IOWA STATE UNIVERSITY Institutional Animal Care and Use Committee Blood Collection Guidelines Purpose To provide Iowa State University (ISU) Institutional Animal Care and Use Committee (IACUC) guidelines for blood collection and dose volume administration. Scope This guidance is applicable to all individuals from all departments performing research and teaching related procedures on laboratory animals. Veterinary medical and animal care procedures are outside the scope of these guidelines when they are unrelated to the research project and when they are performed to improve animal health or well-being or to treat an illness or injury under the direction of the ISU Attending Veterinarian or designee. General Requirements These guidelines are intended to provide general guidance to teachers and researchers planning experimental procedures on animals. Exceptions to these guidelines may be approved by the IACUC based on written scientific justification provided by the investigator. For species or guidelines not listed, the Attending Veterinarian and LAR clinical veterinarians can assist with the selection of appropriate procedures. Procedure A. Circulating blood volume by species (See Table 1.) The volume of blood that can safely be removed from animals without causing hypovolemia or other physiologic stress is related to their circulating blood volume. The following table lists the average blood volume of healthy animals. For IACUC purposes, the volumes listed will be used when calculating whether the proposed amount of blood to be removed falls within appropriate guidelines. Office for Responsible Research Page 1 of 6
Table 1 Circulating Blood Volume in Laboratory Animals Species Mean Blood Volume (ml/kg) mouse 72 rat 64 gerbil 72 guinea pig 70 hamster 72 rabbit 56 dog (beagle) 85 pig 67 cattle (400 kg) 60 sheep (45 kg) 60 horse (500 kg) 75 goat (40 kg) 70 snake (100 g) 50 80 B. Volume withdrawal limits and recovery periods by species (See Tables 2 and 3.) The following table lists the approximate recovery period after some blood is collected before additional samples should be collected. For IACUC purposes, the volumes and times listed will be used when calculating whether the proposed amount of blood and the proposed recovery period fall within appropriate guidelines. Table 2 Appropriate Recovery Periods Single Sampling Multiple Sampling (e.g., pharmacokinetic study) % circulatory blood volume removed Approximate recovery period % circulatory blood volume removed in 24h Approximate recovery period 7.5% 1 week 7.5% 1 week 10% 2 weeks 10 15% 2 weeks 15% 4 weeks 20% 3 weeks Office for Responsible Research Page 2 of 6
To simplify calculations, the following table lists the total blood volume and relevant collection volumes for the species and body weights listed. Table 3 Total Blood Volumes and Relevant Collection Volumes Species (Weight) Total Blood Volume 7.5% 10% 15% 20% mouse (25 g) 1.8 (ml) 0.1 (ml) 0.2 (ml) 0.3 (ml) 0.4 (ml) rat (250 g) 16 (ml) 1.2 (ml) 1.6 (ml) 2.4 (ml) 3.2 (ml) gerbil (50 60 g) 3.6 (ml) 0.3 (ml) 0.4 (ml) 0.6 (ml) 0.7 (ml) guinea pig (900 g) ) 62 (ml) 4.7 (ml) 6.2 (ml) 9.3 (ml) 12.4 (ml) hamster (100 g) 7.2 (ml) 0.5 (ml) 0.7 (ml) 1.0 (ml) 1.4 (ml) rabbit (4 kg) 224 (ml) 17 (ml) 22 (ml) 34 (ml) 45 (ml) dog (10 kg) 850 (ml) 64 (ml) 85 (ml) 127 (ml) 170 (ml) snake (100 g) 7.2 (ml) 0.5 (ml) 0.7 (ml) 1.0 (ml) 1.4 (ml) pig (100 kg) 6.7 (L) 500 (ml) 670 (ml) 1000 (ml) 1340 (ml) cattle (400 kg) 24 (L) 1.8 (L) 2.4 (L) 3.6 (L) 4.8 (L) sheep (45 kg) 2.7 (L) 202 (ml) 270 (ml) 404 (ml) 540 (ml) horse (500 kg) 37.5 (L) 2.8 (L) 3.7 (L) 5.6 (L) 7.4 (L) goat (40 kg) 2.8 (L) 210 (ml) 280 (ml) 420 (ml) 560 (ml) C. Blood sampling sites by species For general guidance, Table 4 lists recommended sites for blood collection by species. Other sites may be used, and a Laboratory Animal Resources (LAR) veterinary clinician may be consulted for assistance in selecting a site. It is important that the minimum number of needle punctures be performed consistent with achieving good data. The same puncture site should not be used, but rather vary the points along the when taking samples. Samples taken from different sites may show variances in clinical pathology values and may cause inconsistencies with respect to historical data. Office for Responsible Research Page 3 of 6
Table 4 Recommended Blood Sampling Sites by Species mouse rat gerbil hamster guinea pig rabbit ferret cat dog pig cattle sheep/goat horse snake lizard turtle fish frog bird all species Species Recommended Sampling Location saphenous, facial lateral tail, jugular, saphenous, facial saphenous, facial retro-orbital sinus, jugular, cardiac puncture, tail, femoral saphenous central ear artery, jugular, marginal ear jugular, cephalic, retro-orbital sinus, tail artery, tarsal (saphenous), cardiac puncture cephalic, jugular cephalic, jugular, saphenous jugular, cranial vena cava (blind stick) tail, jugular jugular, cephalic jugular palatine, ventral tail, and heart ventral tail (large lizards) toe nail clip and micro capillary tube (small lizards) heart, jugular, brachial anterior vena cava cardiac puncture, retro-orbital sinus, median abdominal brachial, cutaneous ulnar, right jugular, cardiac puncture, medial metatarsal cardiac puncture, vena cava using dissection for exposure (terminal samples only under anesthesia) Cannulation is an important technique for repeated bleeding. Butterfly needles and over-the-needle cannulae may be used for the short term (day); however for long-term use, surgical implantation of a tissue-compatible cannula should be considered. This method of cannulation allows for repeated blood drawing with minimal discomfort and distress for the animal. For long-term use, a subcutaneous vascular access port is preferred. Long-term cannulation may lead to thrombosis of the vessel and other medical complications. It is critical that when choosing a technique for blood sampling, the researcher gives careful consideration to possible adverse side effects associated with the procedure and consults a LAR veterinary clinician. Office for Responsible Research Page 4 of 6
D. Dose administration sites by species The following table lists common administration sites by species. Additional sites may be used, and an LAR veterinarian can provide assistance in selecting administration sites. Table 5 Common Administration Sites by Species Species Intramuscular Intravenous Subcutaneous Intraperitoneal Intradermal mouse caudal hind limb lateral tail or saphenous lower right quadrant of rat caudal hind limb lateral tail or saphenous lower right quadrant of guinea pig caudal hind limb ear or saphenous lower right quadrant of dorsal thorax rabbit caudal hind limb or perilumbar area marginal ear or jugular uncommon dog caudal hind limb, perilumbar area, quadriceps, or triceps cephalic, saphenous, or jugular uncommon cattle neck muscle jugular, tail neck or shoulder NA neck goat/sheep neck muscle jugular skin below elbow NA neck pig base of ear ear, anterior vena cava, jugular base of ear NA neck horse neck muscle jugular neck NA neck Office for Responsible Research Page 5 of 6
E. Special circumstances 1. Anesthesia Anesthesia is required if blood collection is being performed either via the retro-orbital sinus or by cardiac puncture due to the distress and pain which can be caused and for the serious complications (injury to the eye, cardiac tamponade and death) associated with these routes. 2. Terminal blood withdrawal All cardiac punctures must be done under anesthesia as a terminal procedure, unless justification for survival is approved by the IACUC. Roles and Responsibilities Roles Research Staff IACUC Members LAR Clinical Veterinarians and/or Attending Veterinarian Responsibilities Researchers provide complete descriptions of proposed animal procedures to the IACUC for review and use these guidelines as appropriate. IACUC members review proposed animal activities to ensure they fall within the listed guidelines unless appropriate scientific justification is provided. Provide guidance regarding administration sites, dosing volumes, blood collection sites, and vehicle selection as needed. Contact Information Direct questions about this document to Mary Sauer, VMD, Attending Veterinarian. References The guidelines in this document were taken from the following references: Diehl, K.L., Hull, D., Phister, R., et al. A Good Practice Guide to the Administration of Substances and Removal of Blood, Including Routes and Volumes, Journal of Applied Toxicology 21: 15-23, 2001. Fowler, M.E., and R.E. Miller. 2008. Zoo and Wild Animal Medicine: Current Therapy. Guidelines for Use of Live Amphibians and Reptiles in Field and Laboratory Research, Second Edition, revised by the Herpetological Animal Care and Use Committee (HACC) of the American Society of Ichthyologists and Herpetologists, 2004. (Committee Chair: Steven J. Beaupre; Members: Elliott R. Jacobson, Harvey B. Lillywhite, and Kelly Zamudio). Lillywhite, H. B., and A. W. Smits. 1984. Liability of blood volume in snakes and its relation to activity and hypertension. Journal of Experimental Biology 110, 267-274. Lillywhite, H. B., R. A. Ackerman, and L. Palacios. 1983. Cardiorespiratory responses of snakes to experimental hemmorage. Journal of Comparative Physiology 152, 59-65. State of Victoria, Department of Primary Industries 2007, Guidelines for the Harvesting of Blood from Livestock for Commercial and Research Purposes. Office for Responsible Research Page 6 of 6