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. 
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...

Heniff, A. C., Gorges, M. A., Lewbart, G. A. et al. 2023. Evaluation of targeted injections of ivermectin or potassium chloride for euthanasia of anesthetized thorny devil stick insects (Eurycantha calcarata). Journal of Zoological and Botanical Gardens 4(1), 231–239.

Insects are commonly utilized in biomedical research and have become increasingly popular in museum collections and as pets. Despite this, objective evaluation of insect euthanasia is scarce. This study investigated the effectiveness of targeted injections...

Koperski, P. 2023. It is not only data—Freshwater invertebrates misused in biological monitoring. Animals 13(16), 2570.

The article presents and discusses the issues of the use of free-living invertebrates to assess the ecological status of freshwater environments with different methods of biological monitoring. Invertebrates are excluded from ethical consideration in the...

Fehrenbach, L. A., Adekanye, A. A., Palillo, M. B. et al. 2022. Internal temperatures of packaging for overnight cross-country shipping of zebrafish (Danio rerio). Comparative Medicine 72(3), 195-203.

As the use of zebrafish (Danio rerio) as a research model continues to rise, so too will the shipping and sharing of zebrafish strains across collaborating institutions. If done incorrectly, shipping can result in significant...

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...

Neil, D. M., Albalat, A., Thompson, J. 2022. The effects of electrical stunning on the nervous activity and physiological stress response of a commercially important decapod crustacean, the brown crab Cancer pagurus L. PLOS ONE 17(7), e0270960.

Increasing attention is being paid to the welfare of decapod crustaceans. Legislation exists for their humane slaughter in several countries and this is being debated in others. Electrical stunning may have potential for humane slaughter...

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...

Wahltinez, S. J., Stacy, N. I., Hadfield, C. A. et al. 2022. Perspective: Opportunities for advancing aquatic invertebrate welfare. Frontiers in Veterinary Science 9, 973376.

Welfare considerations and regulations for invertebrates have lagged behind those for vertebrates, despite invertebrates comprising more than 95% of earth’s species. Humans interact with and use aquatic invertebrates for exhibition in zoos and aquaria, as...

Ramos, J., Balasch, J. C., Tort, L. 2021. About welfare and stress in the early stages of fish. Frontiers in Veterinary Science 8, 634434.

In conclusion, the investigations carried out up to now demonstrate that during the early stages fish show high sensitivity to many types of stressors involving an array of responses to overcome alterations that could affect...

Conte, F., Voslarova, E., Vecerek, V. et al. 2021. Humane slaughter of edible decapod crustaceans. Animals 11(4), 1089.

Vast numbers of crustaceans are produced by aquaculture and caught in fisheries to meet the increasing demand for seafood and freshwater crustaceans. Simultaneously, the public is increasingly concerned about current methods employed in their handling...

Wahltinez, S. J., Kroll, K. J., Nunamaker, E. A. et al. 2021. Practical euthanasia method for common sea stars (Asterias rubens) that allows for high-quality RNA sampling. Animals 11(7), 1847.

Sea stars in research are often lethally sampled without available methodology to render them insensible prior to sampling due to concerns over sufficient sample quality for applied molecular techniques. The objectives of this study were...

Filliettaz, C., Maxwell, L., Dragon, M. et al. 2020. Novel bleeding techniques in hamsters. Laboratory Animal Science Professional 8(1) (January/February), 44-46.

Hamsters have historically been used in our pharmacokinetic (PK) studies using the retro-orbital (RO) bleeding technique to collect blood samples. If performed incorrectly, this technique has the potential for animal welfare complications not usually seen...

Lawrence, M. J., Raby, G. D., Teffer, A. K. et al. 2020. Best practices for non-lethal blood sampling of fish via the caudal vasculature. Journal of Fish Biology 97(1), 4–15.

Blood sampling through the caudal vasculature is a widely used technique in fish biology for investigating organismal health and physiology. In live fishes, it can provide a quick, easy and relatively non‐invasive method for obtaining...

Reinhardt, V. (ed.) 2020. It’s Okay to Cry - Discussions by the Laboratory Animal Refinement & Enrichment Forum [LAREF], Volume V. Animal Welfare Institute, Washington, DC.

This is the 5th volume of selected discussions that took place on the electronic Laboratory Animal Refinement & Enrichment Forum between February 2016 and December 2019. The forum was created in October 2002; it allows...

Shomer, N. H., Allen-Worthington, K. H., Hickman, D. L. et al. 2020. Review of rodent euthanasia methods. JAALAS 59(3), 242–253.

The optimal choice of euthanasia method for laboratory rodents depends on a number of factors, including the scientific goals of the study, the need to minimize animal pain and/or distress, applicable guidelines and laws, the...

Schou, T. M., Palme, R., Malmkvist, J. et al. 2019. Relocation shortly after mating does not have a major impact on stress responses and reproduction in female farm mink. Applied Animal Behaviour Science 214, 89-94.

On farms, female mink are exposed to acute stressors multiple times due to handling, capturing and transport with restricted movement being a typical part of the mating procedure. We hypothesised that no relocation (group NON...

Axiak Flammer, S., Eskes, C., Kohler, I. et al. 2019. Alternatives to carbon dioxide—Taking responsibility for humanely ending the life of animals. Animals 9(8), 482.

Carbon dioxide (CO2) is commonly used to kill rodents. However, a large body of research has now established that CO2 is aversive to them. A multidisciplinary symposium organized by the Swiss Federal Food Safety and...

Adams, R., Stanley, C. E., Piana, E. et al. 2019. Physiological and behavioral indicators to measure crustacean welfare. Animals 9(11), 914.

The purpose of this project was to determine how neural circuits were affected during warming by examining sensory neurons, the neuromuscular junction, and the cardiac function and behavior of the commercially important crustacean species, the...

Darbyshire, A. K., Oliver, K. H., Dupont, W. D. et al. 2019. Anesthesia and euthanasia of brine shrimp (Artemia franciscana). JAALAS 58(1), 58-63.

Invertebrates are often overlooked as laboratory animals, yet they are commonly used in toxicology, developmental, cellular and molecular biology, and radiation studies with euthanasia as an endpoint. Little is known regarding appropriate euthanasia methods for...

Dunford, K., Hakkesteeg, J., Wilson, C. 2019. Stiff as a board: Measuring rigor mortis in zebrafish. Animal Technology and Welfare 18(3), 203-205.

Schedule one killing (S1K) methods require a two-step process: a humane method of death, typically for Zebrafish an anaesthetic overdose and confirmation of death, such as confirmation of rigor mortis. There is widespread variation of...

Robinson, P., Trim, S., Trim, C. 2019. Non-invasive extraction of Cnidarian venom through the use of autotomised tentacles. Animal Technology and Welfare 18(3), 167-173.

The animals contained within the phylum Cnidaria (Sea Anemones, Corals, Sea Pens, Jellyfish, Boxjellies and Hydra) have origins that can be dated back to around 750 million years ago (mya) and as such, they represent...

Thurman, C. E., Rasmussen, S., Prestia, K. A. 2019. Effect of 3 euthanasia methods on serum yield and serum cortisol concentration in zebrafish (Danio rerio). JAALAS 58(6), 823–828.

Zebrafish are an important model in neuroscience and developmental biology and are also an emerging model in hematology and immunology. Little information is available for zebrafish regarding the physiologic impact of different euthanasia methods and...

Wallace, C. K., Bright, L. A., Marx, J. O. et al. 2018. Effectiveness of rapid cooling as a method of euthanasia for young zebrafish (Danio rerio). JAALAS 57(1), 58-63.

Despite increased use of zebrafish (Danio rerio) in biomedical research, consistent information regarding appropriate euthanasia methods, particularly for embryos, is sparse. Current literature indicates that rapid cooling is an effective method of euthanasia for adult...

Baker, S., Knight, E., Pellett, S. et al. 2018. Spider and chips: The use of internal Radio Frequency Identification (RFID) chips as a minimally invasive method to measure internal body temperatures in invertebrates. Animal Technology and Welfare 17(1), 1-7.

Internal RFID transponders have been used in vertebrates for many years, however studies into their use in invertebrates are less well represented in the literature. The use of RFID transponders for internal temperature measurement represents...