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. 
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...

Bradley, H. S., Craig, M. D., Tomlinson, S. et al. 2023. Ecological considerations when designing mitigation translocations: An Australian reptile case study. Animals 13(16), 2594.

Translocation science has made considerable progress over the last two decades; however, reptile translocations still frequently fail around the world. Major knowledge gaps surround the basic ecology of reptile species, including basic factors such as...

Du Plooy, K. J., Swan, G. E., Myburgh, J. G. et al. 2023. Electroencephalogram (EEG) assessment of brain activity before and after electrical stunning in the Nile crocodile (Crocodylus niloticus). Scientific Reports 13(1), 20250.

Electrical stunning is used to capture crocodiles to perform routine management procedures. It is essential from a welfare point that electrical stunning must cause unconsciousness in animals. However, there is no information of whether or...

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...

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...

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...

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...

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...

Martinic, G. 2021. Husbandry and healthcare of the Olive python (Liasis olivaceus). Animal Technology and Welfare 20(1), 44-50.

The article describes the care of the Olive python (Liasis olivaceus) including information on housing, feeding, handling, as well as the general health care of these animals. Some of the information presented is derived from...

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...

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...

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...

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...

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...

Eichner, M., Garcia, K. 2019. Increase in morbidity and mortality in a shipment of red-eared slider turtles (Trachemys scripta elegans). Laboratory Animals 53(6), 634–640.

A cohort of captive-bred red-eared slider turtles, Trachemys scripta, was received from a commercial vendor. Shortly after arrival, several turtles presented as lethargic with subjectively pale skin and multifocal areas of cotton-like tufts in the...

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...

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...

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...

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...

Köhler, A., Collymore, C., Finger-Baier, K. et al. 2017. Report of Workshop on Euthanasia for Zebrafish—A Matter of welfare and science. Zebrafish 14(6), 547-551.

The increasing importance of zebrafish as a biomedical model organism is reflected by the steadily growing number of publications and laboratories working with this species. Regulatory recommendations for euthanasia as issued in Directive 2010/63/EU are...

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...

Lawton, S. 2017. Do Buccal swabs from Zebrafish give enough of a sample of DNA to be used as a viable non-invasive method of genotyping? Animal Technology and Welfare 16(3), 155-164.

The zebrafish has become a valuable research model within the laboratory. Currently the proven method of genotyping involves taking a tail fin clip from the fish to gain the sample. This involves surgically removing a...

Breacker, C., Tilley, C. 2017. DNA sampling by skin swabbing for Zebrafish (Danio rerio) and sticklebacks (Gasterosteus aculeatus). Animal Technology and Welfare 16(3), 211-213.

Fin clipping of live fish under anaesthesia is widely used to collect tissues samples for DNA extraction. However, this technique raises a number of ethical concerns, since the use of anaesthetic and/or physical fin damage...