrit, hemoglobin and total plasma protein, and prior approval. Please submit an addendum to these guidelines describing any alternative bleeding schedules.