Cardiovascular Journal of Africa: Vol 32 No 5 (SEPTEMBER/OCTOBER 2021)

CARDIOVASCULAR JOURNAL OF AFRICA • Volume 32, No 5, September/October 2021 AFRICA 255 and Use of Laboratory Animals 5 was used, and two attendant veterinarians controlled all procedures. The investigators found it helpful to consult with experts regarding statistical analysis for required animal numbers, and database searches to identify potential alternatives to painful or distressing procedures. 6 Retrobulbar injection of no more than 200 μl of injectable anaesthetic solution (ketamine:xylazine) was used, resulting in death within five seconds of cessation of injection, 7 as mentioned among the forms of euthanasia in the AVMA Guidelines for the Euthanasia of Animals , 2020 edition. Twenty-one New Zealand male white rabbits (eight months old, mean weight 3 kg, range 2.6–3.4 kg) were obtained from a private farm, by veterinary faculty authority. The 21 rabbits were divided into three groups: group I received a proximal femoral artery clamp; group II received a proximal thigh tourniquet, and group III was the control group. The rabbits were prepared in the supine position after anaesthesia using 2 mg/kg of intramuscular diazepam and 40 mg/kg ketamine, and they were draped after shaving and cleaning their skin with betadine. In order not to affect tumour necrosis factor alpha (TNF- α ) values, only one limb side was used in all subjects. In the clamp group, a proximal incision was made anteromedially over the femoral neurovascular margin, and the skin, subcutaneous tissue and deep fascia were incised. After dissecting the muscles and exposing the neurovascular bundle, the femoral artery was dissected and clamped with a microvascular Fig. 1. (A) Clamp: CFA, black arrow: femoral vein, nerve, asterisk: rectus femoris muscle. (B) Asterisk: rectus femoris muscle. (C) Black arrow: areas without endothelial cells (H&E stain, x200). (D) Yellow line: fibrin thrombi (elastin van Gieson stain, x200) A C B D