IACUC Administrative Policy P : Blood Collection Guidelines

Transcription

1 1 I. PURPOSE: Institutionl Animl Cre & Use Committee Cmpus Mil: 108C Foust Hll Telephone: Fx: IACUC Administrtive Policy P : Blood Collection Guidelines A. This document is designed to provide generl guidelines on blood collection methods for common lbortory nimls. All procedures must be pproved by the Institutionl Animl Cre nd Use Committee (IACUC), however protocol my reference these guidelines nd describe in detil ny devitions from them. For trining on specific blood collection methods nd techniques, plese contct: B. It is recommended to tke no more blood thn is bsolutely necessry. Remember to clculte beforehnd the minimum mount of blood necessry to perform ll tests nd ssys, nd tht the serum frction is bout ½ of the totl blood volume. When clculting blood volumes bsed on body weights (see below), remember tht body weights in kilogrms (kg) will convert to blood volumes in liters, nd weights in grms will convert to volumes in milliliters (ml). II. PROCEDURES: A. Approximte Blood Volume % (men of 6%) of body weight = totl blood volume b The circulting blood volume cn generlly be estimted s ml/kg of totl body weight. However, cre should be tken in these clcultions s the % of totl blood volume will be lower (-15%) in sick, obese nd older nimls. See Tble 2 t end of document for some specific blood volumes. B. Blood Collection Volumes 1. 1% of body weight = mximum volume per collection every 14 dys, without requiring supplementl fluids. This pplies for single blood collections s well s repeted collections. For irregulr smpling schedules, clculte the totl mount needed over 14 dy spn % of body weight = mount tht cn be tken dily without requiring supplementl fluids 3. 3% of body weight = mount expected t exsnguintions (pproximtely 9ml from 300g rt).

2 2 C. Single Blood Drw 1. Mximum of 1% of body weight cn be removed s single blood drw every 14 dys, without requiring dministrtion of supplementl replcement fluids. Withdrwing the minimum mount of blood necessry is strongly recommended. Exmples: b 0.15 ml from 15 g mouse 3 ml from 300 g rt D. Multiple Blood Drws 1. If the totl volume withdrwn over 14-dy period is less thn 1% BW, then no dditionl ction needs to be tken. 2. If the totl volume withdrwn over 14 dy period will be 2% BW (or over), fluid volume replcement is necessry. Withdrwing the minimum mount of blood necessry is strongly recommended. Exmples: b c d Up to 0.15 ml withdrwn from 15g mouse over 2 weeks is OK Up to 0.3 ml withdrwn from 15g mouse over 2 weeks, replce volume with 0.3 ml sline SQ Up to 3 ml from 300g rt over 2 weeks is OK Up to 6ml from 300g rt over 2 weeks, replce volume with 6 ml sline SQ 3. As helpful guideline, dily blood drws under 0.07% BW will keep the totl 2-week withdrwl under 1% BW. E. Exsnguintion/Terminl Blood Collection 1. Approximtely 50-75% of totl blood volume (3% men, 4-5% mximum of body weight) cn be obtined by terminl exsnguintion. The niml must be deeply nesthetized, or recently euthnized, prior to exsnguintion. Since the mount of blood obtined is substntilly incresed if the hert is beting during the bleeding procedure, use of surgicl plne of nesthesi is recommended. The procedure for nesthesi nd/or euthnsi must be described fully in the pproved IACUC protocol. Exmple mximums: ml from 15g mouse b ml from 300g rt F. Fluid Replcement If the volume of blood removed from n niml exceeds the mximum recommended blood collection volumes (i.e., > 1% body weight every 14 dys), replcement of the removed volume of blood with wrm (30-35ºC) isotonic solution (e.g., 0.9% sline, lctted Ringer s solution) constitute ccepted veterinry prctice. When this volume of blood is collected, it should be withdrwn t slow, stedy rte, nd the volume of solution to be infused should be dministered similrly. Contct ANIMALCARE@cmich.edu for trining on specific methods. Recommended

3 3 route(s) of dministrtion: 50% of volume ech delivered IV nd SubQ or IP. G. Monitoring 1. If too much blood is withdrwn too rpidly or too frequently without replcement, the niml my go into hypovolemic shock. If signs of shock re observed, contct the ttending veterinrin immeditely. Signs of shock include: Fst nd thredy pulse b Ple dry mucous membrnes c Cold skin nd extremities d Restlessness e Hyperventiltion f Sub-norml body temperture 2. If 15-20% of totl blood volume is removed, crdic output nd blood pressure will be reduced. 3. If 30-40% of totl blood volume is removed, deth will result in t lest 50% of nimls. 4. If > 40% of totl blood volume is removed, deth of the niml is expected. 5. Stressed, sick, or otherwise compromised nimls my not tolerte the blood collection criteri noted bove, which is for helthy nimls. 6. By monitoring hemtocrit (Hct or PCV) nd/or hemoglobin (Hb) it is possible to evlute if the niml hs sufficiently recovered from single or multiple blood drws. Remember it my tke up to 24 hours for hemtocrit or hemoglobin to reflect sudden or cute blood loss. In generl, if the niml is nemic (below the norml PCV rnge for the species), or if the hemoglobin concentrtion is less thn 10 g/dl, it is not sfe to remove the volumes of blood listed bove. H. Blood Collection Sites nd Methods Tble 1 lists the blood collection sites for common lbortory niml species. They re listed from most common/desirble to lest common/desirble bsed on ese of collection from the site. For uncommon lbortory niml species, plese contct the ttending veterinrin Dr. Robert Werner t werne2r@cmich.edu or ANIMALCARE@cmich.edu. For smller species, the volume of blood ttinble for ech site is listed; however, the volumes re estimtion nd will lso depend on the size, heth, nd hydrtion sttus of the niml s well s the experience nd skill level of the person collecting the smple. Bsed on the gols nd requirements of the study, certin sites my be preferble. Additionlly, publictions hve indicted tht the results from blood nlysis (especilly cellulr indices) my vry bsed on the site of blood collection; consult the literture for more informtion. In ll cses, crdic puncture my be used to obtin single, lrge volume of blood from hevily nesthetized (terminl procedure only) or euthnized nimls.

4 4 Tble 1 Frequently Used Sites For Blood Drws for Common Lb Animl Species Species Site Anesthesi Repet Bleeds* Expected Volume Mouse Lterl til vein No Yes µl Sphenous vein No Yes µl Submndibulr vein No Yes µl (discourged) Distl til trnsect./lncing Required** Yes- limited µl (discourged) Retro-orbitl sinus Required Yes- limited 200 µl Sublingul vein*** Required Yes 500 µl Jugulr vein Recommended Yes Crdic puncture (terminl Required Terminl ~ 1 ml only) Dorsl mettrsl/pedl No Yes < 100 µl Rt Sphenous vein No Yes µl Lterl til vein*** No Yes µl Dorsl mettrsl/pedl No Yes µl Submndibulr/Fcil vein No Yes µl Jugulr vein Recommended Yes ml (discourged) Sublingul vein*** Required Yes ml Retro-orbitl plexus Required Yes-limited ml Crdic puncture Required Terminl ~3 ml (terminl only) Ventrl til Recommended yes 100 µl til rtery Required yes ml Hmster Lterl trsl vein No Yes µl Toenil clipping No Yes µl Retro-orbitl sinus Required Yes- limited µl Jugulr vein Required Yes ml Crdic puncture Required Terminl ~3 ml (terminl only) Gerbil Lterl til vein No Yes µl Toenil clipping No Yes µl Distl til trnsection (1-3 mm) Required Yes (1-2 times only) µl Retro-orbitl sinus Required Yes- limited µl Jugulr vein Required Yes ml Crdic puncture Required Terminl ~3 ml (terminl only) Guine pig Auriculr vein No Yes µl Cephlic vein No Yes µl Rbbit Sphenous vein No Yes µl Jugulr vein Recommended Yes 2-3 ml Crnil ven cv Recommended Yes 2-3 ml Crdic puncture (terminl only) Required Terminl Mrginl er vein/centrl er Locl nesthesi Yes 1-3 ml rtery recommended Lterl sphenous vein No Yes Cephlic vein No Yes Jugulr vein Recommended Yes Crdic puncture (terminl Required Terminl only) *Plese refer to section D bove for detils. **Distl til trnsection in dult mice requires the use of generl nesthesi unless scientificlly justified nd pproved in the IACUC protocol. ***Anesthesi is recommended for sublingul blood collection. ****Contct ANIMALCARE@cmich.edu for trining.

5 5 Tble 2 Circulting blood volumes in common lb niml species (dopted from Heinz-Diehl, 2001 nd Hwk et l. 2005) Species Men blood volume (ml/g) Mouse Rt Hmster.078 Gerbil.067 Guine pig Rbbit Rnge of men blood volume (ml/g) REFERENCES Clemons DJ, Seemn JL The Lbortory Guine Pig. Boc Rton (FL): CRC Press, p Ebert RV, Sted EA, Gibson JG Response of norml subjects to cute blood loss. Arch Int Med; 68: BVA/FRAME/RSPCCA/UFAW Joint Working Group on Refinement Removl of blood from lbortory mmmls nd birds (first report). Lbortory Animls; 27:1-22. Field KJ, Sibold AL The Lbortory Hmster nd Gerbil. Boc Rton (FL): CRC Press, p Hwk CT, Lery SL, Morris TH Formulry for Lbortory Animls. Ames (IO): Blckwell Publishing. p Heimnn M, Käsermnn HP, Pfister R, Roth DR, nd Bürki K Blood collection from the sublingul vein in mice nd hmsters: suitble lterntive to retrobulbr technique tht provides lrge volumes nd minimizes tissue dmge. Lb Animl; 43: Heinz-Diehl K, Hull R, Morton D, Pfister R, Rbemmpinin Y, Smith D, Vidl JM, Vorstenbosch C A good prctice guide to the dministrtion of substnces nd removl of blood, including routes nd volumes. J Appl Toxicol; 21: Hoff J. Methods of blood collection in the mouse Lb Animl; 29(10): McGuill MW, Rown AN Biologicl effects of blood loss: Implictions for smpling volumes nd techniques. ILAR Journl; 31(4):5-20. Nerenberg ST, Zedler P, Prsd R, Biskup N, Pedersen L Hemtologicl response of rbbits to chronic, repetitive, severe bleedings for the production of ntiser. J Immunol Meth; 24: Otto G, Rosenbld WD, Fox JG Prcticl venipuncture techniques for the ferret. Lb Anim 27:26-29 Scipioni RL, Diters RW, Myers WR, Hrt SM Clinicl nd clinicopthologic ssessment of seril phlebotomy in the Sprgue Dwley rt.

6 6 Lb Anim Sci; 47(3): Scipioni RL, Guidi DA, Stehr JE, Hrt SM, Diters RW Clinicl, hemtologic, nd clinicochemicl ssessment of seril blood smple collection in Sprgue- Dwley rts. Contemp Top Lb Anim Sci; 35(6):90. [Abstrct] Skvlen PA, Bron SJ, Stevens JO Effect of blood collection volumes on the hemogrms of rbbits. Contemp Top Lb Anim Sci; 31(4):23. [Abstrct] Villno JS, Shrp PE The Lbortory Rt. Boc Rton (FL): CRC press, p Yle CE, Torhortst JB Criticl bleeding nd plsm volumes of the dult germfree rt. Lb Anim Sci; 22(4):