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. 
Domínguez-Oliva, A., Hernández-Ávalos, I., Olmos-Hernández, A. et al. 2023. Thermal response of laboratory rats (Rattus norvegicus) during the application of six methods of euthanasia assessed by infrared thermography. Animals 13(18), 2820.

Refinement is one of the principles aiming to promote welfare in research animals. The techniques used during an experimental protocol, including euthanasia selection, must prevent and minimize suffering. Although the current euthanasia methods applied to...

Domínguez-Oliva, A., Olmos-Hernández, A., Hernández-Ávalos, I. et al. 2023. Rat grimace scale as a method to evaluate animal welfare, nociception, and quality of the euthanasia method of wistar rats. Animals 13(20), 3161.

Refinement of experimental procedures in animal research has the objective of preventing and minimizing pain/distress in animals, including the euthanasia period. This study aimed to evaluate pain associated with six methods of euthanasia in Wistar...

Heniff, A. C., Gorges, M. A., Lewbart, G. A. et al. 2023. Evaluation of targeted injections of ivermectin or potassium chloride for euthanasia of anesthetized thorny devil stick insects (Eurycantha calcarata). Journal of Zoological and Botanical Gardens 4(1), 231–239.

Insects are commonly utilized in biomedical research and have become increasingly popular in museum collections and as pets. Despite this, objective evaluation of insect euthanasia is scarce. This study investigated the effectiveness of targeted injections...

Stevens, C., Hammonds, T., Hinchcliffe, J. et al. 2023. Report of the 2022 RSPCA/UFAW Rodent Welfare Group Meeting. Animal Technology and Welfare 22(2), 91–100.

The RSPCA/UFAW Rodent Welfare Group has held a one-day meeting every autumn for the last 29 years, so that its members can discuss current welfare research, exchange views on welfare issues and share experiences of...

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...

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...

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...

Neil, D. M., Albalat, A., Thompson, J. 2022. The effects of electrical stunning on the nervous activity and physiological stress response of a commercially important decapod crustacean, the brown crab Cancer pagurus L. PLOS ONE 17(7), e0270960.

Increasing attention is being paid to the welfare of decapod crustaceans. Legislation exists for their humane slaughter in several countries and this is being debated in others. Electrical stunning may have potential for humane slaughter...

Wahltinez, S. J., Stacy, N. I., Hadfield, C. A. et al. 2022. Perspective: Opportunities for advancing aquatic invertebrate welfare. Frontiers in Veterinary Science 9, 973376.

Welfare considerations and regulations for invertebrates have lagged behind those for vertebrates, despite invertebrates comprising more than 95% of earth’s species. Humans interact with and use aquatic invertebrates for exhibition in zoos and aquaria, as...

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...

Conte, F., Voslarova, E., Vecerek, V. et al. 2021. Humane slaughter of edible decapod crustaceans. Animals 11(4), 1089.

Vast numbers of crustaceans are produced by aquaculture and caught in fisheries to meet the increasing demand for seafood and freshwater crustaceans. Simultaneously, the public is increasingly concerned about current methods employed in their handling...

Wahltinez, S. J., Kroll, K. J., Nunamaker, E. A. et al. 2021. Practical euthanasia method for common sea stars (Asterias rubens) that allows for high-quality RNA sampling. Animals 11(7), 1847.

Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were...

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...

Frost, K., Shah, M., Leung , V. S. Y. et al. 2020. Aversion to desflurane and isoflurane in Sprague-Dawley rats (Rattus norvegicus). Animals 10(6), 950.

Carbon dioxide and isoflurane are widely used for killing rats, yet may not truly achieve “euthanasia”, because they elicit aversion. The inhalant anesthetic desflurane is faster acting than isoflurane, representing a potential refinement. Using an...

Ballantine, A., Rodgers, D., Watts, C. et al. 2020. The cotton rat – a new challenge. Animal Technology and Welfare 19(1), 92-95.

This poster describes how the Animal Technologists at Envigo adapted their current practices and procedures for a new species at their laboratory: the cotton rat. The paper describes cotton rat biology and behaviour; housing; environmental...

Améndola, L., Weary, D. M. 2020. Understanding rat emotional responses to CO2. Translational Psychiatry 10, 253.

The aim of this review is to summarize evidence regarding rat emotional experiences during carbon dioxide (CO2) exposure. The studies reviewed show that CO2 exposure is aversive to rats, and that rats respond to CO2...

Turner, P. V., Hickman, D. L., van Luijk, J. et al. 2020. Welfare impact of carbon dioxide euthanasia on laboratory mice and rats: A systematic review. Frontiers in Veterinary Science 7, 411.

Background: There has been increased concern about the suitability of CO2 as a method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals may experience prior to loss...

Shomer, N. H., Allen-Worthington, K. H., Hickman, D. L. et al. 2020. Review of rodent euthanasia methods. JAALAS 59(3), 242–253.

The optimal choice of euthanasia method for laboratory rodents depends on a number of factors, including the scientific goals of the study, the need to minimize animal pain and/or distress, applicable guidelines and laws, the...

Laferriere, C. A., Pang, D. S. 2020. Review of intraperitoneal injection of sodium pentobarbital as a method of euthanasia in laboratory rodents. JAALAS 59(3), 254–263.

Euthanasia is one of the most commonly performed procedures in biomedical research, involving tens of millions of animals in North America and Europe every year. The use of sodium pentobarbital, injected intraperitoneally, for killing rodents...

Cohen, S., Kwok, M., Huang, J. 2020. Humane euthanasia of guinea pigs (Cavia porcellus) with a penetrating spring-loaded captive bolt. Animals 10(8), 1356.

Guinea pigs (Cavia porcellus) have been used in research since the 19th century to collect data due to their physiological similarities to humans. Today, animals perform a vital role in experiments and concerns for laboratory...

Kulkarni, S., Hickman, D. 2020. Isoflurane and carbon dioxide elicit similar behavioral responses in rats. Animals 10(8), 1431.

Euthanasia in rodents is an ongoing topic of debate due to concerns regarding the aversive nature of gases with anesthetic properties such as carbon dioxide (CO2) and isoflurane. The aim of this study was to...

Axiak Flammer, S., Eskes, C., Kohler, I. et al. 2019. Alternatives to carbon dioxide—Taking responsibility for humanely ending the life of animals. Animals 9(8), 482.

Carbon dioxide (CO2) is commonly used to kill rodents. However, a large body of research has now established that CO2 is aversive to them. A multidisciplinary symposium organized by the Swiss Federal Food Safety and...

Steiner, A. R., Axiak Flammer, S., Beausoleil, N. J. et al. 2019. Humanely ending the life of animals: Research priorities to identify alternatives to carbon dioxide. Animals 9(11), 911.

The use of carbon dioxide (CO2) for stunning and killing animals is considered to compromise welfare due to air hunger, anxiety, fear, and pain. Despite decades of research, no alternatives have so far been found...

Adams, R., Stanley, C. E., Piana, E. et al. 2019. Physiological and behavioral indicators to measure crustacean welfare. Animals 9(11), 914.

The purpose of this project was to determine how neural circuits were affected during warming by examining sensory neurons, the neuromuscular junction, and the cardiac function and behavior of the commercially important crustacean species, the...

Darbyshire, A. K., Oliver, K. H., Dupont, W. D. et al. 2019. Anesthesia and euthanasia of brine shrimp (Artemia franciscana). JAALAS 58(1), 58-63.

Invertebrates are often overlooked as laboratory animals, yet they are commonly used in toxicology, developmental, cellular and molecular biology, and radiation studies with euthanasia as an endpoint. Little is known regarding appropriate euthanasia methods for...