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Refinement Alternatives
Dosing and Blood Sampling
Efforts should constantly be made to refine scientific techniques so as to decrease the volume
of blood samples required. In small animals such as mice, sample volume and frequency of sampling
are of particular importance. If the animal's welfare may be compromised by the volume of the
sample required, either more animals should be used or a compensatory volume replacement
(with warmed saline or a blood transfusion) should be performed. In general, the following points
should be considered:
- rather than multiple sampling carried out by repeated needle punctures, a butterfly needle or a
percutaneous (over the needle) cannula taped in position should be used;
- in removing volumes greater than 0.1 ml, a needle with as large a bore as possible should be
used, in order to ensure rapid blood withdrawal without collapsing the vein or causing formation
of a hematoma;
- before taking a sample, it is important to accurately locate the vein and dilate it by gentle
obstruction or warming (if general body warming is used, the animal must be constantly
observed to prevent hyperthermia, as evidenced by more rapid breathing, panting or salivating); and
- the use of xylene (xylol, dimethylbenzene) as a dilator is not recommended as it causes skin
rashes and is easily misused.
Efforts should also constantly be made to refine techniques for animal dosing (including reducing
volume size; improving solvent formulations and minimizing distress associated with dosing procedures).
For more information on blood sampling and dosing, the following resources may be useful:
CCAC resources
Online resources
- Fisheries and Oceans Canada (DFO) (2004) Blood sampling of finfish. Canada Department of
Fisheries and Oceans animal-user training template 4.
- Hem A., Smith A.J. & Solberg, P. (1998). Saphenous vein puncture for blood sampling of the
mouse, rat, hamster, gerbil, guinea pig, ferret and mink.
Laboratory Animals
32:364-368.
- Diehl K-H., Hull R., Morton D., Pfister R., Rabemampianina Y., Smith D., Vidal J-M. & van
de Vorstenbosch C. (2001) A good practice guide to the administration of substances and removal
of blood, including routes and volumes. Journal of Applied Toxicology 21:15-23.
- Morton D., Jennings M., Buckwell A., Ewbank R., Godfrey C., Holgate B., Inglis I., James R., Page C.,
Sharman I., Verschoyle R., Westall L. & Wilson A.B. (2001) Refining procedures for the administration
of substances. Report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement. Laboratory
Animals 35:1-41.
- National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) (2008).
Blood Sampling Microsite.
- This UK-based website provides information and photographs describing the
techniques for blood sampling from mice, rats, hamsters, guinea pigs, rabbits,
ferrets, dogs, pigs and marmosets. It also contains information about the
general principles of blood sampling and choosing the most appropriate
technique, and references for further information. However, the CCAC
considers that some of the methods described in this site (e.g., retro-orbital
bleeding and decapitation) should only be carried out in exceptional
circumstances, following careful review and approval by an animal care committee,
based on scientifically justified necessity.
- van Herck H., Baumans V., Brandt C.J.W., Hesp A.P.M., Sturkenboom J.H., van Lith H.A.,
van Tintelen G. & Beynen A.C. (1998) Orbital sinus blood sampling in rats as performed
by different animal technicians: the influence of technique and expertise.
Laboratory Animals 32:377-386.
- Zeller W., Weber H., Panoussis B., Burge T. & Bergmann R. (1998) Refinement of blood
sampling from the sublingual vein of rats. Laboratory Animals 32:369-376.
Other publications
- Coleman K., Pranger L., Maier A., Lambeth S.P., Perlman J.E., Thiele E. & Schapiro S.J.
(2008) Training rhesus macaques for venipuncture using positive reinforcement techniques:
A comparison with chimpanzees. Journal of the American Association for Laboratory Animal
Science 47:37-41.
- Christensen S.D., Mikkelsen L.F., Fels J.J., Bodvarsdóttir T.B. & Hansen A.K. (2009) Quality of plasma sampled by different methods for multiple blood sampling in mice. Laboratory Animals 43: 65-71.
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