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Guidelines for the Removal of Blood from Laboratory Animals

Often, as a critical part of the research protocol, blood or usually the serum, plasma, or cells harvested from blood are collected from laboratory animals and used to evaluate various parameters related to the focus of the research project.

Survival Blood Collection – Mammals and Birds

Blood loss can produce subtle to profound physiologic and clinical effects in the animal. Various recommendations regarding the volume of blood that can be safely collected from different species have been published1-3. As part of the IACUC review of a submitted “Protocol for the Use of Live Vertebrates”, the proposed volume and frequency of blood collection is evaluated. OLAC recommends the following simple calculation that results in volumes within the range of volumes suggested for survival blood collections.

During a single collection from a non-compromised, healthy animal, blood can be collected in a volume less than or equal to 1% of body weight. Within a 2 week period, blood can be collected in a volume totaling less than or equal to 1.5% of body weight.

 Survival blood collections in volumes larger than those recommended above may be permitted by the IACUC in cases when adequate scientific justification is defined in the submitted protocol.

To use the calculation, assume 1 mL of blood weighs approximately 1 gram, and thus:

From a 3.8 kg = 3800 gram (18-20 week old New Zealand white rabbit), 38 (3800 x 0.01) mL of blood could be collected during a single collection. Up to 57 (3800 x 0.015) mL could be collected within a two week period of time.

Similarly,

up to 0.3 mL (300 µL) could be collected from a 30 gram mouse during a single collection and 0.45 mL (450 µL) could be collected within a two week period of time.

Since the weights of animals can change with age, using phrases incorporating, “volume totaling less than or equal to 1%” or “volume totaling less than or equal to 1.5% of body weight within a two week period” are appropriate and simple ways to indicate the maximum volume of blood to be collected when submitting a protocol.

Survival Blood Collection – Reptiles

A recomendation4,5 for the volume of blood that can be taken during a single collection has been made and will be used by the UTK-IACUC.

During a single collection from a non-compromised, healthy animal, blood can be collected in a volume less than or equal to 0.7 % of body weight.

Survival blood collections in volumes larger than those recommended above may be permitted by the IACUC in cases when adequate scientific justification is defined in the submitted protocol.

Survival Blood Collection – Fish

The volume of blood/unit of body weight is considerably less in bony fish compared to mammals. Thus the recommendation6 for calculating the volume of blood that can be taken during a single blood collection based on a percentage of body weight is reduced.

During a single collection from a non-compromised, healthy fish, blood can be collected in a volume less than or equal to 0.5 % of body weight.  No more than 1% of the fish’s body weight (i.e. 1mL/kg) should be removed from a fish that will be recovered from blood collection. Survival blood collections in volumes larger than those recommended above may be permitted by the IACUC in cases when adequate scientific justification is defined in the submitted protocol.

Exsanguination

If a large volume of blood is to be collected terminally, the American Veterinary Medicine Association (AVMA) Guidelines for the Euthanasia of Animals: 2013 Edition (https://www.avma.org/KB/Policies/Documents/euthanasia.pdf) states that,

“Exsanguination can be used to ensure death subsequent to stunning, or in otherwise unconscious animals. Because anxiety is associated with extreme hypovolemia, exsanguination must not be used as a sole means of euthanasia.7 Animals may be exsanguinated to obtain blood products, but only when they are sedated, stunned or anesthetized.8

References

1Hawk, C.T. and S.L. Leary. (1995). Formulary for Laboratory Animals. Iowa State University Press, Ames, Iowa. p.79.

2Morton, D.B., et al (1993). Removal of blood from laboratory mammals and birds. Laboratory Animals 27:1-22.

3McGuill, M.W. and A.N. Rowan. (1989). Biological Effects of Blood Loss: Implications for Sampling volumes and Techniques. ILAR News 31(4):5-18.

4Jacobson, E.R. (1993), Blood Collection Techniques in Reptiles: Laboratory Investigations. In (ed: M. E. Fowler) Zoo and Wild Animal Medicine: Current Therapy 3, W.B. Saunders. Com., Denver, CO, pp. 144.

5Guidlelines the 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).

6Canada Department of Fisheries and Oceans. September 2004. 4.0 Blood sampling of Finfish.1-15.

7Blackmore, DK., (1984). Differences in behavior between sheep and cattle during slaughter. Res Vet Sci, 37: 223-226

8Gregory, NG and SB Wotton. (1984). Time to loss of brain responsiveness following exsanguination in calves. Res Vet Sci, 37: 141-143.

 

(Approved by the IACUC on 10/3/96 updated 10/24/1997-ECS and 4/22/2013)

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