SOP: Blood Collection in Cattle

These SOPs were developed by the Office of the University Veterinarian and reviewed by Virginia Tech
IACUC to provide a reference and guidance to investigators during protocol preparation and IACUC reviewers
during protocol review. They can be used as referenced descriptions for procedures on IACUC protocols.
However, it is the sole responsibility of the Principal Investigator to ensure that the referenced SOPs adequately
cover and accurately represent procedures to be undertaken in any research project. Any modification to
procedure as described in the SOP must be outlined in each IACUC protocol application (e.g. if the Principal
Investigator plans to use a needle size that is not referenced in the SOP, simply state that alteration in the
IACUC protocol itself).

Table of Contents

I. Procedure Summary & Goal ......................................................................................... 1

II. Personal Protective Equipment & Hygiene ................................................................... 1
III. Supply List..................................................................................................................... 1
IV. Detailed Procedure ........................................................................................................ 2
V. Variations ...................................................................................................................... 3
VI. Potential Adverse Effects, Mitigation, or Treatment..................................................... 3
VII. References ..................................................................................................................... 4

Version: 1
Original date of approval: 12/12/2017
Current version date: 12/12/2017
 SOP: BLOOD COLLECTION IN CATTLE

I. Procedure Summary and Goal

Describes procedures for blood collection via the jugular vein in cattle.
Considerations
Refer to SOP: Cattle Restraint for methods of restraint
Handlers should be vigilant at all times so as to avoid injury to animals or themselves.
a. Head butting and arc of swing
b. Being caught between animal and solid structure, (e.g., a wall, fence, chute)
c. Avoid being kicked or stepped on:
i. Front foot pawing
ii. Hind foot swings forward and back out to the side (“cow kick”)

II. Personal Protective Equipment (PPE) and Hygiene

a. Ensure appropriate PPE is used to protect handler from accidental injury or exposure to blood and other
body fluids, such as:
i. Scrubs or coveralls
ii. Steel-toed shoes or boots
iii. Optional
a. Disposable gloves (e.g., latex, nitrile)
b. Eye protection
iv. Other PPE as required by protocol/facility
b. Hands should be washed and/or gloves changed between animals; if vacutainer hubs used, ideally, they
should be disinfected between barns or other animal holding areas.
c. Promptly dispose of used sharps in the provided leak-proof, puncture resistant sharps container.

III. Supply List

a. Blood collection vials (e.g., syringes, vacutainer tubes)
b. Needles (18 gauge X 1.5 inch) or vacutainer needles and hub
c. Restraint (e.g., halter, squeeze chute)
d. Clippers (optional)
e. Antiseptic
f. Gauze

1
 SOP: BLOOD COLLECTION IN CATTLE

IV. Detailed Procedure

a. Restrain animal in squeeze chute, halter and secure lead to the stanchion with a quick-release knot
(Figure 1) with head elevated and jugular vein exposed.
b. Clip (optional) and swipe with antiseptic gauze to remove superficial dirt and debris. This may also
assist in visualizing raised vein.
c. Occlude jugular vein by applying pressure at the base of the jugular groove and visualize raised vein
(Figure 2).
d. With bevel up, insert needle firmly into skin and into vein at 20° angle (Figure 2).
e. If using vacutainer, once needle inserted, stabilize needle and push the vacutainer tube into hub. If you
have hit the vein, blood will flow freely into tube. Multiple tubes can be filled by removing filled tube
and replacing with fresh tube.
NOTE: Do not pull needle out of vein with vacutainer tube still attached as this will release vacuum in
vacutainer.
f. If you have missed the vein, carefully reposition needle, with vacutainer attached, until vessel
penetrated. Vessel is fairly deep and may roll away from needle. Typically no more than two to three
attempts should be made at a time to minimize distress to the animal and potential damage to the vein.
g. Alternately, you can use needle and syringe. Break the seal on the syringe by gently pulling back
before using. Clear air, and with needle attached to syringe, insert firmly needle at 20° angle, and
aspirate syringe to confirm insertion and collect blood (Figure 3).
h. Once collection complete, remove vacutainer tube, then, applying pressure over injection site, remove
needle.
i. Dispose of needle in approved Sharps container.
j. In order to ensure adequate hemostasis, apply pressure with gauze for 30 to 60 seconds.
k. Serial samples can be taken by alternating sides, and by moving insertion sites cranially, as long as
there is no hematoma formation.

Figure 1. Pull Head to the Side, and Figure 2. Occlude Jugular Vein by Figure 3. Aspirate Syringe to Collect
Secure Lead with Quick Release Knot Applying Pressure at Base of Jugular Blood
Groove and Insert Needle

2
 SOP: BLOOD COLLECTION IN CATTLE

V. Variations
Blood can be collected via the coccygeal vein (Figure 4).
1. Restrain animal.
2. Raise the tail vertically until it is horizontal to the ground.
3. Locate the groove lying in the ventral midline of the tail.
4. Swab the site with antiseptic.
5. Midway along the body of a coccygeal vertebra, insert the needle perpendicularly to the surface of
the skin to a depth of a few millimeters.
6. Withdraw blood sample and remove needle.
7. Dispose of needle in approved sharps container.
8. In order to ensure adequate hemostasis, apply pressure with gauze for 30 to 60 seconds.

Figure 4. Collecting Blood via the Coccygeal Vein

VI. Potential Adverse Effects, Mitigation, or Treatment

a. Hematoma or thrombus
i. Enter vessel at an angle of 30 degrees or less
ii. Use a gauge of needle smaller than the vein
iii. Apply pressure until bleeding has stopped (1+ minutes)
b. Pain at blood collection site
i. Use a needle of smaller gauge than the vein
ii. Practice on vein models prior to live animal
c. Infection at blood collection site
i. Use sterile single-use devices only
ii. Clean work surfaces with disinfectant

3
 SOP: BLOOD COLLECTION IN CATTLE

iii. Wear gloves, wash hands

iv. Contact a qualified veterinarian for treatment recommendations if any of the following are
noted.
a. Heat, pain, swelling first noted at the insertion site of the blood draw, purulent material
draining from the insertion site.
b. Induration (hardening) of the vessel
c. Pyrexia, local or systemic infections, septic shock

VII. References
American Association of Laboratory Animal Science. Assistant Laboratory Animal Technician Training
Manual. (Memphis, TN: Drumwright and Co, 2012)
Cattle Restraint Laboratory Manual. Foundations in Veterinary Medicine – Restraint of Cattle. Great Plains
Veterinary Educational Center, University of Nebraska-Lincoln. Lincoln, Nebraska
Donovan, J., and Brown, P. Removal of blood from laboratory mammals and birds: First report of the
BVA/FRAME/RSPCA/UFAW Joint working group on refinement. Lab Animal 27, 1-22. (1993)
Hawk, C.T., Leary, S.T., and Morris, T.H. Formulary for Laboratory Animals (3rd ed.). (Ames, Iowa:
Blackwell Publishing, 2005)
Iowa State University Laboratory Animal Resources Cattle Basics
http://www.lar.iastate.edu/index.php?option=com_content&view=article&id=183&Itemid=204
McCurnin, D., and Bassert, J. Clinical Textbook for Veterinary Technicians (5th ed.). (Philadelphia,
PA:Saunders Elsevier 2002)