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. 
Weaver, H. L., Carbaugh, C. M., Madejczyk, M. S. et al. 2024. Comparison of tricaine methanesulfonate (MS-222) and alfaxalone anesthesia in zebrafish (Danio rerio). JAALAS 63(1), 74–80.

The research use of zebrafish has risen exponentially over the past decade while anesthetic options have remained largely unchanged.6 ricaine methanesulfonate (MS-222) is widely accepted as an anesthetic for routine husbandry procedures, however it has...

Jorge, S., Félix, L., Costas, B. et al. 2023. Housing conditions affect adult zebrafish (Danio rerio) behavior but not their physiological status. Animals 13(6), 1120.

Zebrafish is a valuable model for neuroscience research, but the housing conditions to which it is exposed daily may be impairing its welfare status. The use of environmental enrichment and the refinement of methodology for...

Wong, C.-H., Tsai, M.-A., Ko, F.-C. et al. 2023. Skin cortisol and acoustic activity: Potential tools to evaluate stress and welfare in captive cetaceans. Animals 13(9), 1521.

As people’s focus broadens from animals on farms to zoos and aquaria, the field of welfare science and the public’s concern for animal welfare continue to grow. In captive animals, stress and its causes are...

Cameron, A. S., Rub, A. M. W., Sandford, B. P. 2023. Evaluation of healing progression at surgical incision sites and the use of antiseptics for enhancing post-operative survival in subyearling Chinook salmon (Oncorhynchus tshawytscha). PLOS ONE 18(7), e0288056.

In an attempt to develop more effective surgical implantation methods for fish, surgical incisions typical of those made for implanting micro-acoustic transmitters into the peritoneal cavity were evaluated on a weekly basis for healing progression...

Jones, B. L., McClain, A. M., Sportelli, J. J. et al. 2023. Return of sound production as a biomarker of bottlenose dolphin emergence from anesthesia. Animals 13(15), 2531.

(1) Background: When a human or animal is recovering from general anesthesia, their medical team uses several behavioral and physiological parameters to assess their emergence from the unconscious state to complete wakefulness. However, the return...

Leyden, C., Brüggemann, T., Debinski, F. et al. 2022. Efficacy of tricaine (MS-222) and hypothermia as anesthetic agents for blocking sensorimotor responses in larval zebrafish. Frontiers in Veterinary Science 9, 864573.

Tricaine, or MS-222, is the most commonly used chemical anesthetic in zebrafish research. It is thought to act via blocking voltage-gated sodium channels, though its mechanism of action, particularly at the neuronal level, is not...

Midttun, H., Øverli, Ø., Tudorache, C. et al. 2022. Non-invasive sampling of water-borne hormones demonstrates individual consistency of the cortisol response to stress in laboratory zebrafish (Danio rerio). Scientific Reports 12(1) 6278.

Glucocorticoid (GC) stress hormones are well-known for their impact on phenotypic traits ranging from immune function to behaviour and cognition. For that reason, consistent aspects of an individual’s physiological stress response (i.e. GC responsiveness) can...

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...

Jang, J. Y., Liang, T., Kim, M.-K. et al. 2022. A rapid and noninvasive method that extracts polymerase chain reaction-ready genomic DNA from adult zebrafish. Zebrafish 19(4), 160-164.

Genotyping usually entails analysis of the products of polymerase chain reaction (PCR) carried out with genomic DNA (gDNA) as template, and is employed for validation of mutant or transgenic organisms. For genotyping of adult zebrafish...

Mason, G. J., Lavery, J. M. 2022. What is it like to be a bass? Red herrings, fish pain and the study of animal sentience. Frontiers in Veterinary Science 9, 788289.

Debates around fishes’ ability to feel pain concern sentience: do reactions to tissue damage indicate evaluative consciousness (conscious affect), or mere nociception? Thanks to Braithwaite’s research leadership, and concerns that current practices could compromise welfare...

Owen, J. P., Kelsh, R. N. 2021. A suitable anaesthetic protocol for metamorphic zebrafish. PLOS ONE 16(3), e0246504.

Zebrafish are frequently used as a means to investigate development. These studies increasingly require repeated anaesthesia of zebrafish during juvenile (i.e. metamorphic) stages. The effects of anaesthesia during this time remain poorly studied. The aim...

Reckendorf, A., Schmicke, M., Bunskoek, P. et al. 2021. Is harbor porpoise (Phocoena phocoena) exhaled breath sampling suitable for hormonal assessments? Animals 11(3), 907.

Over the last decades, exhaled breath sampling has been established for laboratory analysis in various cetacean species. Due to their small size, the usability of respiratory vapor for hormone assessments was questionable in harbor porpoises...

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...

Kolarevic, J., Calduch-Giner, J., Espmark, Å. M. et al. 2021. A novel miniaturized biosensor for monitoring Atlantic salmon swimming activity and respiratory frequency. Animals 11(8), 2403.

The advanced development of sensor technologies has led to the emergence of fish biosensors that are currently used for research and commercial purposes. AEFishBIT is a miniaturized biosensor attached to fish operculum that measures physical...

Zemanova, M. A. 2021. Noninvasive genetic assessment is an effective wildlife research tool when compared with other approaches. Genes 12(11), 1672.

Wildlife research has been indispensable for increasing our insight into ecosystem functioning as well as for designing effective conservation measures under the currently high rates of biodiversity loss. Genetic and genomic analyses might be able...

Jorge, S., Ferreira, J. M., Olsson, I. A. S. et al. 2021. Efficacy and behavioral recovery of different anesthetics. Zebrafish 18(5), 330-337.

The use of proper anesthesia in zebrafish research is essential to ensure fish welfare and data reliability. However, anesthesia long-term side effects remain poorly understood. The purpose of this study was to assess anesthesia quality...

Byrnes, E. E., Lear, K. O., Morgan, D. L. et al. 2020. Respirometer in a box: Development and use of a portable field respirometer for estimating oxygen consumption of large‐bodied fishes. Journal of Fish Biology 96(4), 1045-1050.

Respirometry has become the standard method for measuring the metabolic rate of fishes. Traditionally, respirometry has required the fish to be kept in captivity and tested under controlled conditions; however, many species do not readily...

Zemanova, M. A. 2020. Towards more compassionate wildlife research through the 3Rs principles: Moving from invasive to noninvasive methods. Wildlife Biology 2020(1), no page numbers.

Research in ecology and wildlife biology remains crucial for increasing our knowledge and improving species management and conservation in the midst of the current biodiversity crisis. However, obtaining information on population status often involves invasive...

Tilley, C. A., Carreño Gutierrez, H., Sebire, M. et al. 2020. Skin swabbing is a refined technique to collect DNA from model fish species. Scientific Reports 10(1), 18212.

Model fish species such as sticklebacks and zebrafish are frequently used in studies that require DNA to be collected from live animals. This is typically achieved by fin clipping, a procedure that is simple and...

Tomasello, D. L., Sive, H. 2020. Noninvasive multielectrode array for brain and spinal cord local field potential recordings from live zebrafish larvae. Zebrafish 17(4), 271-277.

Zebrafish are an important and expanding experimental system for brain research. We describe a noninvasive electrophysiology technique that can be used in living larvae to measure spontaneous activity in the brain and spinal cord simultaneously...

Musk, G. C., Ezzy, B. J., Kenchington, L. M. et al. 2020. A comparison of buffered tricaine methanesulfonate (MS-222) and isoeugenol anesthesia for caudal fin clipping in zebrafish (Danio rerio). JAALAS 59(6), 732-736.

The purpose of this study was to compare the safety and efficacy of buffered tricaine methanesulfonate (MS-222) and isoeugenol for the anesthesia of zebrafish undergoing caudal fin clipping. Eighty 9 mo Danio rerio (AB strain)...

Serres, A., Robeck, T., Deng, X. et al. 2020. Social, reproductive and contextual influences on fecal glucocorticoid metabolites in captive Yangtze finless porpoises (Neophocaena asiaeorientalis asiaeorientalis) and bottlenose dolphins (Tursiops truncatus). Journal of Zoological and Botanical Gardens 1(1), 24–41.

Although the use of fecal glucocorticoid metabolite (FGCM) measurements as non-invasive biomarkers for the stress response in mammals has increased, few studies have been conducted in odontocetes. We investigated if animal sex, age, pregnancy or...

McMurray, C., Allen, N., Hook, S. et al. 2018. I3S software as a refinement technique to identify individuals within a captive Scyliorhinus canula population. Animal Technology and Welfare 17(3), 204-205 (IAT Congress 2018 Poster Presentation).

Demarking individual animals within a group is often required in research. However, the process of invasive tagging induces stress and if a tag becomes detached it may wound the animal and/or prevent identification. Photo identification...

McKenzie, D. J., Sadoul, B., Geffroy, B. 2019. Measuring cortisol, the major stress hormone in fishes. Journal of Fish Biology 94(4), 540-555.

Stress in teleosts is an increasingly studied topic because of its interaction with growth, reproduction, immune system and ultimately fitness of the animal. Whether it is for evaluating welfare in aquaculture, adaptive capacities in fish...

Sloman, K, A., Bouyoucos, I. A., Brooks, E. J. et al. 2019. Ethical considerations in fish research. Journal of Fish Biology 94(4), 556-577.

In this review, we focus primarily on the refinement of common methods used in fish research based on emerging knowledge with the aim of improving the welfare of fish used in scientific studies. We consider...