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. 
Franks, B., Gaffney, L. P., Graham, C. et al. 2023. Curiosity in zebrafish (Danio rerio)? Behavioral responses to 30 novel objects. Frontiers in Veterinary Science 9, 1062420.

Curiosity—the motivation to seek out information—has been studied widely across the animal kingdom. To investigate curiosity in zebrafish we presented 30 novel objects to groups of zebrafish housed in semi-naturalistic tanks (6 tanks; 10 fish/tank...

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...

Marks, C. A., Allen, L., Lindeberg, H. 2023. Non-lethal dose-response models replace lethal bioassays for predicting the hazard of para-animopropiophenone to Australian wildlife. Animals 13(3), 472.

Para-aminopropiophenone (PAPP) is a potent methaemoglobin (MetHb) forming agent used for the lethal control of exotic carnivores and mustelids. To assess the sensitivity of Australian wildlife to PAPP we developed an in vivo assay that...

Sen Sarma, O., Frymus, N., Axling, F. et al. 2023. Optimizing zebrafish rearing−Effects of fish density and environmental enrichment. Frontiers in Behavioral Neuroscience 17, 1204021.

Despite its popularity in research, there is very little scientifically validated knowledge about the best practices on zebrafish (Danio rerio) husbandry, which has led to several facilities having their own husbandry protocols. This study was...

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...

Baskir, E., Parsons, A., Elden, M. et al. 2023. Quantifying acute behavioral reactions of Bali mynas (Leucopsar rothschildi) to environmental and progressively challenging enrichment. Journal of Zoological and Botanical Gardens 4(1), 176–190.

Animals use specific behaviors and skills to overcome challenges and access resources. Environmental enrichment is provided to animals in human care to both promote species-appropriate behaviors and reduce undesired behaviors. Feather pecking in birds is...

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(2), 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...

Mair, A., Dadda, M., Kitaoka, A. et al. 2023. Illu-shoal choice: An exploration of different means for enrichment of captive zebrafish. Animals 13(16), 2640.

Fish of any variety are nowadays being kept captive for several purposes, from recreational to alimentary to research. It is possible that we humans often underestimate or misunderstand the basic, natural needs of the species...

Gatto, E., Bruzzone, M., Dal Maschio, M. et al. 2022. Effects of environmental enrichment on recognition memory in zebrafish larvae. Applied Animal Behaviour Science 247, 105552.

Zebrafish (Danio rerio) constitute a useful model for studying memory function and impairment in vertebrates and are now widely used in translational research. On the one hand, the adoption of simple, fast and reliable tests...

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...

Marques Maia, C., Volpato, G. L., Braithwaite, V. A. 2022. A psychological aversive condition does not change individual zebrafish preference for background color or artificial plant enrichments. Journal of Applied Animal Welfare Science 25(4), 427-438.

Information on fish preference for environmental conditions can be a useful tool to offer them what they want, an important requirement for welfare purposes. Giving that such an approach deals with psychological states of the...

Alaniz, A. J., Astudillo-Hinojosa, K., Soto, A. F. et al. 2022. Rapid behavioral recovery based on environmental enrichment of a white-throated toucan (Ramphastos tucanus: Ramphastidae) affected by collision trauma. Journal of Veterinary Behavior 57, 1-5.

Captive animals that have experienced traumatic events require a high range of care prior to reintroducing them into their natural habitat; and this care should ensure their welfare in captivity. This report shows the ethological...

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...

Saraiva, J. L., Rachinas-Lopes, P., Arechavala-Lopez, P. 2022. Finding the “golden stocking density”: A balance between fish welfare and farmers’ perspectives. Frontiers in Veterinary Science 9, 930221.

The effects of stocking density on fish welfare are complex and involve many interacting parameters. This complex relationship between fish welfare, stocking density and influencing factors make it challenging to define a specific optimal (“golden”)...

Tsang, B., Gerlai, R. T. 2022. Common aquarium plants as an enrichment strategy in zebrafish

An increasing number of laboratories utilize zebrafish as this species is now represented in practically every subfield of biology research. Environmental enrichment has been shown to improve welfare and health of a large number of...

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...

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...

Barretta, L. A., Maloney, S. K., Blachea, D. 2021. Pekin ducks are motivated to access their nest site and exhibit a stress-induced hyperthermia when unable to do so. Animal 15(1), 100067.

The origins of floor-laying in ducks could be low motivation for a nest, or stress related to difficulties with accessing a nest (e.g. competition). Using a behavioural demand test, we investigated if increasing the work...

Marchetto, L., Barcellos, L. J. G., Koakoski, G. et al. 2021. Auditory environmental enrichment prevents anxiety-like behavior, but not cortisol responses, evoked by 24-h social isolation in zebrafish. Behavioural Brain Research 404, 113169.

The zebrafish (Danio rerio) is widely used as a promising translational model organism for studying various brain disorders. Zebrafish are also commonly used in behavioral and drug screening assays utilizing individually tested (socially isolated) fish...

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...

Stevens, C. H., Reed, B. T., Hawkins, P. 2021. Enrichment for laboratory zebrafish—A review of the evidence and the challenges. Animals 11(3), 698.

Good practice for the housing and care of laboratory zebrafish Danio rerio is an increasingly discussed topic, with focus on appropriate water quality parameters, stocking densities, feeding regimes, anaesthesia and analgesia practices, methods of humane...

Martini, A., Huysseune, A., Witten, P. E. et al. 2021. Plasticity of the skeleton and skeletal deformities in zebrafish (Danio rerio) linked to rearing density. Journal of Fish Biology 98(4), 971-986.

The teleost zebrafish (Danio rerio), an established model for human skeletal diseases, is reared under controlled conditions with defined parameters for temperature and photoperiod. Studies aimed at defining the proper rearing density have been performed...

Huber, N., Mahr, K., Tóth, Z. et al. 2021. The stressed bird in the hand: Influence of sampling design on the physiological stress response in a free-living songbird. Physiology & Behavior 238, 113488.

Despite the widely used application of standardized capture-handling protocols to collect blood and assess the physiological stress response, the actual sampling design (e.g., timing and the number of blood samples) often differs between studies, and...

DePasquale, C., Kemerer, N., White, N. et al. 2021. The influence of an enriched environment in enhancing recognition memory in zebrafish (Danio rerio). Frontiers in Veterinary Science 8, 749746.

Environmental enrichment is used to increase social and physical stimulation for animals in captivity which can lead to enhanced cognition. Fundamental to the positive effect enrichment has on the brain is that it provides opportunities...