3 Minute 3Rs

September 2022The papers behind the pod: Pinho RH et al. (2022). Validation of the rabbit pain behaviour scale (RPBS) to assess acute postoperative pain in rabbits (Oryctolagus cuniculus). PLoS ONE 17(5): e0268973. https://doi.org/10.1371/journal.pone.0268973 Stull C, Heagerty A and Coleman K (2022). Video Conference Technology as a Tool for Pair Introduction in Rhesus Macaques. Animals 12(14): e1783. https://doi.org/10.3390/ani12141783  Zentrich E et al. (2022). Postoperative Severity Assessment in Sheep. European Surgical Research, in press. https://doi.org/10.1159/000526058 It’s the third Thursday of September, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce, and refine the use of animals in research. This month we’re focusing on refinements for working with three different non-rodent species. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-september-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

August 2022The papers behind the pod: Arjmand S et al. (2022). Tips and traps for behavioural animal experimentation. Acta Neuropsychiatrica, in press. https://doi.org/10.1017/neu.2022.4Wrede P et al. (2022). Real-time 3D optoacoustic tracking of cell-sized magnetic microrobots circulating in the mouse brain vasculature. Science Advances 8(19). https://doi.org/10.1126/sciadv.abm9132 Russell A et al. (2022). Reduced housing density improves statistical power of murine gut microbiota studies. Cell Reports 39(6): e110783. https://doi.org/10.1016/j.celrep.2022.110783It’s the third Thursday of August, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce, and refine the use of animals in research. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-august-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

July 2022The papers behind the pod: Ferreira DA et al. (2021). Alternative to Soft Lithography for the Fabrication of Organ-on-a-Chip Elastomeric-Based Devices and Microactuators. Advanced Science 8:2003273.  https://doi.org/10.1002/advs.202003273Eigel D et al. (2019). Cryogel scaffolds for regionally constrained delivery of lysophosphatidylcholine to central nervous system slice cultures: A model of focal demyelination for multiple sclerosis research. Acta Biomaterialia 97:216.  https://doi.org/10.1016/j.actbio.2019.08.030It’s the third Thursday of July, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce, and refine the use of animals in research. This month, we’ve got a special double feature highlighting the publications commended in the International 3Rs Prize, awarded by the NC3Rs and sponsored by GSK. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-july-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

June 2022The papers behind the pod: Williams WO and Baneux P (2022). Humane Intervention Points: Refining endpoint terminology to incorporate non-euthanasia intervention options to improve animal welfare and preserve experimental outcomes. Laboratory Animals, in press. https://doi.org/10.1177/00236772221090801 Osorio D et al. (2022). scTenifoldKnk: An efficient virtual knockout tool for gene function predictions via single-cell gene regulatory network perturbation. Patterns 3(3): e100434. https://doi.org/10.1016/j.patter.2022.100434Moudio S et al. (2022). Exposure of α-Synuclein Aggregates to Organotypic Slice Cultures Recapitulates Key Molecular Features of Parkinson's Disease. Frontiers in Neurology 13: e826102. https://doi.org/10.3389/fneur.2022.826102 It’s the third Thursday of June, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-june-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

May 2022 The papers behind the pod:  Wang, Q et al. (2022). Assessment of a 3D neural spheroid model to detect pharmaceutical-induced neurotoxicity. ALTEX 39. https://doi.org/10.14573/altex.2112221 Reynolds P.S. (2022) Between two stools: preclinical research, reproducibility, and statistical design of experiments. BMC Res. Notes 15, 73.  https://doi.org/10.1186/s13104-022-05965-w Tappe-Theodor, A. et al. (2022) The “WWHow” Concept for Prospective Categorization of Post-operative Severity Assessment in Mice and Rats. Front. Vet. Sci. 9, 841431. https://doi.org/10.3389/fvets.2022.841431It’s the third Thursday of May, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. This month, we’re bringing you a paper for each R. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-may-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

April 2022The papers behind the pod:  Nigri M et al. (2022). Role of Environment and Experimenter in Reproducibility of Behavioral Studies With Laboratory Mice. Frontiers in Behavioural Neuroscience 16: e835444. https://doi.org/10.3389/fnbeh.2022.835444 Vámos A et al. (2022) Mitophagy Mediates the Beige to White Transition of Human Primary Subcutaneous Adipocytes Ex Vivo. Pharmaceuticals 15(3): 363. https://doi.org/10.3390/ph15030363 Ratuski AS and Weary DM (2022) Environmental Enrichment for Rats and Mice Housed in Laboratories: A Metareview. Animals 12(4): 414. https://doi.org/10.3390/ani12040414 It’s the third Thursday of April, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. This month, we’re highlighting three papers focusing on replacement. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-april-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

The papers behind the pod: 1.      Baran SW et al. (2022). Perspectives on the Evaluation and Adoption of Complex In Vitro Models in Drug Development: Workshop with the FDA and the Pharmaceutical Industry (IQ MPS Affiliate). ALTEX, in press. https://doi.org/10.14573/altex.21122032.      Borba JVB et al. (2022) STopTox: An in Silico Alternative to Animal Testing for Acute Systemic and Topical Toxicity. Environmental Health Perspectives 130(2). https://doi.org/10.1289/EHP93413.      van der Velden J et al. (2022) Animal models and animal-free innovations for cardiovascular research: current status and routes to be explored. Consensus document of the ESC working group on myocardial function and the ESC Working Group on Cellular Biology of the Heart. Cardiovascular Research, in press. https://doi.org/10.1093/cvr/cvab370 It’s the third Thursday of March, and you’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. This month, we’re highlighting three papers focusing on replacement. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-march-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

February https://doi.org/10.1186/s12915-021-01184-0https://doi.org/10.1177/00236772211065920https://doi.org/10.1038/s41598-021-04502-2It’s the 3rd Thursday of February, and you’re listening to 3 Minute 3Rs. This month, we’re bringing you 3 refinement papers. Let’s start with rodent housing.Research rodents are conventionally housed in shoebox sized cages that limit their ability to perform natural behaviors such as nesting and burrowing. These restrictions are known to impair welfare, but could they even increase disease risk and shorten lifespans? A new meta-analysis compares the morbidity and mortality of rodents in conventional vs enriched housing. Conventional housing was found to significantly worsen disease severity for cancer, cardiovascular disease, stroke, anxiety, and depression. Conventional housing also increased all-cause mortality. In conclusion, conventional housing appears to be distressing enough to compromise rodent health in a biologically significant manner. This lends more evidence to the importance of refining rodent housing for ethical, validity, and translational reasons. To learn more, read the full paper online.Sticking with the rodent theme, if you work with rats, you might find standard cages don’t provide enough room for a fully enriched environment – one where rats can socialize, exercise and express natural behaviours. If larger cages are not currently an option, two solutions with increasing research to back them up are playpens and ball pits, the focus of a recent paper in LA.Justyna Hinchcliffe et al. describe using 50 kHz ultrasonic vocalisations as an objective, quantifiable measure of how rats responded to ball pits and playpens. They found that these vocalisations were more frequent for rats exposed to the enriched environments, when compared with control conditions. They also tested the effect of playpens and ball pits on rats given an aversive drug treatment, finding that exposure to these environments reduced its negative impact. Besides the important welfare benefits of environmental enrichment, there are also scientific benefits in the shape of reduced variability and therefore more reliable results. With new evidence supporting the use of playpens for rats to minimize stress, there has never been a better time to try them out in your facility.And finally, let’s see how sleep could be a helpful tool to help us track laboratory animal welfare.In humans, sleep quality is strongly related to a person’s well-being and recent research suggests the same may be true in laboratory animals, such as dogs. If this is the case, sleep quality may then be useful as a non-invasive measure of animal welfare. A study in Sci. Rep. examined this hypothesis further using an observational approach to characterize sleeping patterns in laboratory dogs and investigate the effects of sleep quality on their daily behaviors. Male and female adult dogs housed in kennels in Brazil were recorded during a continuous 24-h, five-day assessment period. The footage was then analysed for daytime behaviours and sleeping metrics, such as number of sleep bouts and their duration. The study showed that the dogs slept far less than reported previously in the literature and during the day they were less active, ate more, played less and were less alert. Alterations of these daytime behaviors as a result of loss of sleep may indicate compromised welfare highlighting appropriate measures should be taken to ensure lab dogs’ sleep quality and welfare.  Hosted on Acast. See acast.com/privacy for more information.

The papers behind the pod:1. https://doi.org/10.7554/eLife.71601 & https://doi.org/10.7554/eLife.679952. https://doi.org/10.3389/fnins.2021.8056793. https://doi.org/10.1038/s41598-021-98356-3It’s the 3rd Thursday of January – happy new year! You’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. Of course, we focus on those three Rs, but many have suggested adding a fourth R to the list: reproducibility. Designing experiments with reproducibility in mind is a key aspect of reducing unnecessary animal use, as well as being good for advancing science.In 2013 the Center of Open Science and Science Exchange began a collaboration to investigate the reproducibility of 193 experiments from 50 high-impact cancer biology papers. Over eight years of repeated experiments, they found that they could only reproduce 50 experiments from 23 papers, generally due to a lack of detail about the methods used or resources being unavailable. 15 of those 50 repeated experiments used animals, and while just over half of them at least partially confirmed the original results, the repeated results were not always statistically significant. Experimental design was also an issue: only one of the original animal experiments used randomization and none used blinding or calculated a sample size before the study began.Papers describing these results are now available in eLife, with all the relevant data available on the Open Science Framework website and more Replication Studies to come from this collaboration. As the reproducibility crisis continues to rumble on, why not check them out and put designing more robust experiments at the top of your agenda?Next, let’s look at how training rats can help make fMRI a less stressful experience. Functional magnetic resonance imaging, or fMRI is a powerful non-invasive procedure that is used to assess brain function and connectivity. However, fMRI research in animals is often confounded due to the physical restraint and loud noises that occur during recordings as these induce stress which can alter information processing and cognition.An article from Frontiers in Neuroscience describes a protocol for habituating rats to fMRI that also avoids the need for surgical head restraint. Rats were gradually trained via 18 sessions over 3 weeks beginning with basic handling phase. After following this protocol, fMRIs in awake rats were successfully conducted without inducing increased stress and still achieving stable images with very low motion artifacts.To learn more about this rat refinement, read the full paper online. Finally, playpens for mice – could they be a viable option for refinement when home cage space is limited? Good environmental enrichment improves the quality of life for laboratory mice by providing increased opportunities to carry out natural behaviours such as running, climbing and burrowing. However, due to space requirements, cost and sanitation constraints many facilities worldwide still use standard housing, which has been associated with potential welfare problems.  In their publication in Scientific Reports, Ratuski et al show temporary access to playpens could be an effective method to provide mice housed in standard cages with space and structures to facilitate natural behaviors. In this study, female mice were given access to playpens three times a week for several weeks. Mice in the playpens were more active, compared to mice in conventional cages and over time, the animals entered the playpen more quickly and showed increased anticipatory behaviors before accessing the playpen. All indicating the mice found access to playpens rewarding. Want to learn more? Follow the link in the description. Hosted on Acast. See acast.com/privacy for more information.

Sedation is necessary for zebrafish during procedures such as imaging, biopsy, and surgery to ensure animal welfare and high-quality science. But the effects of sedation can last beyond the administration period and should also be carefully considered. A paper by Gressler et al. explores the use of eugenol and propofol during a 3-hour sedation and their subsequent effects after a 1 hour washout period. Both drugs had effects on behavior and physiology even after the washout period. For behavior, in a novel tank test, eugenol was found to amplify diving response while propofol induced anti-anxiety responses. For physiology, both drugs caused alterations in gill structure. Clearly, as with other species, sedation of zebrafish can significantly affect behavior and physiology beyond the administration period. Therefore, sedation procedures must be carefully designed and reported to refine experiments. Read more: https://doi.org/10.1016/j.applanim.2021.105482Next, let’s look at euthanasia, an important consideration for any animal study, and those involving zebrafish are no exception. Despite zebrafish being widely used in research, there is no consensus on which method to use when euthanizing them. The most commonly-used method – an overdose of tricaine, or MS-222 – is versatile, readily available and, in fact, is the only legal option in some areas, but it is now known to be aversive. A new paper by von Krogh et al. describes work to address this discrepancy. The team screened overdoses of 7 common alternative anaesthetics dissolved in water, to determine whether they led adult zebrafish to lose reflexes in a rapid, reliable, and non-aversive manner. Other substances were used to buffer the anaesthetics where needed, adjusting the water pH to reduce irritation. The authors found adding 1g/L lidocaine hydrochloride buffered with 2g/L sodium bicarbonate was particularly effective, reliably inducing loss of all reflexes within 2 mins and provoking little aversive behavior. Adding 50mL/L ethanol further reduced these issues. While the authors recommend this method, they also stress the importance of further investigation, including for zebrafish at different developmental stages. Read the paper: https://doi.org/10.3390/biology10111133Finally, let’s focus on mice and how researchers are working to ensure they are euthanized humanely. Gradual exposure to carbon dioxide remains the most common method for euthanising laboratory rodents. Although CO2 is generally considered an acceptable option when properly administered, there are concerns that CO2 is aversive to rodents and might cause distress and pain above certain concentrations. In a new study, Rodriguez-Sanchez and colleagues investigated using a voluntarily ingested sedative to reduce CO2 aversiveness. Cream cheese mixed with different doses of a rapid-acting anesthetic was provided to C57BL/6 mice before exposing the animals to CO2. Using a broad range of behavioural parameters, the team showed 20 mg/kg of sedative resulted in a mild sedation and likely reduced the aversiveness of CO2. While the investigators acknowledge more work is needed to determine the experience of the mice during sedation, they suggest voluntary oral administration of a sedative is potentially an effective, affordable, and easy way to minimise the stress of mice during CO2 euthanasia. Find out more: https://doi.org/10.3390/ani11102879 Hosted on Acast. See acast.com/privacy for more information.