Gjendal, K., Kiersgaard, M. K., Abelson, K. et al. 2020. Comparison of sublingual, facial and retro-bulbar blood sampling in mice in relation to animal welfare and blood quality. Journal of Pharmacological and Toxicological Methods 103, 106680.

Introduction: Repeated blood sampling is a common procedure in laboratory mice, but at present it is unknown which technique has the least impact on the animals when large or repeated blood samples are required. Retro-bulbar sinus puncture is a reliable technique but has been shown to cause many changes in the animals, why sublingual and facial vein puncture have been suggested as suitable alternatives. This study investigated 1) which of the three blood sampling techniques had the least impact on nest building activity, level of faecal corticosterone metabolites, body weight, fur status, and macroscopic changes, 2) whether the blood sampling techniques gave rise to variation in blood quality between blood samples, and 3) whether sublingual and facial vein puncture should be performed with or without anaesthesia in female C57BL/6 mice. Method: Three hundred and sixty C57BL/6 female mice divided into five batches were included in the study and randomized to a short (blood sampling on Day 8, 9 and 10) or a long protocol (blood sampling on Day 8, 15 and 22). Each protocol consisted of six identical groups: sublingual vein puncture (SVP), sublingual vein puncture in isoflurane (SVPiso), facial vein puncture (FVP), facial vein puncture in isoflurane (FVPiso), retro-bulbar sinus puncture (RBP), and a control group (CONTROL) with only scruffing being performed. At baseline (Day 2) nest building activity (NBA) was assessed and faecal pellets collected for evaluation of faecal corticosterone metabolites (FCM). The day after each blood sampling day NBA and FCM were reassessed. Results and conclusion: None of the blood sampling techniques proved to be superior to the others in any of the measured parameters. Finally, sublingual and facial vein puncture performed under anaesthesia gave rise to variation in the quality of the blood. A refinement of all three techniques are therefore warranted.

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