Blood Collection Guidelines

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					Adopted: September 1, 2010

                                    IOWA STATE UNIVERSITY
                         Institutional Animal Care and Use Committee

                                  Blood Collection Guidelines

To provide Iowa State University (ISU) Institutional Animal Care and Use Committee (IACUC)
guidelines for blood collection and dose volume administration.

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

    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

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Adopted: September 1, 2010

                     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

                                                                         Multiple Sampling
                     Single Sampling                              (e.g., pharmacokinetic study)
                                                          % circulatory blood
    % circulatory blood           Approximate recovery     volume removed          Approximate recovery
     volume removed                      period                 in 24h                    period

           7.5%                          1 week                 7.5%                      1 week
           10%                           2 weeks               10–15%                     2 weeks
           15%                           4 weeks                 20%                      3 weeks

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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

                             Total Blood
   Species (Weight)           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)   50. (ml)       67 (ml)           100(ml)           134 (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)    2.7 (L)        3.7 (L)           5.4 (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 vein when taking samples. Samples taken from different sites may
         show variances in clinical pathology values and may cause inconsistencies with respect to
         historical data.

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Adopted: September 1, 2010

    Table 4 Recommended Blood Sampling Sites by Species

                    Species                             Recommended Sampling Location
      mouse                         saphenous vein, facial vein
      rat                           lateral tail vein, jugular vein, saphenous vein, facial vein
      gerbil                        saphenous vein, facial vein
                                    retro-orbital sinus, jugular vein, cardiac puncture, tail vein, femoral
      hamster                       vein
      guinea pig                    saphenous vein
      rabbit                        central ear artery, jugular vein, marginal ear vein
                                    jugular vein, cephalic vein, retro-orbital sinus, tail artery, tarsal vein
      ferret                        (saphenous), cardiac puncture
      cat                           cephalic vein, jugular vein
      dog                           cephalic vein, jugular vein, saphenous vein
      pig                           jugular vein, cranial vena cava (blind stick)
      cattle                        tail vein, jugular vein
      sheep/goat                    jugular vein, cephalic vein
      horse                         jugular vein
      snake                         palatine vein, ventral tail vein, and heart
      lizard                        ventral tail vein (large lizards)
                                    toe nail clip and micro capillary tube (small lizards)
      turtle                        heart, jugular vein, brachial vein
      fish                          anterior vena cava
      frog                          cardiac puncture, retro-orbital sinus, median abdominal vein
                                    brachial vein, cutaneous ulnar vein, right jugular vein, cardiac
      bird                          puncture, medial metatarsal vein
                                    cardiac puncture, vena cava using dissection for exposure
      all species                   (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.

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Adopted: September 1, 2010

    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 vein    caudo-dorsal       lower right       thorax or
                                      or saphenous         neck or thorax     quadrant of       abdomen
                                      vein                                    abdomen

  rat            caudal hind limb     lateral tail vein    caudo-dorsal       lower right       thorax or
                                      or saphenous         neck or thorax     quadrant of       abdomen
                                      vein                                    abdomen

  guinea pig     caudal hind limb     ear vein or          caudo-dorsal       lower right       dorsal thorax
                                      saphenous vein       neck or thorax     quadrant of

  rabbit         caudal hind limb     marginal ear         caudo-dorsal       uncommon          thorax or
                 or perilumbar area   vein or jugular      neck or thorax                       abdomen

  dog            caudal hind limb,    cephalic vein,       caudo-dorsal       uncommon          thorax or
                 perilumbar area,     saphenous vein,      neck or thorax                       abdomen
                 quadriceps, or       or jugular vein

  cattle         neck muscle          jugular vein, tail   neck or shoulder   NA                thorax or neck

  goat/sheep     neck muscle          jugular vein         skin below elbow   NA                thorax or neck

  pig            base of ear          ear vein,            base of ear        NA                thorax or neck
                                      anterior vena
                                      cava, jugular

  horse          neck muscle          jugular vein         neck               NA                thorax or neck

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Adopted: September 1, 2010

    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                            Responsibilities
 Research Staff                   Researchers provide complete descriptions of proposed animal procedures to
                                  the IACUC for review and use these guidelines as appropriate.

 IACUC Members                    IACUC members review proposed animal activities to ensure they fall within
                                  the listed guidelines unless appropriate scientific justification is provided.
 LAR Clinical Veterinarians       Provide guidance regarding administration sites, dosing volumes, blood
 and/or Attending Veterinarian    collection sites, and vehicle selection as needed.

Contact Information
Direct questions about this document to Mary Sauer, VMD, Attending Veterinarian.

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.

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