LabAnimal

February 2023 The papers behind the pod: Hobbiesiefken U et al. (2023). Rating enrichment items by female group-housed laboratory mice in multiple binary choice tests using an RFID-based tracking system. PloS one 18(1): e0278709. doi: 10.1371/journal.pone.0278709Petkov C et al. (2022). Unified ethical principles and an animal research ‘Helsinki' declaration as foundations for international collaboration. Current Research in Neurobiology 3:100060. doi: 10.1016/j.crneur.2022.100060Weber E et al. (2023). Aggression in Group-Housed Male Mice: A Systematic Review. Animals 13(1):143. doi: 10.3390/ani13010143It's the third Thursday of February and you're listening to the final episode of 3 minute 3Rs, recapping the latest efforts to replace, reduce and refine the use of animals in research. Follow this link for the full transcript: https://www.nc3rs.org.uk/3-minute-3rs-podcast-february-2023-transcript Hosted on Acast. See acast.com/privacy for more information.

January 2023 The papers behind the pod: Cassotta M et al. (2022). A worldwide survey on the use of animal-derived materials and reagents in scientific experimentation. Engineering in Life Sciences 22(9):561-604. doi: 10.1002/elsc.202100167 Nuber M et al. (2022). Development of a 3D simulator for training the mouse in utero electroporation. PLOS One 17(12): e0279004. doi: 10.1371/journal.pone.0279004Dickmann J et al. (2022). Clicker Training Mice for Improved Compliance in the Catwalk Test. Animals 12(24): 3545. doi: 10.3390/ani12243545Happy New Year to all 3 Minute 3Rs listeners. It's 2023, and we're back with three more papers highlighting efforts to replace, reduce and refine the use of animals in research. Follow this link for the full transcript: https://www.nc3rs.org.uk/3-minute-3rs-podcast-january-2023-transcript Hosted on Acast. See acast.com/privacy for more information.

December 2022The papers behind the pod: Menéndez ABC et al. (2022). Creating a kidney organoid-vasculature interaction model using a novel organ-on-chip system. Scientific Reports 12:20699. doi: 10.1038/s41598-022-24945-5Karp N et al. (2022). A qualitative study of the barriers to using blinding in in vivo experiments and suggestions for improvement. PLOS Biology 20(11): e3001873. doi: 10.1371/journal.pbio.3001873Talbot S et al. (2022). RELSA—A multidimensional procedure for the comparative assessment of well-being and the quantitative determination of severity in experimental procedures. Frontiers in Veterinary Science 9:937711. doi: 10.3389/fvets.2022.937711 It's the third Thursday of December, and you're listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. To round off 2022 we are highlighting a paper for each R.Follow this link for the full transcript: https://www.nc3rs.org.uk/3-minute-3rs-podcast-december-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

November 2022The papers behind the pod: Aulehner K et al. (2022). Grimace scale, burrowing, and nest building for the assessment of post-surgical pain in mice and rats—A systematic review. Frontiers in Veterinary Science 9:930005. doi: 10.3389/fvets.2022.930005Hair K et al. (2022). ‘Living' evidence frameworks for in vivo animal research: towards translational evidence-based medicine. BMJ Evidence-Based Medicine 2022;27:A17. doi: 10.1136/ebm-2022-EBMLive.31 Wong J et al. (2022). Electrochemically induced in vitro focal hypoxia in human neurons. Frontiers in Cell and Developmental Biology 10:968341. doi: 10.3389/fcell.2022.968341It's the third Thursday of November, 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://www.nc3rs.org.uk/3-minute-3rs-podcast-november-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

October 2022The papers behind the pod: Piper, SK et al. (2022). Statistical review of animal trials—A guideline. Biometrical Journal https://doi.org/10.1002/bimj.202200061 Gargaté MJ et al. (2022). Parallel Propagation of Toxoplasma gondii In Vivo, In Vitro and in Alternate Model: Towards Less Dependence on the Mice Model. Pathogens https://doi.org/10.3390/pathogens11091038 Mieske P et al. et al. (2022). Bored at home?—A systematic review on the effect of environmental enrichment on the welfare of laboratory rats and mice. Frontiers in Veterinary Science https://doi.org/10.3389/fvets.2022.899219 It's the third Thursday of October, 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 have a paper on each R. Follow this link for the full transcript: https://nc3rs.org.uk/3-minute-3rs-podcast-october-2022-transcript Hosted on Acast. See acast.com/privacy for more information.

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 Our GDPR privacy policy was updated on August 8, 2022. Visit 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 See acast.com/privacy for privacy and opt-out 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 See acast.com/privacy for privacy and opt-out 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 See acast.com/privacy for privacy and opt-out 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 See acast.com/privacy for privacy and opt-out 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 See acast.com/privacy for privacy and opt-out 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. See acast.com/privacy for privacy and opt-out 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. See acast.com/privacy for privacy and opt-out information.

Dec 21 For our last episode of 2021, we're highlighting two important topics in refinement: sedation and euthanasia. 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 in Applied Animal Behavior Science.Next, 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. It describes how the team screened overdoses of seven 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 pH of the water to avoid causing irritation. The authors found adding one gram of lidocaine hydrochloride per litre buffered with two grams of sodium bicarbonate per litre, was particularly effective, reliably inducing loss of all reflexes within two minutes and provoking little aversive behavior. Adding fifty milliliters of ethanol per liter 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. You can read this important work now in Biology.Finally, let's shift focus to mice and how researchers are working to ensure they are euthanized humanely.Gradual exposure to carbon dioxide remains the most common method in use today 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 about the study in Animals. See acast.com/privacy for privacy and opt-out information.

Nov 21The papers behind the pod:1. A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates. npj Vaccines, January 2021. https://doi.org/10.1038/s41541-020-00263-72. Inducible dominant negative ErbB2 rat spermatogonial line for generation of transgenic rat model and dissecting ERBB2 tyrosine kinase mediated pathways. Experimental Oncology, June 2019. https://doi.org/10.32471/exp-oncology.2312-8852.vol-41-no-2.130263. Characterization of drug responses of mini patient-derived xenografts in mice for predicting cancer patient clinical therapeutic response. Cancer Communications, September 2018. https://doi.org/10.1186/s40880-018-0329-5Visit the AAALAC website to learn more about this year's Global 3Rs Award winners: https://www.aaalac.org/awards/global-3rs-winners/It's the third Thursday of November, 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 celebrating our fourth birthday, and we're also celebrating this year's Global AAALAC/IQ Award winners. Three papers were recognized this year, by scientists in Europe, North America and the Pacific Rim. First let's hear about the European winner: Dr Rachel Tanner at the University of Oxford.Tuberculosis is a serious global health problem with rising cases and deaths, in part, due to the current vaccine being insufficient. To develop a new vaccine, non-human primates are necessary relevant animal model, but efforts to refine their use are needed. Recently, a team has developed an in vitro assay to refine and expedite early tuberculosis vaccine testing. This assay uses non-human primate blood or cell samples to measure the ability to control mycobacterial replication. Importantly, it allows vaccine candidates to be tested without infecting non-human primates with tuberculosis directly. Rather, blood samples can be taken before and after vaccination and tested in vitro to examine efficacy. This work is a promising 3Rs tool to facilitate early testing of new vaccine candidates.To learn more, read the full paper online. Next, let's move to North America and Dr Andrew Syvyk from Texas A&M University, who won the prize for work on a simpler way to generate transgenic animal models. Such models can be tailored to study specific genes, for example by introducing a gene construct into a zygote or embryonic stem cell, but this is a long process that requires numerous steps and manipulations, and therefore a lot of animals.In his winning paper, Syvyk describes a simpler approach using immature sperm cells, or spermatogonia. The model involves manipulating spermatogonial stem cells from a donor male, which are then transplanted into recipient males. These can be used to breed transgenic animals directly, eliminating mosaicism and reducing the number of animals required.The paper discusses using this approach to study ERBB2, a protein involved in multiple biological processes and forms of cancer. As well as being used to generate transgenic animals, the cells can be used for in vitro research, further reducing reliance on in vivo models for investigating molecular pathways.Want to learn more about the model? Find the paper in Experimental Oncology via the link in the description.And finally, the winner... See acast.com/privacy for privacy and opt-out information.

Oct 2021The papers behind the pod:1. Professional Quality of Life in Research Involving Laboratory Animals. Animals 11(9), 2639 (2021) https://doi.org/10.3390/ani110926392. Improving culture of care through maximising learning from observations and events: Addressing what is at fault. Laboratory Animals (2021). https://doi.org/10.1177%2F002367722110371773. Epidemiology and reporting characteristics of preclinical systematic reviews. PLOS Biology (2021). https://doi.org/10.1371/journal.pbio.3001177 Transcript:It's the 3rd Thursday of October, 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 the people and strategies behind the science.First up, new research on compassion fatigue and its impact on those working with laboratory animals.Most professionals work in animal research because they want to help both animals and people. Their jobs can bring them satisfaction, but can also result in workplace stress. After all, their work can be challenging and is subject to social stigma. Recently, a team investigated laboratory animal personnel's professional quality of life in Spain. They found that less positive professional quality of life was associated with several factors: (1) receiving less social support, (2) being a PhD student, and (3) working as a researcher versus as animal-facility personnel. Their work could assist the design of interventions to help those working with laboratory animals.Want to learn more? Read the full paper open access in Animals. Next, let's talk about culture of care: what does it mean for animal facilities, and how can it become established practice? Sally Robinson and colleagues at AstraZeneca draw on their experience of making this happen in a new review paper.In an animal research setting, an establishment with a positive culture of care is committed to improving animal welfare, scientific quality, transparency and staff wellbeing. However, if things go wrong this culture of care can be derailed if lessons are not learned. The authors describe a system for logging events and observations to enable better tracking, analysis and oversight of both negative and positive occurrences. Using the philosophy of human and organisational performance, the system enabled them to move away from focusing on individual errors and towards addressing any issues in a more constructive way – for example, by recognizing that human error is inevitable, but the consequences of an error depend on how an organization is set up.This shift in focus, along with positive reinforcement of good practice through awards and wider dissemination, helped create an environment where staff felt more able to report issues. The paper includes examples of how the system was used and the factors that led to success.Inspired to strengthen your facility's culture of care? Check out the paper in Laboratory Animals.Finally, let's shift over to systematic reviews and how researchers can make sure they're doing them right.If you're looking to understand what research has already been published to answer a hypothesis, systematic reviews can be really helpful. Comprehensively identifying previous studies, their key findings and any reasons for differences between results can also have 3Rs benefits by helping to inform model choice and optimise sample size.However, if key details are not reported the transparency and... See acast.com/privacy for privacy and opt-out information.

Sept 2021 You're listening to the September episode of 3 Minute 3Rs. The papers behind the pod: Performance of preclinical models in predicting drug-induced liver injury in humans: a systematic review. Scientific Reports 11, 6403 (2021) https://doi.org/10.1038/s41598-021-85708-2 Ratlas-LH: An MRI template of the Lister hooded rat brain with stereotaxic coordinates for neurosurgical implantations Brain and Neuroscience Advances (2021) https://doi.org/10.1177/23982128211036332 O mouse, where art thou? The Mouse Position Surveillance System (MoPSS)—an RFID-based tracking system. Behavior Research Methods (2021) https://doi.org/10.3758/s13428-021-01593-7 [Transcript] It's the 3rd 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've got a paper for each R. Let's get started with a replacement. NA3RsC Drug induced liver injury in clinical trial and for newly approved drugs is a serious adverse reaction that causes massive financial loss and preventable human suffering. But what if we could more accurately predict these adverse reactions before they happened? A systematic review in Scientific Reports compared two anti-diabetic drugs, one that was withdrawn from the market due to liver injuries and another that remains. They reviewed in vivo studies, in vitro data, and reported liver injury cases. While the animal and human trials failed to predict liver injuries, the in vitro assays showed the withdrawn drug had twice the activity than the other. Overall, in vitro assays may offer a new paradigm to predict drug induced liver injuries thereby improving drug safety and development. Want to learn more? Read the full paper online. Next let's reduce... NC3Rs If you want to know where you're going, an atlas can help – and now, if you're performing rodent stereotaxic surgery, Ratlas can help! Lister hooded rats are widely used in behavioural neuroscience studies in the UK and beyond. When these studies involve stereotaxic surgery, accurate coordinates are vital for precisely targeting the relevant areas of the brain – otherwise, additional animals may be used for pilot surgeries to establish the correct coordinates. To reduce the need for these pilot surgeries, ­researchers at the University of Nottingham and Leibniz Institute for Neurobiology have developed Ratlas-LH, a user-friendly template combining in vivo MRI images and ex vivo micro-CT images of young adult male Lister hooded rats. The team have published a paper describing the development and validation of Ratlas-LH, and aim to expand their work to other commonly used rat and mouse strains. Interested? Read the paper in Brain and Neuroscience Advances and download Ratlas-LH for free. And finally, let's refine: Lab Animal Most mice will spend most of their time just hanging out in the comfort of their home cages. Providing simple husbandry refinements there can go a long way for the animals' welfare, but what do mice want? Why not ask them? A new home cage device from researchers at the German Center for the Protection of Laboratory Animals aims to do just that. Dubbed the Mouse Position... See acast.com/privacy for privacy and opt-out information.

You're listening to the August episode of 3 Minute 3Rs!The papers behind the pod: Refinement of the spinal cord injury rat model and validation of its applicability as a model for memory loss and chronic pain. Heliyon 7, E07500 (2021) https://doi.org/10.1016/j.heliyon.2021.e07500 Strategies to encourage the adoption of social housing during cardiovascular telemetry recordings in non-rodents. Journal of Pharmalogical & Toxicological Methods 108, 106969 (2021). https://doi.org/10.1016/j.vascn.2021.106959 Refine acquisition of high-resolution chest CTs in macaques by free breathing. Laboratory Animals https://doi.org/10.1177%2F00236772211026562 Transcript:It's the 3rd Thursday of August, and you are 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 three refinement papers. Let's jump right in with a laminectomy. NC3Rs Rats are widely used in spinal cord injury research where the spinal cord is bruised surgically to represent the injury. Accessing the spinal cord requires parts of the vertebrae to be removed, which can result in post-operative complications, but these laminectomy procedures, descriptive protocols and potential complications are not discussed in depth in the literature. A recent paper in Heliyon builds upon the evidence-base that using a dental burr assisted technique reduces the impact of laminectomy. Rats lost less weight, were more active during the dark phase and scored less points on the Rat Grimace scale indicating the animals were in less pain. The mechanised laminectomy also reduced surgical bleeding during the procedure and did not impact the spinal cord injury or its healing process. To learn more, check out the link in the description. Next up, how about some social housing. NA3RsC Social housing is critical for optimal welfare of social species. However, exceptions can be made if necessary for solid science. Traditionally, researchers have made the case that dogs, pigs, and nonhuman primates should be singly housed during cardiovascular telemetry recordings, which are needed to assess the safety of a novel drug. But could that be changing? The NC3Rs conducted surveys and working groups to benchmark practices and address barriers to social housing in this situation. Results showed that industry use of social housing is increasing – but still not standard. Commonly cited barriers relate to data cross-contamination and sensitivity of different study designs. However, shared data indicates that these barriers can be effectively managed t0 be compatible with social housing. To learn more about strategies to encourage this social housing refinement read the full paper online. Lab Animal And finally, take a deep breath, and hold… If you or I were getting a CT scan, the doctor would likely ask to us to hold our breath for a moment while the machine did its thing. That's because movement, like breathing, is the bane of most imaging modalities. An easy request for us, however, isn't necessarily as simple for animals. Nonhuman primates like macaques are... See acast.com/privacy for privacy and opt-out information.

It's the 3rd 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. The papers behind the pod: SARS-CoV-2 infects an upper airway model derived from induced pluripotent stem cells. Stem Cells https://stemcellsjournals.onlinelibrary.wiley.com/doi/full/10.1002/stem.3422A break from the pups: The effects of loft access on the welfare of lactating laboratory rats. PLos One https://doi.org/10.1371/journal.pone.0253020 3D printed rodent skin-skull-brain model: A novel animal-free approach for neurosurgical training. PLoS One https://doi.org/10.1371/journal.pone.0253477 This month, we'll be covering welfare for rat moms and an animal-free option for practicing your surgical skills. But first, let's hear about an in vitro model for COVID-19 research. [NC3Rs] As researchers around the world fight against COVID-19, in vitro models continue to gain traction. As well as reducing reliance on animals, they offer a cost-effective approach for generating models on a large scale. This is particularly useful for high-throughput drug screening, making in vitro models a key tool in the search for effective COVID-19 treatments. A recent paper in Stem Cells by Ivo Djidrovski and colleagues describes an upper airway model derived from induced pluripotent stem cells. These allow a large number of cells of different types to be generated without relying on primary samples, which can vary considerably between donors. The model can produce mucus and functional cilia and replicates the behaviour observed in vivo when infected with SARS-CoV-2. The team's next step will be to incorporate immune cell components into the model to further increase its relevance. Want to learn more about how stem cells could play a key role in fighting COVID? Follow the link in the description. [Speaking of COVID...] [NA3RsC] Were you locked down in a small apartment with multiple young children during the pandemic? If so, you probably wanted a break from them. New research shows that rat moms also want a break from their pups. In standard housing, rat dams are unable to spend time away from their pups due to caging set-up. But a recent study by researchers at University of British Columbia explores how providing a loft effects dam welfare. They found that as pups age, dams with lofts spend more time in their lofts, less time nursing, and, specifically, less time passively nursing. By the time that pups are 3 weeks old, dams will actually spend 50% of their time in their lofts. Furthermore, dams without a loft may show signs of negative affect. Want to learn more about giving rat dams a break from their pups? Read the full paper online in Plos One. [Lab Animal] And finally, let's talk about rodent surgery. Building up those surgical skills takes practice. The more complicated the procedure, the more time you'll need to spend perfecting your technique before you're ready to move on to a live... See acast.com/privacy for privacy and opt-out information.

You're listening to the June 2021 episode of 3 Minute 3Rs The papers behind the pod: Winning paper: Pellegrini L et al. (2020). Human CNS barrier-forming organoids with cerebrospinal fluid production. Science 369(6500): eaaz5626. doi.org/10.1126/science.aaz5626. Highly commended: Ashworth J et al. (2020). Peptide gels of fully-defined composition and mechanics for probing cell-cell and cell-matrix interactions in vitro. Matrix Biology 85-86: 15-33. doi.org/10.1016/j.matbio.2019.06.009. [Transcript]It's the 3rd 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. This month, we've got a special double feature to highlight the results of the 2020 3Rs Prize, awarded by the NC3Rs and sponsored by GSK. Let's start with the winner: Laura Pellegrini at the MRC Laboratory of Molecular Biology, for her animal-replacing organoid work. NA3RsC Deep within each of our brains, there's a vital fluid providing essential nutrients and signaling molecules while protecting the brain from toxic compounds. It's called the cerebrospinal fluid which is produced by the choroid plexus. Our understanding of this fluid and organ is limited due to the difficulty in studying them. In turn, many new drugs developed for the central nervous system have failed because of lack of efficacy, inability to cross the blood-brain barrier, or limited translation from animals. But now, Dr. Pellegrini and her colleagues have developed a human choroid plexus organoid. This organoid has a selective barrier that quantitatively predicts the permeability of small molecules into the brain. It even secretes a liquid very much like cerebrospinal fluid. Overall, this model holds great promise for deeper study into this incredible part of our bodies. To learn more, read the full paper in Science. 2020 also brought us highly commended replacement to mouse-derived Matrigel. NC3Rs A cell's environment has a lot of influence on their behavior so for cells to be the best experimental representative their environment needs to be as similar to in vivo as possible. In many research areas, there is now a move towards 3D culture techniques using biomaterials as a matrix, providing both structure and cell-matrix interactions. Often these matrices are animal-derived, such as Matrigel which is derived from mouse sarcomas. Approximately one hundred mice can be required to produce the Matrigel needed by a single research institute every year. However, the chemical components of Matrigel are not well defined affecting the reproducibility of studies and given the number of different cell types, organs and tissues, a one-size-fits all approach is unlikely to be the best representation of an in vivo situation. Dr Jennifer Ashworth was highly commended in the 2020 3Rs Prize for her work published in Matrix Biology detailing a non-animal derived synthetic hydrogel that can replace the use of Matrigel. The hydrogels are derived from a commercially available precursor peptide. This ‘blank slate' can then be ‘tuned' altering a hydrogel's stiffness and composition by adding proteins and sugars to replicate different in vivo environments. Check out Jenny's paper to learn more. And with... See acast.com/privacy for privacy and opt-out information.

You're listening to the April episode of 3 Minute 3Rs.The papers behind the pod:1. PCR Testing of Media Placed in Soiled Bedding as a Method for Mouse Colony Health Surveillance. JAALAS https://doi.org/10.30802/AALAS-JAALAS-20-0000962. New mosquito repellency bioassay for evaluation of repellents and pyrethroids using an attractive blood-feeding device. Parasites & Vectors https://doi.org/10.1186/s13071-021-04656-y3. Lung cancer organoids analysed on microwell arrays predict drug responses within a week. Nature Communications https://doi.org/10.1038/s41467-021-22676-1 Transcript: It's the 3rd 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 looking at animal free options for testing mosquito repellents and cancer therapies. But first, can we replace sentinel animals for health monitoring? [NA3RsC]To support high-quality research data, it's important to ensure that colony animals are healthy. Traditionally, rodent health surveillance is done through dirty bedding sentinel animals that act as a sort of canary in the coal mine. But what if, we could replace these animals, reduce husbandry time, and get even better data? New research shows that we can, regardless of the type of caging we use.Researchers from Emory University developed an effective protocol to conduct environmental monitoring in both individually ventilated racks filtered at the cage level and static cages. The most effective method was to simply replace dirty bedding sentinel rodents with 10 flocked swabs and cage shaking. This method was actually MORE effective than traditional sentinel animals. To learn how to replace your sentinel rodents with environmental monitoring, read the full paper online.On to repelling mosquitoes [NC3RS] The threat of mosquito-borne infectious diseases is rising, fueled in part by climate change and increases in international travel. Consequently, there is a growing interest in developing textiles treated with mosquito repellents and a growing need to test potential products.These test methods must be standardized, but this process requires a regular supply of animal blood for the mosquitos to feed on. Using living animals for this purpose raises animal welfare concerns. To address these issues, Morimoto and colleagues have developed a new attractive blood-feeding device consisting of a feeding unit with a reservoir of equine blood overlaid with a plastic membrane. The mosquitos can artificially feed on equine blood by penetrating the membrane, avoiding the use of live animals. The authors describe using the device to evaluate how mosquitos responded to three chemical repellents, producing highly accurate and reproducible results. The attractive blood-feeding device could also be used to develop new repellent chemicals and learn more about mosquito physiology.Got an itch to learn more about this refined test system – follow the link in the description. [LA] And finally, we've all heard of patient-derived xenograft mouse models. To many in the cancer field, these patient-personalized mice have been important tools for testing patient responses to treatments. PDX models are however expensive, and they take a long time and a lot of mice to produce. Mice, however, aren't the only option for personalized medicine. As in vitro technology advance, patient derived organoids are emerging alternatives. In a new paper published in the journal Nature... See acast.com/privacy for privacy and opt-out information.

You're listening to the April episode of 3 Minute 3Rs.The papers behind the pod:1. Identification of individual zebrafish (Danio rerio): a refined protocol for VIE tagging whist considering animal welfare and the principles of the 3RS. Animals 11, 616 (2021) https://doi.org/10.3390/ani110306162. Enrichment for laboratory zebrafish – a review of the evidence and challenges. Animals 11, 698 (2021) https://doi.org/10.3390/ani110306983. Research relevant background lesions and conditions: ferrets, dogs, swine, sheep, and goats. ILAR J ilab005 (2021) https://doi.org/10.1093/ilar/ilab0054. *Bonus* Research-relevant background lesions and conditions in common avian and aquatic species. ILAR J ilab008 (2021) https://doi.org/10.1093/ilar/ilab008Transcript: It's the 3rd 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've got some zebrafish refinements, plus a refresher on some basics for a bunch of different animals. But first, ID please. [NC3Rs]If you've got a tank full of fish that look identical, how do you make sure you can tell them apart? The zebrafish research community have tried various tagging methods, but until now there hasn't been a refined and standardized method that can be used on a large scale.In a new paper in Animals, Anita Rácz and colleagues describe such a method for tagging zebrafish using visible implant elastomer, or VIE. This method is widely used for marking large fish and other aquatic organisms and involves injecting a small amount of coloured elastomer under the fish's skin.As well as explaining their refined protocol, including appropriate anaesthesia and analgesia, Rácz and colleagues compare it to a previous protocol and show that it offers significant welfare improvements, including a ten-fold reduction in mortality following the tagging procedure. With zebrafish being used in thousands of labs around the world, particularly in genetic research, this work is bound to make a big splash. While we're thinking about zebrafish, have you thought about their tanks look like lately? [NA3RsC] Zebrafish have rapidly become one of the most common research animal species with over 5 million fish used worldwide. Currently, most of these fish are housed in barren tanks with no environmental enrichment.But in a recent review in Animals, the RSPCA Science Department describes current evidence supporting zebrafish enrichment. Its provision improves zebrafish welfare, behavior, and physiology, which can lead to better science. One well-supported and fairly easily implemented enrichment is using an image of gravel on the bottom of tanks. The paper also discuss evidence for social housing, plastic plants, live food, and more. To learn more about the current evidence for zebrafish enrichment, read the full paper online. [LA] And finally, a double feature. A lot of different animals turn up in research labs beyond rodents. But regardless of what species you are using, you need to know what's normal and what's not for a given animal in order to accurately interpret any experimental results. How well do you know an animals' anatomy and how it differs from the humans they are intended to model? Is that lesion intentional, or a spontaneous... See acast.com/privacy for privacy and opt-out information.

You're listening to the March episode of 3 Minute 3Rs.The papers behind the pod:1. Novel three-dimensional biochip pulmonary sarcoidosis model. PLoS One https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0245805 2. A new group housing approach for non-human primate metabolism studies. Journal of Pharmacological and Toxicological Methods https://doi.org/10.1016/j.vascn.2020.1069473. The contribution of environmental enrichment to phenotypic variation in mice and rats. eNeuro https://doi.org/10.1523/ENEURO.0539-20.2021Transcript: It's the 3rd 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've got two refinements to cover – a paper for primates and one for rodents too. But first, let's have some lung chips. [NC3Rs]Despite being first described towards the end of the 19th century, a lot about sarcoidosis remains unknown. The granulomas characteristic of the disease can affect almost any organ in the body, but the lung is most commonly affected, and associated with the highest morbidity and mortality in patients. Pulmonary granulomas can be modelled in animals, but as there is no known specific genetic component, transgenic animals representing the human condition cannot be easily created. Instead, animals are typically pre-sensitised to environmental agents, such as bacteria, and then repeatedly exposed to the same antigens to form granulomas.Using organ-on-a-chip technology, Calcagno et al introduced granulomas, isolated from patient blood samples, to its air-lung interface. They were able to detect macrophages and lymphocytes from the developed granulomas in the interface and inflammatory cytokines being released into the culture media. As well as better understanding the mechanisms behind sarcoidosis, the system can also be used to develop new treatments, with high-throughput potential and AI compatibility. You can read more about the method by following the link in the description.Next up, thinking beyond single housing[NA3RsC] Before new drugs are approved by regulatory bodies, they must undergo testing to understand their absorption, distribution, metabolism, and excretion. This is a key step in assuring their safety and efficacy. Sometimes, when there is no scientific alternative, non-human primates are used for these tests.Traditionally, for metabolism studies animals are singly-housed in cages that limit normal vertical movement & social behaviors which negatively impacts welfare. However, recently, Novo Nordisk, Covance, & other collaborators worked together to design a refined group housing metabolism cage. This cage has extensive advantages such as more space, better socialization, and less stress. Importantly, excretion data from this cage is comparable to the singly housed cages which supports its suitability for future metabolism studies.To read more, see the full paper online.And finally, a reminder that enrichment is a good thing. [LA] Researchers often to want to control as many variables as they can in their experiments. However, providing environmental enrichment is beneficial for the welfare of... See acast.com/privacy for privacy and opt-out information.

You're listening to the February episode of 3 Minute 3Rs.The papers behind the pod:1. In Vitro Tests for Assessing the Neutralizing Ability of Snake Antivenoms: Toward the 3Rs Principles. Frontiers in Immunology https://doi.org/10.3389/fimmu.2020.6174292. The Impact of Acute Loud Noise on the Behavior of Laboratory Birds. Frontiers in Veterinary Science https://doi.org/10.3389/fvets.2020.6076323. A multicomponent screen for feeding behavior and nutritional status in Drosophila to interrogate mammalian appetite-related genes. Molecular Metabolism https://doi.org/10.1016/j.molmet.2020.101127Transcript: It's the 3rd Thursday of February 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 work on finches and flies to feature, but first, let's review what we know about applying the 3Rs to antivenom research.[NA3RsC] Every year, venomous snakebites kill over 100,000 people and maim over 400,000 more. Treatment for these bites comes in the form of antivenom, which is typically made from the plasma of horses immunized with snake venom. Before being used in humans, this plasma is tested in mice to ensure that it's safe and effective. But unfortunately, these tests require large number of mice and can cause significant pain & distress.A new paper in Frontiers in Immunology calls for an urgent need to strengthen the implementation of the 3Rs in antivenom research. This is a challenging task due to the complexity of snake venom, but there has been progress. For replacement & reduction, the authors recommend correlated in vitro surrogate assays. For refinement, they recommend routine analgesia, improved housing, and re-designed protocols. To learn more about how the 3Rs can be implemented in antivenom research, read the full paper online. [LA] Next up, we all know that noise can be a cause for concern for laboratory rodents, but what about for the other species we keep in the lab? Mitigation measures for mice and rats – for example, those low frequency, quote unquote silent fire alarms – might not be so silent for other animals. A study published in Frontiers in Veterinary Science considers whether weekly fire alarm tests might be stressing out a colony of zebra finches. Pairs of finches were recorded before and after they were exposed to the fire alarm and again during alarm-free control sessions. Even though the alarm only lasted a few seconds and the zebra finches had been born and raised with the noise, they still took note. Movement decreased, and the usually chatty birds fell a bit silent for at least fifteen minutes following the disruption. Instead, they sat perched in the middle of their cages and increased preening of one another, a possible fear response. The results suggest more work needs to be done to assess the welfare of birds in response to noise disturbances. And finally, can flies replace mice for obesity research? [NC3Rs] Obesity is a serious public health issue due to its link with other co-morbidities such as type 2 diabetes and cardiovascular disease. Someone's response to the modern “obesogenic” lifestyle is largely due to their genetics and murine models can be used to explore these effects. But studying genes of interest requires extensive breeding programmes and significant resources to generate and phenotype... See acast.com/privacy for privacy and opt-out information.

You're listening to the January episode of 3 Minute 3Rs.The papers behind the pod:1. Internal consistency and compatibility of the 3Rs and 3Vs principles for project evaluation of animal research. Laboratory Animals https://doi.org/10.1177/00236772209685832. Noise and Vibration in the Vivarium: Recommendations for Developing a Measurement Plan. JAALAS https://doi.org/10.30802/AALAS-JAALAS-19-0001313. A human tissue screen identifies a regulator of ER secretion as a brain-size determinant. Science https://doi.org/10.1126/science.abb5390 Transcript: It's the 3rd Thursday of January and you're listening to the first 3 Minute 3R of the new year. Each month, we recap three efforts to replace, reduce, and refine the use of animals in research. To kick off 2021, we'll be discussing the issue of noise in the vivarium, and consider an organoid rather than animal-based approach for studying brain development. But first let's hear about another letter-based trio: the Three Vs. [NC3Rs] Anyone involved in animal research must consider how to both minimise harm to animals and maximise scientific progress. The 3Rs are one framework for doing this, but they are not the only one. There's also harm-benefit analysis, as well as the 3Vs of scientific validity – though modern definitions of the 3Rs, specifically reduction, do incorporate validity as a way to avoid unnecessary animal use. A review by Matthias Eggel and Hanno Würbel in Laboratory Animals explores further how these three ideas can work together.They explain that while there is little internal conflict between the 3Vs, there is potential for some in the 3Rs. Specifically, between refinement and reduction, where judgements may have to be made based on “more animals experiencing less harm” versus “fewer animals experiencing more harm”. Ethical review based on harm-benefit analysis can help solve such questions. Eggel and Würbel also look at the compatibility between the 3Vs and the 3Rs, finding that any potential conflicts can be resolved either by harm-benefit analysis or by good experimental design – for example, using randomised block designs to minimise sample size while maximising reproducibility. Ultimately the authors show that these three concepts can be integrated, forming a logical structure for better animal welfare and better science.Next, let's make some noise – or actually, don't. [NA3RsC] Did you know that mice and rats can hear sounds that humans can't? We can only hear up to 20 kHz, but mice and rats can up to 90 kHz. Furthermore, as they are nocturnal animals with relatively poor vision, noise and vibration are heavily used by rodents to navigate their environment. Unfortunately, these factors are rarely measured, can vary greatly from facility to facility or even room to room, and have the potential to cause both animal distress and be a confounding variable in scientific studies.A new paper in the Journal of the American Association for Laboratory Animal Science outlines these concerns and presents recommendations for developing a measurement plan. For example, facilities should develop a written plan, assess noise & vibration annually and as needed, and, importantly, maintain chronic noise below the threshold of negative animal welfare or scientific impacts. To learn more, read the full paper online. And finally, we'll finish with a replacement paper. [Lab... See acast.com/privacy for privacy and opt-out information.

You're listening to the December episode of 3 Minute 3Rs.The papers behind the pod:1. Protective cranial implant caps for macaques. Journal of Neuroscience Methods https://doi.org/10.1016/j.jneumeth.2020.1089922. The ‘Cage Climber' – A new enrichment for use in large-dimensioned mouse facilities. Applied Animal Behaviour Science https://doi.org/10.1016/j.applanim.2020.1050783. Skin swabbing is a refined technique to collect DNA from model fish species. Scientific Reports https://www.nature.com/articles/s41598-020-75304-1Transcript: It's the 3rd Thursday of December and you're listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce, and refine the use of animals in research. For the last episode of 2020, we're focusing on refinements for three different animals. Let's start big.[NC3Rs]Measuring the electrical activity of neurons during sensory or motor activities can reveal how the brain works.Macaques are often used in these studies as their brains most closely resemble those of humans. Devices to access the brain and to fix the head for stable electrophysical recordings are surgically implanted under general anaesthesia. These implants are designed to integrate with the skull however the surgical wound can be slow to heal. The animals are also prone to picking at the sutures increasing the likelihood of infection.A new paper published in Journal of Neuroscience Methods from technical and research staff at the University of Oxford and Newcastle University details how a protective cap can be used to promote wound healing. The plastic cap is adjustable to cover most primate cranial implants and can be affixed whilst the animal is under anaesthetic. Across the two facilities, the protective head cap reduced wound opening, the need to re-suture and the length of time animals needed to be administered analgesia and antibiotics.You can find out more about the cranial caps by following the link in the description.Next, a refinement for mice: [NA3RsC]Proper enrichment of mice in their home cages is important to decrease mouse stress, reduce stereotypic behaviors, and improve well-being. When developing new enrichments it's important to ensure they benefit both males & females and do not have unintended experimental effects or increase data variability.A new paper in Applied Animal Behavior Science describes the development and testing of a new enrichment made from recycled cage lids. Results showed that naive mice were extremely interested in these enrichments. Furthermore in a test battery assessing locomotion, anxiety, sociability, and stress physiology there was no impact on data results or variability. Furthermore, the enrichments reduced aggression.Ultimately the authors recommend the use of structural enrichments and nesting material to satisfy mouse physical and thermal needs. To find out more, read the full paper online.[Lab Animal]And finally, let's not forget about our fish. When a zebrafish or stickleback needs to be genotyped, that's usually accomplished via fin clipping. But, a growing body of literature suggests that fish might not be too fond of being removed from their tanks and waking up some time later with a small bit of their caudal fin missing. The tissue grows back, but studies have shown increases in cortisol and anxiety-like behavior in fin clipped fish, which may affect their welfare and the scientific results obtained with them.A new study led by William Norton at... See acast.com/privacy for privacy and opt-out information.

You're listening to the November episode of 3 Minute 3Rs.The papers behind the pod:1. Three-Dimensional Human Alveolar Stem Cell Culture Models Reveal Infection Response to SARS-CoV-2. Cell Stem Cell https://doi.org/10.1016/j.stem.2020.10.0042. Improving reproducibility in animal research by splitting the study population into several ‘mini-experiments'. Sci Rep https://doi.org/10.1038/s41598-020-73503-43. Three Pillars of Automated Home-Cage Phenotyping of Mice: Novel Findings, Refinement, and Reproducibility Based on Literature and Experience. Front Behav Neurosci https://doi.org/10.3389/fnbeh.2020.575434Transcript: It's the 3rd Thursday of November 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 one paper for each R – let's jump right in with a replacement. [NC3Rs]Severe acute respiratory coronavirus 2 (SARS-CoV-2), the cause of the current pandemic, infects human lung alveolar type 2 (hAT2) cells. To develop strategies for efficient prevention, diagnosis and treatment, we need to understand the disease pathogenesis. Using human cells in a physiologically relevant system has advantages over using cells lines and animal models including allowing viral replication. Organoid systems, using cells from the kidney, liver and airway, have been used in SARS-CoV-2 research however there have been technical challenges in using hAT2 cells in this way. Scientists in Korea in collaboration with research groups in the UK have successfully developed a 3D culture technique for primary human hAT2 cells that has been used to investigate the infection response to SARS-CoV-2. This 3D culture has provided an enhanced understanding of how SARS-CoV-2 causes such severe disease, whilst reducing reliance on animal models. The culture can also be combined with other techniques, such as co-culture experiments with immune cells and high throughput in vitro screening of antiviral agents, crucial to select successful treatments. Finally, it can be used for research for other respiratory diseases, reducing the reliance on animal models in respiratory research further.Want to know more? Check out the paper in the journal Cell Stem Cell.On to the next R: reduction [LA] No matter how carefully you try to control every little detail in your experiments, variables in your lab are going to differ from those elsewhere. As a result, a positive result in your hands may fail to replicate in the hands of another – with wasted animals in between. Rather than trying to standardize things further, a growing body of literature suggests a little more heterogeneity may be in order to improve reproducibility and reduce overall animal use. Multi-lab study designs are promising, but these can be logistically challenging. For the single lab that wants to introduce heterogeneity into their studies, researchers at the University of Munster suggest a simple solution: split that experiment up into several miniature versions. Rather than testing every animal all at once, the proposed mini-experiment design spreads those animals out across several time points. The idea being, things will change a bit in the lab in between testing rounds, similar to what might be expected if animals were instead being tested in separate facilities. In comparisons of four mouse strains across four replicate behavioral &... See acast.com/privacy for privacy and opt-out information.

You're listening to the October episode of 3 Minute 3Rs.The papers behind the pod:1. The epidemiology of fighting in group-housed laboratory mice. Scientific Reports https://www.nature.com/articles/s41598-020-73620-02. Development of a miniaturized 96-Transwell air–liquid interface human small airway epithelial model. Scientific Reports https://www.nature.com/articles/s41598-020-69948-23. Publication rate in preclinical research: a plea for preregistration. BMJ Open Science https://openscience.bmj.com/content/4/1/e100051Transcript: It's the 3rd Thursday of October 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'll take a look at preclinical publications and a microplate system to study small airway epithelium. But first, can we stop the fighting? [NA3Rs] Mice, the most commonly used mammal in scientific research, can sometimes be quite aggressive to each other in their home cages which can lead to serious injuries. In fact, mouse aggression is arguably the most common preventable morbidity factor. But its risk factors are little understood.However, a new epidemiological publication looked at mouse aggression in 60,000 cages across 1 year. They found that the biggest risk factor was housing. On average, 15% more fights were seen in individually ventilated cages with corn cob bedding than compared to static cages with wood chip bedding. Furthermore, they found that industry-standard cage-side checks for mouse aggression missed the majority of fights. They also found that fighting peaked during the summer months, on the top rack, and when racks were parallel versus perpendicular.To learn more about their findings, read the full paper in Scientific ReportsNext, an animal free epithelium model. [NC3Rs] The pharmaceutical industry is increasingly turning to human cell-based in vitro methods as a screening tool to identify promising drug candidates during the drug development process. Not only do these models reduce reliance on animals, they can also reduce drug attrition and provide a more cost-effective approach while allowing for higher throughput. Research into drugs for respiratory diseases is no exception to this trend, but small airway epithelial cells are particularly tricky to obtain from clinical sampling and struggle to divide ex vivo.Overcoming these issues means maximising the information you can get out of a limited number of cells. In a new paper, Teresa Bluhmki and colleagues at Boehringer Ingelheim describe a new miniaturised method of culturing small airway epithelial cells within a 96-well plate. After four weeks, an epithelium was produced with a similar composition to those created in 24-wells, which was then validated for use in drug discovery. Another exciting feature of the method is that it can be automated, allowing for even more efficient research into treatments for Idiopathic Pulmonary Fibrosis and other respiratory diseases.Interested in learning more about this small but mighty technique? Check out the paper in Scientific Reports. [Lab Animal] And finally, let's shine some light on preclinical publication. It's no secret that a lot of different animals are used in biomedical research each year. But how many of those animals end up in a publication describing the scientific results obtained with them? If University Medical Center Utrecth is any example, not many. A look at a selection of animal study protocols approved... See acast.com/privacy for privacy and opt-out information.

You're listening to the September episode of 3 Minute 3Rs.The papers behind the pod:1. Multicellular 3D Neurovascular Unit Model for Assessing Hypoxia and Neuroinflammation Induced Blood-Brain Barrier Dysfunction. Scientific Reports https://www.nature.com/articles/s41598-020-66487-82. Automated and rapid self-report of nociception in transgenic mice. Scientific Reports https://www.nature.com/articles/s41598-020-70028-83. Zebrafish as an alternative animal model in human and animal vaccination research Laboratory Animal Research https://labanimres.biomedcentral.com/articles/10.1186/s42826-020-00042-4 Transcript: It's the 3rd 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'll let mice do the talking and hear about using zebrafish to study vaccines. But let's start with an update on an organoid.[NC3Rs] Back in 2018, Goodwell Nzou and colleagues from the Wake Forest Institute for Regenerative Medicine in the United States published their microscopic replica of the human brain, formed from the six major neural cell types including neurons and immune cells. This miniature organ, or organoid, not only promoted the formation of a blood brain barrier, the resultant barrier was also functional.Fast forward to today, and in a publication in Scientific Reports, Nzou et al have demonstrated how this platform could be used in drug screening. Disruption of the blood brain barrier in neurological disorders, such as ischaemic stroke, is common, exacerbating the injury to the brain and contributing to cognitive impairment. By culturing the organoid in hypoxic conditions, replicating low oxygen resultant from a stroke, they were able to show expression of proteins critical for blood brain barrier function were altered, leaving the barrier disrupted and leaky. Inducing ischaemic stroke in rodents is associated with significant welfare concerns, including death, weight loss, sensorimotor defects and seizures. Using organoids can replace some of these experiments in disease modelling and therapeutic development, so this new publication could have big implications for the 3Rs. You can find out more by following the link in the description.[Lab Animal] Next, some self-reporting of pain. Mice are inevitably used to study the mechanisms underlying nociception, with the goal of better understanding pain in people and how to treat it. Many studies rely on reflex assays, but interpreting these can be subjective and there's uncertainty about what the mice are feeling and reacting to. This can limit study of the neuronal pathways involved and the affective components of pain perception.A new study in the journal Scientific Reports presents an assay in which transgenic mice learn to self-report exposure to a nociceptive stimulus. The mice, head fixed in the current study, were trained to lick a water spout in response to optogenetic stimulation of heat-sensing neurons in their hind paw. The authors suggest that self-reporting may provide a quicker read-out of nociception that may better reflect the animals' affective state, and hopefully help make results with the mice more translationally relevant. And we'll finish with a topic that's likely on many of our minds: vaccines.[NA3RsC] You are probably aware of the recent push to develop a vaccine for COVID-19. Before these vaccines are given to people... See acast.com/privacy for privacy and opt-out information.

You're listening to the August episode of 3 Minute 3Rs.The papers behind the pod:1. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PloS Biology https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.30004102. Acute and chronic stress prevents responses to pain in zebrafish: evidence for stress-induced analgesia. Journal of Experimental Biology. https://jeb.biologists.org/content/223/14/jeb2245273. 3Rs-based optimization of mice behavioral testing: The habituation/dishabituation olfactory test. Journal of Neuroscience Methods. https://doi.org/10.1016/j.jneumeth.2019.108550 Transcript:It's the 3rd 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. This month we've got papers on fish pain and a refined mouse olfaction task. But let's kick the episode off with the arrival of ARRIVE 2.0[NC3Rs] Improving the reproducibility of biomedical research is a major challenge. A lack of experimental detail in publications reporting in vivo experiments, such as experimental design and statistical analysis used, means researchers cannot adequately scrutinise the experimental results or repeat experiments. Consequently, data from these published experiments do not contribute to the knowledge base as researchers need to perform or to develop experimental procedures themselves. To highlight the information required the ARRIVE guidelines were developed in 2010. Although these guidelines were widely endorsed by journals and funders they have not led to improvements in reporting.To address this, ARRIVE 2.0 has been launched and published in PLOS Biology. The recommendations have been organised into two sets, so scientists can more easily incorporate them into their reporting. The first set, the essential 10 constitute the minimum reporting requirement for reviewers and readers to assess the reliability of the findings presented. The second set, the recommended set, provides further context to the study described. An Explanation and Elaboration document giving further information about each of the 21 items in ARRIVE 2.0 has also been published in PLOS Biology.To learn more about ARRIVE 2.0, explore the recommendations and find resources, visit its new home on the web at arriveguidelines.org. [Lab Animal] Next up, let's think about pain processing. In mammals, studies have shown that stress can alter pain-detection thresholds. Acute stress can have an analgesic effect, while chronic sources can enhance responses to pain, an effect known as hyperalgesia. Both can potentially confound experiments.Scientists at the University of Liverpool recently explored whether such stress-induced responses are also evident in zebrafish.The short answer? Yes. The team looked at the behavior of acutely and chronically stressed fish, as well as stress-free controls, after fin clipping. Following the potentially painful procedure, unstressed fish swam more slowly and lingered near the bottom of their tanks.No such change in behavior was observed in... See acast.com/privacy for privacy and opt-out information.

You're listening to the July episode of 3 Minute 3Rs.The papers behind the pod:1. Reproducibility of animal research in light of biological variation. Nature review Neuroscience https://www.nature.com/articles/s41583-020-0313-32. Inactivated rabies vaccines: Standardization of an in vitro assay for residual viable virus detection. Plos Neglected Tropical Diseases https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.00081423. A novel biosafety level 2 compliant tuberculosis infection model using a ΔleuDΔpanCD double auxotroph of Mycobacterium tuberculosis H37Rv and Galleria mellonella. Virulence https://doi.org/10.1080/21505594.2020.1781486Transcript: [NA3RsC] Reproducibility is a cornerstone of the scientific method. But unfortunately, it's considered to be quite poor for all research findings, and especially animal studies. Experts in animal biology, experimental design, and statistics argue that one major cause of poor reproducibility is inappropriate standardization in the face of biological variation. In nature, individual animals and humans vary greatly as a result of the interaction between their genes and the environment. So if studies standardize this natural variation too much, then they risk producing findings that are not reproducible outside of a very specific set of conditions. As a solution, experts recommend that researchers should deliberately diversify their study subjects such as by varying sex, genotype, age, and environmental conditions. This does not necessarily need to increase sample size or cost. And ultimately, this diversification can reduce long-term research costs, increase efficiency, and reduce overall animal suffering. To learn more, go online to read the full paper in Nature Perspectives. [NC3Rs] Vaccines are a vital tool in the fight against disease. This includes neglected tropical diseases such as rabies, which kills around 60,000 people each year, mainly in Africa and Asia.When inactivated rabies vaccines are produced, they go through many stages of quality control, including testing for any remaining live virus. This process traditionally uses a lot of animals, but a recent paper by Beatriz Lourenço Moreira and colleagues explores an alternative in vitro assay that could replace some of this animal useThe new method combines two in vitro techniques to create an assay that's three times more sensitive than in vivo testing and five times quicker than other in vitro assays. What's more, it could replace the use of animals in residual live virus detection, decreasing animal use in quality control by two thirds overall.While further validation is needed, and regulatory challenges still exist, this study shows what the 3Rs have to offer in vaccine development. Read the paper now in PLoS Neglected Tropical Diseases.[LA]... See acast.com/privacy for privacy and opt-out information.

You're listening to the June episode of 3 Minute 3Rs.The papers behind the pod:A novel, high-welfare methodology for evaluating poultry red mite interventions in vivo. https://www.sciencedirect.com/science/article/pii/S030440171930041XPluripotent state transitions coordinate morphogenesis in mouse and human embryos. https://www.nature.com/articles/nature24675TranscriptIt'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. Instead of the usual 3 segments this month, we're featuring two, in recognition of the two winners of the 2019 3Rs Prize, awarded by the NC3Rs and sponsored by GlaxoSmithKline. We'll start with a recap of Dr. Francesca Nunn's paper on red mite interventions in poultry. [NA3RsC] Laying hens can be infested with poultry red mites which causes major economic and animal welfare issues. These mites can cause anemia, reduced production, and even death. Current treatments are often inadequate and research methods to find new treatments typically have poor translation to field trials. A recent paper in Veterinary Parasitology describes a novel, high-welfare method for evaluating poultry red mite interventions in vivo. This paper is the winner of 2020 NC3Rs 3Rs Prize award. Nuun and colleagues developed and optimized a sealed mesh device that attaches to a hen's thigh. The depth and width of the mesh is precisely sized to both contain the mites and allow them to feed on the hens. This device will both reduce and refine the use of animals in research by allowing efficient pre-screening of new treatments before they enter large field trials. To learn more, read the full paper online. [NC3Rs] The second winner of the 2019 3Rs Prize award is Dr Marta Shahbazi from the MRC Laboratory of Molecular Biology for her work reducing the number of mice needed for embryonic developmental research. Historically embryos have been difficult to culture beyond the implantation stage of pregnancy. But with up to 40 per cent of pregnancies ending by implantation, it's a critical stage to understand. So researchers typically use extensive breeding programmes to generate transgenic mice and retrieve embryos needed for experiments with invasive surgeries. Now, building upon previous research, Marta and colleagues have established advanced 3D cultures of human and mouse embryonic stem cells to mimic the embryo at implantation. Using these cultures, they've been able to study the cellular events triggering implantation in greater detail than ever before. Not only that, they've also replaced the use of 500 mice. Groups worldwide are now adopting the cultures to answer their own biological questions about embryonic development. Why not join them, and check out Marta's prize-winning research by following the link in the description.With congratulations to Dr. Nunn and Dr. Shahbazi, that'll do it for June! 3 Minute 3Rs is brought to you by the NC3Rs, the North American 3Rs Collaborative, and Lab Animal. Come back in July for three more 3rs papers. See acast.com/privacy for privacy and opt-out information.

You're listening to the May episode of 3 Minute 3Rs. The papers behind the pod: 1 Refining rodent models of spinal cord injury: https://doi.org/10.1016/j.expneurol.2020.1132732 Preclinical validation of the micropipette-guided drug administration (MDA) method in the maternal immune activation model of neurodevelopmental disorders https://www.sciencedirect.com/science/article/pii/S08891591203025433 Identifying obstacles preventing the uptake of tunnel handling methods for laboratory mice: An international thematic survey https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0231454TranscriptIt'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 episode, we'll be talking about that last R, starting with a refined rodent model of spinal cord injury.[NC3Rs] Spinal cord injury research relies heavily on the use of rodent models, but these models can cause significant suffering. To address this, the UK's RSPCA convened an expert working group whose practical experience and expertise have informed a new set of recommendations on applying the 3Rs to rodent models of spinal cord injury. The 13 recommendations include reducing the severity of cervical level injuries or applying them to just one side of the body; providing adequate pain relief as standard; and planning and publishing research in accordance with good practice guidelines. A common theme is finding a balance between refining protocols and reducing animal use – for example, the appropriate use of both male and female subjects to reduce sex bias and improve translatability. If you work on spinal cord injury, these recommendations can help guide your 3Rs initiatives and ultimately improve treatments for those affected. Read them online now in Experimental Neurology. [Lab Animal] Next, what if there was a better way to administer treatments to your mice? Oral gavage can cause stress and minipumps require surgery, while providing a drug in food or water for the animals to consume at their leisure can be imprecise. To refine oral delivery, Joseph Scarborough and colleagues at the University of Zurich present their micropipette-guided drug administration, or MDA, method. They trained their mice to voluntarily consume what was on offer from a micropipette by sweetening the deal, adding a touch of sweetened condensed milk mixed with water as a delivery vehicle. The mice found things palatable indeed but to validate the approach, the researchers compared pharmokinetic profiles of risperidone administered via traditional gavage and the new MDA method. Those profiles were similar, except for stress levels. Cortisone was lower in the animals who consumed the risperidone willingly. To check out MDA protocol and see additional results with mouse models of maternal immune activation, see the publication in Brain, Behavior, and Immunity. And finally, let's take a look at... See acast.com/privacy for privacy and opt-out information.

You're listening to the April episode of 3 Minute 3Rs, brought to you by the NC3Rs, North American 3Rs Collaborative, and Lab Animal. The papers behind the pod: 1. In Vitro Antiviral Activity and Projection of Optimized Dosing Design of Hydroxychloroquine for the Treatment of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) (https://www.ncbi.nlm.nih.gov/pubmed/32150618)2. Engineering a 3D functional human peripheral nerve in vitro using the Nerve-on-a-Chip platform. https://www.nature.com/articles/s41598-019-45407-5 3. Laboratory Animal Welfare Meets Human Welfare: A Cross-Sectional Study of Professional Quality of Life, Including Compassion Fatigue in Laboratory Animal Personnel https://www.frontiersin.org/articles/10.3389/fvets.2020.00114/fullTranscript:Still social distancing, 3 Minute 3Rs is back with your April recap of efforts to replace, reduce, and refine the use of animals in research. Let's get right to it, starting this month with a replacement paper related to a topic that's probably near the top of many of our minds. [NC3Rs] Understandably, within recent weeks the focus of scientific research has predominantly shifted to COVID-19. While many experiments investigating treatment options or vaccines may require animals, there are opportunities where the 3Rs could be implemented in COVID-19 research.Researchers in Beijing have been doing just that by using in vitro and in silico methods to investigate the impacts of Chloroquine and Hydroxycholoroquine, two drugs that have recently received media attention. In their publication in Clinical Infectious Diseases, Yao et al used an established cell line to determine both drugs reduced the replication of COVID-19 in vitro. When administered prior to viral challenge, Hydroxychloroquine provided increased protection over Chloroquine. The group then used existing data from clinical trials and in vivo studies to develop an in silico physiologically based pharmacokinetic model and investigate potential effective dosing regimens in humans. The data from this publication are now being used to inform the design of a future clinical trial. While further experiments will be needed to confirm whether Hydroxycholoroquine is able to help patients with COVID-19, the publication demonstrates there continues to be opportunities to apply the 3Rs in scientific research. And where these opportunities exist, the 3Rs principles should always be considered. [NA3RsC] Neurodegenerative diseases such as Alzheimer's and ALS are often serious, life-threatening, and without a cure. Treatments can help, but finding drugs that work in final human clinical trials has been challenging. In part, this is due to the difference in biology between human and animal models. Microphysiological systems hold potential to increase translation but developing neural related models... See acast.com/privacy for privacy and opt-out information.

The March 2020 episode of 3 Minute 3Rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban) The papers: 1. https://pubmed.ncbi.nlm.nih.gov/32076633 2. https://www.sciencedirect.com/science/article/pii/S0273230020300507 3. https://www.nature.com/articles/s41598-020-59012-4 Transcript [NC3Rs] Nanomedicines have many potential advantages, including their ability to home to specific diseased tissues and exert a localised effect. Due to their relatively large size, the biodistribution of nanomedicines does not follow the same rules as small molecules and is largely dictated by the diameter of the blood vessels they pass through en route to their target tissue. This means that although current methods to assess biodistribution rely heavily on the use of animals including rodents and non-human primates, the results can vary greatly between preclinical species and man. Using an in vitro cell based model of nanoparticle uptake to build a computer simulation, Edward Price and colleagues at the University of Central Florida have developed a predictive system to replace animal use for determining nanomedicine PKPD. The model takes account of blood vessel size, tissue-specific porosity and the kinetics of distribution to cellular and paracellular compartments. By being able to better predict where nanomedicines will end up in the body, scientists can design safer and more effective drugs based on nanomaterials without using animals to assess biodistribution. For more information on their study, see the paper linked in the podcast description. [NA3RsC] Human safety is a key priority when developing new drugs. Toxicity testing often requires drugs to be used in animal models before human clinical trials. Historically, toxicology testing has involved two species: one rodent and one non-rodent. However, in some cases, only 1 animal species is used successfully. This month, an international working group of 37 diverse organizations led by Dr. Helen Prior published a paper on this topic in Regulatory Toxicology and Pharmacology. They reviewed data from 172 potential new drugs, across a wide range of small molecules and vaccines. These potential new drugs were tested in either 1 or 2 species. The working group found that often when 2 species were used they showed similar toxicity profiles in both. Therefore, it may be possible to reduce the number of species new drugs are tested in while maintaining human safety. [LA]. Social housing is generally recommended for social species, such as the mouse, whenever possible. But housing animals together can lead to incidences of aggression – an unwanted outcome for the welfare of the animals involved. Female mice tend to get along just fine, but males can sometimes be another story. In a new 14 week study measuring aggression and physical signs of well-being in male Cd-1 mice, a stock with an aggressive reputation, Paulin Jirkof and colleagues consider three variables that might influence how well those animals get along: group size, the age at which males are allocated to their groups, and handling frequency. The full results can be found in the journal Scientific Reports but overall levels of aggression were actually low. And for these mice, three was not a crowd. Trios got on better than pairs, with solo males scoring the lowest on measures of well-being. See acast.com/privacy for privacy and opt-out information.

The February 2020 episode of 3 Minute 3Rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban). 1. https://www.sciencedirect.com/science/article/pii/S0092867419313236 2. https://www.sciencedirect.com/science/article/pii/S0165027019303097 3. https://www.ncbi.nlm.nih.gov/pubmed/31852563 [NC3Rs] If you are bitten by a snake today, your anti venom treatment would be largely similar to if you were bitten 100 years ago. Anti venom is created by isolating antibodies from a large animal, typically a horse, after the animal has been given repeated injections of small amounts of venom. But due to issues with anti venom production, efficacy and safety standards, over 100,000 people worldwide will die from snake bites each year. Venom is a complex mix of molecules, which vary between different snake species and can only be obtained by milking snakes. This has been an obstacle in anti venom research. But now scientists from the Netherlands have generated an in vitro method for producing venom from snake venom gland cells cultured in 3d structures. In a publication Cell, Post et al. successfully cultured snake venom gland organoids from nine different snake species including the Cape Coral Cobra. The organoids growing definitely and produce functioning toxins, which could be used to further anti venom research. The organoids could also have further reaching consequences by enabling the development of new treatment options that might avoid the need for animals altogether. You can read more about snake venom gland organoids by following the link in the description. [LA] Neurons are often studied in cell cultures, but those cultures have to come from an animal originally. Nor do they last all that long – primary neuron cultures generally need to be used in a few weeks. That can put a rush on reproducing experiments, and often means extra animals need to be maintained for future use. To make cultures from a given cohort of mice last, researchers from University College London present a protocol to cryopreserve brain tissue and prepare neuronal cultures from it. Neurons retrieved from cryopreserved mouse hippocampi perform on par with freshly dissected cultures, with most physiological properties appearing unaffected by the process. Properly stored in liquid nitrogen, tissues can be frozen for at least two years. The results and step-by-step details can be found in the Journal of Neuroscience Methods. [NA3RsC] In a manuscript recently published in JAALAS, Dr. Brianna Gaskill and Dr. Joe Garner address considerations for publishing negative results and make practical recommendations for conducting sufficiently powered research. This includes challenging the traditional use of a priori power analyses and pilot studies for justifications is animal numbers and instead using more sophisticated experimental designs and calculating the least significant number of experimental units necessary to produce sufficient power. The authors suggest that researchers collaborate with experts in biostatistics, comparative medicine and other disciplines early in the research planning process in order to consider improvements in experimental design and select of appropriate statistical tests. Such collaboration will ensure the creation of high-quality experiments, helping researchers to address the 3Rs while improving reproducibility and translational value of animal studies. Finally, the authors offer recommendations for reporting to improve the interpretation of results. See acast.com/privacy for privacy and opt-out information.

The January 2020 episode of 3 Minute 3Rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban). Papers: 1. https://doi.org/10.15252/embr.201949709 2. https://www.nature.com/articles/s41598-019-56860-7 3. https://doi.org/10.1177/0023677219879455 [NC3Rs] Academic scientists produce many of the clever ideas which end up as medicines. However, the results in the papers reporting the work are almost always positive, and null or negative results rarely get a look in – a phenomenon known as publication bias. This is problematic as the negative or null data could be useful to other researchers. It can also lead to a culture of striving for positive results, potentially increasing poor experimental design or the inappropriate use of statistics. Overall this contributes to the so-called ‘reproducibility crisis' and means that animals are used in studies that are not correctly designed to demonstrate an effect. In response to this, scientists at the German Centre for the Protection of Laboratory Animals have set up a global ‘Animal Studies Registry' where a researcher can preregister their study and following completion the Registry is updated with the results. This publicly accessible database allows scrutiny of the planning and execution of animal studies. The Registry could become an important addition to the preregistration landscape - time will demonstrate the impact this can have on reproducibility and the use of animals in research. You can read more about the registry in this month's EMBO Reports via the link in the description. [LA] Next, how do you handle your mice? There's growing evidence that picking up the animals with cupped hands or a tunnel is less aversive than picking them up by the tail, but mice still need to be familiarized with the process. Across an entire animal facility, that time can add up. What's practical? Kelly Gouveia and Jane Hurst from the University of Liverpool have some suggestions. They recently evaluated BALBc mice handled by the tail, cupped hands, or a tunnel in three experiments: the first looked at the impact of holding duration; the second, the frequency of handling; and the third, responses to repeated immobilization for necessities such as subcutaneous injections. Tunnels rose to the top. Mice became familiar with tunnel handling from use during routine animal husbandry – a couple seconds each cage change was sufficient, and tunnel-acclimated animals remained less stressed than their tail-handled peers, even after the repeated immobilizations,. For all the details, see the publication in the Journal Scientific Reports. And finally, let's hit the wheel running with another refinement. [NA3RsC] Objectively measuring well-being in laboratory mice can be challenging. In a study published in the journal Laboratory Animals, voluntary wheel running behavior was evaluated as a measure of discomfort and disease severity in a mouse model of human Inflammatory Bowel Disease. While voluntary wheel running has been successfully used to assess severity levels in singly housed mice, mice are social animals. Here, voluntary wheel running was assessed in group housed mice undergoing DSS-induced colitis and in group housed mice that underwent a blood collection procedure at 3 different time points throughout the study. While a there was a significant reduction in wheel running and body weight, and a significant elevation in histological scores as disease severity increased, there were few signs of disturbed well-being based on clinical scores alone. Wheel running was also decreased in mice that underwent blood collection while other measures remained unaffected. This study suggests that voluntary wheel running may be a sensitive and suitable tool for individual severity assessment in group housed mice and offers a refinement to welfare by enabling social housing See acast.com/privacy for privacy and opt-out information.

The December episode of 3 Minute 3Rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban). Papers: 1. Cage aggression in group-housed laboratory male mice: an international data crowdsourcing project. https://www.nature.com/articles/s41598-019-51674-z 2. Towards animal-free neurotoxicity screening: Applicability of hiPSC-derived neuronal models for in vitro seizure liability assessmenthttps://www.altex.org/index.php/altex/article/view/1321 3. Optimal solid state neurons https://www.nature.com/articles/s41467-019-13177-3 [NC3Rs] Mice are social creatures. It's why we house them in groups in the lab to improve their welfare and allow the animals to make social interactions that simply aren't possible if they're individually housed. However, the social groupings mice make in the wild are complex, with hierarchies of social dominance. In the lab group has mice are kept in cages, which means dominant and subordinate males must stay in close quarters, which can lead to aggression related stress and injuries. To better understand this welfare concern, the NC3Rs have used a crowdsourcing approach, engaging with lab technicians worldwide to estimate the prevalence of male mouse aggression. Data from over 100,000 mice in over 45,000 cages was submitted for analysis, which has been published last month in Scientific Reports.The publication by Lidster et al. recommends small practical husbandry changes such as spot cleaning cages or transferring nesting material to minimize male mouse aggression. You can find out more in the link in the description. [NA3RsC] While drug approval rates are on the rise, only about 10% of drugs in phase 1 development will gain full approval. One of the causes of drug attrition is drug-induced seizures; There is a need for more translational in vitro alternatives to predict seizure liability. Here, the authors evaluated three human induced pluripotent stem cell (hiPSC)-derived neuronal models. All models demonstrated development of functional neuronal networks that exhibited parameters linked to seizure-like events in vivo. These parameters were then modulated with three compounds known to induce seizures. Differences in sensitivity were found between the models. Yet, compared to rat primary cortical neurons, the in vitro models were equally, or better able to detect seizureogenicity. While the authors recommend more compounds be tested and the available models further characterized before serving as full replacement, this data indicates the potential of these in vitro models as initial screening tools for seizure liability assessment which could lead to reduced animal use requirements. [LA] Neurons transmit electrical signals – so too do silicon chips. Moving that in vivo function in silico has long been a goal of many neuroscientists, but biological neurons are complex, and their electrical properties and nuances have been difficult to predict and mimic artificially. That hasn't stopped researchers from trying. A team led by Alain Nogaret at the University of Bath recently made progress, presenting artificial neurons on a silicon chip that matched the patterns of rat neurons with over 90% accuracy. It took lots of mathematical modeling, derived equations, and tweaking, but the team was able to optimize the chips so that they relayed electrical signals in the non-linear fashion that's characteristic of biological neurons, all with considerably less power required than prior attempts. The design details and rat neuron comparisons can be found in the journal Nature Communications. See acast.com/privacy for privacy and opt-out information.

The November episode of 3 Minute 3rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban). Papers: 1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6662399/ 2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5371849/ 3. https://pubs.rsc.org/en/content/articlelanding/2019/LC/C9LC00160C [NA3RsC] Evaluation of developmental and reproductive toxicity is a key aspect of drug development. Each year, 5-7% of newborns worldwide are born with serious birth defects, a number correlated to increased use of pharmaceutical drugs for managing health conditions. Reproductive toxicology studies account for roughly 90% of all animal use in toxicological safety assessment. The FDA requires that these animal studies be performed in two species, often rodents and rabbits. The embryonic stem cell test (EST) is an alternative in vitro assay that does not require the use of pregnant animals but is not yet powerful enough to provide a complete alternative, as it only considers the direct toxicity of compounds. Consequently, in vivo animal models are still required to evaluate effects on drug metabolism. This research established an in vitro platform (metaEST) which uses 3D embryoid bodies and 3D primary human liver microtissues on a chip to evaluate alterations in metabolism. Not only does this significantly reduce the number of animals required, these results demonstrate the platform's ability to evaluate metabolic effects that are missed using the standard EST assay and mimic physiologically relevant conditions in vitro. [NC3Rs] Botulinum neurotoxin can cause fatal paralysis following ingestion or infection of a wound and is a potential bioterrorism agent. Therefore, the ability to quickly and accurately detect the toxin in a sample is crucial. The current method used to detect the toxin is the mouse lethality bioassay, where mice are injected with neurotoxin to determine the LD50 value, or the dose that kills half the animals. This uses approximately 70,000 mice per year in the UK and more than 600,000 mice annually worldwide and causes severe suffering prior to death. Researchers at the Centers for Disease Control and Prevention in the US have developed a mass spectrometry-based method that can successfully identify botulinum neurotoxin activity for the four human-toxic serotypes. It is faster - the test can be done in just 4- 6 hours on the spectrometer rather than the 1- 4 days typically required by the mouse bioassay. The test also requires samples with volumes as low as 100 µL per toxin type, instead of the 1 mL required for the mouse bioassay, avoiding the problem of insufficient sample volumes. This mass spectrometry- based assay is a fast and accurate alternative to the mouse bioassay and further implementation of the technique has the potential to reduce the number of animals used for this purpose. [LA] Tissues do not exist in isolation from one another. As increasingly sophisticated organ-on-chip technologies continue to emerge, many researchers want to link those chips together to mimic multi-organ systems. That's been easier said than done. Yi-Chin however has been working on making better connections. The new microfluidic platform she helped developed consists of modular tissue and fluid control units, referred to as TILE modules. With the help of magnetic connectors, these modules can be arranged and re-arranged into different perfusion circuits as needed. It's backwards compatible too, meaning existing microfluidic devices can be used. The team involved demonstrated the platform in action with 2 and 3 tissue modules that mimic liver-mediated bio activation in response to a nutraceutical and a cancer prodrug. All using human cells, rather than animal models. See acast.com/privacy for privacy and opt-out information.

The October episode of 3 Minute 3rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban). Papers 1. Modelling human hepato-biliary-pancreatic organogenesis from the foregut–midgut boundary https://www.nature.com/articles/s41586-019-1598-0 2. Automated Platform for Long-Term Culture and High-Content Phenotyping of Single C. elegans Worms https://www.nature.com/articles/s41598-019-50920-8 3. Analysis of vertebrate vision in a 384-well imaging system. Thorn RJ. Sci Rep. https://www.ncbi.nlm.nih.gov/pubmed/31562366 Transcript [LA] Organoids are three dimensional structures derived from human cells. Unlike traditional cell cultures that are grown flat on petri dishes, multicellular organoids can offer researchers a little extra physiological context. But, an organoid of one organ still exists in isolation. A new paper published last month in the journal Nature starts to put some organoids together. The team, led by Takanori Takebe at Cincinnati Children's Hospital, wanted to study organagenesis, the process by which organs form. They took human skin cells, reverted them to pluripotent stem cells, and meticulously guided those to become small balls called spheroids that mimicked foregut and midgut. The team then put the two kinds of spheroids together in a special medium and let them grow. The structure that ultimately formed recapitulated functions of the liver, bile ducts, and pancreas. Such an integrated system is another step forward for organoid research. [NA3RsC] The worm, C. elegans, is an increasingly popular model for drug discovery and screening. For example, adverse effects of compounds can easily be detected by looking for changes in how fast the C. elegans grow, how big they get, and once they are larvae - how they move. Atakan and colleagues present a platform for automating the process of phenotyping single C elegans from the embryo to the adult. The platform overcomes many of the challenges of prior systems. It automates the painstaking process of distributing embryos into individual chambers and can expose the worms to up to 8 different chemical conditions in a single experiment. The phenotypes, including growth and movement, are measured automatically through analysis of 10 second videos captured every four hours across the worms' life cycle. To validate the platform, C. elegans were exposed to increasing concentrations of tetramisole and the known negative effects on growth and motility were demonstrated. The continued progress toward automating C. elegans studies with platforms, such as this one, will allow scientists to discover new opportunities for using the worm in early drug discovery. [NC3Rs] Automated behavioural monitoring can reveal subtle changes in an animal's vision before they can be detected physically. While these tests can be applied to rodents in the lab, each animal must be individually trained to use the system and only one animal can be monitored per cage, making it impractical for high throughput studies. In search of an alternative approach, Robert Thorn and colleagues at Brown University turned to zebrafish larvae at five days post-fertilisation. In their recent Scientific Reports paper, the team describe imaging 384 larvae at once in four 96-well plates, monitoring their response to moving visual stimuli. They found that diazepam significantly reduced their response, while flumazenil, a diazepam antidote, reversed this effect, illustrating that the larvae can be used to detect clinically relevant changes in the visual system, With three percent of the world's population affected by visual impairment, this new tool could speed up the screening and development of treatments while also replacing mammalian models. See acast.com/privacy for privacy and opt-out information.

The September episode of 3 Minute 3rs, brought to you by the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org), and Lab Animal (www.nature.com/laban) Papers behind the pod: 1. http://jpet.aspetjournals.org/content/371/1/15.long 2. https://www.sciencedirect.com/science/article/pii/S001216061830006X 3. https://www.nature.com/articles/s41467-019-11259-w [NC3Rs] Accurate measurements of drug and metabolite concentrations in the blood are vital to estimate exposure to the target in humans and animals during drug development. Zebrafish larvae are increasingly used for pharmacological research, but measurements of blood drug concentrations in these small animals have been technically challenging. A team at Leiden University have developed a method for nanoscale blood sampling from the posterior cardinal vein of zebrafish larvae at five days post fertilisation. A median volume of 1.12 nL of blood can be collected from each embryo and samples pooled to form a single replicate, which can be analysed by Liquid chromatography–mass spectrometry. While drug and metabolites could be successfully measured in the pooled samples, the authors suggest improvements to the sensitivity of the technique, which could reduce the number of embryos needed for each replicate. In addition, using microfluidic embryo handling techniques, blood sampling could be further automated and yield improved while reducing the amount of drug required. Further development of this microsampling technique has the potential to increase the use of the zebrafish embryo model to define drug pharmacokinetic properties. [NA3RsC] Josephine Morris and her colleagues at Bristol University used transgenic lines of zebrafish to study mechanisms of collagen formation and repair. This work was published in the journal Developmental Biology, Vol 441. They crossed transgenic zebrafish lines which integrated green fluorescent protein, expressed in the epidermis and mCherry collagen which is specifically expressed in the basal epidermis which allowed them to understand the dynamic nature of collagen 1 fibril deposition. The authors used Transmission Electron microscopy to demonstrate the intricate pattern integration of the fluorescent proteins in embryonic development. In other studies, a wound was created with a 30 g needle on the flank of 4 day old larvae and collagen repair was documented pictorially. By using the GFP collagen and mCherry collagen lines together, they were able to exploit the unique live imaging in larval fish to probe the process of collagen deposition and wound remodeling specifically in the basal epidermis and deeper layers. It is hoped that these transgenic lines will enable live imaging of collagen deposition and remodeling in various other organs and diseases. [LA] And finally, say hello to LipoGlo, a new reporter system for keeping track of lipoproteins in vivo. These proteins ferry fats throughout the body – you may be familiar with HDLs and LDLs, the latter of which can contribute to cardiovascular disease in people. A particular particle, Apolipoprotein B-containing lipoprotein, is particularly problematic and while it has been studied in mammals, such animals don't lend themselves to the large numbers needed for high-throughput drug discovery work. Enter the larval zebrafish. In order to visualize APoB-lipoproteins across the whole translucent organism, the research team took advantage of a glowing enzyme called NanoLuc, which they attached to APoB in the larvae. This bioluminescent reporter is highly sensitive and quite bright, allowing the team to follow the distribution and concentrations of the lipoprotein as it traversed the vascular system of the tiny little zebrafish. Additional details about the new tool can be found in the journal Nature Communications. See acast.com/privacy for privacy and opt-out information.

The August episode of 3-Minute 3Rs. Papers: 1. https://advances.sciencemag.org/content/5/7/eaaw4099 2. https://www.ncbi.nlm.nih.gov/pubmed/31026040 3. https://www.ncbi.nlm.nih.gov/pubmed/31343844 [LA] Nociception is the sense that tells animals they are encountering a potentially damaging stimulus. In people, we call it pain. Most pain research is done with rodents, but nociception is pretty well conserved beyond mammals. That includes invertebrates, such as the fruit fly. Some work has been done in larval drosophila, but larvae exist in a transient state; any quote unquote pain is therefore transient too. To study longer lasting changes in nociception, University of Sydney researcher Greg Neely recent;y described his lab's efforts to develop an adult fly model. They took advantage of the fly's natural aversion to surface temperatures above 42 degrees Celsius, and show that after peripheral injury-in this case, a leg amputation-the flys thermal sensitivity changes in a chronic manner. Looking deeper, they saw evidence of a complex mechanism known as central disinhibition at work, along with a role for a conserved gene called Twist in mediating that chain of events. You can read more about this potential fly replacement for nociception studies in the journal Scientific Advances. [NC3Rs] Before starting an experiment, a researcher should always have its end in mind. What is the hypothesis? The experimental objectives? And crucially for animal experiments, what signs indicate the experiment should be stopped to prevent unnecessary suffering? It's a challenging task though, identifying appropriate humane endpoints, as these should be specific to the animal and model but aren't always described in detail in the literature. Computers, however, are notoriously good at identifying patterns so researchers at the University of Berlin have investigated whether machine learning could be used to define humane endpoints in mouse models of stroke and sepsis. Data was collated from previous studies, focussing on commonly applied humane endpoints such as loss of body weight, changes in body temperature and the use of a sickness severity score. Using these measures, the machine learning based approach was able to predict whether an animal had a high risk of death, with up to 93% accuracy in the mouse model for stroke and 96% accuracy for sepsis. Notably, machine learning could make these predictions one to two days before a humane endpoint would be reached. And the faster we can identify an animal suffering or in pain, the faster we can intervene. [NA3RsC] A paper in Journal of BioPhotonics sheds some light on the value of fibered confocal fluorescence microscopy. Fibered confocal fluorescence microscopy is a confocal microscope equipped with a fiber-optic bundle which provides light to organs and tissues, enabling collection of real-time images of biologic processes in live animals. With conventional confocal microscopes, cell imaging is limited due to the difficulty with accessing deep tissues and organs. However, advancements in fiber-optic imaging technologies enables improved visualization of tissues and organs, including blood vessels, vasculature and nerves, and mucosal surfaces of the colon. In addition to the many scientific advantages, fibered confocal fluorescence microscopy also enables 3Rs impact. Each animal can be used as its own control with more information obtained from fewer animals, reducing the total number of animals required and inter-animal variability while increasing statistical validity. Also, in vivo imaging is non-invasive providing a valuable refinement to enabling identification of tissues under pathological conditions and responses to drug treatments. Finally, imaging of human organs and tissues could ultimately replace the use of animals, leading to improved clinical relevance making this a valuable translational tool expected to have increased impact in drug research and development. See acast.com/privacy for privacy and opt-out information.

The July episode of 3-Minute 3Rs from the North American 3Rs Collaborative (www.na3rsc.org), the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) Papers; 1. https://advances.sciencemag.org/content/5/5/eaav6528 2. https://www.nature.com/articles/s41598-019-44705-2 3. https://www.nature.com/articles/s41598-019-45464-w [NC3Rs] Colorectal cancer is particularly hard to treat, and only 11% of patients whose tumours have metastasised survive five years after diagnosis. Mouse models that incorporate patient tissue are widely used in drug discovery, but they're both expensive and time consuming, and often don't fully represent human disease. A team at the Mount Sinai Hospital in New York, led by Erdem Bangi, developed a screening platform using Drosophila to test potential therapies for a patient whose cancer had proved resistant to treatment. The team identified nine genes that were driving the progression of the disease, so they modified orthologs of these genes in the flies. They were able to use the flies to screen a raft of drug combinations, discovering that the most effective treatment combined trametinib, a chemotherapy drug, and zoledronate, usually used to treat bone diseases. When the patient was treated with this novel combination, their tumours shrank, then remained stable for a total of 11 months. This platform could reduce the number of mice used to find effective cancer treatments, not to mention getting them to patients quicker and cheaper. Personalised medicine is often said to be the next big thing in healthcare, but this new approach might just live up to the buzz. [NA3RsC] Too much standardization of experimental design may be unintentionally contributing to the reproducibility crisis in animal-based research. Today, rigorously standardized behavioral studies may be so well controlled that statistically significant findings may actually be irrelevant and not reproducible across laboratories. Bodden and colleagues at the University of Münster, in Germany, explore whether systematically varying study conditions can improve reproducibility. They specifically investigated the impact of the time of day the testing occurred on the outcomes of five different behavioral tasks across several replicates. They found that behavioral task results differed significantly depending on the time of day animals were tested, morning, noon, or afternoon. Follow-up simulations using the data generated suggest that systematically introducing two testing times, both morning and afternoon, into the same experimental design can significantly improve reproducibility of results within the same lab and, more importantly, is predicted to lead to better reproducibility across laboratories. This study describes a novel, easy to implement methodological approach to refining study design that has the potential to improve reproducibility of animal-based research. [LA] Researchers will want to know that their animals are in good health. But how can you tell if your zebrafish are in pain or stressed out from a procedure? Recent studies say that such fish tend to be less active and linger near the bottoms of their tanks. Perhaps easy enough to gauge by eye if your numbers are small, but some facilities maintain hundreds of tanks at a time. To help make those assessments more feasible for large-scale operations, and to reduce the risk of human bias, researchers from Lynne Sneddon's lab at the University of Liverpool developed the automated Fish Behaviour Index, or FBI. The FBI is based on swimming activity and swimming distance, recorded by cameras. The researchers developed the FBI with female AB strain zebrafish after different invasive procedures, such fin clipping and PIT tagging, and to evaluate the effects of different analgesics. Against human assessment, the FBI correlated well. You can read more and find links to the software in the journal Scientific Reports. See acast.com/privacy for privacy and opt-out information.

The June episode of 3-Minute 3Rs from the North American 3Rs Collaborative (www.na3rsc.org), the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) Papers; 1. https://bit.ly/2UH4hZt 2. https://go.nature.com/2W5CIea 3. https://bit.ly/2Fi4td4 [LA] On the road to the clinic, immunocompromised mice transplanted with patient-derived xenografts have been a frequent step for testing a drug's efficacy and safety. However, there may soon be fisher options afoot that might help replace some of those mice. Writing in the June issue of the journal Cell, David Langenau and colleagues describe an adult zebrafish xenograft model, into which they successfully engrafted several different solid pediatric tumors. Larval versions had existed before, but an adult model offers a few notable advantages. Bigger fish can bear a larger tumor burden for a longer period of time than the small fry. Adults can be orally gavaged, meaning researchers can know exactly how much of a drug they are getting. And with acclimation, they can handle the heat – the Langenau lab kept them at 37 degrees Celsius, the same temperature a tumor will find in the human body. In the paper, the zebrafish model performed comparably to a murine one. As such, xenografted zebrafish might be valid alternatives to mice, at least at the early stages of drug testing. [NA3RsC] What if it were possible to screen for compounds with cardiotoxic side effects in less than a few hours? Dr. Farraj and colleagues from the National Health and Environmental Effects Research Laboratory explore the possibility of using zebrafish embryos to accurately predict cardiotoxicity quickly. In their publication in Scientific Reports, Farraj and colleagues describe a multi-well platform with the ability to automatically quantify the heart rate of multiple zebrafish embryos per imaging field using an algorithm they call FisHRateZ. To demonstrate the utility of this approach, embryos were exposed to compounds known to either increase or decrease the heart rate. FisHRateZ accurately detected expected changes in heart rate with high sensitivity. The whole process is also quite fast. The assessment of a full 96-well plate can be conducted in less than 50 minutes. One limitation, they note, is the inability of FisHRateZ to measure arrhythmias. Check out this paper to learn more about the platform and download the free video processing algorithms. [NC3Rs] Scientists often use indirect measures such as changes in behavior or physiology to assess whether the welfare of the animal has been compromised, but methods which have been applied to larger non-aquatic animals are often not appropriate for fish. Recently, a mathematical method has come to the rescue, allowing the amount a fish moves to be used as a marker for welfare. Dr Anthony Deakin and team at the University of Liverpool developed the method, which used cameras to track the movement of individually housed zebrafish before and after undergoing potentially painful procedures. The trajectories created by the movement of the fish were analysed using the fractal dimension technique, which is a method used to reduce the complexity of data and produce a simple scale on which to compare the adversity of different procedures. This revealed significant reductions in movement complexity in procedural groups in comparison to the control. In particular, fin-clipped and PIT-tagged groups showed reduced movement complexity, highlighting the likelihood that these routine procedures are indeed painful. The addition of lidocaine reduced the impact of the fin clipping, demonstrating that this analgesia is an advantageous refinement in fish. Follow the link in the description to find out more about this non-invasive tool to monitor zebrafish welfare. See acast.com/privacy for privacy and opt-out information.

The May episode of 3-Minute 3Rs from the North American 3Rs Collaborative (www.na3rsc.org), the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) Papers: 1. https://bit.ly/2vYuUPR 2. https://go.nature.com/2w2tzre 3. https://go.nature.com/2VEjEHz [NA3RC] Early mortality of pups in breeding colonies can reach upwards of 50% or higher depending upon the strain. While high pup mortality is often considered "normal", it instead may be indicative of an underestimated animal welfare issue that requires attention. Leidinger and colleagues investigate the influence of environmental enrichment as one way to mitigate early pup mortality rates. Enrichment, with nesting material and a shelter, protected against early mortality. Survival rates of pups born into enriched environments were more than double that of pups born into impoverished environments. The impoverished environment also had a negative impact on the developmental course of the remaining pups. Enrichment, among many potential benefits, may have reduced maternal stress and improved thermoregulation, which is particularly important for newborn pups. The findings suggest that early pup mortality is an animal welfare problem that can be improved through the characterization and selection of appropriate enrichment strategies. The refinement in breeding conditions can improve pup welfare and survival and lead to the need for smaller breeding colonies. [LA] Enrichment helps pups, check. But how can you tell how they're doing in the first place? Invasive measures are challenging in such small animals, and pups lack features like fur and certain behaviors that can visually indicate health in adults. Dorothee Viemann and colleagues at Hannover Medical School recently proposed a new clinical health scoring system for neonatal mice to solve the pup assessment problem. They developed a score sheet based on observations of movement, nursing, and skin color and from quick examinations of capillary refill time, dehydration, reaction to tactile stimulus, and abdominal palpitation. They put their scoring to the test with healthy Black6 pup and two sepsis models known to cause mortality. The scoring correlated well with diseases, and was even more sensitive than blood-based biomarkers... [NC3Rs]…most creatures take great efforts to separate their ‘doings' from anything else they do. Rodents especially will almost always have a dedicated latrine area away from their nest site. Laboratory mice, typically housed in small, open chambers are prevented from performing this instinctual behavior, and their nesting material is often found in close contact with their droppings. Joanna Makowska from the University of British Columbia and her colleagues have investigated how this housing arrangement affects the behavior of these animals. Groups of mice were kept in one of two housing systems. A standard single cage or a rather more luxurious series of three interconnected cages set up in a similar manner to rodent burrows in the wild. Mice in the triple cages were observed to segregate their nesting and latrine areas between different cages. Single cage mice attempted to separate these sites with their cage, urinating and defecating away from their nests. However the day to day movements of the animals invariably scattered the contents of the enclosure. Notably, mice housed in triple cages showed more affiliative behaviors, such as grooming or resting with one another than those in single enclosures. indicating a stronger social bond between the animals. They also were less disturbed by weekly cage cleaning, a typically stressful and highly disruptive event for lab mice. In the paper, published in Scientific Reports this April the authors conclude that mice find being housed in contact with their waste aversive, and that facilitating separation of nesting and latrine areas could be a simple, effective way of improving the welfare. See acast.com/privacy for privacy and opt-out information.

This is the April episode of 3-Minute 3Rs, brought to you by the North American 3Rs Collaborative (www.na3rsc.org), the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) Source papers: 1. Characterizing Sources of Variability in Zebrafish Embryo Screening Protocols. https://bit.ly/2VP8njP 2. Effective chair training methods for neuroscience research involving rhesus macaques. https://bit.ly/2V7NOSv 3. AutonoMouse. https://bit.ly/2VP8tbb. Video: https://www.youtube.com/watch?v=6d7ctvj953M [NA3RsC] The zebrafish embryo, considered to experience little or no pain or distress, has been investigated as a humane replacement to the adult fish and adopted for chemical characterization and hazard assessments. The Systematic Evaluation of the Application of Zebrafish in Toxicology (SEAZIT) initiative was developed by the NTP NICEATM to address inconsistent test guidelines and identify sources of variability for zebrafish embryo-based assays. The goals included gathering input from zebrafish investigators on sources of outcome variability and facilitating a multi-laboratory evaluation of protocol variables. The SEAZIT team identified key factors producing variability which are highlighted in a report in a recent publication in ALTEX. While the current effort focused on defining a set of parameters that all researchers should consider, further advancements would improve the ease with which zebrafish embryo screening data could be used to inform a broad range of health-related research areas, including chemical hazard assessments. As the SEAZIT effort progresses, connections continue to be made with zebrafish researchers to collect a broad range of opinions and foster greater consistency in experimental approaches. [LA]... To move an animal as large as a macaque, many primate labs use special chair-like devices in which the animals are restrained. It can take considerable time to gets them trained and comfortable in the chairs, and the process can be stressful to all the primates involved. A team at Oxford, which uses macaques for neuroscience research, set out to refine their transportation training procedures. Details about their approach were published in the Journal of Neuroscience Methods at the beginning of the month. In short summary, they found that short but frequent training sessions started soon after animals arrived at the facility and using mostly positive re-enforcement in the form of tasty treats was most effective. They recommend that animals be trained in pairs or small groups rather than alone, and that negative reinforcement be limited as much as possible. They found that they could get their macaques into their transport devices in as few as 10 training sessions. [NC3Rs] Behavioral tasks in rodents have been an important tool in neuroscience research, but they are not without limitations. Traditional approaches are labor-intensive and require handling of the animals to move them to the testing arena, which can cause stress and affect the experimental outcomes. Erskine and colleagues...developed a fully automated system that addresses these issues. AutonoMouse... houses cohorts of up to 25 mice and allows continuous training and testing in enriched home cage environment, over a period of several months. Controlled by an ID microchip in each mouse, the cage is designed so that the animals can enter a behavioral test area individually, access water and engage in the tasks whenever they chose. The automated weighing module can enable monitoring to make sure the mice receive the amount of fluid they require. The system is also designed for minimal disturbance during cleaning, including bedding exchange without direct human contact. Detailed instructions and software are included in the paper to facilitate setup in other labs. The technology is less time-consuming and offers benefits to animal welfare, as the animals are kept in their home cage and don't have to be picked up as often. See acast.com/privacy for privacy and opt-out information.

This is the March episode of 3-Minute 3Rs, brought to you by the North American 3Rs Collaborative (www.na3rsc.org, the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) The papers behind the pod: 1. Progressive Motor Neuron Pathology and the Role of Astrocytes in a Human Stem Cell Model of VCP-Related ALS. https://bit.ly/2uiF64X 2. A critical evaluation of TRPA1-mediated locomotor behavior in zebrafish as a screening tool for novel anti-nociceptive drug discovery. https://go.nature.com/2Cu7u8C 3. No-touch measurements of vital signs in small conscious animals. https://bit.ly/2WdN5Mw [NC3Rs] A team led by Dr Rickie Patani has developed a human-derived model of ALS that could bring us a step closer to treating the disease effectively while avoiding the use of animals altogether. Animal models are widely used to study ALS, also known as motor neurone disease, but current therapies can only slow its progression – and even then, the effect is modest. Instead, Dr Patani's team, based at UCL and the Francis Crick Institute, used human induced pluripotent stem cells to study how ALS causes motor neurones to degenerate. They investigated the molecular processes that lead to the death of motor neurones, which are kickstarted by the loss of a protein called TDP-43 from the cell nucleus. They also discovered that ALS makes astrocytes degenerate too, so they can't play their usual role in helping motor neurones survive, compounding the effects of the disease. For this work, Dr Patani was awarded the NC3Rs' International 3Rs Prize earlier this month. The prize is sponsored by GSK and celebrates outstanding 3Rs science every year. Read the paper in Cell Reports or visit the NC3Rs website to learn more about the 3Rs Prize. [LA] Rodents remain popular for in vivo validation of novel drugs. But screening candidates is costly to do in rodents, which has researchers looking for alternatives to evaluate compounds in a higher throughput manner. Zebrafish are increasingly used for such screening purposes. A new paper from Richard van Rijn's lab at Purdue published in the journal Scientific Reports evaluates a zebrafish screen for drugs that Transient Receptor Potential A1, or TRPA1. TRPA1 encodes a calcium ion channel and has been shown to be involved in pain perception in rodent models. In zebrafish, activating TRPA1 causes hyperlocomotion, which the researchers hypothesized could be a useful phenotypic readout of drug efficacy. They tested compounds known to activate and inactivate TRPA1 in human cells, mice, and zebrafish larvae and found that the compounds affect all three models in a dose-dependent manner. Evaluation can be somewhat tricky in the zebrafish because they have a second copy of TRPA1 to contend with, but the fish could still help screen initial compounds before researchers take them onward. [LA] What if you could measure the vital signs of your animals without having to prep or handle them? Engineers at Cornell recently described the use of radio frequency near-field coherent sensing to do just that in a paper published in Science Advances. They developed the technology first for humans, but have now shown its potential for use with small animals in real-time. The technology uses radio waves. These penetrate the body and can be used to detect the motion of internal organs. When the signal is processed, parameters like heart rate and respiration rate can be captured. The system can be wired or wireless, and was shown to work with an anesthetized rat and freely moving hamster, Russian tortoise, and betta fish. There was some variability and further comparisons with existing methods would help clarify the accuracy and robustness of the new system, but near-field coherent sensing could be a promising new way to keep an eye on animal vital signs. See acast.com/privacy for privacy and opt-out information.

This is the February episode of 3-Minute 3Rs, brought to you by the North American 3Rs Collaborative (www.na3rsc.org, the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) The papers behind the pod: 1. 3D-printed Wash Station with Integrated Anesthesia Delivery Manifold for High-throughput Depilation of Laboratory Mice. https://bit.ly/2IpGrAP 2. DeepSqueak: a deep learning-based system for detection and analysis of ultrasonic vocalizations .https://go.nature.com/2tupSJK 3. Super-Mendelian inheritance mediated by CRISPR–Cas9 in the female mouse germline. https://go.nature.com/2RgnoYw [NA3RsC] Depilation (hair removal) is needed for many procedures done with laboratory mice, including surgery and imaging. Commercial depilatory creams are a safe, efficacious, and often-used method to achieve this hair removal. However, the use of these creams tends to be messy and time consuming, and wide procedural variation exists between laboratories. Furthermore, mice are generally under anesthesia during depilation, so increasing efficiency of the process is a valuable refinement to minimize anesthesia-associated adverse effects. Researchers from the University of Notre Dame have designed a 3D-printed device to simplify, standardize, and streamline the hair removal process. The Mouse Depilation Station (MDS) consists of an elevated stage with flow-through slats and is designed for integration with a self-scavenging anesthesia manifold and waste collection tray. It allows three mice to be depilated in parallel, thus improving efficiency compared to processing mice in sequence. The design files for the MDS stage are available online for free download – read the full article in JAALAS for the link. [NC3Rs] Imagine if laboratory animals could tell us how they were feeling. Scientists at the University of Washington in Seattle have moved this a step closer by developing Deepsqueak, a system to decipher rodent ultrasonic vocalisations. Usually outside the human range of hearing, rats and mice use a characteristic set of squeaks to communicate with one another. Writing in the journal Neuropsychopharmacology Coffey et al have used deep learning algorithms similar to those which power Alexa's speech recognition abilities to identify and categorise rodent ‘syllables'. The rodent chatter can be linked to behaviours and physiological parameters, allowing an electronic dictionary of rodent calls to be built. One particular area of interest is in neuroscience research, where rodent behavioural tests are routinely applied to study a diverse collection of outcomes and diseases. By using fully automated monitoring of ultrasonic vocalisations during experiments an additional level of behavioural information can be collected, providing a non-invasive method for monitoring rodent welfare. [LA] All animals inherit their genes from their parents, but there's a 50/50 chance whether a particular copy of a gene will come from the mother or from the father. When you want to make a mouse model of a particular genetic mutation, that often means a lot of animals are bred that don't have the desired change. But what if you could weigh that probability? It's been demonstrated already in insects, but Kimberly Cooper from the University of California San Diego and her colleagues bring active genetics to mammals for the first time in a new publication in Nature. They developed a CRISPR/Cas9 construct that could produce of litter of mice in which over 70% of the pups had the desired mutation. It only worked in females so far, but the approach proves that it is feasible to weight the genetic coin in mice. See acast.com/privacy for privacy and opt-out information.

This is the January episode of 3-Minute 3Rs, brought to you by the North American 3Rs Collaborative (www.na3rsc.org, the NC3Rs (www.nc3rs.org.uk), and Lab Animal (www.nature.com/laban) The papers behind the pod: 1. An Atypical Parvovirus Drives Chronic Tubulointerstitial Nephropathy and Kidney Fibrosis https://bit.ly/2FA6dPA 2. Remote Welfare Monitoring of Rodents Using Thermal Imaging https://bit.ly/2suGlgm 3. No experimental evidence of stress-induced hyperthermia in zebrafish (Danio rerio)https://bit.ly/2Db8m2P [NA3RsC] Chronic kidney disease results from fibrosis of the kidney parenchyma due to a variety of inciting factors. Existing models of tubulointerstitial nephropathy and fibrosis involves administration of nephrotoxic compounds or surgical interventions, but these models often fail to recapitulate the chronic, insidious nature of fibrotic disease. In a recent publication from the October 4, 2018 issue of Cell, Ben Roediger and his colleagues at the University of Sidney Australia observed a high incidence of kidney disease in 3 separate colonies of immunodeficient mice; disease was characterized by enlarged nuclei in tubular epithelial cells in addition to intranuclear inclusions and marginated chromatin. RNA sequencing identified a complete novel viral genome that was given the name ‘mouse kidney parvovirus.' Resulting pathology included extensive fibrotic changes to the kidneys with significantly reduced renal mass. Virus was absent in mouse colonies free of kidney disease, transmissible following co-housing and infected mice exhibited similar transcriptional changes to humans with fibrotic disease. Ultimately, mouse kidney parvovirus may represent a new tool for understanding viral nephropathy as well as the fibrotic processes and pathogenesis of chronic kidney disease. [NC3RS] To assess the well-being of an animal, it is useful to be able to continuously monitor their vital signs, such as heart rate and breathing rate, and the amount they move. Vital signs can be assessed by implanting a telemetry sensor or using electrocardiography. However, these methods can lead to unnecessary discomfort and stress for the animal and can compromise their welfare. A recent study by Pereira et al. addresses these issues by using thermal imaging to monitor animal welfare. Thermal imaging is a remote and passive monitoring technique that can continuously record the heat emitted from the animal's body. Using this technology, the researchers were able to remotely monitor breathing rate of anaesthetised rats and mice, and the position and velocity of active rodents during open field tests. This promising technique has the potential to refine current monitoring practice and improve animal welfare. Follow the link in the description to find out more. [LA] When endothermic animals, like ourselves or our laboratory mammals, get stressed, we often get hot. Literally. But not all animals control their internal body temperature. A 2015 paper published in the Proceedings of the Royal Society B indicated that zebrafish stressed out from temporary confinement will deliberately move to a warmer area of their tank in order to raise their body temperature. This suggests that they too exhibit stress-induced hyperthermia. But questions have lingered about whether the fish were actively seeking heat or rather, just moving away from the chamber where they had been confined. Writing in the Journal of Experimental Biology, researchers from the University of St. Andrews repeated that experiment, but included control tanks without a temperature gradient. They saw no discernible effect. They also developed a second tank set up - in that case, stressed fish actually spent a little less time in warmer water. Don't neglect zebrafish stress, the authors stress, but their results suggest that the fish don't experience stress-induced hyperthermia in the same as other animals. See acast.com/privacy for privacy and opt-out information.

It's the December episode of 3-Minute 3Rs, brought to you by , the NC3Rs (www.nc3rs.org.uk), the North American 3Rs Collaborative (www.na3rsc.org) and Lab Animal (www.nature.com/laban) The papers behind the pod: 1. Optimizing the design of population-based patient-derived tumor xenograft studies to better predict clinical response https://bit.ly/2GwGJ7N 2. An In Vitro Model of Hematotoxicity https://bit.ly/2Ezgaw3 3. Found In Translation https://go.nature.com/2UVJ0g6 [NC3Rs] Every cancerous tumor is different and treating two tumors with the same drug may result in two very different outcomes, which contributes to the high failure rate during cancer drug discovery. One experiment type gaining interest to model this tumor heterogeneity is screening a population of patient-derived tumor xenograft models, or PDX models. However, there is little guidance available for these experiments. This risks poor experimental design that could result in a study being under or overpowered. Both these scenarios waste animals. A new freely available interactive tool from a team at AstraZeneca, published in Disease Models & Mechanisms, enables scientists to design robust and reproducible population PDX studies. The tool developed by Floc'h et al explores the design of population PDX studies and how this can impact the risk of both false-negative and false-positive results. Each experiment for a specific drug or cancer type is unique so the tool cannot provide exact recommendations, but what it can do is support you in selecting the optimal design for your experiment whilst ensuring the 3Rs are also considered. Details for how to access the tool can be found via the link in the description. [NA3RsC] Mature blood cells and bone marrow are common targets of drug toxicity. In addition to leading to significant complications for patients receiving antineoplastic agents for cancer treatment, these hematotoxic effects can be limiting, preventing administration of the doses needed for therapeutic efficacy. Screening for potential toxicity to the hematopoietic system in animals and in human cell-based in vitro approaches is critical in early drug development. The CFU assay has been validated as the traditional screen for hematotoxicity, but has limitations. A publication in Current Protocols in Toxicology by Mahalingaiah and colleagues describes the next-generation, high-throughput assay as a refined approach. A liquid culture, lineage-specific, in vitro cell differentiation model offers multiple advantages over the traditional assay, making it a versatile and useful tool to screen compounds. In addition to decreasing the number of animals required for hematoxicity determination, this human, cell-based in vitro assay is a useful alternative to in vivo studies for investigative work in understanding mechanisms of lineage-specific hematotoxicity at different stages of differentiation. [LA] So you've made a mouse model of disease, applied a treatment, and measured gene expression. Now what? You might think the gene with the greatest changes in expression is going to be the most important for the disease. But there's a pretty big evolutionary gap between mouse and man. From just your experiment, do you really know how relevant that gene will be? To better bridge the species gap, researchers from Technion have developed a new statistical model that takes decades of previously recorded gene expression data in mouse and human into account. They call it Found in Translation, or FIT for short. For new mouse data, FIT calculates a per-gene effect size and predicts which gene in your mouse might be most relevant to humans. It can potentially rescue genes that might not otherwise make the cut for further studies and de-prioritize those important to a mouse but perhaps less so to a person. No new mice experiments required. You can find all the details about FIT in Nature Methods, and you can try it out yourself with your own gene expression data at mouse2man.org. See acast.com/privacy for privacy and opt-out information.