Refinement Database

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

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

Links to the full text for publications that appear in open access journals or are published on the AWI website are provided under the abstract.

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 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. Most publications older than 2010 can only be searched by keyword. 
Baker, P. E., Nicol, C. J., Weeks, C. A. 2022. The effect of hard pecking enrichment during rear on feather cover, feather pecking behaviour and beak length in beak-trimmed and intact-beak laying hen pullets. Animals 12(6), 674.
Injurious pecking, commonly controlled by beak trimming (BT), is a widespread issue in laying hens associated with thwarted foraging. This controlled study compared the effect in intact and beak-trimmed pullets of providing pecking pans to...
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...
Davis, A. K., Garner, J. P., Chu, D. K. et al. 2022. Propofol immersion as a euthanasia method for adult zebrafish (Danio rerio). Comparative Medicine 72(3), 204-209.
The exponential rise of the zebrafish (Danio rerio) as a model organism in biomedical research has far outstripped our un- derstanding of basic husbandry and welfare for this species. As a case in point, here...
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...
Lennox, R. J., Stöger, E., Dahlmo, L. S. et al. 2022. Effects of tag type and surgery on migration of Atlantic salmon (Salmo salar) smolts. Journal of Fish Biology 101(3), 515-521.
Tagging salmon smolts to provide information about the timing of outmigration has been a common approach to monitor phenology and model the risk of encountering stressors. However, the validity of tagging has come under scrutiny...
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...
Ferreira, J. M., Félix, L., Jorge, S. et al. 2022. Anesthesia overdose versus rapid cooling for euthanasia of adult zebrafish. Zebrafish 19(4), 148-159.
The rapid increase in zebrafish use needs to be accompanied by research into the refinement of procedures. The European (EU) Directive lists three possible euthanasia methods for fish: anesthetic overdose, electrical stunning, and concussion. However...
Wotton, S., Grist, A., O’Callaghan, M. et al. 2021. Head-only stunning of turkeys Part 2: Subjective and objective assessment of the application of AC and DC waveforms. Animals 11(2), 286.
Electrical stunning is likely to remain an important stunning method for turkeys at slaughter. The purpose of this study is to understand the application of various waveforms of alternating current (AC) and pulsed direct currents...
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...
Rácz, A., Allan, B., Dwyer, T. et al. 2021. Identification of individual zebrafish (Danio rerio): A refined protocol for VIE tagging whilst considering animal welfare and the principles of the 3Rs. Animals 11(3), 616.
In aquatic ecology, studies have commonly employed a tagging technique known as visible implant elastomer (VIE). This method has not been widely adopted by the zebrafish research community and also lacks refinement with regard to...
Burn, C. C., Mazlan, N. H. B., Chancellor, N. et al. 2021. The pen is milder than the blade: Identification marking mice using ink on the tail appears more humane than ear-punching even with local anaesthetic. Animals 11(6), 1664.
Identification marking mice commonly involves ear-punching with or without anaesthetic, or tail-marking with ink. To identify which is most humane, we marked weanling male BALB/c mice using ear-punching (EP), ear-punching with anaesthetic EMLATM cream (EP+A)...
Grün, S., Damme, K., Müller, M. et al. 2021. Welfare and performance of three turkey breeds—Comparison between infrared beak treatment and natural beak abrasion by pecking on a screed grinding wheel. Animals 11(8), 2395.
Feather pecking and cannibalism are behavioral disorders that cause animal-welfare-relevant and economic problems. To mitigate these problems, the beaks of conventionally reared turkeys are usually already trimmed in the hatcheries. To find an alternative to...
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...
Schwarzer, A., Plattner, C., Bergmann, S. et al. 2021. Feather pecking in non-beak-trimmed and beak-trimmed laying hens on commercial farms with aviaries. Animals 11(11), 3085.
Severe feather pecking (SFP) is a major animal welfare problem in layers. It results in pain and injuries in the affected animal. It was the aim of this study to gain insight into the actual...
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...
Guerri, G., Cerasoli, I., Straticò, P. et al. 2021. The clinical effect of xylazine premedication in water buffalo calves (Bubalus bubalis) undergoing castration under general anaesthesia. Animals 11(12), 3433.
Castration is one of the most frequently performed surgical procedures in livestock. All methods of castration are associated with a significant degree of pain, affecting the patients’ welfare. Limited information on species-specific protocols in buffaloes...
Guinebretière, M., Mika, A., Michel, V. et al. 2020. Effects of management strategies on non-beak-trimmed laying hens in furnished cages that were reared in a non-cage system. Animals 10(3), 399.
Beak trimming in laying hens limits the negative consequences of injurious pecking, but could be prohibited by future regulations. This study assessed a combination of management strategies during the rearing period (objects, perches, music, human...
Guijarro, A., Mauri, S., Aviles, C. et al. 2020. Effects of two CO2 stunning methods on the efficacy of stunning and blood stress indicators of turkeys under commercial processing conditions. Journal of Applied Animal Welfare Science 23(2), 231-243.
The effect of CO2 gas-stunning methods (G1: 30% CO2 15 sec, 55% CO2 40 sec, 70% CO2 45 sec; G2: 30% CO2 15 sec, 80% CO2 85 sec) on the efficacy of stunning, blood stress...
Waiblinger, S., Hajek, F., Lambacher, B. et al. 2020. Effects of the method of restraint for shearing on behaviour and heart rate variability in alpacas. Applied Animal Behaviour Science 223, 104918.
Alpacas are increasingly kept in Europe for different purposes including fibre production. Yearly shearing is necessary to harvest fibre and for welfare reasons. Different methods of restraint are used during shearing, which may affect the...
Young, T., Whiteside, T., Locklear, J. 2020. What is your tattoo ink telling you? Laboratory Animal Science Professional 8(2) (March/April), 62-65.
Tattooing traumatizes the skin, which can result in microbial infections with the severity ranging from minor to life-threating septicemia. Additionally, the metals in colored tattoo ink are known to cause dermal inflammation in some people...
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...

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