Refinement Database

Database on Refinement of Housing, Husbandry, Care, and Use of Animals in Research

This database, created in 2000, is updated every four months with newly published scientific articles, books, and other publications related to improving or safeguarding the welfare of animals used in research.

Tips for using the database:

  • This landing page displays all of the publications in the database.
  • Use the drop-down menus to filter these publications by Animal Type, Setting, and/or Topic.
  • Clicking on a parent category (e.g., Rodent) will include publications relating to all the items in that category (e.g., Chinchilla, Gerbil, Guinea Pig, etc.).
  • You may also add a keyword to further narrow your search.
  • Please note that at this time, only publications dated 2010 or later (with some exceptions) can be filtered by Animal Type and Topic, and only publications dated 2020 or later (with some exceptions) can be filtered by Setting. Most publications older than 2010 can only be searched by keyword. 
McNamara, A., Ballantine, A. R., Deuchande, R. et al. 2023. Comparison of fixed and rising displacement rates of CO2 for euthanasia of aged rats. JAALAS 62(5), 453–463.

This study compared euthanasia induced by rising concentrations of CO2 in aged rats (n = 59) using different gas displacement rates. Rats were preimplanted with cardiovascular telemetry devices and had been previously used for short...

Stuckey, J. E., Makhija, S. D., Reimer, D. C. et al. 2023. Effects of different grades of carbon dioxide on euthanasia of mice (Mus musculus). JAALAS 62(5), 430–437.

Carbon dioxide (CO2) is a conditionally acceptable method for mouse euthanasia, according to the American Veterinary Medical Association Guidelines for the Euthanasia of Animals. In a 2012 Office of Laboratory Animal Welfare (OLAW) webinar, representatives...

Butcher, G., Davidson, A., Sloanc, A. et al. 2021. An apparatus for automatically training and collecting individualized behavioral data with socially housed rodents. Journal of Neuroscience Methods 365, 109387.

Background: Conventional methods for individually housing, training, and testing rodents in behavioral assays can impose constraints that may limit some kinds of experimental external validity, preempt environmental enrichment, impose heavy experimenter time burdens that limit...

Hickman, D. L. 2022. Evaluation of carbon dioxide euthanasia of female Sprague Dawley rats alone or with unfamiliar conspecifics. JAALAS 61(2), 195-200.

Most studies evaluating methods of euthanasia to date have focused on the euthanasia of individual animals. However, larger chambers are commonly used to euthanize multiple cages of animals at once. This study evaluated the use...

Reuser, A., Wenzel, K., Felix, S. B. et al. 2022. Simultaneous assessment of spontaneous cage activity and voluntary wheel running in group-housed mice. Scientific Reports 12(1), 4444.

Small animal models are frequently used to improve our understanding of the molecular and biological signaling pathways underlying the beneficial effects of physical activity and exercise. Unfortunately, when running wheels are employed, mice and rats...

Kohler, J., Mei, J., Banneke, S. et al. 2022. Assessing spatial learning and memory in mice: Classic radial maze versus a new animal-friendly automated radial maze allowing free access and not requiring food deprivation. Frontiers in Behavioral Neuroscience 16, 1013624.

The radial arm maze (RAM) is a common behavioral test to quantify spatial learning and memory in rodents. Prior attempts to refine the standard experimental setup have been insufficient. Previously, we demonstrated the feasibility of...

Hickman, D. L. 2022. Minimal exposure times for irreversible euthanasia with carbon dioxide in mice and rats. JAALAS 61(3), 283-286.

When using an anesthetic overdose to euthanize laboratory rodents, a secondary method of euthanasia is recommended to ensure that the apparent death is irreversible. This secondary method usually is accomplished through the collection of tissues...

McKinney, M. M., Dupont, W. D., Corson, K. J. et al. 2022. Physiologic and behavioral effects in mice anesthetized with isoflurane in a red-tinted or a traditional translucent chamber. JAALAS 61(4), 322-332.

Isoflurane has been characterized as a distressing agent for rodents, causing both physiologic and behavioral effects. Using a "darkened home cage" has been recommended during CO2 administration for rodent euthanasia; this is arguably a similar...

Barkus, C., Bergmann, C., Branco, T. et al. 2022. Refinements to rodent head fixation and fluid/food control for neuroscience. Journal of Neuroscience Methods 381, 109705.

The use of head fixation in mice is increasingly common in research, its use having initially been restricted to the field of sensory neuroscience. Head restraint has often been combined with fluid control, rather than...

Clarkson, J. M., Martin, J. E., McKeegan, D. E. F. 2022. A review of methods used to kill laboratory rodents: Issues and opportunities. Laboratory Animals 56(5), 419-436.

Rodents are the most widely used species for scientific purposes. A critical pre-requisite of their use, based on utilitarian ethical reasoning, is the provision of a humane death when necessary for scientific or welfare grounds...

Dickmann, J., Gonzalez-Uarquin, F., Reichel, S. et al. 2022. Clicker training mice for improved compliance in the catwalk test. Animals 12(24), 3545.

The CatWalk test relies on the run of mice across the platform to measure a constant speed with low variation. Mice usually require a stimulus to walk to the end of the catwalk. However, such...

Nuber, M., Gonzalez-Uarquin, F., Neufurth, M. et al. 2022. Development of a 3D simulator for training the mouse in utero electroporation. PLOS ONE 17(12), e0279004.

In utero electroporation (IUE) requires high-level training in microinjection through the mouse uterine wall into the lateral ventricle of the mouse brain. Training for IUE is currently being performed in live mice as no artificial...

Brough, D. W., Murkin, J. T., Amos, H. E. et al. 2022. Comparing variability in measurement of subcutaneous tumors in mice using 3D thermal imaging and calipers. Comparative Medicine 72(6), 364–375.

Repeatable tumor measurements are key to accurately assessing tumor growth and treatment efficacy. A preliminary study that we conducted showed that a novel 3D and thermal imaging system (3D-TI) for measuring subcutaneous tumors in rodents...

Hepps Keeney, C., Harrison, T. M. 2022. Euthanasia complications in non-domestic species. Journal of Zoological and Botanical Gardens 3(4), 616–623.

There are very few studies describing euthanasia complications in non-domestic species. The goal of this study was to survey veterinarians to determine what complications may commonly occur during the euthanasia of non-domestic species. An online...

Zeng, Y., Jin, H., Wang, J. et al. 2022. An optimized method for intratracheal instillation in mice. Journal of Pharmacological and Toxicological Methods 118, 107230.

Non-invasive intratracheal instillation is an important method for direct exposure of the respiratory tract which is commonly used in toxicology, environmental science, and other research fields. However, there is no standard operating process for non-invasive...

Hickman, D. L. 2021. Wellbeing of mice euthanized with carbon dioxide in their home cage as compared with an induction chamber. JAALAS 60(1), 72-76.

The AVMA Guidelines on Euthanasia state that, to decrease potential distress of animals, the home cage should be used for the euthanasia of mice. The current study evaluated this recommendation by comparing behavioral and physiologic...

Améndola, L., Ratuski, A., Weary, D. M. 2021. Individual differences in rat sensitivity to CO2. PLOS ONE 16(1), e0245347.

Feelings of fear, anxiety, dyspnea and panic when inhaling carbon dioxide (CO2) are variable among humans, in part due to differences in CO2 sensitivity. Rat aversion to CO2 consistently varies between individuals; this variation in...

Wooden, J. I., Spinetta, M. J., Nguyen, T. et al. 2021. A sensitive homecage-based novel object recognition task for rodents. Frontiers in Behavioral Neuroscience 15, 680042.

The recognition of novel objects is a common cognitive test for rodents, but current paradigms have limitations, such as low sensitivity, possible odor confounds and stress due to being performed outside of the homecage. We...

Rodriguez-Sanchez, R., Barnaby, E., Améndola, L. et al. 2021. Voluntary oral ingestion of a sedative prior to euthanasia with CO2: Behavioural responses of mice. Animals 11(10), 2879.

Laboratory mice are commonly euthanised with carbon dioxide (CO2); however, there is ample evidence that this gas is aversive. Previous work suggests that sedation achieved via injection with benzodiazepines prior to CO2 administration could reduce...

Brunt, M. W., Améndola, L., Weary, D. M. 2021. Attitudes of laboratory animal professionals and researchers towards carbon dioxide euthanasia for rodents and perceived barriers to change. Laboratory Animals 55(6), 531-539.

Evidence indicates that carbon dioxide (CO2) induces negative affective states (including anxiety, fear and distress) in laboratory rodents, but many countries still accept it for euthanasia. Alternative methods (e.g. inhalant anaesthetic) may represent a refinement...

Verjat, A., Devienne, P., Rödel, H. G. et al. 2021. More exploratory house mice judge an ambiguous situation more negatively. Animal Cognition 24(1), 53–64.

Exploration tendency, one of the most investigated animal personality traits, may be driven by either positive (when seeking interesting information) or negative (to reduce the uncertainty of the environment) affective/emotional profiles. To disentangle the valence...

Persson, K., Selter, F., Neitzke, G. et al. 2020. Philosophy of a “good death” in small animals and consequences for euthanasia in animal law and veterinary practice. Animals 10(1), 124.

Moral stress is a major concern in veterinary practice. Often, it is associated with the challenges in end-of-life situations. Euthanasia, however, is also meant to bring relief to animal patients and their owners. The reasons...

Gades, N. M., Thompson, K. L., Mi, L. et al. 2020. Impact of euthanasia on compassion fatigue in personnel working in animal research. Laboratory Animal Science Professional 8(1) (January/February), 22-23.

We conducted a cross-sectional study to investigate the association between euthanasia and compassion fatigue among employees working in animal research at a large academic medical center. In summary, animal research workers who euthanize animals reported...

Gehlen, H., Loschelder, J., Merle, R. et al. 2020. Evaluation of stress response under a standard euthanasia protocol in horses using analysis of heart rate variability. Animals 10(3), 485.

The effects of a standard protocol for euthanasia on heart rate variability (HRV) as a consequence of stress response were analyzed in this prospective clinical study. The HRV was determined in 40 horses undergoing euthanasia...

Mei, J., Kohler, J., Winter, Y. et al. 2020. Automated radial 8-arm maze: A voluntary and stress-free behavior test to assess spatial learning and memory in mice. Behavioural Brain Research 381, 112352.

The radial arm maze (RAM) is a common behavioral test to assess spatial working and reference memory in mice. However, conventional RAM experiments require a substantial degree of manual handling and animals are usually subjected...