Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

Katharina Hohlbaum; Bettina Bert; Silke Dietze; Rupert Palme; Heidrun Fink; Christa Thöne-Reineke

doi:10.3791/57046

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Method Article

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

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DOI:

10.3791/57046

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March 20th, 2018

Katharina Hohlbaum¹^,² , Bettina Bert²^,³ , Silke Dietze² , Rupert Palme⁴ , Heidrun Fink² , Christa Thöne-Reineke¹

¹Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Department of Veterinary Medicine, Freie Universität Berlin, ²Institute of Pharmacology and Toxicology, Department of Veterinary Medicine, Freie Universität Berlin, ³German Federal Institute for Risk Assessment (BfR), ⁴Unit of Physiology, Pathophysiology and Experimental Endocrinology, Department of Biomedical Sciences, University of Veterinary Medicine

Summary

We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner.

Abstract

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal experimentation whenever possible. When there is no alternative to animal experimentation, the total number of laboratory animals used should be the minimum needed to obtain valuable data. Moreover, appropriate refinement measures should be applied to minimize pain, suffering, and distress accompanying the experimental procedure. The categories used to classify the degree of pain, suffering, and distress are non-recovery, mild, moderate, or severe (EU Directive 2010/63). To determine which categories apply in individual cases, it is crucial to use scientifically sound tools.

The well-being-assessment protocol presented here is designed for procedures during which general anesthesia is used. The protocol focuses on home cage activity, the Mouse Grimace Scale, and luxury behaviors such as burrowing and nest building behavior as indicators of well-being. It also uses the free exploratory paradigm for trait anxiety-related behavior. Fecal corticosterone metabolites as indicators of acute stress are measured over the 24-h post-anesthetic period.

The protocol provides scientifically solid information on the well-being of mice following general anesthesia. Due to its simplicity, the protocol can easily be adapted and integrated in a planned study. Although it does not provide a scale to classify distress in categories according to the EU Directive 2010/63, it can help researchers estimate the degree of severity of a procedure using scientifically sound data. It provides a way to improve the assessment of well-being in a scientific, animal-centered manner.

Introduction

EU Directive 2010/63¹ stipulates that the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch² is to be applied whenever animal experimentation is necessary. The ultimate goal of the EU Directive is to phase out all animal testing, but the Directive acknowledges that, for the time being, some animal experiments are still needed to conduct research that will protect human and animal health. Thus, if an animal experiment cannot be replaced by any alternative method, only the minimum number of laboratory animals is to be used to obtain reliable results. In addition, the amount of pain, suffering,....