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. 
Rault, J.-L., Lai, A., Hemsworth, L. et al. 2020. Wireless ‘under the skull’ epidural EEG and behavior in piglets during nitrous oxide or carbon dioxide gas euthanasia. Physiology & Behavior 227, 113142.

Consciousness is central to animal welfare concerns. Its assessment is most often conducted based on behavior, with a poor understanding of the correspondence between behavior and the neurobiological processes that underlie the subjective experience of...

Arruda, A. G., Beyene, T. J., Kieffer, J. et al. 2020. A systematic literature review on depopulation methods for swine. Animals 10(11), 2161.

Swine mass depopulation refers to the destruction of large numbers of pigs and may include not only animals affected with a disease but also healthy pigs in a facility or surrounding areas. Emerging applications of...

Atkinson, S., Algers, B., Pallisera, J. et al. 2020. Animal welfare and meat quality assessment in gas stunning during commercial slaughter of pigs using hypercapnic-hypoxia (20% CO2 2% O2) compared to acute hypercapnia (90% CO2 in Air). Animals 10(12), 2440.

This study assessed aversion, stunning effectiveness, and product quality of nitrogen and carbon dioxide (CO2) mixtures used for stunning pigs. A total of 1852 slaughter pigs divided into two similar batches was assessed during routine...

Lindahl, C., Sindhøj, E., Brattlund Hellgren, R. et al. 2020. Responses of pigs to stunning with nitrogen filled high-expansion foam. Animals 10(12), 2210.

Nitrogen gas (N2) delivered in high expansion foam in a closed container could be a feasible method for humanely stunning pigs. This study aimed to evaluate potential aversion in pigs to the N2 foam method...

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)...

Stanger, K. J., Kells, N. J., Fisher, A. D. et al. 2019. Evaluation of euthanasia of sheep with intravenous saturated salt solutions to enable the collection of whole, intact brains. Animal Welfare 28(4), 397-406.

Captive bolts or firearms are unsuitable for euthanasia of livestock when an intact brain is required for diagnostics. Injectable barbiturates can be used, but this method carries risk of poisoning animals eating the carcase. Intravenous...

Bouwsema, J. A., Lines, J. A. 2019. Could low atmospheric pressure stunning (LAPS) be suitable for pig slaughter? A review of available information. Animal Welfare 28(4), 421-432.

Low atmospheric pressure stunning (LAPS) is a slaughter technique which may be less stressful for pigs (Sus scrofa domestica) than current commercial stunning and slaughter methods. The main methods used currently for slaughtering pigs are...

Steiner, A. R., Axiak Flammer, S., Beausoleil, N. J. et al. 2019. Humanely ending the life of animals: Research priorities to identify alternatives to carbon dioxide. Animals 9(11), 911.

The use of carbon dioxide (CO2) for stunning and killing animals is considered to compromise welfare due to air hunger, anxiety, fear, and pain. Despite decades of research, no alternatives have so far been found...

Lutz, J., Burla, J.-B., Gygax, L. et al. 2019. Horned and dehorned dairy cows differ in the pattern of agonistic interactions investigated under different space allowances. Applied Animal Behaviour Science 218, 104819.

Limited space in dairy cattle farming is associated with frequent encounters between cows, often resulting in agonistic interactions and injuries, especially in horned cows. Previous studies with dairy cows investigated space allowances in the waiting...

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...

Cornale, P., Macchi, E., Miretti, S. et al. 2015. Effects of stocking density and environmental enrichment on behavior and fecal corticosteroid levels of pigs under commercial farm conditions. Journal of Veterinary Behavior 10(6), 569-576.

In intensive pig farming of Western and Southern Europe, welfare concerns are still often related to barren environments and crowded conditions. Pig producers need to balance the requirements to improve welfare conditions at farm with...

Musk, G. C., Kershaw, H., He, B. 2019. Multiple refinements to the anaesthetic protocol for laparoscopic orthotopic kidney auto-transplantation in a pig model. Laboratory Animals 53(1), 72–78.

Fifty-four pigs (Sus scrofa, Large White cross) from a commercial pig farm were anaesthetised over a six-year period from October 2011. The procedural stages included initial instrumentation for intra-operative monitoring, laparoscopic nephrectomy, preparation of the...

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...

Cox, R. J., Nol, P., Ellis, C. K. et al. 2019. Research with agricultural animals and wildlife. ILAR Journal 60(1), 66-73.

In fiscal year 2016, agricultural animals such as swine, sheep, goats, and cattle represented 10% of the 820 812 animals used in USDA-regulated research. In addition to traditional agricultural animals, research studies using captive wildlife...

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...

Kells, N. J., Beausoleil, N. J., Sutherland, M. A. et al. 2018. Electroencephalographic responses of anaesthetised pigs to intraperitoneal injection of sodium pentobarbital. Animal Welfare 27(3), 205-214.

Small laboratory animals are commonly euthanased via intraperitoneal (IP) injection of sodium pentobarbital. However, there is concern that animals may experience pain prior to loss of consciousness with this delivery route. The present study investigated...

Gilliam, J. N., Woods, J., Hill, J. et al. 2018. Evaluation of the CASH Dispatch Kit combined with alternative shot placement landmarks as a single-step euthanasia method for cattle of various ages. Animal Welfare 27(3), 225-233.

Humane euthanasia of cattle under field conditions presents special challenges for veterinarians and producers. The purpose of this study was to evaluate the effectiveness of the CASH Dispatch Kit captive-bolt system combined with improved shot...

Grist, A., Lines, J., Knowles, T. et al. 2018. The use of a non-penetrating captive bolt for the euthanasia of neonate piglets. Animals 8(4), 48.

The most common method for the on-farm euthanasia of neonate piglets is reported to be manual blunt force trauma. This paper presents the results of research to evaluate a mechanical non-penetrating captive bolt (the Accles...

Grist, A., Lines, J., Knowles, T. et al. 2018. The use of a mechanical non-penetrating captive bolt device for the euthanasia of neonate lambs. Animals 8(4), 49.

A non-penetrating captive bolt device, powered by a 1-grain 0.22″ cartridge delivering a calculated kinetic energy of 47 Joules was tested as a euthanasia method on 200 neonate lambs (Ovis aries) of 4.464 kg (SD...

Smith, R., Rault, J., Gates, R. et al. 2018. A two-step process of nitrous oxide before carbon dioxide for humanely euthanizing piglets: On-farm trials. Animals 8(4), 52.

Current methods of euthanizing piglets are raising animal welfare concerns. Our experiment used a novel two-step euthanasia method, using nitrous oxide (N2O) for six minutes and then carbon dioxide (CO2) on compromised 0- to 7-day-old...

Shearer, J. 2018. Euthanasia of cattle: Practical considerations and application. Animals 8(4), 57.

Acceptable methods for the euthanasia of cattle include overdose of an anesthetic, gunshot and captive bolt. The use of anesthetics for euthanasia is costly and complicates carcass disposal. These issues can be avoided by use...

Grist, A., Lines, J., Knowles, T. et al. 2018. Use of a non-penetrating captive bolt for euthanasia of neonate goats. Animals 8(4), 58.

A non-penetrating captive bolt device, powered by a 1 grain 0.22″ cartridge delivering a calculated kinetic energy of 47 Joules was tested as a euthanasia method on 200 neonate goats (Capra aegagrus hircus) of mean...

Kells, N., Beausoleil, N., Johnson, C. et al. 2018. Evaluation of different gases and gas combinations for on-farm euthanasia of pre-weaned pigs. Animals 8(3), 40.

The aim of this research was to evaluate the welfare of pre-weaned piglets euthanised using three different gas treatments: 100% carbon dioxide (CO2), 100% argon (Ar) or a mixture of 60% Ar/40% carbon dioxide (Ar/CO2)...

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...