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. 
Lambert, H., Cornish, A., Waley, D. 2024. The value of regulating stocking densities in aquaculture must not be dismissed: A reply to Saraiva et al. 2022. Frontiers in Veterinary Science 10.

We would like to rebut the argument made by Saraiva et al. (1) in their paper “Finding the “golden stocking density”: a balance between fish welfare and farmers' perspectives,” that stocking densities should not be...

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...

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...

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...

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...

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”)...

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...

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...

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...

da Silva Medeiros Elidio, H., Coelho, J. W. R., Cavalcanti Pereira da Silva, L. C. et al. 2021. Housing density and aggression in Syrian hamsters. JAALAS 60(5), 506-509.

The Syrian hamster (Mesocricetus auratus) is a solitary and naturally territorial animal, with female hamsters being more aggressive than males. This behavior makes handling difficult because they are usually housed in groups, which can lead...

Andersson, M., Kettunen, P. 2021. Effects of holding density on the welfare of zebrafish: A systematic review. Zebrafish 18(5), 297-306.

The zebrafish is becoming an increasingly popular research animal around the world. Its welfare is affected by an array of environmental factors, such as food access and water quality. Holding density is an important welfare...

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...

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...

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)...

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...

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...

Ehrlich, O., Karamalakis, A., Krylov, A. J. et al. 2019. Clove oil and AQUI-S efficacy for zebrafish embryo, larva, and adult anesthesia. Zebrafish 16(5), 451-459.

Since the use of the zebrafish Danio rerio genetic model organism within the scientific research community continues to grow rapidly, continued procedural refinement to support high-quality, reproducible research and improve animal welfare remains an important...

Lambert, C. J., Freshner, B. C., Chung, A. et al. 2018. An automated system for rapid cellular extraction from live zebrafish embryos and larvae: Development and application to genotyping. PLOS ONE 13(3), e0193180.

Zebrafish are a valuable model organism in biomedical research. Their rapid development, ability to model human diseases, utility for testing genetic variants identified from next-generation sequencing, amenity to CRISPR mutagenesis, and potential for therapeutic compound...

Santurtun, E., Broom, D. M., Phillips, C. J. C. 2018. A review of factors affecting the welfare of Atlantic salmon (Salmo salar). Animal Welfare 27(3), 193-204.

In the expanding salmon industry, many farmers use production methods that could result in poor welfare of the fish at various points of their lifecycle. We have reviewed methods used for producing salmon for food...

Kennedy, R. J., Allen, M., Wilson, R. 2018. Tag retention and mortality of adult Atlantic salmon Salmo salar gastrically tagged with different sized telemetry transmitters. Journal of Fish Biology 92(6), 2016-2021.

A batch of 1 sea winter pre-spawning adult Salmo salar from the Bush river in Northern Ireland, U.K., were gastrically tagged with large (13-mm diameter) and small (9-mm diameter) dummy acoustic telemetry tags alongside untagged...

Riley, W. D., Ibbotson, A. T., Gregory, S. D. et al. 2018. Under what circumstances does the capture and tagging of wild Atlantic salmon Salmo salar smolts affect probability of return as adults? Journal of Fish Biology 93(3), 477-489.

Adult return rates for wild Atlantic salmon Salmo salar smolts captured in a rotary screw trap and tagged with coded wire (CW) tags were compared with a control group, using detections from passive integrated transponder...

Chance, R. J., Cameron, G. A., Fordyce, M. et al. 2018. Effects of repeated anaesthesia on gill and general health of Atlantic salmon, Salmo salar. Journal of Fish Biology 93(6), 1069-1081.

Fish are the second most widely utilized vertebrate group used for scientific procedures in the United Kingdom, but the development and application of 3Rs (the principles of replacement, reduction, and refinement) in aquaculture disease research...

Ruberto, T., Clément, R. J. G., Spinello, C. et al. 2018. The tagging procedure of visible implant elastomers influences zebrafish individual and social behavior. Zebrafish 15(5), 433-444.

One commonly used method to preserve individual identity in the study of social behavior of zebrafish is through silicone-based visible implant elastomers (VIEs), which represent a safe and durable tagging procedure. While the effects of...

Breacker, C., Barber, I., Norton, W. H. J. et al. 2017. A low-cost method of skin swabbing for the collection of DNA samples from small laboratory fish. Zebrafish 14(1), 35-41.

Fin clipping of live fish under anesthesia is widely used to collect samples for DNA extraction. An alternative, potentially less invasive, approach involves obtaining samples by swabbing the skin of nonanesthetized fish. However, this method...