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...

Diao, W., Jia, R., Hou, Y. et al. 2023. Effects of stocking density on the growth performance, physiological parameters, antioxidant status and lipid metabolism of Pelteobagrus fulvidraco in the integrated rice-fish farming system. Animals 13(11), 1721.

Pelteobagrus fulvidraco is a freshwater fish commonly raised in rice fields, yet the optimal stocking density for this species remains unknown. Therefore, this study aimed to investigate the appropriate stocking density of P. fulvidraco in...

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...

Rucinque, D. S., van de Vis, H., Reimert, H. et al. 2023. Pre-slaughter stunning of farmed Atlantic halibut in CO2-saturated seawater: Assessment of unconsciousness by electroencephalography (EEG). Animals 13(12), 1993.

As fish welfare becomes a growing concern, it is important to ensure humane treatment during slaughter. This study aimed to assess the onset of unconsciousness in Atlantic halibut immersed in CO2-saturated seawater through electroencephalography (EEG)...

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...

Watanabe, S. 2023. Infrared thermography for non-invasive measurement of social inequality aversion in rodents and potential usefulness for future animal-friendly studies. Frontiers in Behavioral Neuroscience 17, 1131427.

Infrared thermography is a method that detects thermal radiation energy and can measure the body surface temperature of animals from a distance. While rectal temperature has traditionally been used to measure animals’ core temperature, thermal...

Buenhombre, J., Daza-Cardona, E. A., Sousa, P. et al. 2023. Structural environmental enrichment and the way it is offered influence cognitive judgement bias and anxiety-like behaviours in zebrafish. Animal Cognition 26, 563–577.

Environmental enrichment in zebrafish generally reduces anxiety-related behaviours, improves learning in maze trials and increases health and biological fitness. However, certain types of enrichment or certain conditions induce the opposite effects. Therefore, it is essential...

Baßmann, B., Hahn, L., Rebl, A. et al. 2023. Effects of stocking density, size, and external stress on growth and welfare of African catfish (Clarias gariepinus Burchell, 1822) in a commercial RAS. Fishes 8(2), 74.

The effects of semi-intensive (100 kg m−3), intensive (200 kg m−3), and super-intensive (400 kg m−3) stocking densities on the growth and welfare of African catfish (Clarias gariepinus) were investigated under commercial production conditions. Plasma...

Fontana, B. D., Alnassar, N., Parker, M. O. 2023. Transgenerational effects of early-life stress on anxiety behavior in zebrafish (Danio rerio). Behavioural Processes 208, 104874.

Early-life adversity impacts on anxiety-related behaviors in adulthood. The effects of such adversity not only affect the animal itself, but can be passed on transgenerationally. Pervasive effects of experimentally-induced early-life stress (ELS) have been documented...

Gauy, A. C. dos S., Bolognesi, M. C., Gonçalves-de-Freitas, E. 2023. Body tactile stimulation reduces the effects of high stocking density on the welfare of Nile tilapia (Oreochromis niloticus). Fishes 8(6), 320.

High stocking density is used in aquaculture to optimize farming. However, such strategies can stress territorial fish by increasing aggression, thus harming the fish welfare and productive performance. Here, we tested the effect of long-term...

Moëne, O. L., Larsson, M. 2023. A new tool for quantifying mouse facial expressions. ENeuro 10(2) ENEURO.0349-22.2022.

Facial expressions are an increasingly used tool to assess emotional experience and affective state during experimental procedures in animal models. Previous studies have successfully related specific facial features with different positive and negative valence situations...

Brown, C., Schluessel, V. 2023. Smart sharks: A review of chondrichthyan cognition. Animal Cognition 26(1), 175–188.

450 million years of evolution have given chondrichthyans (sharks, rays and allies) ample time to adapt perfectly to their respective everyday life challenges and cognitive abilities have played an important part in that process. The...

Clemente, G. A., Tolini, C., Boscarino, A. et al. 2023. Farmed fish welfare during slaughter in Italy: Survey on stunning and killing methods and indicators of unconsciousness. Frontiers in Veterinary Science 10.

Information on slaughter procedures for farmed fish in aquaculture is limited, both in Europe and in Italy, due to a general lack of field data. The aim of this study was to gather information on...

Stevens, C., Hammonds, T., Hinchcliffe, J. et al. 2023. Report of the 2022 RSPCA/UFAW Rodent Welfare Group Meeting. Animal Technology and Welfare 22(2), 91–100.

The RSPCA/UFAW Rodent Welfare Group has held a one-day meeting every autumn for the last 29 years, so that its members can discuss current welfare research, exchange views on welfare issues and share experiences of...

Tanaka, Y., Nakata, T., Hibino, H. et al. 2023. Classification of multiple emotional states from facial expressions in head-fixed mice using a deep learning-based image analysis. PLOS ONE 18(7), e0288930.

Facial expressions are widely recognized as universal indicators of underlying internal states in most species of animals, thereby presenting as a non-invasive measure for assessing physical and mental conditions. Despite the advancement of artificial intelligence-assisted...

Wiese, T. R., Rey Planellas, S., Betancor, M. et al. 2023. Qualitative Behavioural Assessment as a welfare indicator for farmed Atlantic salmon (Salmo salar) in response to a stressful challenge. Frontiers in Veterinary Science 10.

Animal welfare assessments have struggled to investigate the emotional states of animals while focusing solely on available empirical evidence. Qualitative Behavioural Assessment (QBA) may provide insights into an animal’s subjective experiences without compromising scientific rigor...

Perret-Thiry, C., Raulic, J., Vergneau-Grosset, C. 2022. Evaluation of prolonged immersion in tricaine methanesulfonate for juvenile goldfish (Carassius auratus) euthanasia. Journal of the American Veterinary Medical Association 260(8), 911-915.

OBJECTIVE: Doses of buffered tricaine methanesulfonate (MS-222) up to 1000 mg/L for 15 minutes are reported inefficient to produce euthanasia in goldfish. The goal of this study was to determine if goldfish can be euthanized...

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...

Hickman, D. L. 2022. Minimal exposure times for irreversible euthanasia with carbon dioxide in mice and rats. JAALAS 61(3), 283-286.

When using an anesthetic overdose to euthanize laboratory rodents, a secondary method of euthanasia is recommended to ensure that the apparent death is irreversible. This secondary method usually is accomplished through the collection of tissues...

McKinney, M. M., Dupont, W. D., Corson, K. J. et al. 2022. Physiologic and behavioral effects in mice anesthetized with isoflurane in a red-tinted or a traditional translucent chamber. JAALAS 61(4), 322-332.

Isoflurane has been characterized as a distressing agent for rodents, causing both physiologic and behavioral effects. Using a "darkened home cage" has been recommended during CO2 administration for rodent euthanasia; this is arguably a similar...

Scariot, P. P., Gobatto, C. A., Polisel, E. E. et al. 2022. Early-life mice housed in standard stocking density reduce the spontaneous physical activity and increase visceral fat deposition before reaching adulthood. Laboratory Animals 56(4), 344-355.

Laboratory rodents spend the entire day housed in standard cages that provide a restricted area for movements and might, therefore, limit physical activity. However, it has not been tested in immature rodents of ages ranging...

Clarkson, J. M., Martin, J. E., McKeegan, D. E. F. 2022. A review of methods used to kill laboratory rodents: Issues and opportunities. Laboratory Animals 56(5), 419-436.

Rodents are the most widely used species for scientific purposes. A critical pre-requisite of their use, based on utilitarian ethical reasoning, is the provision of a humane death when necessary for scientific or welfare grounds...

MacLellan, A., Nazal, B., Young et al. 2022. Waking inactivity as a welfare indicator in laboratory mice: Investigating postures, facial expressions and depression-like states. Royal Society Open Science 9(11), 221083.

Animal welfare assessment relies on valid and practical indicators of affect. In mice, the most widely used research vertebrates, lying still with eyes open, inactive-but-awake (IBA) in the home cage, has potential to be one...

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...

Novak, J., Jaric, I., Rosso, M. et al. 2022. Handling method affects measures of anxiety, but not chronic stress in mice. Scientific Reports 12(1), 20938.

Studies in mice have shown that less aversive handling methods (e.g. tunnel or cup handling) can reduce behavioural measures of anxiety in comparison to picking mice up by their tail. Despite such evidence, tail handling...