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. 
Eeza, M. N. H., Singer, R., Ding, Y. et al. 2023. Probing microstructural changes in muscles of leptin-deficient zebrafish by non-invasive ex-vivo magnetic resonance microimaging. PLOS ONE 18(4), e0284215.

Leptin is a hormone that plays a key role in controlling food intake and energy homeostasis. Skeletal muscle is an important target for leptin and recent studies have shown that leptin deficiency may lead to...

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...

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)...

Alef, R., Blaser, R. E. 2023. Social group during housing and testing modulates the effect of ethanol on zebrafish (Danio rerio) behavior. Behavioural Processes 209, 104877.

Zebrafish (Danio rerio) are a popular model organism in behavioral pharmacology research due to many genetic and neurological similarities with humans. As a social species, the presence (or absence) of conspecifics during housing and testing...

Deng, J., Zhang, H., Wang, Q. et al. 2023. An optimized environmental DNA method to improve detectability of the endangered Sichuan taimen (Hucho bleekeri). Fishes 8(7), 339.

Environmental DNA (eDNA) techniques have emerged as a cost-effective and non-invasive strategy for monitoring the distribution of endangered aquatic species. Despite their numerous advantages, operational uncertainty at each step of the process represents a significant...

Cameron, A. S., Rub, A. M. W., Sandford, B. P. 2023. Evaluation of healing progression at surgical incision sites and the use of antiseptics for enhancing post-operative survival in subyearling Chinook salmon (Oncorhynchus tshawytscha). PLOS ONE 18(7), e0288056.

In an attempt to develop more effective surgical implantation methods for fish, surgical incisions typical of those made for implanting micro-acoustic transmitters into the peritoneal cavity were evaluated on a weekly basis for healing progression...

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...

Boardman, R. M., Pinder, A. C., Piper,A. T. et al. 2022. Non-lethal sampling for the stable isotope analysis of the critically endangered European eel Anguilla anguilla: How fin and mucus compare to dorsal muscle. Journal of Fish Biology 100(3), 847-851.

Ecological studies on the critically endangered European eel Anguilla anguilla often incorporate stable isotope analysis that typically uses dorsal muscle sampled from euthanised eels. To minimise the lethal sampling of imperilled populations, fin tissue and/or...

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...

Schilling, A.-K., Mazzamuto, M. V., Romeo, C. 2022. A review of non-invasive sampling in wildlife disease and health research: What’s new? Animals 12(13), 1719.

In the last decades, wildlife diseases and the health status of animal populations have gained increasing attention from the scientific community as part of a One Health framework. Furthermore, the need for non-invasive sampling methods...

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...

Santymire, R. M., Young, M., Lenihan, E. et al. 2022. Preliminary investigation into developing the use of swabs for skin cortisol analysis for the ocean sunfish (Mola mola). Animals 12(20), 2868.

The ocean sunfish (mola; Mola mola) is the heaviest bony fish in the world. This slow-moving fish often is injured by fishing boats that use drift gillnets attributing to its listing as Vulnerable by the...

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...

Camargo-dos-Santos, B., Carlos, C. L., Favero-Neto, J. et al. 2021. Welfare in Nile tilapia production: Dorsal fin erection as a visual indicator for insensibility. Animals 11(10), 3007.

In aquaculture, to ensure animal welfare in pre-slaughter and slaughter stages, it is fundamental that fish are insensible. A method for evaluating fish insensibility is based on visual sensibility indicators (VSI) assessment (i.e., self-initiated behavior...

Prior, H., Blunt, H., Crossman, L. et al. 2021. Refining procedures within regulatory toxicology studies: Improving animal welfare and data. Animals 11(11), 3057.

During the development of potential new medicines or agrochemicals, an assessment of the safety profile to humans and environmental species is conducted using a range of different in silico and in vitro techniques in conjunction...

Zemanova, M. A. 2020. Towards more compassionate wildlife research through the 3Rs principles: Moving from invasive to noninvasive methods. Wildlife Biology 2020(1), no page numbers.

Research in ecology and wildlife biology remains crucial for increasing our knowledge and improving species management and conservation in the midst of the current biodiversity crisis. However, obtaining information on population status often involves invasive...

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...

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...

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...

Association of Primate Veterinarians. 2019. Association of Primate Veterinarians’ Position Statement: Cerebrospinal fluid aspiration for nonhuman primates in biomedical research. JAALAS 58(4), 428-429.

The Association of Primate Veterinarians (APV) recognizes that CSF collection may be required for both clinical and research purposes in nonhuman primates (NHP). Because there are inherent risks associated with the technique, the laboratory animal...

Winter, E. R., Nyqvist, M., Britton, J. R. 2019. Non-lethal sampling for stable isotope analysis of pike Esox lucius: How mucus, scale and fin tissue compare to muscle. Journal of Fish Biology 95(3), 956-958.

Stable isotope analysis (SIA) was used to examine the isotopic relationships between dorsal muscle and fin, scale and epidermal mucus in pike Esox lucius. δ13C and δ15N varied predictably within each tissue pairing, with conversion...

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...