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Blood Collection (Guideline)

IACUC Guidelines:

Blood Collection

 

Guidelines: The IACUC has provided a set of guidance documents (Policies, Guidelines, and Informational Sheets) for use when planning animal procedures at the University of Iowa. An exception to a Guideline must be described and justified in the Animal Protocol and approved during the normal review process.

 

Purpose:

This document provides direction and guidance on appropriate blood collection methods and volumes for animals used in research at the University of Iowa.  These guidelines are intended for use by qualified personnel performing blood collection as described on an IACUC-approved Animal Protocol.

 

There are several factors to consider when determining the appropriate blood collection volume and technique. These include:

  • The species to be sampled
  • The size of the animal to be sampled
  • The age and health of the animal to be sampled
  • The minimum volume required for analysis
  • The frequency of sampling necessary
  • The training and experience of the personnel performing the collection
  • The suitability of sedation and/or anesthesia

The sample volume selected should always be the minimum volume of blood which satisfies experimental needs.  Appropriate restraint (physical or chemical) should be employed to minimize risk of injury to the animal and personnel.

Guidelines for calculation of collection volume:

  • The maximum permitted blood volume includes blood lost during collection.
  • As a general rule, 20 drops = 1 mL  (i.e. 5 drops = 250 uL)

Maximal blood collection limits are as follows:

  • No more than 1% of the animal’s body weight in one collection or over a 24 hour period
    • For example: 25g mouse x 1% = 0.25mL or 250uL maximum blood removal
  • No more than 1.5% of the animal’s body weight in two weeks (14 days)
    • For example: 200g rat x 1.5% = 3.0mL maximum over 14 days

Frequent Rodent Calculations

Species

Weight

Maximum blood loss at one time/ in 24 hours

Maximum blood loss  over 14 days

Mouse

20 g

200 uL

300 uL

 

25 g

250 uL

375 uL

 

30 g

300 uL

450 uL

Rat

200 g

2.0 mL

3 mL

 

250 g

2.5 mL

3.75 mL

 

300 g

3.0 mL

4.5 mL

 

 

Species

Common Blood Collection Route(s)

Sedation Recommended

Anesthesia Required

Mouse

Submandibular vein

 

 

Tail vein

 

 

Saphenous vein

 

 

Retro-orbital sinus (see below)

 

Yes

Cardiac (non-survival)

 

Yes

Rat

Tail vein

 

 

Saphenous vein

 

 

Jugular vein

Yes

 

Retro-orbital plexus (see below)

 

Yes

Sublingual vein

Yes

 

Cardiac (non-survival)

 

Yes

Ferret

Cephalic vein

 

 

Saphenous vein

 

 

Jugular vein

Yes

 

Cranial vena cava

Yes

 

Rabbit

Marginal ear vein

 

 

Central auricular artery

 

 

Saphenous vein

Yes

 

Jugular vein

Yes

 

Cardiac (non-survival)

 

Yes

Hamster

Saphenous vein

 

 

Cephalic vein

 

 

Jugular vein

Yes

 

Cranial vena cava

Yes

 

Cardiac (non-survival)

 

Yes

Guinea Pigs

Ear vein (droplet)

 

 

Saphenous vein

 

 

Cranial vena cava

Yes

 

Cardiac (non-survival)

 

Yes

Gerbils

Lateral saphenous vein

 

 

Cranial vena cava

Yes

 

Cardiac (non-survival)

 

Yes

Xenopus

Dorsal tarsal vein

 

Yes

Cardiac (survival)

 

Yes

Cardiac (non-survival) (also tadpoles)

 

Yes

Pigeon

Brachial wing vein

Yes

 

Dog, Cat

Cephalic vein

 

 

Saphenous vein

 

 

Jugular vein

 

 

Cardiac (non-survival)

 

Yes

Pig

Ear vein

Yes

 

Cranial vena cava

Yes

 

Jugular vein

Yes

 

Cardiac (non-survival)

 

Yes

Ruminants

Jugular vein

 

 

Lateral saphenous vein

 

 

Tail vein

 

 

Ear vein

 

 

 

Retro-Orbital Sampling:

Retro-orbital blood collection in rodents can provide moderate to large amounts of blood when performed by well-trained personnel. However, severe injuries may occur to the animal if this procedure is not done properly, and available alternatives should be used whenever possible.

The use of retro-orbital bleeding must be described in the protocol and approved by the IACUC. Because rats have a venous plexus rather than a sinus (as in the mouse), the use of this method may result in greater tissue damage and alternative collection sites are strongly recommended.

If retro-orbital collection is necessary, the following guidelines apply:

  • General anesthesia is required
  • Microhematocrit tubes that hold 50 - 75 microliters are recommended to minimize risk of injury
  • Only one eye may be sampled at any time
    • If attempted collection from one eye is unsuccessful, an alternate method approved in the Animal Protocol (e.g. submandibular or saphenous route) must be used, rather than reattempting retro-orbital collection from the same or opposite eye
  • Alternate between left and right eyes per session
  • No more than 1 collection performed per 7 days (alternate eyes). therefore 14 days between collections in the same eye
    • Exception:  If repeated sampling within 8 hours is necessary and approved in the Animal Protocol, the retro-orbital sinus may be re-sampled by disrupting the blood clot (from the original collection site) without repeated damage to the sinus, provided the 24 hour maximum blood collection limits are not exceeded
      • Please consult with veterinary staff for demonstration and training of proper technique to reduce risk of trauma
  • A maximum of 3 procedures may be performed per eye (up to 6 collections total)
  • If injury and/or rupture of the eye or surrounding tissues occurs due to this method, the animal must be immediately euthanized or an OAR veterinarian consulted for guidance

Application of a topical ophthalmic anesthetic during/after collection should be considered to provide post-procedural analgesia.

 

Last Reviewed by the IACUC 12/12/18