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. 
Buenhombre, J., Daza-Cardona, E. A., Sousa, P. et al. 2023. Structural environmental enrichment and the way it is offered influence cognitive judgement bias and anxiety-like behaviours in zebrafish. Animal Cognition 26, 563–577.

Environmental enrichment in zebrafish generally reduces anxiety-related behaviours, improves learning in maze trials and increases health and biological fitness. However, certain types of enrichment or certain conditions induce the opposite effects. Therefore, it is essential...

Fontana, B. D., Alnassar, N., Parker, M. O. 2023. Transgenerational effects of early-life stress on anxiety behavior in zebrafish (Danio rerio). Behavioural Processes 208, 104874.

Early-life adversity impacts on anxiety-related behaviors in adulthood. The effects of such adversity not only affect the animal itself, but can be passed on transgenerationally. Pervasive effects of experimentally-induced early-life stress (ELS) have been documented...

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...

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...

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...

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...

Gilmour, K. M., Bard, B. 2022. Social buffering of the stress response: Insights from fishes. Biology Letters 18(10), 20220332.

Social buffering of stress refers to the effect of a social partner in reducing the cortisol or corticosterone response to a stressor. It has been well studied in mammals, particularly those that form pair bonds...

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...

Sørensen, D., Cloutier, S., Gaskill, B. (Eds.). 2021. Animal-centric Care and Management - Enhancing Refinement in Biomedical Research. CRC Press, Boca Raton, FL.

The concept of the 3Rs (Refinement, Reduction and Replacement) has been used as a framework for improving the welfare of laboratory animals for the last half century. By establishing an animal-centric view on housing and...

Burbano Lombana, D. A., Macrì, S., Porfiri, M. 2021. Collective emotional contagion in zebrafish. Frontiers in Behavioral Neuroscience 15, 730372.

Seeking to match our emotional state with one of those around us is known as emotional contagion-a fundamental biological process that underlies social behavior across several species and taxa. While emotional contagion has been traditionally...

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...

Reese, L., Ladwig-Wiegard, M., von Fersen, L. et al. 2020. Deflighting zoo birds and its welfare considerations. Animal Welfare 29(1), 69-80.

For over a century, the practice of deflighting has taken place in zoological collections in order to ensure birds remain in open-topped enclosures. Over time, efforts have been made to improve or develop new (surgical)...

Angiulli, E., Pagliara, V., Cioni, C. et al. 2020. Increase in environmental temperature affects exploratory behaviour, anxiety and social preference in Danio rerio. Scientific Reports 10(1), 5385.

The aim of this work is to investigate the effect of a temperature increase on the behaviour of adult zebrafish (Danio rerio) maintained for 21 days at 34 °C (treatment) and 26 °C (control). The temperatures chosen...

Yaw, T. J., Mans, C., Johnson,S. et al. 2020. Evaluation of subcutaneous administration of alfaxalone-midazolam and dexmedetomidine-midazolam for sedation of ball pythons (Python regius). Journal of the American Veterinary Medical Association 256(5), 573-579.

OBJECTIVE: To evaluate SC administration of alfaxalone-midazolam and dexmedetomidine-midazolam for sedation of ball pythons (Python regius). ANIMALS: 12 healthy juvenile ball pythons. PROCEDURES: In a randomized crossover study, each snake was administered a combination of...

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...

Aleström, P., D’Angelo, L., Midtlyng, P. J. et al. 2020. Zebrafish: Housing and husbandry recommendations. Laboratory Animals 54(3), 213–224.

This article provides recommendations for the care of laboratory zebrafish (Danio rerio) as part of the further implementation of Annex A to the European Convention on the protection of vertebrate animals used for experimental and...

Kane, L. P., Chinnadurai, S. K., Vivirito, K. et al. 2020. Comparison of isoflurane, sevoflurane, and desflurane as inhalant anesthetics in prairie rattlesnakes (Crotalus viridis). Journal of the American Veterinary Medical Association 257(9), 945-949.

OBJECTIVE: To characterize induction and recovery characteristics of 3 commonly used inhalant anesthetics in prairie rattlesnakes (Crotalus viridis): isoflurane, sevoflurane, and desflurane. ANIMALS: 12 healthy adult prairie rattlesnakes. PROCEDURES: In a randomized crossover design, snakes...

Natusch, D. J. D., Aust, P. W., Khadiejah, S. et al. 2020. Behavioral and corticosterone responses to carbon dioxide exposure in reptiles. PLOS ONE 15(10), e0240176.

The use of carbon dioxide (CO2) exposure as a means of animal euthanasia has received considerable attention in mammals and birds but remains virtually untested in reptiles. We measured the behavioral responses of four squamate...

Kane, L. P., Chinnadurai, S. K., Vivirito, K. et al. 2020. Comparison of isoflurane, sevoflurane, and desflurane as inhalant anesthetics in prairie rattlesnakes (Crotalus viridis). Journal of the American Veterinary Medical Association 257(9), 945-949.

OBJECTIVE: To characterize induction and recovery characteristics of 3 commonly used inhalant anesthetics in prairie rattlesnakes (Crotalus viridis): isoflurane, sevoflurane, and desflurane. ANIMALS: 12 healthy adult prairie rattlesnakes. PROCEDURES: In a randomized crossover design, snakes...

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)...

Tan, S. L. T., Handasyde, K. A., Rault, J.-L. et al. 2020. Insensitivity to reward shifts in zebrafish (Danio rerio) and implications for assessing affective states. Animal Cognition 23(1), 87–100

Theory and empirical findings predict that individuals in a negative affective state are more sensitive to unexpected reward loss and less sensitive to unexpected reward gain compared to individuals in a neutral or positive affective...

Fife-Cook, I., Franks, B. 2019. Positive welfare for fishes: Rationale and areas for future study. Fishes 4(2), 31.

Traditional animal welfare paradigms have focused on maintaining physical health and mitigating negative impacts to wellbeing. Recently, however, the field has increasingly recognized the importance of positive welfare (i.e., mental and physical states that exceed...

Silva, P. F., de Leaniz, C. G., Luchiari, A. C. 2019. Fear contagion in zebrafish: A behaviour affected by familiarity. Animal Behaviour 153, 95-103.

Emotional contagion has recently been described in fish but whether it is affected by familiarity is not known. We tested whether the sight of a distressed conspecific elicited fear in zebrafish, Danio rerio, and whether...

Lambert, H., Carder, G., D'Cruze, N. 2019. Given the cold shoulder: A review of the scientific literature for evidence of reptile sentience. Animals 9(10), 821.

We searched a selection of the scientific literature to document evidence for, and explorations into reptile sentience. The intention of this review was to highlight; (1) to what extent reptile capability for emotions have been...

Baker, S. W., Tucci, E. R., Felt, S. A. et al. 2019. A bird's-eye view of regulatory, animal care, and training considerations regarding avian flight research. Comparative Medicine 69(3), 169–178.

A thorough understanding of how animals fly is a central goal of many scientific disciplines. Birds are a commonly used model organism for flight research. The success of this model requires studying healthy and naturally...