Large Volume Blood Collection from Swine for In Vitro Applications: Use of Intracardiac Cannulation and a Vacuum Pump as a Terminal Procedure

Summary

Here we present a method to collect a large volume of blood from swine using percutaneous intracardiac cannulation connected to a vacuum pump, as a terminal procedure.

Abstract

Routine blood collection is required in many basic and applied research studies. Multiple methods for swine blood collection are well described. These include using the auricular veins for the collection of small blood volumes, and the jugular veins or cranial vena cava for the collection of larger blood volumes. Some experiments require large volumes of blood from one individual animal at one time for analysis or use in primary cell cultures, which can best be achieved through exsanguination. Blood collected for primary cell culture must be freshly and carefully processed to maintain cellular integrity. We describe a method of aseptically harvesting blood from swine under a surgical plane of anesthesia, using a large-bore intravascular catheter stylet inserted percutaneously into the heart and a vacuum pump, which is efficient, provides a large yield, and minimizes cell lysis. This technique is performed once on an animal, at a surgical plane of anesthesia, as a terminal procedure, providing minimal risk for pain or distress.

Introduction

Pigs have been used in research in studies ranging from cardiovascular studies to wound healing and toxicology, as well as agricultural-based research, immunology, and disease studies¹. Blood collected in research can be used to study diseases, pharmacology, genetics, immune system research, and biomarker discovery. Cell culture is one of the most basic procedures done in research and allows researchers to study cell behavior and function, as well as disease models and vaccine development². Cultivation of disease-causing pathogens, such as African Swine Fever Virus, is frequently done in a predetermined culture medi....

Protocol

This study was approved by the Midwest Veterinary Services Inc. Animal Care and Use Committee. As the IACUC-approved, primary method of euthanasia during this procedure is exsanguination, it is expected that some pigs will reach cardiac arrest due to blood loss. When this occurs, a secondary euthanasia method, such as bilateral thoracotomy, is performed. After the required blood volume is collected, if there is a palpable heartbeat, a syringe with euthanasia solution is attached to the stylet and administered intracardiac as the primary euthanasia method, followed by a secondary physical method. Swine are maintained in accordance with the Guide for the Care and Use of....

Results

Prior to blood collection, sows were prescreened for Mycoplasma hyopneumoniae, porcine adenovirus, porcine parvovirus, swine coronaviruses (transmissible gastroenteritis, porcine epidemic diarrhea), porcine hemoagglutinating encephalomyelitis, porcine reproductive and respiratory syndrome, influenza group A, circovirus 1 & 2, and Seneca virus A. We have repeated this procedure multiple times in female Yorkshire cross sows weighing 300-600 lbs. Blood collection yielded between 3 L from the smaller swine to 8 .......

Discussion

The procedure here describes a catheter-stylet-to-vacuum-pump assembly for collecting a one-time large volume of blood from the heart of sows. Because of the potential for pain and distress, this procedure must only be performed in animals that are in a surgical plane of anesthesia. Immobility from the anesthesia allows this procedure to be learned by beginners and mastered quickly. Multiple veins can be used to collect blood from animals. The most common and accessible routes are the jugular veins, cranial vena cava, an.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
1/8 HP Lab Model Diaphragm Pump – 115 vac, 60 max psi	Base Supply Center	80058845
10 G x 6 in catheter-over-needle	Covetrus North American Veterinary Supplies	024616
50 mL syringe, luer lock	Fisher Scientific	13-689-8
Aquasonic 100 ultrasound gel blue	Midwest Veterinary Supply	001.02109.2
Argyl bubble tubing, Nonconductive (¼")	Fisher Scientific	NC2282605 (Vendor catalog # 8888280412)
Chlorhexidine Q Scrub 2%	Midwest Veterinary Supply	562.30804.3
Corning Disposable Roller Bottles.	Fisher Scientific	06-413
Corning PYREX narrow mouth heavy-duty glass Erlenmeyer flask or similar creating suction	Fisher Scientific	10-040F
Eisco nylon test tube brush	Fisher Scientific	S13962
Fisherbrand two-hole rubber stoppers	Fisher Scientific	Dependent on size
Ketamine (10 mg/mL)	Midwest Veterinary Supply	275.22100.3
MLL to 6 mm hose end (stainless steel)	Cadence Science	6547IND
MLL to 8mm hose end (stainless steel)	Cadence Science	6549IND
MVet gauze sponge non-woven 4 x 4 4 ply	Midwest Veterinary Supply	000.30156.2
MVet isopropyl alcohol 70%	Midwest Veterinary Supply	000.50054.3
MVet Kelly hemostatic forceps straight 5.5”	Midwest Veterinary Supply	000.36800.2
Sodium Heparin (1000 U/mL)	Midwest Veterinary Supply	191.46701.3
Sodium Pentobarbital (390 mg/mL)	Midwest Veterinary Supply	749.20000.3
Stainless steel straws	Hobby Lobby	1893635
Telazol (100 mg/mL)	Midwest Veterinary Supply	275.40000.3
Xylazine (100 mg/mL)	Midwest Veterinary Supply	310.01150.3