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3 Minute 3Rs

The NC3Rs, the North American 3Rs Collaborative, and Lab Animal

3 Minute 3Rs is your monthly recap of efforts to replace, reduce, and refine the use of animals in research, brought to you by the NC3Rs, the North American 3Rs Collaborative, and Lab Animal.

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Issues with conventional rodent housing, playpens for rats, and using sleep to assess welfare
Feb 17 2022
Issues with conventional rodent housing, playpens for rats, and using sleep to assess welfare
February https://doi.org/10.1186/s12915-021-01184-0https://doi.org/10.1177/00236772211065920https://doi.org/10.1038/s41598-021-04502-2It’s the 3rd Thursday of February, and you’re listening to 3 Minute 3Rs. This month, we’re bringing you 3 refinement papers. Let’s start with rodent housing.Research rodents are conventionally housed in shoebox sized cages that limit their ability to perform natural behaviors such as nesting and burrowing. These restrictions are known to impair welfare, but could they even increase disease risk and shorten lifespans? A new meta-analysis compares the morbidity and mortality of rodents in conventional vs enriched housing. Conventional housing was found to significantly worsen disease severity for cancer, cardiovascular disease, stroke, anxiety, and depression. Conventional housing also increased all-cause mortality. In conclusion, conventional housing appears to be distressing enough to compromise rodent health in a biologically significant manner. This lends more evidence to the importance of refining rodent housing for ethical, validity, and translational reasons. To learn more, read the full paper online.Sticking with the rodent theme, if you work with rats, you might find standard cages don’t provide enough room for a fully enriched environment – one where rats can socialize, exercise and express natural behaviours. If larger cages are not currently an option, two solutions with increasing research to back them up are playpens and ball pits, the focus of a recent paper in LA.Justyna Hinchcliffe et al. describe using 50 kHz ultrasonic vocalisations as an objective, quantifiable measure of how rats responded to ball pits and playpens. They found that these vocalisations were more frequent for rats exposed to the enriched environments, when compared with control conditions. They also tested the effect of playpens and ball pits on rats given an aversive drug treatment, finding that exposure to these environments reduced its negative impact. Besides the important welfare benefits of environmental enrichment, there are also scientific benefits in the shape of reduced variability and therefore more reliable results. With new evidence supporting the use of playpens for rats to minimize stress, there has never been a better time to try them out in your facility.And finally, let’s see how sleep could be a helpful tool to help us track laboratory animal welfare.In humans, sleep quality is strongly related to a person’s well-being and recent research suggests the same may be true in laboratory animals, such as dogs. If this is the case, sleep quality may then be useful as a non-invasive measure of animal welfare. A study in Sci. Rep. examined this hypothesis further using an observational approach to characterize sleeping patterns in laboratory dogs and investigate the effects of sleep quality on their daily behaviors. Male and female adult dogs housed in kennels in Brazil were recorded during a continuous 24-h, five-day assessment period. The footage was then analysed for daytime behaviours and sleeping metrics, such as number of sleep bouts and their duration. The study showed that the dogs slept far less than reported previously in the literature and during the day they were less active, ate more, played less and were less alert. Alterations of these daytime behaviors as a result of loss of sleep may indicate compromised welfare highlighting appropriate measures should be taken to ensure lab dogs’ sleep quality and welfare.  Hosted on Acast. See acast.com/privacy for more information.
Reproducibility in cancer biology, training rats for refined fMRI, and playpens for mice
Jan 20 2022
Reproducibility in cancer biology, training rats for refined fMRI, and playpens for mice
The papers behind the pod:1. https://doi.org/10.7554/eLife.71601 & https://doi.org/10.7554/eLife.679952. https://doi.org/10.3389/fnins.2021.8056793. https://doi.org/10.1038/s41598-021-98356-3It’s the 3rd Thursday of January – happy new year! You’re listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. Of course, we focus on those three Rs, but many have suggested adding a fourth R to the list: reproducibility. Designing experiments with reproducibility in mind is a key aspect of reducing unnecessary animal use, as well as being good for advancing science.In 2013 the Center of Open Science and Science Exchange began a collaboration to investigate the reproducibility of 193 experiments from 50 high-impact cancer biology papers. Over eight years of repeated experiments, they found that they could only reproduce 50 experiments from 23 papers, generally due to a lack of detail about the methods used or resources being unavailable. 15 of those 50 repeated experiments used animals, and while just over half of them at least partially confirmed the original results, the repeated results were not always statistically significant. Experimental design was also an issue: only one of the original animal experiments used randomization and none used blinding or calculated a sample size before the study began.Papers describing these results are now available in eLife, with all the relevant data available on the Open Science Framework website and more Replication Studies to come from this collaboration. As the reproducibility crisis continues to rumble on, why not check them out and put designing more robust experiments at the top of your agenda?Next, let’s look at how training rats can help make fMRI a less stressful experience. Functional magnetic resonance imaging, or fMRI is a powerful non-invasive procedure that is used to assess brain function and connectivity. However, fMRI research in animals is often confounded due to the physical restraint and loud noises that occur during recordings as these induce stress which can alter information processing and cognition.An article from Frontiers in Neuroscience describes a protocol for habituating rats to fMRI that also avoids the need for surgical head restraint. Rats were gradually trained via 18 sessions over 3 weeks beginning with basic handling phase. After following this protocol, fMRIs in awake rats were successfully conducted without inducing increased stress and still achieving stable images with very low motion artifacts.To learn more about this rat refinement, read the full paper online. Finally, playpens for mice – could they be a viable option for refinement when home cage space is limited? Good environmental enrichment improves the quality of life for laboratory mice by providing increased opportunities to carry out natural behaviours such as running, climbing and burrowing. However, due to space requirements, cost and sanitation constraints many facilities worldwide still use standard housing, which has been associated with potential welfare problems.  In their publication in Scientific Reports, Ratuski et al show temporary access to playpens could be an effective method to provide mice housed in standard cages with space and structures to facilitate natural behaviors. In this study, female mice were given access to playpens three times a week for several weeks. Mice in the playpens were more active, compared to mice in conventional cages and over time, the animals entered the playpen more quickly and showed increased anticipatory behaviors before accessing the playpen. All indicating the mice found access to playpens rewarding. Want to learn more? Follow the link in the description. Hosted on Acast. See acast.com/privacy for more information.
Refining anaesthesia and euthanasia for zebrafish and mice
Dec 16 2021
Refining anaesthesia and euthanasia for zebrafish and mice
Sedation is necessary for zebrafish during procedures such as imaging, biopsy, and surgery to ensure animal welfare and high-quality science. But the effects of sedation can last beyond the administration period and should also be carefully considered. A paper by Gressler et al. explores the use of eugenol and propofol during a 3-hour sedation and their subsequent effects after a 1 hour washout period. Both drugs had effects on behavior and physiology even after the washout period. For behavior, in a novel tank test, eugenol was found to amplify diving response while propofol induced anti-anxiety responses. For physiology, both drugs caused alterations in gill structure. Clearly, as with other species, sedation of zebrafish can significantly affect behavior and physiology beyond the administration period. Therefore, sedation procedures must be carefully designed and reported to refine experiments. Read more: https://doi.org/10.1016/j.applanim.2021.105482Next, let’s look at euthanasia, an important consideration for any animal study, and those involving zebrafish are no exception. Despite zebrafish being widely used in research, there is no consensus on which method to use when euthanizing them. The most commonly-used method – an overdose of tricaine, or MS-222 – is versatile, readily available and, in fact, is the only legal option in some areas, but it is now known to be aversive. A new paper by von Krogh et al. describes work to address this discrepancy. The team screened overdoses of 7 common alternative anaesthetics dissolved in water, to determine whether they led adult zebrafish to lose reflexes in a rapid, reliable, and non-aversive manner. Other substances were used to buffer the anaesthetics where needed, adjusting the water pH to reduce irritation. The authors found adding 1g/L lidocaine hydrochloride buffered with 2g/L sodium bicarbonate was particularly effective, reliably inducing loss of all reflexes within 2 mins and provoking little aversive behavior. Adding 50mL/L ethanol further reduced these issues. While the authors recommend this method, they also stress the importance of further investigation, including for zebrafish at different developmental stages. Read the paper: https://doi.org/10.3390/biology10111133Finally, let’s focus on mice and how researchers are working to ensure they are euthanized humanely. Gradual exposure to carbon dioxide remains the most common method for euthanising laboratory rodents. Although CO2 is generally considered an acceptable option when properly administered, there are concerns that CO2 is aversive to rodents and might cause distress and pain above certain concentrations. In a new study, Rodriguez-Sanchez and colleagues investigated using a voluntarily ingested sedative to reduce CO2 aversiveness. Cream cheese mixed with different doses of a rapid-acting anesthetic was provided to C57BL/6 mice before exposing the animals to CO2. Using a broad range of behavioural parameters, the team showed 20 mg/kg of sedative resulted in a mild sedation and likely reduced the aversiveness of CO2. While the investigators acknowledge more work is needed to determine the experience of the mice during sedation, they suggest voluntary oral administration of a sedative is potentially an effective, affordable, and easy way to minimise the stress of mice during CO2 euthanasia. Find out more: https://doi.org/10.3390/ani11102879 Hosted on Acast. See acast.com/privacy for more information.
Discover the winners of this year's Global AAALAC/IQ Awards
Nov 18 2021
Discover the winners of this year's Global AAALAC/IQ Awards
The 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. Hosted on Acast. See acast.com/privacy for more information.
People and strategies behind 3Rs science: compassion fatigue, culture of care and systematic reviews
Oct 21 2021
People and strategies behind 3Rs science: compassion fatigue, culture of care and systematic reviews
The 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.3001177Transcript: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... Hosted on Acast. See acast.com/privacy for more information.
3 Minute 3Rs September 2021
Sep 16 2021
3 Minute 3Rs September 2021
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... Hosted on Acast. See acast.com/privacy for more information.
3 Minute 3Rs August 2021
Aug 19 2021
3 Minute 3Rs August 2021
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... Hosted on Acast. See acast.com/privacy for more information.
3 Minute 3Rs July 2021
Jul 15 2021
3 Minute 3Rs July 2021
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... Hosted on Acast. See acast.com/privacy for more information.