Hormonal stress response of laboratory mice to conventional and minimally invasive bleeding techniques

Voigt, Christian C.; Klöckner, Pia; Touma, Chadi; Neuschl, Christina; Brockmann, Gudrun A.; Göritz, Frank; Palme, Rupert; Thomsen, Ruth

doi:10.7120/09627286.22.4.449

Cited by 9 publications

(13 citation statements)

References 29 publications

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“…The same method for FCM analysis was previously used to identify a correlation between an increase in FCM levels and various stressors in mice [ 4 ], e.g. train-induced vibrations [ 63 ], oral gavage [ 64 ], and blood sampling [ 65 ].…”

Section: Discussionmentioning

confidence: 99%

Impact of repeated anesthesia with ketamine and xylazine on the well-being of C57BL/6JRj mice

et al. 2018

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Within the scope of the 3Rs of Russel and Burch, the number of laboratory animals can be reduced by repeated use of an animal. This strategy only becomes relevant, if the total amount of pain, distress or harm the individual animal experiences does not exceed the severity of a single manipulation. For example, when using imaging techniques, an animal can be examined several times during a study, but it has to be anesthetized each time imaging is performed. The severity of anesthesia is thought to be mild according to the Directive 2010/63/EU. However, the Directive does not differentiate between single and repeated anesthesia, although repeated anesthesia may have a greater impact on well-being. Hence, we compared the impact of single and repeated anesthesia (six times at an interval of three to four days) by injection of ketamine and xylazine (KX) on the well-being of adult female and male C57BL/6JRj mice. After anesthesia, well-being of mice was assessed according to a protocol for systematic assessment of well-being including nesting, the Mouse Grimace Scale (MGS), a test for trait anxiety, home cage activity, and the rotarod test for motor activity, food intake, and body weight, as well as corticosterone (metabolite) analysis. Repeated anesthesia increased the MGS in mice of both sexes and caused short-term effects on well-being of female mice in the immediate post-anesthetic period, indicated by longer lasting effects on trait anxiety-related behavior. However, corticosterone metabolite concentrations suggested that mice habituated to the stress induced by repeated KX administration. Hence, the mildly negative effects on well-being of repeated KX anesthesia do not seem to accumulate over time using the respective regimen. However, further observations for severity classification are warranted in order to more specifically determine the duration of mild distress and trait anxiety.

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Section: Discussionmentioning

confidence: 99%

Impact of repeated anesthesia with ketamine and xylazine on the well-being of C57BL/6JRj mice

et al. 2018

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“…However, only a limited number of publications describe the impacts of retro-bulbar blood sampling in mice [12], [33] and only one study compares this method to venipuncture of the facial vein (sometimes incorrectly referred to as the submandibular vein) [34]. Occasionally, the superficial temporal vein is used rather than the facial vein.…”

Section: Discussionmentioning

confidence: 99%

Physiological and Pathological Impact of Blood Sampling by Retro-Bulbar Sinus Puncture and Facial Vein Phlebotomy in Laboratory Mice

et al. 2014

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Retro-bulbar sinus puncture and facial vein phlebotomy are two widely used methods for blood sampling in laboratory mice. However, the animal welfare implications associated with these techniques are currently debated, and the possible physiological and pathological implications of blood sampling using these methods have been sparsely investigated. Therefore, this study was conducted to assess and compare the impacts of blood sampling by retro-bulbar sinus puncture and facial vein phlebotomy. Blood was obtained from either the retro-bulbar sinus or the facial vein from male C57BL/6J mice at two time points, and the samples were analyzed for plasma corticosterone. Body weights were measured at the day of blood sampling and the day after blood sampling, and the food consumption was recorded automatically during the 24 hours post-procedure. At the end of study, cheeks and orbital regions were collected for histopathological analysis to assess the degree of tissue trauma. Mice subjected to facial vein phlebotomy had significantly elevated plasma corticosterone levels at both time points in contrast to mice subjected to retro-bulbar sinus puncture, which did not. Both groups of sampled mice lost weight following blood sampling, but the body weight loss was higher in mice subjected to facial vein phlebotomy. The food consumption was not significantly different between the two groups. At gross necropsy, subcutaneous hematomas were found in both groups and the histopathological analyses revealed extensive tissue trauma after both facial vein phlebotomy and retro-bulbar sinus puncture. This study demonstrates that both blood sampling methods have a considerable impact on the animals' physiological condition, which should be considered whenever blood samples are obtained.

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“…FCM reliably indicate stress in mice 33,34,35 . The peak FCM concentrations typically occur 8-10 h after a stressor but depend on the intestinal transit time…”

confidence: 91%

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

Hohlbaum¹,

Bert²,

Dietze³

et al. 2018

JoVE

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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.

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Hormonal stress response of laboratory mice to conventional and minimally invasive bleeding techniques

Cited by 9 publications

References 29 publications

Impact of repeated anesthesia with ketamine and xylazine on the well-being of C57BL/6JRj mice

Impact of repeated anesthesia with ketamine and xylazine on the well-being of C57BL/6JRj mice

Physiological and Pathological Impact of Blood Sampling by Retro-Bulbar Sinus Puncture and Facial Vein Phlebotomy in Laboratory Mice

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

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