In recent years, “tunnelling” and “cupping” have been recommended to eliminate the drawbacks of tail handling 28, 29. Studies on behavioural phenotypes in mice have identified stress handling as one of the most likely causes of failure to replicate phenotypes within and between experiments 27. However, lifting the mouse by its tail is aversive to the mouse, causing stress and anxiety. This is specified in standardized protocols for the handling of experimental mice 24– 26. In many experiments, including behavioural tests, the most common method used to capture and carry mice is to pick their tail at its base between the thumb and index finger.
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In behavioural tests, it is important to understand the types of these tests susceptible to background stress and how to minimize its influence. Consequently, the results of behavioural tests often differ among researchers. However, it is often unclear how a laboratory or researcher handles the mice 20– 23. Thus, differences in handling techniques can greatly affect the results of behavioural tests. In many behavioural tests, the experimenter frequently needs to come into contact with the mouse. In particular, conducting a behavioural test battery is extremely important for detecting abnormal behaviour in mice 17– 19. Behavioural tests in mice are important tests that have been performed since a long time and are widely used worldwide 13– 16.
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Often, laboratory mice are evaluated for depression, aggression, activity, and anxiety-like behaviour through a series of behavioural tests to analyse the effects of drugs or other stimuli 12. Insufficient description of this process can cause experimenter errors and biases 11. To improve the reproducibility of animal studies, we need to consider all stressors that may influence the experimental animals, and handling is one of them. For practical reasons, animals are usually subjected to stress between periods of inactivity during the light cycle 10. injections and blood sampling) procedures. cage cleaning and breeding) and research (e.g. Care and handling are integral parts of an animal’s daily life 8, 9, and handling is the most common procedure experienced by laboratory mice because it is necessary for routine (e.g. Lack of consideration for handling techniques can impede reproducibility and cast doubt on the relevance of otherwise valuable scientific research. A factor often overlooked is how researchers handle animals. Environmental conditions in animal facilities can affect the results of tests that measure natural behaviour in particular 7. The assessment of subjective animal welfare components relies primarily on physiological 2, 3 and behavioural 4– 6 measurements. Environmental factors in the home cage play an important role in the health of these animals. Laboratory mice spend most of their lives in home cages before being used in experimental procedures. To successfully transfer the results obtained in mice to human studies, it is necessary to determine appropriate treatment and handling methods for experimental mice since handling of normal mice may affect the obtained experimental results. Mice have been the most widely used laboratory animals for disease, behaviour, and pharmacology studies over the past century 1. Our findings demonstrated that mice repeatedly handled by the experimenter before behavioural tests showed reduced anxiety about high altitudes and improved spatial cognition, suggesting that repeated contact can affect the results of some behavioural tests.
Grip strength, rotarod, sociability, tail suspension, Porsolt forced swim, and passive avoidance tests revealed no significant differences between repeatedly handled and control mice. However, there was no change in anxiety-like behaviour in the light/dark transition test or open-field test.
In behavioural tests, repeatedly handled mice showed improved spatial cognition in the Y-maze test and reduced anxiety-like behaviour in the elevated plus-maze test.
Repeatedly handled mice gained slightly more weight than control mice. To investigate these effects, we used a behavioural test battery to assess differences between mice accustomed to the experimenter’s handling versus control mice. To minimise mice stress, methods of moving mice using transparent tunnels or cups have been recommended but are impractical in behavioural tests. However, the effects of repeated handling on mice behaviour remains unclear. During behavioural experiments, mice frequently experience human contact. Behavioural experiment battery aids in evaluating abnormal behaviour in mice. Mice are the most commonly used laboratory animals for studying diseases, behaviour, and pharmacology.
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