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

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.

[1]  R. Palme,et al.  Analyzing corticosterone metabolites in fecal samples of mice: a noninvasive technique to monitor stress hormones , 2004, Hormones and Behavior.

[2]  S. Rivest,et al.  Subchronic memantine administration on spatial learning, exploratory activity, and nest-building in an APP/PS1 mouse model of Alzheimer’s disease , 2011, Neuropharmacology.

[3]  K. Craig,et al.  Coding of facial expressions of pain in the laboratory mouse , 2010, Nature Methods.

[4]  G. Sales,et al.  Behaviour of rats in their home cages: daytime variations and effects of routine husbandry procedures analysed by time sampling techniques , 1996, Laboratory animals.

[5]  P. H. Griffiths,et al.  Guidelines on the recognition of pain, distress and discomfort in experimental animals and an hypothesis for assessment , 1985, Veterinary Record.

[6]  Nikola Cesarovic,et al.  Assessment of postsurgical distress and pain in laboratory mice by nest complexity scoring , 2013, Laboratory animals.

[7]  J. Hurst,et al.  Taming anxiety in laboratory mice , 2010, Nature Methods.

[8]  L. Keeling,et al.  Assessment of positive emotions in animals to improve their welfare , 2007, Physiology & Behavior.

[9]  J. Parmigiani,et al.  Characterization of Train-Induced Vibration and its Effect on Fecal Corticosterone Metabolites in Mice. , 2015, Journal of the American Association for Laboratory Animal Science : JAALAS.

[10]  M. Leach,et al.  The effect of isoflurane anaesthesia and buprenorphine on the mouse grimace scale and behaviour in CBA and DBA/2 mice , 2015, Applied animal behaviour science.

[11]  I. Cuthill,et al.  Reporting : The ARRIVE Guidelines for Reporting Animal Research , 2010 .

[12]  Paulin Jirkof,et al.  Burrowing and nest building behavior as indicators of well-being in mice , 2014, Journal of Neuroscience Methods.

[13]  C. Belzung,et al.  Further pharmacological validation of the BALB/c neophobia in the free exploratory paradigm as an animal model of trait anxiety , 1997, Behavioural pharmacology.

[14]  R. Palme,et al.  Severity classification of repeated isoflurane anesthesia in C57BL/6JRj mice—Assessing the degree of distress , 2017, PloS one.

[15]  S. Wells,et al.  Joint Working Group on Refinement A guide to defining and implementing protocols for the welfare assessment of laboratory animals , 2011 .

[16]  P. Bugnon,et al.  What the literature tells us about score sheet design , 2016, Laboratory animals.

[17]  J. Rawlins,et al.  Hippocampal cytotoxic lesion effects on species-typical behaviours in mice , 2002, Behavioural Brain Research.

[18]  Rowland R. Sillito,et al.  Analysis of Individual Mouse Activity in Group Housed Animals of Different Inbred Strains using a Novel Automated Home Cage Analysis System , 2016, Front. Behav. Neurosci..

[19]  R. Lister.  Ethologically-based animal models of anxiety disorders. , 1990, Pharmacology & therapeutics.

[20]  V. Perry,et al.  Burrowing into prion disease , 2001, Neuroreport.

[21]  K. Unno,et al.  Social stress increases biopyrrins, oxidative metabolites of bilirubin, in mouse urine. , 2006, Biochemical and biophysical research communications.

[22]  V. Perry,et al.  MCP-1 and murine prion disease: Separation of early behavioural dysfunction from overt clinical disease , 2005, Neurobiology of Disease.

[23]  V. Perry,et al.  Hippocampal scrapie infection impairs operant DRL performance in mice , 2005, Behavioural Brain Research.

[24]  R. Palme,et al.  A less stressful alternative to oral gavage for pharmacological and toxicological studies in mice. , 2012, Toxicology and applied pharmacology.

[25]  R. Deacon.  Assessing nest building in mice , 2006, Nature Protocols.

[26]  M. Arras,et al.  EHP: history and objectives. , 1974, Environmental Health Perspectives.

[27]  R. Deacon.  Burrowing in rodents: a sensitive method for detecting behavioral dysfunction , 2006, Nature Protocols.

[28]  R. Deacon,et al.  Assessing Burrowing, Nest Construction, and Hoarding in Mice , 2012, Journal of visualized experiments : JoVE.

[29]  Hanno Würbel,et al.  Facial Indicators of Positive Emotions in Rats , 2016, PloS one.

[30]  B. Seifert,et al.  Burrowing Behavior as an Indicator of Post-Laparotomy Pain in Mice , 2010, Front. Behav. Neurosci..

[31]  R. Palme,et al.  Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice. , 2003, General and comparative endocrinology.

[32]  Amy L. Miller,et al.  The Influence of Isoflurane Anaesthesia on the Rat Grimace Scale , 2016, PloS one.

[33]  V. Perry,et al.  Early behavioural changes in scrapie‐affected mice and the influence of dapsone , 2001, The European journal of neuroscience.

[34]  A. Rex,et al.  Evaluation of cage leaving behaviour in rats as a free choice paradigm. , 2013, Journal of pharmacological and toxicological methods.