Abstract

Simple SummaryBlood sampling is often performed in laboratory mice. Whilst the techniques are likely to cause only momentary pain or distress, given their frequency of performance, it is essential that the method which best safeguards welfare is used. The small size of mice makes sampling challenging, and use of some routes is controversial due to perceived impact on animal welfare. However, to date, no summary of the evidence relating to welfare impacts arising from these techniques has been presented. This paper presents a systematic review of the literature, with quality appraisal of the studies and an assignment of certainty in the evidence. We conclude that there is not enough high-quality evidence available to make a determination on optimal blood sampling route. We provide recommendations for improving future laboratory animal welfare research through standardisation of outcome measures and enhanced adherence to experimental design and reporting guidelines. Blood sampling is often performed in laboratory mice. Sampling techniques have the potential to cause pain, distress and impact on lifetime cumulative experience. In spite of institutions commonly providing guidance to researchers on these methods, and the existence of published guidelines, no systematic evaluation of the evidence on this topic exists. A systematic search of Medline, Scopus, and Web of Science was performed, identifying 27 studies on the impact of recovery blood sample techniques on mouse welfare and sample quality. Studies were appraised for quality using the SYstematic Review Centre for Laboratory animal Experimentation (SYRCLE) risk of bias tool. In spite of an acceptable number of studies being located, few studies examined the same pairwise comparisons. Additionally, there was considerable heterogeneity in study design and outcomes, with many studies being at a high risk of bias. Consequently, results were synthesised using the Synthesis Without Meta-analysis (SWiM) reporting guidelines. Grading of Recommendations, Assessment, Development and Evaluation (GRADE) was utilised for assessment of certainty in the evidence. Due to the heterogeneity and GRADE findings, it was concluded that there was not enough high-quality evidence to make any recommendations on the optimal method of blood sampling. Future high-quality studies, with standardised outcome measures and large sample sizes, are required.

Highlights

  • It is common in biomedical research for protocols to require blood collection from mice (Mus musculus) in order to measure a range of circulating products

  • Included techniques were: sublingual, retrobulbar sinus, facial, tail sampling methods, and saphenous vein. Studies that examined both one-off and serial sampling were eligible for inclusion; (C) studies were included that compared the intervention to no blood sample, or other included recovery sample method

  • Whilst summary documents or guidelines exist on this topic, a number of these are older documents and it is not clear whether they have been based on a systematic review of the available evidence

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Summary

Introduction

It is common in biomedical research for protocols to require blood collection from mice (Mus musculus) in order to measure a range of circulating products. Retrobulbar bleeding (retroorbital) enables acquisition of larger blood volumes (e.g., 0.2–1 mL) [3], but has been controversial due to the risk of substantial tissue damage to the eye [3,4] It appears that this technique has fallen out of favour, in some jurisdictions, such as Australia. Despite the rise in popularity of this method, perhaps driven by the aesthetically repugnant use of the ocular area, veins in the facial region lie beneath other important tissues such as glandular tissue [6]. This method poses a risk of causing secondary complications via tissue damage. In contrast to the submandibular technique, sublingual sampling is generally performed under anaesthesia to immobilise the animal [4]

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