Impact of blood sampling technique on blood quality and animal welfare in haemophilic mice

Abstract

Blood collection in mice can be a challenge, in particular for samples used for coagulation analysis as initiation of coagulation during the procedures can influence the results. Blood collection from the retrobulbar venous plexus is commonly used but the method remains controversial. Several alternatives exist but not all are applicable to mice with a compromised coagulation system because of subsequently excessive bleeding. We therefore wanted to explore whether blood collection by puncture of the submandibular vein could replace blood collected from the retrobulbar venous plexus during pharmacokinetic and pharmacodynamic studies in mice lacking coagulation factor VIII (FVIII). The plasma concentrations of recombinant activated factor VII were independent of the blood collection method in a pharmacokinetic study. The same applied to the thromboelastographic profile of mice with normal coagulation in a pharmacodynamic study. However, excessive haemorrhages were observed in all FVIII knockout mice after a single puncture of the submandibular vein and 60% of the mice were euthanized 2-4 h after the blood collection. In contrast, no or only slight haemorrhage was observed in animals subjected to blood collection from the retrobulbar venous plexus. No signs of distress determined by blood glucose level or clinical abnormalities of the eye were observed after puncture of the retrobulbar venous plexus. In conclusion, blood collected by puncture of the submandibular vein and retrobulbar venous plexus has a quality which allows it to be used in coagulation assays. However, because of excessive bleedings, puncture of the submandibular vein is not recommended in mice lacking FVIII.

Comparison of serological assessments in the diagnosis of liver fibrosis in bile duct ligation mice

“…FIB-4 was initially evaluated in human immunodeficiency virus (HIV)/HCV co-infected patients, 27 and Forns 24 HIV/HCV co-infected patients. 32 In our study, the AUROC (95% confidence interval) values for the detection of liver cirrhosis in BDL mice were 0.867 (0.725-0.953) and 0.879 (0.739-0.960) for FIB-4 and Forns Index, respectively.…”

“…The operations were performed as previously described in another study. 24 Platelets (PLT) were counted using a manual counting method by microscope in strict accordance with the provisions of the operation as previously described. 25 Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), and total cholesterol were measured using commercially available kits (Jiancheng Institute of Biotechnology, Nanjing, China) according to the manufacturer's instructions.…”

Blood Collection from Mice and Hematological Analyses on Mouse Blood

“…Blood collection from the submandibular vein was reported as a suitable alternative method to retro-bulbar puncture for collecting larger blood samples from anesthetized or conscious mice (Golde et al, 2005). However, several studies indicated problems that can occur with this technique, including difficulties in hitting the correct point of puncture in the first attempt, especially in conscious mice, which might be struggling heavily, as well as excessive bleeding and subcutaneous hemorrhages, mainly due to accidental puncture of the accompanying artery, especially in mice with impaired blood clotting (Heimann et al, 2010;Holmberg et al, 2011). The latter event also leads to the collection of a mix of arterial and venous blood.…”

Impact of three commonly used blood sampling techniques on the welfare of laboratory mice: Taking the animal’s perspective

“…To make a step towards Refinement concerning blood sampling in laboratory mice, our in-depth study comprehensively investigated the stress response caused by the three mentioned frequently used blood sampling techniques by also considering the animals' perspective. Earlier studies have shown that the different sampling techniques affect the quality of the blood sample, change several biochemical parameters, and cause varying tissue damage at the sampling site [7,[12][13][14][15][16][17][18]. However, the assessment of animal welfare in response to stressful sampling procedures in an objective and scientifically reliable manner is challenging.…”