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Open Access Physiologic, Behavioral, and Histologic Responses to Various Euthanasia Methods in C57BL/6NTac Male Mice

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Rodent euthanasia using exposure to increasing concentrations of CO2 has come under scrutiny due to concerns of potential pain during the euthanasia process. Alternatives to CO2, such as isoflurane and barbiturates, have been proposed as more humane methods of euthanasia. In this study, we examined 3 commonly used euthanasia methods in mice: intraperitoneal injection of pentobarbital–phenytoin solution, CO2 inhalation, and isoflurane anesthesia followed by CO2 inhalation. We hypothesized that pentobarbital–phenytoin euthanasia would cause fewer alterations in cardiovascular response, result in less behavioral evidence of pain or stress, and produce lower elevations in ACTH than would the isoflurane and CO2 methods, which we hypothesized would not differ in regard to these parameters. ACTH data suggested that pentobarbital–phenytoin euthanasia may be less stressful to mice than are isoflurane and CO2 euthanasia. Cardiovascular, behavioral, and activity data did not consistently or significantly support isoflurane or pentobarbital–phenytoin euthanasia as less stressful methods than CO2. Euthanasia with CO2 was the fastest method of the 3 techniques. Therefore, we conclude that using CO2 with or without isoflurane is an acceptable euthanasia method. Pathologic alterations in the lungs were most severe with CO2 euthanasia, and alternative euthanasia techniques likely are better suited for studies that rely on analysis of the lungs.

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Document Type: Research Article

Affiliations: 1: Departments of Pathology, Wright State University, Dayton, Veterans Affairs Medical Center, Cincinnati, Ohio;, Email: [email protected] 2: Departments of Mathematical and Microbiologic Sciences, Wright State University, Dayton, Ohio 3: Departments of Psychology, Wright State University, Dayton, Ohio 4: Departments of Neuroscience, Cell Biology and Physiology, Boonshoft School of Medicine, Wright State University, Dayton, Ohio 5: Departments of Pharmacology and Toxicology, Boonshoft School of Medicine, Wright State University, Dayton, Ohio

Publication date: 2017-01-01

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  • The Journal of the American Association for Laboratory Animal Science (JAALAS) serves as an official communication vehicle for the American Association for Laboratory Animal Science (AALAS). The journal includes a section of refereed articles and a section of AALAS association news. The mission of the refereed section of the journal is to disseminate high-quality, peer-reviewed information on animal biology, technology, facility operations, management, and compliance as relevant to the AALAS membership. JAALAS accepts research reports (data-based) or scholarly reports (literature-based), with the caveat that all articles, including solicited manuscripts, must include appropriate references and must undergo peer review.

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