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. 
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...

Galex, I. A., Gallant, C. M., D'Avignon, N. et al. 2020. Evaluation of effective and practical euthanasia methods for larval African clawed frogs (Xenopus laevis). JAALAS 59(3), 269–274.

Larval, or tadpole-stage Xenopus laevis frogs are a popular research model for developmental biology and disease studies. Existing euthanasia guidance documents offer recommendations for both eggs and adult stages, yet do not specifically address the...

Baskir, E. A., Kucharski, S., Powell, D. M. 2020. Chinchilla (Chinchilla lanigera) behavioral responses to a visual signal preceding handling. Zoo Biology 39(6), 391-396.

Zoos use ambassador animals in educational programs featuring close contact with humans. Chinchillas (Chinchilla lanigera) at the Saint Louis Zoo are retrieved for programs by a keeper wearing brown handling gloves, but green cleaning gloves...

Dorey, N. R., Blandina, A., Udell, M. A. R. 2020. Clicker training does not enhance learning in mixed-breed shelter puppies (Canis familiaris). Journal of Veterinary Behavior 39, 57-63.

Clicker training has been a popular form of training for decades and is used in zoos, aquariums, and shelters. Only a handful of studies have investigated the efficacy of the clicker-training method itself despite its...

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...

Vieira de Castro, A. C., Fuchs, D., Morello, G. M. et al. 2020. Does training method matter? Evidence for the negative impact of aversive-based methods on companion dog welfare. PLOS ONE 15(12), e0225023.

Dog training methods range broadly from those using mostly positive punishment and negative reinforcement (aversive-based) to those using primarily positive reinforcement (reward-based). Although aversive-based training has been strongly criticized for negatively affecting dog welfare, there...

Pedrazzani, A. S., Quintiliano, M. H., Bolfe, F. et al. 2020. Tilapia on-farm welfare assessment protocol for semi-intensive production systems. Frontiers in Veterinary Science 7, 606388.

The aim of this study was to develop and test a tilapia on-farm welfare assessment protocol, based on Brazilian semi-intensive production systems. The study included two mains steps: the elaboration of tilapia welfare protocol and...

Butler, J. L., Kennerley, S. W. 2019. Mymou: A low-cost, wireless touchscreen system for automated training of nonhuman primates. Behaviour Research Methods 51, 2559–2572.

Training nonhuman primates (NHPs) to perform cognitive tasks is essential for many neuroscientific investigations, yet laboratory training is a time-consuming process with inherent limitations. Habituating NHPs to the laboratory staff and experimental equipment can take...

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...

Domingues, R. R., Garrone-Neto, D., Hilsdorf, A. W. S. et al. 2019. Use of mucus as a non-invasive sampling method for DNA barcoding of stingrays and skates (batoid elasmobranchs). Journal of Fish Biology 94(3), 512-516.

In this study we tested the use of mucus from five species of Neotropical marine batoid elasmobranchs to extract genomic DNA for barcoding and phylogenetic analysis. The DNA from all individuals sampled was successfully amplified...

Anders, N., Roth, B., Grimsbø, E. et al. 2019. Assessing the effectiveness of an electrical stunning and chilling protocol for the slaughter of Atlantic mackerel (Scomber scombrus). PLOS ONE 14(9), e0222122.

Inducing unconsciousness in fish using electrical stunning prior to slaughter may improve fish welfare and fillet quality if such practises can be disseminated into wild capture fisheries. The objectives of this study were to: 1)...

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...

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...

Golledge, H., Carter, E. 2018. Welfare of fish farmed in the EU during transport and at slaughter. Animal Welfare 27(3), 287-288.

This Report, commissioned by the European Commission, summarises the current state of transport and slaughter practices in the aquaculture industry within eleven case-study countries in the European Economic Area (EAA). Comparisons are also made with...

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...

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...

Balko, J. A., Oda, A., Posner, L. P. 2018. Use of tricaine methanesulfonate or propofol for immersion euthanasia of goldfish (Carassius auratus). Journal of the American Veterinary Medical Association 252(12), 1555-1561.

OBJECTIVE To substantiate current AVMA guidelines for immersion euthanasia of goldfish (Carassius auratus) with tricaine methanesulfonate (TMS), determine whether immersion in propofol at 5 times its immersion anesthesia concentration for 30 minutes is sufficient for...

Riemer, S., Ellis, S. L. H., Thompson, H. et al. 2018. Reinforcer effectiveness in dogs—The influence of quantity and quality. Applied Animal Behaviour Science 206, 87-93.

Reinforcer effectiveness refers to the reinforcer’s ability to control the subject’s target behaviour and is therefore critical to training success. Yet animals’ preferences, and the effectiveness of different rewards to function as reinforcers, are often...

González-Pech, P. G., Marín-Tun, C. G., Valladares-González, D.A. et al. 2018. A protocol of human animal interaction to habituate young sheep and goats for behavioural studies. Behavioural Processes 157, 632-637.

Animal habituation is key to obtain reliable data on behavioural studies but detailed procedures to achieve it are scarce. This study designed a set of actions to habituate sheep and goats to human observers. Pelibuey...

Sutherland, M. A., Watson, T. J., Millman, S. T. 2017. Technical contribution: evaluation of the efficacy of a non-penetrating captive bolt to euthanase dairy goat kids up to 30 days of age. Animal Welfare 26(3), 277-280.

The objective of this study was to evaluate the effectiveness of a non-penetrating captive bolt (NPCB) to euthanase previously disbudded Saanen dairy goat kids (Capra hircus) up to 30 days of age and 9 kg...

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...