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1August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareEditorial Jas BarleyGuest Editorial Vicky Robinson Handling mice using gloves sprayed with alcohol-based hand sanitiser: acute effects on mouse behaviour Noelia Lopez-Salesansky, Dominic J Wells, Natalie Chancellor, Lucy Whitﬁ eld and Charlotte C Burn Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting Chloe Stevens, Penny Hawkins, Tom V Smulders, Aileen Maclelan, Lars Lewejohann, Paulin Jirkof,Jackie Boxall, Helen Murphy, Carley M Moody, Patricia V Turner, I J Makowska and Charlotte InmanEmotional challenges in our work with laboratory animals: tools that support caring for others and yourselfAngela Kerton and Jordi L Tremoleda Husbandry and healthcare of the Olive python (Liasis olivaceus)Gary MartinicPAPER SUMMARY TRANSLATIONSFrench, German, Italian, Spanish LASA, RSPCA guiding principles to help deliver the ethics learning outcomes of module 2 for personal licenseesM Jennings and M Berdoy (editors), A-M Farmer, P Hawkins, E Lilley, A Kerton,B Law, Jordi L Tremoleda, C Stanford, L Whitﬁ eld and K RyderTECH-2-TECH ASRU-RSU workshop: impact of COVID-19 on present and future training for animal research – issues and opportunities for training and CPD in light of COVID-19Linda Horan and Ngaire DennisonBOOK REVIEWSIndex to Advertisers214481735196Vol 20 No 1 April 2021EditorialJas Barley, Chair of the Editorial BoardReport of the 2019 RSPCA/UFAW RodentWelfare Group meetingChloe Stevens, Emily Finnegan, Jasmine Clarkson,Charlotte Burns, Sonia Bains, Colin Gilbert,Caroline Chadwick, Samantha Izzard, Charlotte Inman,Penny Hawkins (Secretary) and Huw GolledgeReduction of the negative effects ofmethionine on bone parameters in broilers’embryos by intra-egg injection of Vitamin B12Mohammad Naser Nazem, Shima Tasharofi,Negin Amiri and Sepideh SabzekarThe care of the Children’s Python(Antaresia children)Alexander Hosking and Gary MartinicFeline-assisted therapy: a promising part of animal assisted therapy (AAT)Eliska Mičková and Krityna MachovaThe care of Central and Pygmy Bearded DragonsAlexander Hosking and Gary MartinicPAPER SUMMARY TRANSLATIONSFrench, German, Italian, SpanishLOOKING BACKPhysical hazards in the laboratory animal houseR.T. CharlesThe incidence of a pathogenic strain of pseudomonas in a rabbit colonyG.R. Alpen and K. MaerzTECH-2-TECHDevelopment of a sifting cage change method for rats to improve welfar eSeonagh HendersonVol 1 9 No 2 A ugust 2020CONTENTSiAugust20:Animal Technology and Welfare 4/8/20 10:48 Page i1135634

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3August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarevOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richard Gardner MA PhD FRSBFIAT (Hon) FRSVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS,Stephen Barnett BA MSc FIAT (Hon) CBiol FRSBRAnTech, Miles Carroll PhD, Paul Flecknell MA Vet MBPhD DLAS DipLECVA MRCVS, FIAT (Hon), PennyHawkins PhD BSc, Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.), DipECLAMFRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&SMSc MRCVS, Tim Morris BVetMed PhD DipACLAMDipECLAM CBiol FRSB CertLAS MRCVS, Clive PageOBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky RobinsonCBE BSc PhD, Paul Sanders MIAT RAnTech, DavidSpillane FIAT, Gail Thompson RLATG, RobertWeichbrod PhD RLATGLife MembersKen Applebee OBE FIAT CBiol FRSB RAnTech,Charlie Chambers MIAT RAnTech, Roger Francis MScFIAT RAnTech, Pete Gerson MSc FIAT RAnTech,Cathy Godfrey FIAT RAnTech, John Gregory BSc (Hons)FIAT CBiol FRSB RAnTech, Patrick Hayes FIAT DipBARAnTech, Robert Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon)RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Sarah Lane MSc FIAT,Sue McHugh BSc FIAT, Norman Mortell BA (Hons)MIAT RAnTech, Wendy Steel BSc (Hons) FIATMembers of CouncilMatthew Bilton, Kally Booth, Steven Cubitt,Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Alan Graham, Linda Horan, Sam Jameson,Elaine Kirkum, Adele Kitching, Theresa Langford,Sylvie Mehigan, Steve Owen, Alan Palmer, AllanThornhill, John Waters, Lynda Westall, Car ole W ilson,Adrian WoodhouseCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary:Simon Cumming BSc FIAT RAnTechTreasurer: Glyn Fisher FIAT RAnTechChair of Board of Educational Policy:Steven Cubitt MSc FIAT RAnTechChair Registration & Accreditation Board:Glyn Fisher FIAT RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATATW/Bulletin Editorial Board:Jas Barley (Chair), Matthew Bilton, Nicky Gent,Patrick Hayes, Elaine Kirkum, Carole Wilson,Lynda WestallBranch Liaison Officer:Kally Booth MIAT RAnTechEFAT Representatives:Glyn Fisher, Alan PalmerWebsite Coordinator:Allan Thornhill FIAT RAnTechAnimal Welfare Officers and LABARepresentatives:Matthew Bilton (Chair), Kally Booth, Lois Byrom,Simon Cumming, Nicky Gent, Sylvie Mehigan,John WatersBoard of Educational Policy:Steven Cubitt (Chair), Adele Kitching (Secretary)Communications Gr oup:Adrian Woodhouse (Chair), Elaine Kirkum,Teresa Langford, Sylvie Mehigan, Allan Thornhill,Lynda WestallIAT REPRESENTATIVESAugust20:Animal Technology and Welfare 4/2/21 13:19 Page v

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4Animal Technology and Welfare August 2020BRANCH SECRETARIES 2021Cambridge: Tony Davidge cambridgebranch@iat.org.ukEdinburgh: Kery-Anne Lavin-Thomson edinburghbranch@iat.org.ukHuntingdon, Suffolk & Norfolk: Jo Martin hssbranch@iat.org.ukIreland: Lisa Watson irelandbranch@iat.org.ukLondon: Rebecca Towns londonbranch@iat.org.ukMidlands: Ian Fielding midlandsbranch@iat.org.ukNorth East England: Zoe Smith and John Bland northeastbranch@iat.org.ukNorth West: Nicky Windows cheshirebranch@iat.org.ukOxford: Adam Truby oxfordbranch@iat.org.ukSurrey, Hampshire & Sussex: Francesca Whitmore shsbranch@iat.org.ukWest Middlesex: Josefine Woodley westmiddxbranch@iat.org.ukWales & West: Rhys Perry waleswestbranch@iat.org.ukWest of Scotland: Joanne King westscotlandbranch@iat.org.ukIAT OFFICERS M AY BECONTACTED VIA:IAT Administrator:admin@iat.org.ukOR VIA THE IAT WEBSITE AT :www.iat.org.ukOR THE REGISTERED OFFICE:5 South Parade, Summertown,Oxford OX2 7JLAdvertisement Managers:PRC Associates LtdEmail: mail@prcassoc.co.ukAlthough every effort is made to ensure that no inaccurate or misleading data, opinion or statement appear in thejournal, the Institute of Animal Technology wish to expound that the data and opinions appearing in the articles,poster presentations and advertisements in ATW are the responsibility of the contributor and advertiser concerned.Accordingly the IAT, Editor and their agents, accept no liability whatsoever for the consequences of any suchinaccurate or misleading data, opinion, statement or advertisement being published. Furthermore the opinionsexpressed in the journal do not necessarily reflect those of the Editor or the Institute of Animal Technology.© 2021 Institute of Animal TechnologyAll rights reserved. No part of this publication may be reproduced without permission from the publisher.CPD Officer: Alan Palmer MIAT RAnTechRegistration and Accreditation Board:Glyn Fisher (Chair), John Gregor y,Cathy Godfrey, Kathy Ryder (Home Office),Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee:Alan Graham (Chair), Haley Daniels, Adele Kitching,Allan Thornhill, John WatersDiversity Officer:Haley Daniels MBA MSc MIAT RAnTech CIPDUK Biosciences ASG Representative/Home Office:Alan Palmer MIAT RAnTechviAugust20:Animal Technology and Welfare 12/8/20 07:54 Page viCPD Ofﬁcer: Alan Palmer MIAT RAnTechRegistration and Accreditation Board:Glyn Fisher (Chair), John Gregory,Cathy Godfrey, Kathy Ryder (Home Ofﬁce),Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee:Alan Graham (Chair), Haley Daniels, Adele Kitching,Allan Thornhill, John WatersDiversity Ofﬁcer:Haley Daniels MBA MSc MIAT RAnTech CIPDUK Biosciences ASG Representative/Home Ofﬁce:Alan Palmer MIAT RAnTechIndex to AdvertisersAAALAC ..........................................................83AS-ET .............................................................83Avid plc ............................................................2Datesand Ltd .................................................IFC Institute of Animal Technology .........34,42-43, OBC IPS Product Supplies Ltd ................................IBCLBS Serving Biotechnology Ltd ...........................5Somni Scientiﬁc ...............................................8Tecniplast UK Ltd ............................................10

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6Animal Technology and Welfare August 2020August 2020 Animal Technology and WelfareEditorialJas BarleyChair of the Editorial BoardLooking back over issues of the Journal through its various identities, one thing is appar ent and that is the contribution thatoverseas authors have made to the content. Topics have varied from dealing with exotic species, lack of sophisticated equipment,different attitudes to everyday problems, staff training and education and disease outbreaks. However, the resolute that has beenconstant throughout, despite the differences across the world, is the love and concern for the animals being cared for.Many include interesting photographs but I unfortunately am unable to use them as the quality of images is so poor whenrepr oduced, to the extent in some cases, they become worthless.Obviously, things have changed over seven decades and the technology described in contributions from overseas is less differentfrom what we use in the UK. This issue welcomes contributions from Australia, the Czech Republic and Iran as well, of coursefrom the UK. Since ATW became an Open Access publication and is being published electronically, it is enjoying a wider audienceand is attracting mor e contributions than usual. Not all are relevant to our profession, but knowledge is transferable so whatseems ‘off beat’ today may become useful in the future. However, as Editor I will always strive to maintain the quality of ourpublications and the usefulness to our readers.In this issue we include the RSPCA 2019 Rodent and Rabbit Welfare group meeting report. The 26th meeting that the RSPCA haveorganised focussed on ‘sentience, positive welfare and psychological well being’. The repor t contains contributions from 11presenters as well as notes on the interactive discussion session on sentience that closed the meeting.A paper from Iran, a first as far as I can see for the Journal, on reducing the negative effects of methionine on bone parametersin broilers’ embryos may seem of little relevance but it offers a better understanding of how methionine affects bone structurewhich is important to most species. Similarly, Feline Assisted Therapy as described by the team at the University of Life SciencesPrague does not appear to fall into the r ealms of Animal Technology but it gives us a better understanding of how animals can havea positive effect on some people, which in the current situation may be of significant benefit to a wider population. Our final paperfrom the team at Western Sydney University, details the care of the Children’ Python and two species of Bearded Dragons. Notperhaps the run of the mill laboratory animals but just as important to many Animal Technologists globally as mice and rats. If youkeep reptiles at home or know of someone who is contemplating one as a pet these papers make useful reference documents. Wealso offer two papers from previous issues of the Journal which were very different in appearance and content than today’s Journalof Animal Technology and Welfare and not only because of the change of title. Issues were printed in black and white and in the veryearly days were produced by hand. The paper from France on Physical Hazards in the laboratory animal house will bring back manymemories for some of the older technicians, myself included, but not necessarily good ones. The use of ether as an anaestheticwhich I know is still used in some countries where resources are limited, for human surgery, presented a very real danger to bothanimals and staff. Disease in laboratory animal units was often a recurring problem, bacterial infections such as Pseudomonas asdescribed in the reprint of the article were still presenting Animal Technologists with problems as late as the end of the 1980s. Whenimporting animals and tissues from overseas it is important to realise that they may be carrying disease not seen in the UK forseveral decades. In recent times, Ectromelia was introduced into a unit in the USA via antibodies produced overseas. Precautionsmust be taken until such time as you are sure that the animals and tissues are clear of any underlying infections.We are also able to offer in this issue an interesting Tech-2-Tech article by Seonagh Henderson of the University of Glasgow, ona novel technique of cage cleaning which hasa positive effect on the welfare of laborator y rats. Finally, we included several postersprepared for AST2020 but sadly at the moment remain unpresented.Thanks again to all of our authors, past and pr esent, both internationally and here in the UK. There would not have been 70 yearsof the Journal without you. Here is to the next seven decades and beyond.THE INSTITUTE OF ANIMAL TECHNOLOGYETHICAL STATEMENT“In the conduct of their Professional duties, Animal Technologists have a moral and legalobligation, at all times, to promote and safeguard the welfare of animals in their care,recognising that good laboratory animal welfare is an essential component of goodlaboratory animal technology and science.The Institute recognises and supports the application of the principles of the 3Rs(Replacement, Reduction, Refinement) in all areas of animal research.”ixAugust20:Animal Technology and Welfare 12/8/20 07:54 Page ixAnimal Technology and Welfare April 2021Welcome to the ﬁrst issue of 2021 and the 71st year of the formation of the ﬁrst professional association of what we know today as Animal Technologists. As always, the Institute of Animal Technology (IAT) is quick to respond to a changing world and following the difﬁcult year the world has experienced due to the advent of COVID-19 and the consequential disruption to what we consider normal life. Just as it has adapted to changes in requirements of our industry, the IAT has innovated, introducing virtual meetings of Council so that the ‘business’ of the IAT can continue, allowing Council to protect members’ interests, in addition online interviews for Registered Animal Technician (RAnTech) status are amongst many other adaptations. By the time you read this issue of Animal Technology and Welfare (ATW) we will have held our ﬁrst virtual Congress which gave access to many more delegates including those outside the British Isles who in the past may not have been able to attend a physical Congress. Over the course of this year’s volume of ATW I hope to bring you papers from Congress along with workshop reports and the usual Congress posters. The change of ATW to an electronic Journal has attracted more contributions from the wider scientiﬁc community. Some of these have dealt with issues that are outside our regular sphere of interest and in these instances the authors, following peer review, have been directed to other publications. Other submissions deal with topics that we would not recognise as obvious Animal Technology perhaps because of the unusual species involved or because the cover problems that do affect us. However, they do deal with animal welfare and/or technology and offer an insight into what working with animals may involve in the wider world. Consequently, a new special interest section will be introduced in this volume, the papers and articles included are certainly interesting and hopefully help those readers who are not working with the standard laboratory species or facing problems that may be unique to their situation. Formal papers will be peer reviewed as usual but I hope that we will also be able to include informal material such as Tech-2-Tech and opinion articles about areas of interest that many of us will be less familiar with. Posters are always welcome but if they have been exhibited elsewhere please ensure you send details of the meeting organisers so that checks can be made regarding copyright and that appropriate acknowledgement is made to the original meeting. Included in this issue I am delighted to publish a guest editorial from Vicky Robinson, chief executive of the National Centre for the Replacement, Reduction and Reﬁnement for Animals in Research (NC3Rs). In her editorial Vicky acknowledges the role of Animal Technologists especially during the COVID-19 crisis and the impact it has made on them. Vicky also mentions the award of Outstanding Technician of the Year at the Times Higher Education Awards recepient John Waters for his work on reﬁning the way mice are handled to reduce the stress they experience. John’s paper was reprinted in the December 2020 (v19.3) issue of Animal Technology and Welfare (ATW) and I would recommend that if you are familiar with John’s work, that you read it as soon as possible. Several years of back issues of ATW are available on the Journals website www.atwjournal,com. Handling of mice also features in a paper from the team at the Royal Veterinary College (RVC) discussing how hand sanitiser on gloves may be affecting mouse behaviour. This is obviously important given the increased used of hand disinfectants during the COVID-19 pandemic especially as handling mice is such a fundamental part of Animal Technology. The impact of COVID-19 is also considered in the paper from Angela and Jordi Lopez-Tremoleda but this time it is the emotional consequences on Animal Technologists and what we can do to support Animal Technologists that are dealt with. EditorialJas BarleyChair of the Editorial Board

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9August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareIt really goes without saying what a challenging year, personally and professionally, 2020 was for so many people as a result of the COVID-19 pandemic. When I look back though, one thing that stood out was the essential role of Animal Technologists up and down the country at a time of crisis. Difﬁ cult decisions had to be made last March (and then again for subsequent lockdowns) about the continuation of in vivo studies and maintenance of breeding colonies. Many Animal Technicians continued to go to work to make sure that the animals in their care remained a priority, at a time when many of us had the safety of being able to work from home. Technicians were required to cull large numbers of animals because of concerns about contingencies in the case of a lack of staff to provide care. A love of animals is why most technicians enter the profession and I know from talking to many over the years how traumatic this part of their role can be and speciﬁ cally the emotional distress that the cull as a result of the pandemic caused. There are lots of resources provided by the IAT and other organisations to help manage and address ‘compassion fatigue’ and I would strongly urge you to use them. But 2020 was not all bad and there was a real boost for the technician community when in November John Waters a senior Animal Technologist and Named Animal Care and Welfare Ofﬁ cer (NACWO) at the University of Liverpool was recognised as the Outstanding Technician of the Year at the Times Higher Education Awards. The competition was tough with a shortlist of stellar candidates from across the higher education sector. This is the ﬁ rst time an Animal Technologist has received this prestigious award and while it is an amazing achievement for John, importantly I think it is also public recognition for the vital work that Animal Technologists do to champion animal welfare and support high quality science. John’s work will be familiar to most – the demonstration with Professor Jane Hurst, that picking mice up by the tail causes aversion and anxiety and that there are better, more reﬁ ned approaches such as using a tunnel or cupped hands that beneﬁ t the mice as well as minimising the data variability that can compromise studies. I do not think I am overstating when I say that this is the biggest improvement in laboratory animal welfare in a decade. I wonder what other welfare innovations will come from the technician community in the next decade.John has taken a very active role in supporting his fellow technicians nationally and internationally to introduce the reﬁ ned handling methods. I have been hugely impressed with how many technicians have embraced this simple reﬁ nement but also disappointed that there has been pushback from some who do not want to change practice or do not believe the substantial evidence base that supports this welfare improvement. Neither objection seems reasonable to me, not just thinking about this speciﬁ c example but what it says more broadly about the role of technicians. The proﬁ le of technicians is now much more visible than perhaps it was 10 or 15 years ago as John’s award illustrates. And with this comes additional responsibility to champion animal welfare and embrace the latest opportunities, recognising that knowledge of what animals experience and what matters to them is increasing rapidly and it needs to be translated into practice. The role of technicians has evolved signiﬁ cantly, and it is important that this continues and that the sector utilises the considerable talent of the technician community. Day-to-day care of the animals is the priority but there are also other ways in which technicians can support best practice in animal research. Many animal experiments are poorly designed and I am keen to see technicians get involved in helping researchers “blind” their experiments so that the ﬁ ndings are robust and animals are not “wasted” in studies that do not provide meaningful data. The NC3Rs has been running training for technicians on this and we are planning to do more so make sure you sign up to the Tech3Rsnewsletter to ﬁ nd out more. I have always felt proud to support the technician community and one of the features we have included in Tech3Rs is a focus on technician champions who have made a real difference to the animals in their care. If you are working on a reﬁ nement approach you would like us to feature, please get in touch – I would love to hear from you! Stay safe and thank you for all that you are doing. Vicky Robinson, Chief Executive, NC3Rs Guest editorialVicky RobinsonNational Centre for the Replacement, Reﬁ nement and Reduction of Animals in ResearchApril 2021 Animal Technology and WelfareWASTE ANAESTHETICGAS EXPOSURE? Get Active.SOMNI EPS-3Active Induction ChamberActive Uni-Flow NoseconeSOMNI PROVIDES UNPARALLELED CUSTOMER SERVICE, CLINICAL AND TECHNICAL SUPPORT.

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11August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAbstract Alcohols are commonly used in laboratory animal facilities to disinfect hands, equipment and laboratory environments. The effect on mice is unknown, so we observed male and female C57BL/6J and BALB/c mice during and after handling with nitrile gloves that were either sprayed with 70% alcohol sanitiser (~67% ethanol, ~3% methanol, and 30% water), or not sprayed. We hypothesised that, if mice perceived this hand sanitiser as aversive, its application to gloves before handling would increase behavioural indicators of fear or defence; it could also affect social interactions and grooming. Handling mice with sanitised gloves increased wall rearing, self-grooming, allogrooming, snifﬁng of cagemates and eating/drinking in one or both strains of mice. In males, it also reduced initial home-cage aggression, replaced by grooming but it is unclear whether aggression was truly decreased or simply delayed. There were no statistically signiﬁcant effects of treatment on avoidance behaviours shown in a hand interaction test. Defensive burying occurred with both sanitised and control gloves during the ﬁrst-hand interaction test and signiﬁcantly declined over the 4-week study, suggesting a novelty effect. Findings indicate that handling mice with alcohol-based hand sanitiser affects mouse behaviour, including social interactions, although replication is required because we could not blind the observer to the treatment. Further research is required to assess the long-term effects of using alcohol-based hand-sanitiser and alternative disinfectants when handling laboratory mice in order to make recommendations for reﬁnement. Keywords: animal behaviour; animal welfare; disinfectant; handling; hygiene; miceIntroductionTo ensure that mice are free from undesirable or pathogenic microorganisms, laboratory animal units put in place strict biosecurity practices for example Shek et al (2015).1 These procedures vary between facilities depending on the level of microbiological exclusion but they generally involve keeping mice in micro-isolation cages, controlling animal imports and routinely monitoring the health status of the colony. Additionally, personal protective equipment is worn, and consumables, equipment and surfaces are decontaminated.2,3 Although these practices are important to safeguard the health of laboratory animals, their impact on animal behaviour and welfare is rarely investigated. Additionally, recent reports suggest that keeping laboratory animals in extremely hygienic facilities impairs their immunological response and compromises the reproducibility and translation of the results to humans.4,5 Garner et al (2017)6 outline the importance for experimental designs to take into account animal biology, husbandry, and welfare (representing three of six key considerations) if research is to produce valid and reproducible results.In a survey of 51 UK mouse facilities, 22-30% of respondents reported using ‘alcohol’ to disinfect a variety of items such as work surfaces, anaesthetic equipment, behavioural apparatus and surgical equipment, and 12% speciﬁcally reported using it as a hand sanitiser.3 In that survey, a subset of respondents suggested that alcohol-based disinfectants caused skin problems (4/9 respondents) respiratory problems (1/9) and behaviour changes (1/9) in mice, with 1/9 suggesting there were no adverse effects. This demonstrates that some mouse facility staff are concerned by the use of Handling mice using gloves sprayed with alcohol-based hand sanitiser: acute effects on mouse behaviourNOELIA LOPEZ-SALESANSKY,1 DOMINIC J WELLS,1 NATALIE CHANCELLOR,2 LUCY WHITFIELD,1 and CHARLOTTE C BURN21 Royal Veterinary College, Royal College Street, London NW1 0TU UK 2 Royal Veterinary College, Hawkshead Lane, North Mymms, Hertfordshire AL9 7TA UKCorrespondence: cburn@rvc.ac.uk April 2021 Animal Technology and WelfareISSUE2 - VISION+ - 210X297.indd 1 05/11/2018 18:14:29find out more on www.tecniplastuk.com Or call us on 0345 050 4556

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12Animal Technology and Welfare August 2020alcohols as sanitisers. Here, we focus on the acute behavioural effects of handling mice using alcohol-based hand sanitiser which usually contains one or more types of alcohol: ethanol, methanol, isopropanol, and/or n-propanol. In the authors’ experience of working in multiple animal facilities in England, it is common practice to use 70% ethanol hand sanitiser immediately before handling rodents. Providing alcohol-based sanitisers has been recommended for use when handling laboratory animals to help prevent infections, even when using gloves because gloves are permeable and can otherwise easily become contaminated.7 LeMoine and colleagues (2015)8 found that 48% of non-sterilised standard nitrile or latex gloves (the two most common glove types in UK laboratory animal facilities)3 tested positive for microbial growth after donning; however, a 30 seconds soak with 70% isopropyl alcohol reduced microbial contamination to 25%. Keen et al (2010)9 also found that using 70% isopropyl alcohol was signiﬁcantly more effective at preventing microbial contamination of gloves during mouse laparotomy than not using it. However to our knowledge, no research has been carried out to assess the impact of this practice on the animals themselves. In humans, frequent use of alcohol-based hand sanitisers by health professionals can cause transient, low, but detectable concentrations of ethanol in the breath, due to inhalation of the vapour.10,11 Such use does not seem to increase alcohol concentrations in the blood,12 nor does it appear to detrimentally affect the skin.11 Overall, the internal concentrations observed via inhalation and dermal routes are well within the range of those occurring with ingestion of non-alcoholic foods, such as fruit juices or ‘alcohol-free’ beers and are well within safe limits.13 However oral ingestion of larger doses of alcohol-based sanitiser can cause intoxication (‘drunkenness’), alcohol poisoning, coma or even death.14-16 It is difﬁcult to extrapolate these ﬁndings to mice, especially because of potential species differences in pharmacology, and in body size; mice are orders of magnitude smaller than an adult human relative to the volume of sanitiser likely to be applied to a human hand. In rodents, effects of alcohol have mainly been investigated in the context of modelling alcoholism. Most experiments have thus investigated the effects of oral ingestion of ethanol but when rodents are unwilling to ingest it, forced inhalation of ethanol vapour intermittently over a 2 week period can increase voluntary consumption of ethanol by rats 2-8h following withdrawal of the vapour.17 Oral ingestion of large enough doses of ethanol by mice can cause ataxia, aggression, cognitive deﬁcits and other effects consistent with intoxication and alcoholism in humans for example.18This alcoholism-related research has limited relevance with respect to the effects that alcohol-based sanitiser could have on mice in applied contexts, for a number of reasons. Firstly, the quantities that mice are likely to inhale or ingest after handling by sanitised gloves are likely to be much lower than those administered to animals modelling alcoholism. However hand sanitisers contain much higher concentrations of alcohol (usually 70%) compared with those used in alcoholism research (10-20%), so it is unclear how the exposure would compare. Secondly, whilst alcohol-based sanitisers in the laboratory commonly contain ethanol, this may be mixed with other alcohols unsuitable for consumption such as methanol or propanol, or other substances entirely. Mice19 and rats20 are less sensitive to the toxic effects of methanol than are humans.20 Nevertheless relatively high doses of methanol can cause skeletal and neural defects in rodents exposed as embryos or juveniles, and chronic methanol ingestion and – to a lesser extent – inhalation can cause pathology of the liver, pancreas and possibly other organs (reviewed in21). Finally, the route of exposure to hand sanitiser is complex with inhalation and oral consumption inﬂuenced by factors such as: the amount of product used; the delay between dispensing the product onto the hand and handling the mouse; and the animals’ behaviour following contact (e.g. whether or not they sniff or lick the product from the hand or their fur). We thus aimed to investigate whether use of an alcohol- based hand spray when handling mice has any acute behavioural effects on the animals in an applied context with a view to reﬁning husbandry protocols. We hypothesised that mice handled using gloves sprayed with 70% alcohol could show differences in behaviour compared to handling without the sanitiser, indicating effects on mouse welfare. We used both sexes, and two strains, of mice to increase external validity of any ﬁndings;22 C57BL/6 mice will drink ethanol relatively willingly, whereas BALB/c mice have high avoidance of it.18,23,24 We used the ‘cupping’ method of handling, to follow the example of reﬁned practice and to represent a handling method that would involve contact between the hand and the mouse’s body, with relevance to activities such as health checking, cage-cleaning, or manual restraint.25MethodsAnimalsWe used off-study stock or breeding mice that were lent by colleagues at the Royal Veterinary College. The resulting sample comprised 22 cages containing: – Adult C57BL/6J mice aged 20-24 weeks (n = 13 cages: 16 males, two cages holding two mice each and two cages holding three mice each; 14 females, four cages holding two mice each and two cages holding three mice each), and – Adult BALB/c mice aged 9-28 weeks (n = 9 cages: ten males, ﬁve cages holding two mice each and two cages holding three mice each; 12 females, three Handling mice using gloves sprayed with alcohol-based sanitiser

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13August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfarecages holding two mice each and two cages holding three mice each). Mice were kept in standard open-top cages (Tecniplast, 32cm x 16cm x 14cm). Cages contained bedding (Litaspen premium, Datesand Ltd), a cardboard tube and additional nesting material (Sizzlenest, Datesand Ltd). Animals had ad libitum access to water and food (Rat & Mouse # 1 Diet, Special Diet Services) and were maintained at constant room temperature (~21°C) and humidity (~45%) and were kept under a regular light/dark schedule with lights on from 08:00 to 20:00 h (light = 270 lux). Cage cleaning was carried out by a member of the animal unit staff once a week on a different day than behavioural observations were recorded. Housing and care were in compliance with the Code of Practice for housing and care of laboratory animals used in scientiﬁc procedures and the experiment was approved by the Royal Veterinary College Ethical Committee (URN 2014 1261). Experimental proceduresTwo experimenters (both female) carried out all the observations in this experiment between 9:00 and 12:00h. Only one of the experimenters (NLS) handled the animals. We used two treatments representing realistic husbandry alternatives: handling using nitrile gloves that were either left unsprayed (Control), or ﬁrst sprayed with 70% alcohol (Sanitised). The 70% alcohol (methylated spirits, Fisher Scientiﬁc Ltd26) comprised 66-68% ethanol, 2-4% methanol, and 30% distilled water. A new pair of gloves was used for each cage to prevent contamination.The mice in our sample were unlikely to have previously experienced glove sanitation with alcohol, so 1 week before the experiment, all mice were gently handled in cupped hands using alcohol-sanitised gloves, to help reduce novelty-induced behaviours the following week. Mice within each cage were then exposed to one of the treatments per week, alternating them each week for a period of 4 weeks; mice therefore experienced both treatments twice during the experimental period. Testing order was randomly allocated to cages balancing across strains and sexes (using an online random number generator), such that half the cages experienced the sequence: Week 1 = Control, Week 2 = Sanitised, Week 3 = Control, Week 4 = Sanitised; the other half received the opposite order, starting with Sanitised in Week 1 and alternating thereafter. Handling effects were observed in four stages: (1) Voluntary interaction with the sanitised or control gloved-hand, similar to the hand interaction test used by Hurst and West;25 (2) Handleability during handling; (3) Voluntary interaction with the hand after handling; and (4) Home-cagebehaviour both immediately after returning to the rack and 20 minutes later.Figure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensive burying of the hand.Voluntary interaction with the hand before handlingEach cage was placed on a bench and aligned such that an adhesive tape on the bench delineated the cross-sectional midline of the cage. The cage was opened on the bench by the non-handling observer, who removed housing and all nesting material, allowing mice to habituate for 1 minute before behavioural observations. After donning a fresh pair of nitrile gloves, the handling experimenter either sprayed the gloves with 70% alcohol or left them unsprayed. The experimenter then placed one hand inside the near corner of the cage and kept it motionless to allow mice to voluntarily interact (Figure 1; the other hand was used to write down behavioural observations). Observations were recorded by both experimenters for 3 minutes to measure any difference in approach/avoidance or defensive behaviour towards the hand.HandleabilityAfter recording hand interaction, for the sanitised treatment, the handling experimenter re-sprayed the glove with the alcohol because many volatiles would have dissipated during the previous 3 minutes observation. To mimic normal handling in applied situations, when there can be little delay between sanitation and handling, the mouse was then almost immediately lifted using its home-cage tunnel following re-spraying and loosely cupped in the hands for 20-30 seconds to measure handleability. The respraying and handling were repeated for each mouse in the cage. Cage was the experimental unit, so all mice within a cage experienced the same treatment on a given day.Figure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveburying of the hand.Figure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveburying of the hand.Figure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveburying of the hand.Figure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveFigure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveFigure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveFigure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveFigure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveFigure 1. The hand interaction test. This screenshot shows a C57BL/6J mouse displaying defensiveHandling mice using gloves sprayed with alcohol-based sanitiser

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14Animal Technology and Welfare August 2020Voluntary interaction with the hand after handlingOnce all mice in the cage had been handled, the observer returned their hand to the corner of the cage and kept it motionless, whilst the behaviour of the mice was observed for a further 2 minutes. Home-cage behaviour after handlingFinally, the cage was relocated to the rack and home-cage behaviour was observed for an initial 2 minute period. Approximately 20 minutes later, home-cage behaviour was again observed for a ﬁnal 2 minutes. Behavioural observationsBehaviours were deﬁned according to an Ethogram (Table 1) and recorded in real time by the two observers. It was not possible for us to blind the observers to the treatment; consideration was given to using water spray as a control but this would have been an unrealistic treatment within this applied study and the volatile odour of the alcohol would have continued to make the treatment obvious. (Blinding would still have been possible, via video recordings analysed by an additional treatment-blind observer but these options were unavailable to us during this study, which had limited coverage of research costs.)Voluntary interaction with the hand before and after handlingThe number of mice were located in the half of the cage nearest to, and furthest from, the hand according to the midline of the cage was noted simultaneously by both observers at 15 defensive instances. All other behaviours were allocated to either one of the observers (because there were too many behaviours for a single observer to record); these were recorded on a one-zero schedule, i.e. whether or not they occurred during each 15 seconds interval. Additionally, when mice showed defensive burying, the height of the resulting sawdust ‘wall’ was measured with a ruler by the non-handling observer after the observation was complete.HandleabilityThe non-handling observer recorded handleability on a subjective scale (no struggle; minor struggle; vigorous struggle or escape), vocalisation or biting, and whether urination or defecation occurred. Home-cage behaviour after handlingEach observer was allocated a time point (NLS: immediately after return to the rack; NC: 20 minutes later). All behaviours (Table 1) were recorded on a one-zero schedule every 15 seconds for a period of 2 minutes.Table 1. Ethogram of the behaviour recorded.Behaviour Deﬁnition Recording stage(s)AggressionBiting (using the teeth to pierce the skin), pinning (grabbing recipient mouse’s ﬂank and holding down), boxing (movements of the body towards the opponent combined with alternated kicking of the forepaws), tail rattling (fast waving movements of the tail).HomecageAllogroomingLicking the fur of another mouse or using the forepaws to smooth it.Voluntary interaction with hand and HomecageBar-biting* Chewing the cage grid. HomecageChasingRapidly following a ﬂeeing mouse.HomecageChewing gloveUsing the teeth as if to pierce the glove material.Voluntary interaction with handClimbing barsHanging from the cage grid, without chewing the bars.HomecageDefensive buryingDisplacing bedding material towards the gloved hand with alternating forward pushing movements of their forepaws and shovelling movements of their heads. Voluntary interaction with handEating or drinkingConsuming food or water. The animal rears up and licks the nozzle of the drinker or gnaws at food pellets through the bars of the food hopper. HomecageGrooming (caudal)Self-cleaning of the body, legs and tail/genitals. Voluntary interaction with hand and HomecageGrooming (rostral)Self-cleaning of the paws, snout and head. Voluntary interaction with hand and HomecageKick diggingDisplacing bedding material with fore paw movements alternated by backwards kicking of both hind legs simultaneously.Voluntary interaction with hand and HomecageLocation: close to hand Mouse is in the half of the cage closer to the hand, relative to the midline of the cage.Voluntary interaction with handLocation: far from handMouse is in the half of the cage further to the hand, relative to the midline of the cage.Voluntary interaction with handNesting Manipulating nesting material HomecagePaws on handMouse places one or both paws on the gloved hand. Voluntary interaction with handRearingStanding upright on hind legs, without the two front paws touching any surfaces.Voluntary interaction with hand and HomecageSleeping or restingLying immobile for at least 5 seconds. HomecageSnifﬁng cagemate (anogenital)Rapid twitching movements of the nose towards the anogenital area of another mouse.Voluntary interaction with hand and HomecageSnifﬁng cagemate (body)Rapid twitching movements of the nose towards the head or body of another mouse.Voluntary interaction with hand and HomecageSnifﬁng handRapid twitching movements of the nose towards the gloved hand, with the nose at < 1 body length from the gloved hand.Voluntary interaction with handVocalizingEmitting vocal sounds audible to humans.Voluntary interaction with hand and HomecageWall rearingStanding upright on the hind legs and resting one or both front paws on a cage wall.Voluntary interaction with hand and HomecageHandling mice using gloves sprayed with alcohol-based sanitiserThe behaviours are adapted from an existing mouse ethogram.27 The behaviour categories are arranged in alphabetical order and the stage at which they were recorded is given. *Other escape or stereotypic behaviours (circling, jumping, barbering and somersaulting) were included but are not shown here because they were never observed.

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15August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareStatistical analysis IBM SPSS Statistics 22 was used to perform the statistical analyses. Cage was the experimental unit. Behavioural data across the 15 second time intervals were summed for each cage at the relevant time points. A mean handleability score was calculated per cage.Normally distributed data were analysed using a Linear Mixed Effects Model with Treatment, Strain, Sex and Week set as ﬁxed factors and Cage as a random factor. The two-way interactions between Treatment and Strain, Treatment and Sex and Treatment and Week were initially included but removed if not statistically signiﬁcant. If interactions were statistically signiﬁcant, post-hoc pairwise comparisons were conducted to discover which categories the signiﬁcant differences lay between. Normality was checked by visual inspection of the histograms of the residuals of the models and by carrying out Kolmorov-Smirnov and Shapiro-Wilk tests of normality. When normality assumptions were not met, the original outcomes were transformed and assumptions checked again. If the residuals did not ﬁt a normal distribution after transformation, the relevant behaviour was converted to a binary variable (presence/absence of the behaviour). Binary responses were analysed using a Generalised Linear Effect Mixed Model (Binary Logistic Regression) with the same random and ﬁxed factors as described above. The standard errors of the resulting coefﬁcients were checked for inﬂation that could indicate multicollinearity.As both observers recorded the location of mice during voluntary interaction, an interrater reliability analysis using the Kappa statistic was performed to determine consistency among raters. The interrater reliability was ‘substantial’ (Kappa = 0.697; P <0.001)28, allowing only one set of observations (the one with fewer missing values) to be analysed for this variable. ResultsStatistically signiﬁcant effects are shown in Table 2. In the hand interaction test, the sanitiser signiﬁcantly increased wall-rearing (during Week 1), self-grooming and – in BALB/c mice – snifﬁng and grooming of the cagemate (Figure 2). There were no signiﬁcant effects observed during handling itself and urination/defecation during handling was recorded three times only, in BALB/c mice (twice with sanitiser and once with the control). Only two mice squeaked or bit with sanitiser versus three with control gloves. Six mice struggled vigorously with sanitiser, versus three with the control.Immediately upon return to the home-cage, aggression frequency showed a statistically signiﬁcant reduction after handling with sanitised gloves compared with control gloves, being observed at least once in four cages immediately after handling with sanitiser, versus seven Figure 2. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser (blue bars) when handling mice increased (a) snifﬁng the cagemate and (b) grooming the cagemate compared with controls (orange bars). This reached statistical signiﬁcance in BALB/c mice: *P<0.05; ***P<0.001. X indicates mean values, the boxes indicate the inter-quartile range, and the whiskers indicate the 95% range. (a)Figure 2. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (awith controls (orange bars). This reachedindicates mean values, the boxes indicate the inter. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (awith controls (orange bars). This reachedindicates mean values, the boxes indicate the inter. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (awith controls (orange bars). This reachedindicates mean values, the boxes indicate the inter*. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (a) sniffing the cagemate and (b) grooming the cagematewith controls (orange bars). This reachedstatistical significance in BALB/c miceindicates mean values, the boxes indicate the inter(b). Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematestatistical significance in BALB/c miceindicates mean values, the boxes indicate the inter-quartile range, and the whiskers indicate the 95% range.. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematestatistical significance in BALB/c micequartile range, and the whiskers indicate the 95% range.***. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematestatistical significance in BALB/c mice: *P<0.05; ***P<0.001.quartile range, and the whiskers indicate the 95% range.. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematecompared: *P<0.05; ***P<0.001.quartile range, and the whiskers indicate the 95% range.. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitisercompared: *P<0.05; ***P<0.001.Xquartile range, and the whiskers indicate the 95% range.(a)(a)Figure 2. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (awith controls (orange bars). This reachedindicates mean values, the boxes indicate the inter. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (awith controls (orange bars). This reachedindicates mean values, the boxes indicate the inter. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (awith controls (orange bars). This reachedindicates mean values, the boxes indicate the inter*. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser(blue bars) when handling mice increased (a) sniffing the cagemate and (b) grooming the cagematewith controls (orange bars). This reachedstatistical significance in BALB/c miceindicates mean values, the boxes indicate the inter(b). Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematestatistical significance in BALB/c miceindicates mean values, the boxes indicate the inter-quartile range, and the whiskers indicate the 95% range.. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematestatistical significance in BALB/c micequartile range, and the whiskers indicate the 95% range.***. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematestatistical significance in BALB/c mice: *P<0.05; ***P<0.001.quartile range, and the whiskers indicate the 95% range.. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitiser) sniffing the cagemate and (b) grooming the cagematecompared: *P<0.05; ***P<0.001.quartile range, and the whiskers indicate the 95% range.. Treatment and strain effects on social behaviour in the hand interaction test. Use of hand sanitisercompared: *P<0.05; ***P<0.001.Xquartile range, and the whiskers indicate the 95% range.(b)cages in the control treatment. This difference disappeared 20 minutes later. Handling with sanitiser increased food/water consumption in C57BL/6J mice at the 20 minute time point. Bar-biting was seen at least once in ﬁve of the cages immediately after handling with sanitiser, compared with just one cage after control handling but this difference was not statistically signiﬁcant (P = 0.343). Some of the effects were interactive, reaching statistical signiﬁcance in only one of the mouse strains or sexes. Alcohol spray signiﬁcantly increased wall rearing and home-cage eating/drinking compared with the control in C57BL/6 mice only. In the case of caudal grooming 20 minutes after being returned to the home-cage, the sanitiser seemingly masked a sex difference whereby males groomed more than females only in control conditions.Handling mice using gloves sprayed with alcohol-based sanitiser

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16Animal Technology and Welfare August 2020Table 2. Statistically signiﬁcant differences in behaviour between mice when handled using gloves sanitised with 70% alcohol versus unsprayed gloves.Handling stage Behaviour affectedEffect direction Odds ratio/ ✝Coefﬁcient (95% CI)Statistic P-ValueHand interaction before handlingWall rearing Sanitised > Control only during the ﬁrst week of the study (statistical interaction)✝5.842 (1.432-10.252)F3, 76 = 5.281 0.002Hand interaction after handlingSnifﬁng of cage-mateSanitised > Control in BALB/c only (statistical interaction)✝0.645 (0.061-1.228)F1, 80 = 4.836 0.031Grooming of cage-mateSanitised > Control in BALB/c only (statistical interaction)✝2.903 (1.453-4.354)F1, 79 = 15.869 <0.001Self-grooming Sanitised > Control✝0.660 (0.370-0.949)F1, 81 = 20.557 <0.001Proportion of rostral versus total groomingSanitised > Control in C57BL/6J only (statistical interaction)✝0.142 (0.005- 0.278)F1, 80 = 4.276 0.042Wall rearing Sanitised > Control in the 1st and 3rd weeks only (statistical interaction)✝Week 1: 5.459 (1.770-9.149); Week 3: 4.904 (1.214 - 8.594)F1,78 = 3.594 0.017 (Week 1: Post-hoc P = 0.004; Week 3 Post-hoc P = 0.010)Home-cage immediately after returnAggression (seen in males only)Control > Sanitised0.251 (0.109-0.574)F1, 58 = 11.605 0.001Self-grooming (caudal)Sanitised > Control2.318 (1.440-3.732)F1, 58 = 9.141 0.004Home-cage 20 minutes after returnSelf-grooming (caudal)Males > Females only in Control conditions (statistical interaction)2.921 (1.366-6.245)F1, 58 = 4.737 0.034 (Post-hoc P < 0.001 in females; P = 0.014 in males)Consumption of food/waterSanitised > Control only in C57BL/6J (statistical interaction)3.142 (2.106-4.688)F1, 57 = 4.467 0.039 (Post-hoc P < 0.001)Handling mice using gloves sprayed with alcohol-based sanitiser†Indicates that, rather than an odds ratio, a coefﬁcient from a Linear Mixed Effects Model is provided, i.e. where the response was able to produce normally distributed residuals. The Control treatment was used as the reference category, so when a behaviour increased with the alcohol spray, the odds ratio is >1 and when a behaviour decreased with alcohol, the odds ratio is <1. The post-hoc p-value is given for statistically signiﬁcant pairwise comparisons when overall two-way interactions were statistically signiﬁcant.

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18Animal Technology and Welfare August 2020scent marking behaviour (e.g. urination frequency and patterning) associated with the establishment of hierarchies in group housed male mice.41,42Lastly, the increase in feeding/drinking observed in C57BL/6J after being handled with sanitised gloves could indicate that the mice settled to normal behaviour more quickly than when handled with control, unsprayed gloves. Alternatively, it could have been a displacement activity. For example, after acute restraint, rats increased their drinking behaviour in the ﬁrst 15 minutes, followed by increased feeding.43 Furthermore, if the alcohol had tasted bitter to the mice during grooming, eating/drinking could have served to rid the mice of the unpalatable taste, and the bitterness would have increased salivation, so it could have increased thirst.36 Again analysis of the microstructure of the oral behaviours could help elucidate whether the mice were gaping as they do with bitter substances.35,36Alternative hand sanitisersThe current study appears to be the ﬁrst investigation of the behavioural effects of hand sanitisers in an applied context on animals, so if users wish to avoid the behavioural effects of methanol-ethanol-based sanitisers, it is difﬁcult to suggest an alternative at present. Alcohol-based sanitisers that contain isopropanol and/or n-propanol instead of methanol are unlikely to be beneﬁcial because toxicity tests reveal that adverse effects occur at lower doses (reviewed in Patocka J and Kuca K. 2012).44Many alcohol-free hand sanitisers also exist. A UK survey revealed that at least seven different hand sanitisers were used for handling laboratory mice across 51 different facilities, with 46% of respondents reporting generically that they used ‘soap’, followed by 24% reporting that they used Hibiscrub™ and 12% reporting ‘alcohol’.3 Hibiscrub™ is thus the most widely reported single brand of sanitiser reported for mouse handling in the UK. Its active ingredient is chlorhexidine gluconate (4.0%) but it also contains a low concentration of n-propanol (4.0%) as a solvent. Whilst use of 80% ethanol as an antiseptic on the ear of an allergic dermatitis mouse model signiﬁcantly worsened inﬂammation of the skin, use of 0.5% chlorhexidine gluconate showed no signiﬁcant difference from the control which suggests that the latter may be less irritant.45 No compound will be entirely free of adverse effects, depending on factors such as dose, form and characteristics of the animals themselves. For example, chlorhexidine compounds can occasionally cause allergic reactions in humans and are irritants of the eye in humans and rabbits at least (reviewed in46) whilst another alcohol-free alternative, benzalkonium chloride, is an irritant of the eyes, skin and mucosa of many species (including some limited data on mice reviewed in47).Further research will be necessary to ascertain, under treatment-blind conditions, the hand sanitiser that causes least harm to animals whilst being effective, practical and safe for humans. In the meantime the current results lead us to make the following recommendations. Researchers and other staff working with animals should:– Consider whether hand sanitisers need to be used at all, given that some gloves are initially sterile when ﬁrst worn and/or, given that mouse immune systems are more normal with a full microbiome.4,5– If alcohol-based sanitisers are required, wait as long as feasible for alcohol to dry before handling mice. – When using a hand-sanitiser for rodent handling, monitor the effects on animals carefully, initially checking at the very least for avoidance, defensive behaviour, excessive grooming and effects on aggression. As several products have irritant properties, it will also be necessary to monitor skin condition and scratching behaviour, eye irritation such as excessive blinking and respiratory signs such as nose-rubbing and sneezing.ConclusionsThis experiment suggested changes in mouse behaviour resulting from sanitising gloves with 70% alcohol. Although the reduction in aggression when the cage was returned to the rack could be interpreted as a positive ﬁnding, it was temporary and is likely to be the result of the increase in grooming to remove the sanitiser. Finally, the increased snifﬁng and grooming following alcohol sanitisation implies that the mice both inhaled and ingested some of the methylated spirits. Rodents are less sensitive to methanol than humans are, so long-term effects may be negligible but are currently unknown. Further work should be carried out under treatment-blind conditions to investigate the suitability of alternative glove sanitisers with different mouse strains using longer term animal health and welfare indicators. AcknowledgementsNoelia Lopez-Salesansky was supported by a Went Scholarship at the Royal Veterinary College (RVC). This experiment was part of a series supported by a Universities Federation for Animal Welfare (UFAW) Small Project Award. We would also like to thank Ruby Yu-Mei Chang for her statistical advice and support and Laura Smith and members of the RVC Biological Services Unit for discussions and information about alternative antiseptic hand sanitisers. We are grateful to colleagues for lending their mice to us for this study. The authors declare no competing interests. The RVC approved this manuscript for submission (PPS_02044).Handling mice using gloves sprayed with alcohol-based sanitiser

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19August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareReferences1 Shek WR, Smith AL and Pritchett-Corning KR. (2015) Chapter 11 - Microbiological Quality Control for Laboratory Rodents and Lagomorphs. In: Fox JG, Anderson LC, Otto GM, Pritchett-Corning KR and Whary MT, (eds.). Laboratory Animal Medicine (Third Edition). Boston: Academic Press, 2015, p. 463-510.2 López-Salesansky N, Mazlan NH, Whitﬁeld LE, Wells DJ and Burn CC.(2015) Olfaction variation in mouse husbandry and its implications for reﬁnement and standardization: UK survey of animal scents. Laboratory Animals. 2015; 50: 362-9.3 López-Salesansky N, Mazlan NH, Whitﬁeld LE, Wells DJ and Burn CC. (2015) Olfactory variation in mouse husbandry and its implications for reﬁnement and standardization: UK survey of non-animal scents. Laboratory Animals. 2015; 50: 286-95.4 Beura LK, Hamilton SE, Bi K, et al. (2017) Normalising the environment recapitulates adult human immune traits in laboratory mice. 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Handbook of clinical neurology. Elsevier, 2014, p. 71-86.25 Hurst JL and West RS.(2010) Taming anxiety in laboratory mice. Nature Methods. 2010; 7: 825-6.26 Fisher Chemical. (2016) Safety data sheet: Methylated spirit industrial. Loughborough,: Fisher Scientiﬁc UK, 2016, p. 2.27 Garner JP, Gaskill BN, Rodda C, et al.(2014) An ethogram for the laboratory mouse. Stanford: Stanford School of Medicine, 2014.28 Landis JR and Koch GG.(1977) The measurement of observer agreement for categorical data. Biometrics. 1977; 33: 159-74.29 Zalaquett C and Thiessen D. (2006) The effects of odors from stressed mice on conspeciﬁc behavior. Physiology and Behavior. 1991; 50: 221-7.Handling mice using gloves sprayed with alcohol-based sanitiser

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20Animal Technology and Welfare August 202030 Lever C, Burton S and O’Keefe J. (2006) Rearing on Hind Legs, Environmental Novelty, and the Hippocampal Formation. Reviews in the Neurosciences. 2006; 17: 111.31 Kemble ED, Garbe CM and Gordon C. (1995) Effects of novel odors on intermale attack behavior in mice. Aggressive Behavior. 1995; 21: 293-9.32 Kalueff AV and Tuohimaa P. (2004) Grooming analysis algorithm for neurobehavioural stress research. Brain Research Protocols. 2004; 13: 151-8.33 Kalueff AV, Aldridge JW, LaPorte JL, Murphy DL and Tuohimaa P. (2007) Analyzing grooming microstructure in neurobehavioral experiments. Nature Protocols. 2007; 2: 2538.34 Smolinsky AN, Bergner CL, LaPorte JL and Kalueff AV. (2009) Analysis of Grooming Behavior and Its Utility in Studying Animal Stress, Anxiety, and Depression. In: T. G, (ed.). Mood and Anxiety Related Phenotypes in Mice. Totowa: Humana Press, 2009, p. 21-36.35 Berridge KC. (2000) Measuring hedonic impact in animals and infants: microstructure of affective taste reactivity patterns. Neuroscience & Biobehavioral Reviews. 2000; 24: 173-98.36 Matsuo R, Yamamoto T, Ikehara A and Nakamura O. (1994) Effect of salivation on neural taste responses in freely moving rats: analyses of salivary secretion and taste responses of the chorda tympani nerve. Brain Research. 1994; 649: 136-46.37 Cooper RG. (2008) Renal Function in Male Sprague-Dawley Rats Concurrently Exposed to Long-Term Nicotine (3-{1-Methyl-2-Pyrrolidinyl}Pyridine) and Methylated Spirits (Methyl Alcohol). Renal Failure. 2008; 30: 107-14.38 Meyer RJ, Beard ME, Ardagh MW and Henderson S. (2000) Methanol poisoning. New Zealand medical journal. 2000; 113: 11-3.39 Van Loo PLP, Van Zutphen LFM and Baumans V. (2017) Male management: coping with aggression problems in male laboratory mice. Laboratory Animals. 2003; 37: 300-13.40 Weber EM, Dallaire JA, Gaskill BN, Pritchett-Corning KR and Garner JP.(2017) Aggression in group-housed laboratory mice: why can’t we solve the problem? Lab Animal. 2017; 46: 157.41 Arakawa H, Arakawa K, Blanchard DC and Blanchard RJ. (200) A new test paradigm for social recognition evidenced by urinary scent marking behavior in C57BL/6J mice. Behavioural Brain Research. 2008; 190: 97-104.42 Nevison CM, Barnard CJ, Beynon RJ and Hurst JL. (2000) The consequences of inbreeding for recognizing competitors. Proceedings of the Royal Society of London Series B. 2000; 267: 687-94.43 Badiani A, Jakob A, Rodaros D and Stewart J. (1996) Sensitization of stress-induced feeding in rats repeatedly exposed to brief restraint: the role of corticosterone. Brain Research. 1996; 710: 35-44.44 Patocka J and Kuca K. (2012) Toxic alcohols: Aliphatic saturated alcohols. Military Medical Science Letters. 2012; 81: 142-63.45 Sadakane K and Ichinose T. (2015) Effect of the hand antiseptic agents benzalkonium chloride, povidone-iodine, ethanol, and chlorhexidine gluconate on atopic dermatitis in NC/Nga mice. International journal of medical sciences. 2015; 12: 116-25.46 Hazardous Substances Databank. Chlorhexidine. Bethesda2015, p. https://toxnet.nlm.nih.gov/cgi-bin/sis/search/a?dbs+hsdb:@term+@DOCNO+7196.47 Hazardous Substances Databank. Benzalkonium Chloride Compounds. Bethesda2010, p. https://toxnet.nlm.nih.gov/cgi-bin/sis/search2/r?dbs+hsdb:@term+@DOCNO+234#permalink.Handling mice using gloves sprayed with alcohol-based sanitiser

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21August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2021 Animal Technology and WelfareIntroductionThe RSPCA/UFAW rodent and rabbit welfare group has held a one-day meeting every autumn for the last 27 years, so that its members can discuss current welfare research, exchange views on welfare issues and share experiences of the implementation of the 3Rs of replacement, reduction and reﬁnement with respect to rodent use. A key aim of the Group is to encourage people to think about the whole lifetime experience of laboratory rodents, ensuring that every potential negative impact on their wellbeing is reviewed and minimised.This year’s meeting was held online for the ﬁrst time and was attended by over 400 delegates from almost 40 countries. The theme was ‘cumulative experiences’ with sessions on ‘the science of cumulative severity’ and ‘practical reﬁnements to reduce severity and promote wellbeing’. The meeting opened with an introductory talk which explained why cumulative experiences are important and how both positive and negative experiences can accumulate over an animal’s lifetime to have long-term impacts on welfare. Further talks discussed different ways to recognise and assess cumulative severity, the cumulative impacts of small reﬁnements and the concept of a ‘good life’ and what this means for laboratory rodents. This was followed by an update from the Home Ofﬁce Animals in Science Regulation Unit (ASRU), which focussed on how cumulative experiences inﬂuence the severity experienced by animals in science. The day closed with an interactive discussion session on ways to identify cumulative suffering in rodents cage side. This report summarises the meeting and ends with a list of action points for readers to consider raising at their own establishments.Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meetingCHLOE STEVENS,1 PENNY HAWKINS,1 TOM V SMULDERS,2 AILEEN MACLELAN,3 LARS LEWEJOHANN,4,5 PAULIN JIRKOF,6 JACKIE BOXALL,7 HELEN MURPHY,7 CARLEY M MOODY,8 PATRICIA V TURNER,8,9 I J MAKOWSKA,10 and CHARLOTTE INMAN11 1 Animals in Science Department, Science Group, RSPCA, Wilberforce Way, Southwater, West Sussex RH13 9RS UK2 Newcastle University, Newcastle upon Tyne NE1 7RU UK3 Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, Ontario, Canada4 Freie Universität Berlin, Kaiserswerther Str. 16-18, 14195 Berlin, Germany5 German Federal Institute for Risk Assessment (BfR), German Centre for the Protection of Laboratory Animals (Bf3R), Postfach 12 69 42, D - 10609 Berlin, Germany 6 Department of Animal Welfare and 3Rs, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland7 GSK, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY UK8 Global Animal Welfare and Training, Charles River Laboratories, Wilmington, MA USA9 Department of Pathobiology, University of Guelph, Guelph, ON Canada10 Animal Welfare Program, University of British Columbia, 2357 Main Mall, Vancouver, BC, V6T 1Z4 Canada11 Home Ofﬁce Animals in Science Regulation Unit, 14th Floor, Lunar House, 40 Wellesley Road, Croydon CR9 2BY UK

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22Animal Technology and Welfare August 2020Cumulative experiences – why are they important?Penny HawkinsRSPCAThe experiences animals have throughout their lives can inﬂuence the way they perceive later events, both positively and negatively. It is important to recognise this, from animal welfare, ethical, legal and scientiﬁc perspectives. The concept of ‘cumulative severity’ or ‘cumulative experiences’ appears in various laws regulating animal experiments, and in guidance on implementing these. However, cumulative experiences can be difﬁcult to predict and it is unclear how they can best be detected and assessed. A wide range of factors may inﬂuence a laboratory animal’s cumulative experiences, including the species of animal, the individual’s personality, the procedures involved, housing and husbandry practices, the empathy of handlers and any prior training the animal has experienced. An animal’s cumulative experiences may also be affected by both habituation (which may reduce the negative impacts of an experience) or sensitisation (which may increase the negative impacts).1 Taking these factors into account leads to some important questions relating to the impacts of cumulative experiences, for example:– Might non-regulated studies involving repeated or chronic sub-threshold harms end up above threshold?– Might some procedures go beyond their severity limits due to a lack of recognition of cumulative severity?– How can the concept of cumulative severity be used to better care for animals and improve their lives?Detecting and predicting cumulative suffering is not easy but is essential for understanding whether severity limits may have been approached or exceeded. Noticing if an animal has become sensitised or is showing an exaggerated response to a ‘routine’ procedure, or that an animal no longer appears to be coping with life in the laboratory (e.g. they may show depressive behaviours or stop using enrichment) can provide some signs of this. Animal welfare science can also provide possible practical indicators of cumulative severity, such as anhedonia (no longer taking pleasure in pleasurable stimuli), ‘inactive but awake’ behaviour (see last year’s meeting report2), or nest quality in mice.3 Any welfare assessment system should include a number of welfare indicators like these to ensure it provides an accurate picture of the animal’s welfare state.Although indicators like those mentioned above are useful, it can still be difﬁcult to fully understand the experiences of the animals in question. The principle of ‘critical anthropomorphism’ must therefore be applied - combining empathy with an objective, knowledge-based consideration of what is likely to be signiﬁcant to an animal. This can be informed by thinking about how animals perceive and interpret their world – for example, mice have poor eyesight but good hearing, so may be sensitive to laboratory noises; are nocturnal and so are likely to be disturbed if used during the working day without a reversed light cycle; and are prey animals so can experience stress during capture and restraint. Attempting to consider laboratory practices like marking for identiﬁcation, genotyping, early maternal separation and scientiﬁc procedures from the animal’s point of view can give us a better understanding of the animal’s whole-life experience.In summary, there are a number of key principles underlying approaches to better understanding of cumulative experiences. Firstly, the precautionary principle should be applied, with the assumption being that if something can affect an animal’s ability to cope, that it will. Next, it must be emphasised that there is always more that can be done to improve animals’ lives - and this can be helped by fostering a culture of support for people who want to address animal welfare issues. Support can also come from the Animal Welfare and Ethical Review Body (AWERB), Animal Welfare Body (AWB) or Institutional Animal Care and Use Committee (IACUC). Finally, it is important that all of those involved with the care and use of animals to engage with animal welfare science, engage in critical anthropomorphism and work together to reduce the impact of research on animals.Neural indicators of cumulative severityTom V Smulders, Newcastle UniversityMany laws and guidelines relating to animal experiments refer to ‘cumulative severity’ or ‘cumulative suffering’ as critical in assessing animal welfare. Indeed, the cumulative experience of a number of mild events can be quite severe under some circumstances, so it is very important to be able to detect whether this is happening. Good indicators of cumulative severity should respond to the individual’s experience of the event (not the objective event itself), increase or decrease in value in response to positive and negative experiences, and integrate the response to those positive and negative experiences over time (Figure. 1).4,5 But do such indicators exist?Some potential biomarkers of cumulative experience have already been identiﬁed – for example, telomeres, which are the caps at the ends of chromosomes, shorten in response to chronic stress. However, telomeres do not appear to lengthen in response to positive experiences, so can only be used as a biomarker of Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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23August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfarehow many negative events an animal has experienced. Alternative biomarkers, which integrate positive and negative experiences, are found in the brain, and these may be useful for understanding an animal’s cumulative experience.One brain structure that consistently varies in animals exposed to unpredictable chronic stress is the hippocampus, which plays a major role in learning and memory as well as regulating stress, anxiety and emotional responses. Unlike most of the rodent brain, it adds new neurons throughout adult life. The formation of new neurons is sensitive to both positive and negative experiences. This also occurs in humans – for example, the hippocampus reduces in size in people with major depression – and in animal ‘models’ of major depression. It has also been shown that rats given access to running wheels – a resource they value highly – had signiﬁcantly larger hippocampal volume than unexercised controls.6 On the other hand, rats exposed to chronic immobilisation stress showed a signiﬁcant decrease in hippocampal volume.7 These results show that hippocampal volume can respond to both positive and negative experiences and so these effects can be integrated to provide an insight into cumulative experiences. Similar evidence exists for the formation of new neurons (neurogenesis), which is signiﬁcantly lower in stressed mice than in unstressed mice, whilst the number of new neurons increases in animals provided with environmental enrichment and voluntary exercise.8 This suggests neurogenesis in the hippocampus can indicate positive emotional (affective) states over time. It has also been observed that some anti-depressant drugs that recover animals from a depressive state to a non-depressive state will also increase hippocampal neurogenesis over the same time course. That is, it takes just as long for hippocampal neurogenesis to recover as it does for behaviours to return to normal, suggesting that there may be a link between these new neurons and behavioural indicators of better welfare. In conclusion, it seems possible that ‘biomarkers’ such as the volume of the hippocampus, and the formation of new neurons, could both have potential as ways of assessing cumulative experience. However, there are some limitations: estimation of hippocampal volume can be performed repeatedly in vivo, but only with the use of MRI scans. This process is expensive and involves repeatedly anaesthetising animals which is stressful (and would, ironically, add to cumulative severity). However, it could be done experimentally to help validate potential behavioural indicators of cumulative severity or to help understand the welfare impact of a particular procedure. Neurogenesis can only be assessed at the end of life, so cannot be used as a monitoring tool over time but could be used for experimentally comparing different treatments to allow users to make more informed decisions about the procedures they use with respect to the animal’s welfare, or to help validate estimates of actual severity. Hence, these kinds of tools can help inform better prediction and assessment of cumulative severity.Can home cage behaviour be used to assess cumulative welfare in laboratory mice?Aileen MacLellan, Andrea Polanco, Georgia Mason, University of GuelphCumulative welfare has become a topic of concern for research animals and may be particularly important for fragile strains, animals used in long term studies or research into ageing and breeding stock. Identifying indicators of cumulative welfare, or severity, is therefore an important goal for animal welfare researchers. Although some potential biomarkers of cumulative welfare currently show promise, such as telomere length or hippocampal volume (see above), they also have limitations for day-to-day use cage-side, as they may be expensive, invasive, involve restraint and handling, and prone to false positives or false negatives e.g. Malmkvist et al 2012.9Figure 1. Two patterns of what potential indicators of cumulative experience could look like. Pattern A shows a cumulative indicator that integrates all negative experiences (red arrows) over a lifetime. Pattern B shows an indicator that integrates both negative and positive (blue arrows) experiences. The advantage of pattern A is that it allows one to measure the total negative experiences. However, positive experiences are not recorded. One indicator that might follow this pattern is the changes in telomere length. The advantage of pattern B is that it takes into account the total net experience, but it cannot distinguish between a life with barely any positive or negative experiences, and one that has large negative, but also large positive experiences. Hippocampal neurogenesis may follow the latter pattern. Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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24Animal Technology and Welfare August 2020We aimed to identify some practical potential indicators of cumulative welfare. Our ﬁrst area of focus was home-cage behaviours such as stereotypic behaviour and ‘inactive-but-awake’ behaviour. These behaviours are considered to indicate poor welfare but might also provide simple, non-invasive markers of cumulative experiences. For example, levels of stereotypic behaviour increase with repeated negative events in some species and also may decrease with positive experiences, such as the provision of environmental enrichment.10–12 However, stereotypic behaviours are also prone to false negatives as indicators of cumulative welfare. In part, this may be because stereotypic behaviour appears to be subject to ceiling effects e.g. Bechard et al 2016, that is, stereotypic behaviours may increase in frequency with increasing stressful events up to a point, but then cease to increase in frequency with additional stressors.10 Also, not all species or strains engage in stereotypic behaviour – for example, C57BL/6 mice, rats and guinea pigs all appear to have a low likelihood of developing stereotypic behaviours.13,14 In such animals, other behavioural indicators such as time spent ‘inactive-but-awake’ may be more useful (for more information on inactive-but-awake behaviour, please see last year’s meeting report). We have also explored the potential for colony morbidity and mortality data to be used as an indicator of cumulative welfare. High morbidity and mortality rates are often associated with negative emotional states, and it is possible that negative emotions play a direct or causal role in affecting morbidity, mortality and longevity by contributing to prolonged activation of physiological systems involved in responses in stressful stimuli.15 In a range of species, including humans and laboratory rodents, higher stress levels and negative emotions are associated with increased mortality16–18 while greater longevity is associated with exposure to positive experiences.19,20 To explore this link further, 165 female mice were reared to adulthood (55 C57BL/6; 55 DBA/2 and 55 Balb/c) in environmentally enriched or non-enriched cages. After being used in behavioural research, they were then allowed to live into middle age and beyond for approximately 570 days. Over time, we found that 23% of mice (38/165) had died unexpectedly or prematurely by 570 days (including animals euthanised in response to health issues).1* This was predicted by housing conditions: of the mice that were still alive at 570 days, less than 65% were from non-enriched cages, compared with over 80% of the enriched mice. We also found that stereotypic behaviour predicted early death. However we found no link between inactive-but-awake behaviour and early death. We concluded that all-cause morbidity and mortality data can therefore be used as a potential indicator of cumulative welfare. However, again these results should be cautiously interpreted. Morbidity and mortality rates can be prone to false positives (e.g. species and strain differences in lifespan exist that are not necessarily correlated with welfare). There is also a risk of false negatives since mild stressors may not affect morbidity and mortality and can therefore be missed. For instance, differences might not be detected in populations not allowed to live their full lifespan since cumulative effects of stress only start affecting senescence, morbidity and mortality after middle age. It is also important to note that morbidity and mortality data can only be used as a retrospective indicator to improve future practices, rather than for current interventions.The relationship between potential indicators of cumulative welfare is complex, variable and needs more research to help develop more useful indicators for laboratory and other settings. It is likely that there is no ‘one-size-ﬁts-all’ indicator due to species, strain, sex and individual differences. However, colony ‘all-cause’ morbidity and mortality data does indicate cumulative stress and therefore morbidity and mortality data that is already collected in facilities can be used by colony managers as an assessment tool and means of improvement, with the principal aim of minimising preventable deaths. Increases in stereotypic behaviour are also a warning indicator of cumulative stress and increases in inactivity may also indicate cumulative stress. It should also be noted that some indicators may have opposing patterns, e.g., stereotypic behaviour may ‘protect’ against cumulative welfare biomarkers like shortened telomeres and decreased hippocampal volume. Therefore, consideration of multiple indicators and recognition of potentially opposing patterns is key when monitoring cumulative welfare. Using welfare science to understand animal’s experiences and needsLars Lewejohann, Freie Universität Berlin The vast majority of laboratory animals in Europe are mice, with millions used or housed as stock animals and many more humanely killed because they are considered ‘surplus’ animals.21 Mice are usually housed in small cages which do not offer much variety, despite the fact that laboratory mice are capable of a wide behavioural 1* Note: This was not a ‘severe’ study and death was not used as an endpoint. The mice were simply allowed to live out their lives into late middle age (as happens with pets or zoo animals) and sick animals were always treated and/or euthanised. Findings indicate that senescence (as indicated by a fall in survivorship) began earlier in conventionally housed than enriched animals. Under UK and EU legislation regulating the use of animals for scientiﬁc purposes, actual severity is presumed to be ‘severe’ if an animal is found dead, unless there is evidence otherwise. Death as an endpoint must be avoided as far as possible. In the UK, causes of death must be noted and mortality reported to the Secretary of State if severity limits have been exceeded as a result. Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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25August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfarerepertoire similar to that seen in their wild counterparts. Housing conditions for laboratory mice have been improved over time, so that items considered ‘enriching’ twenty years ago, such as nesting material and mouse houses, are now part of a ‘standard’ cage. One of the key ways to make the lives of laboratory mice better is to aim to continually improve their housing and living conditions.Observing laboratory mice during the working day in a facility with a ‘standard’ light cycle may give the impression that the mice are not experiencing problems but observations conducted during the dark phase at night – when mice are most active – are more likely to pick up signs of poor welfare such as stereotypic behaviour. This kind of behaviour may be reduced with the provision of environmental enrichment and lots of items are now commercially available, such as different types of mouse houses, climbing structures and platforms to increase available space. Enrichment aimed at providing cognitive stimulation can also help to alleviate boredom – these kinds of items usually require the mice to interact with an object in order to obtain a reward. In our laboratory, we have introduced boxes with lids which the mice must lift to access millet seeds, hollow balls stuffed with nesting material and millet seeds for the mice to remove, tunnels containing pebbles which the mice can dig out and balls containing millet seeds which will fall out through a small hole if the ball is rolled around the cage. We have also noted that mice like to engage with running wheels or discs and that running discs seem preferable as the mice can run without having their spine bent as it would be in a running wheel.To establish which of these types of enrichment items are best for promoting good welfare, we compared different housing types: a conventional cage containing a mouse house and nesting material; an enriched cage containing platforms: different types of housing, a running disc and different cognitive enrichment items, regularly exchanged to provide novelty. We found that behaviours associated with poor welfare, such as inactive behaviour and stereotypies, were signiﬁcantly reduced in enriched cages compared to controls. We were also interested in rating the different enrichment items from the mouse’s perspective, so we conducted a large number of preference tests. To do this, we tagged mice in the neck region with radio-frequency identiﬁcation (RFID) tags so their locations in a cage could be tracked using our newly-developed surveillance system.22 We then presented mice with different combinations of enrichment items in order to develop a rank order of preference for these items (Lewejohann and Talbot, in prep). We found that mice showed the greatest preference for a plastic ﬂoor house on which they could climb in comparison with other types of mouse house and a ball-shaped house was least preferred. Structural items with a ﬂat surface on top were the most preferred of all the climbing elements we presented with a plastic suspended tube the least preferred. Finally, we found that the most preferred form of cognitive enrichment was the latticed ball containing removable nesting material and millet seeds and a puzzle box which required mice to slide open a lid to access millet seeds was least preferred. Our next step will be to conduct consumer demand tests, which are tests which can be used to assess how hard mice are willing to work for access to a reward or condition.23 We have previously shown that mice will work harder (press a lever more times) to access an enriched cage than an additional non-enriched cage, suggesting that the enriched cage is more highly valued by mice.Beyond the forms of enrichment described here, we have found other ways that help to improve the welfare of our mice include provision of treats like millet seeds and providing opportunities for exercise by adding running discs to cages (Figure 2.). We have noticed that aged mice provided with running discs looked healthier and more active after two weeks, suggesting that this provision may be important for limiting the welfare impacts of aging. Continuing to trial and evaluate these kinds of interventions are important ways to keep improving the welfare of research animals, even outside of an experimental context.21Figure 2. A mouse using a running disc. Credit: Lars Lewejohann. Small reﬁnements to improve lifetime welfarePaulin Jirkof, University of ZurichReﬁning experimental procedures to reduce pain, stress or other negative emotional (affective) states is a crucial tool to improve experimental animal welfare. However, laboratory rodents spend much of their lives in their cages, outside the experiment and many are Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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26Animal Technology and Welfare August 2020not even used for experiments but maintained for breeding. To ensure the lifetime welfare of all animals bred and housed for scientiﬁc purposes, all aspects of husbandry, breeding, housing and research procedures must be considered.Mice account for the majority of research animals globally and are usually housed in groups as they are social animals. However, inter-male aggression in group-housed mice is very common, and can lead to stress, severe injuries and death – especially as ﬁghting wounds may not be noticed until it is too late.24 This means that the severity of ﬁghting in male mice is often under-estimated. A potential solution to this problem is to house male mice singly, but this intervention is not ideal as it deprives mice of their social needs and also makes mice more vulnerable to cold stress as they cannot huddle together with others for warmth. In general, male mice prefer to be group housed so it is important to seek a better solution than individual housing.25 Although there is some ambiguity in the literature regarding alternative interventions, some show consistent and promising results. For example, some laboratory mouse strains are less prone to aggression than others, which can provide a useful starting point; grouping siblings together, grouping mice when young and keeping these groups stable once established can also help reduce aggression. The ideal group size for male mice has not yet been agreed upon, as some research has suggested that smaller groups may be better than larger groups, nevertheless recent research resulted in ambiguous results.25–28 Aggression also tends to be lower when steps are taken to reduce stress – for example, moving used (but not soiled) nest material (not litter) into a new cage when cages are changed and choosing less stressful handling techniques such as tunnel handling and predictable handling.28 If none of these interventions work and aggression persists, mice may have to be housed singly, but extra nesting material should be provided in order to reduce the risk of cold stress. Another area which can be reﬁned to improve the experience of animals and may contribute to improving an animal’s overall experience is drug administration. Typically, this procedure is stressful for rodents as it may involve restraint and unpleasant or aversive experiences like injection or oral gavage. With well-trained personnel and habituation, stress can be reduced somewhat but this process is still likely to be stressful. However, it is possible to train rodents to ingest substances, either directly from a syringe or by mixing with preferred foods. Some restraint may be initially necessary, but if the carrier substance is palatable, and as the animal habituates to the experience, less restraint will be needed, possibly to the point where no restraint is needed at all.29 This technique works with both rats and mice – and could even become a positive experience for the animal. As another alternative, drugs can be mixed with palatable substances and provided in the animal’s home cage so that no handling is required. Nutella®, honey, strawberry jam, baby food and condensed milk are all good options to try and sterile or calorie-free jellies are commercially available if they are needed, as are emulsiﬁers which may be needed to mix the substance with the carrier. However, note that methods based on uncontrolled voluntary ingestion (e.g. via drinking water or ad libitum food) may not be suitable for protocols which require the animals to ingest a controlled amount, especially as eating or drinking events may vary greatly in frequency between the light and the dark phase.30There is great potential for improving the lives of laboratory animals both in and outside of experiments. If you wish to apply these, or any other reﬁnements in your facility, consider putting together an action plan which takes into account the latest advances in the ﬁeld, challenges the status quo and aims to ﬁnd creative ways to solve any problems that may arise. Train collaborators in the reﬁnement procedures you wish to use and test options systematically, with alternative methods in place in case they are needed. Finally, share your experiences with internal and external colleagues – letting others know what works and what does not, is key for promoting better welfare for a greater number of laboratory animals. Development of a visual approach to severity assessmentJackie Boxall and Helen Murphy, GSK Guidance on severity assessment, such as the EU Severity Assessment Framework,31 states that the duration of adverse effects should be considered when assessing harms to animals resulting from procedures – but how do we decide when a transient effect becomes persistent? Do we all think the same way? Good communication between all stakeholders is key when making decisions about animals used in a study, but it can be challenging to ensure a consistent approach between research projects. Our Animal Welfare and Ethical Review Body (AWERB) set a 2020 objective to review internal guidance on severity assessment, clarify the transitions from mild, to moderate, to severe for commonly observed clinical signs and consider cumulative severity.Our approach was to develop a ‘heat map’ for each individual clinical sign, with the descriptor of the sign along one axis and the duration of the sign along the other axis. This would allow a colour-coded severity classiﬁcation to be assigned for each clinical sign that takes both factors into account, so that a sign which appears fairly mild, but lasts for a long time, may Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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27August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfareactually be considered a sign of moderate severity, and a long-lasting moderate sign may be considered severe. The guidance we currently use leaves decisions about how the duration of a sign should be interpreted up to the observer, whereas this heat map approach can help to remove some of the ambiguity. To develop our guidance, we formed a working group, which included animal care staff, researchers, veterinary surgeons and a statistician. The group members came from diverse areas of our animal research community and worked on a range of different species to provide a broad basis of knowledge and help achieve consensus regarding how to classify the different clinical signs. We started by forming a list of the clinical signs we wanted to develop guidance for, and discussed each sign and how to interpret it in detail. We agreed upon the basic structure of the map for each sign by deciding on what the descriptors and timelines were going to be. Next, each group member was asked to ﬁll in the heat map independently from other Working Group members although they could consult other colleagues if they wished. Data from this was then collated and visualised using mosaic plots. The mosaic plots showed where there was a strong consensus over how a sign should be interpreted and this information was used to begin assigning colours to the boxes within the heat map. For areas without a strong consensus, we used the current guidance to help inform our decisions and engaged in further discussions within the group to better understand each other’s viewpoints. This has resulted in usable heat maps for several generalised clinical signs including hunched posture, subdued behaviour and piloerection and are working on maps for body weight changes (Figure. 3). When considering how individual clinical signs may affect cumulative severity, we must consider the total number of clinical signs as well as the magnitude and duration (Figure. 4). To use the heat maps for multiple clinical signs, we assign the severity level for each sign individually, then take the highest severity level as the minimum actual severity experienced by that animal. Where all the signs fall into one band, we can look at how close the signs are to the threshold for the next band and consider whether a higher overall severity needs to be assigned to take into account the cumulative experience of the animals.Although the development of our heat maps has been a positive step forward, there are some limitations to our approach and some next steps we are taking to develop the guidance further. On the positive side, the effects of duration are now well-deﬁned, severity can be assessed on a continuous spectrum and the heat maps are based on a wide consensus with transparency over all stages of the decision-making process which should mean there will be good consistency between users. However, assessment of animals still tends to be based on professional experience and opinions, rather than animal welfare or behaviour science, especially as some of the descriptors are still open to interpretation. We also have yet to deﬁne how to interpret intermittently displayed clinical signs. Our next steps will be to develop guidance for further clinical signs and procedural effects, further develop our method for interpreting multiple clinical signs and to seek further consensus and feedback to continue to improve our approach. Figure 4. Factors affecting cumulative severity considered for the ‘heat map’ approach to actual severity assessment.Figure 3. An illustration of a heatmap. A similar table for each clinical sign covered by the new guidance will be shared once completed. Severity bands, descriptors and timelines would be speciﬁc for each clinical sign.Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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28Animal Technology and Welfare August 2020Establishing trust with laboratory rats: how long does it really take?Carly M Moody, Patricia V Turner, Charles River Laboratories University of GuelphLaboratory rats and mice are handled frequently in research settings for example, during cage change, for physical examination and for various study procedures. Suboptimal handling and restraint procedures can cause prolonged stress responses caused by negative reactions to people, which may have further negative effects such as delayed wound healing, reduced learning and cognitive abilities and reduced animal health and wellbeing. On the other hand, the use of low-stress handling techniques like cup and tunnel capture and handling to improve human-animal interactions has a number of beneﬁts.32,33 Reduced fear in laboratory rodents reduces the risk of injuries to both animals and staff, makes the interactions more enjoyable which can beneﬁt animal welfare, staff job satisfaction and the overall human-animal bond and helps to minimise bias in the study data. Despite these highly publicised beneﬁts, there is still poor uptake of low-stress handling practices, and a common reason given is that habituation takes too long. We therefore conducted a study to investigate how long it takes to improve rat-human interactions.The aim was to evaluate whether short periods of habituation and counter-conditioning would reduce measures of fear, stress and anxiety in handled rats. Habituation (the gradual exposure of an animal to a stimulus) and counter-conditioning (where a negatively-perceived event is paired with a positive stimulus to reduce the negative effects) are both training techniques that could be incorporated into the regular husbandry of animals to reduce their negative responses to people, procedures or the general laboratory environment. We carried out the study over a two-week period and included three groups of male Sprague-Dawley rats. The control group received no handling over the study period, the ‘low handling’ group received 15 seconds of gentle handling three times a week and the ‘moderate handling’ group received 45 seconds of gentle handling three times a week. The handling consisted of gentle body restraint and stroking of the head, body, tail and limbs on a soft handling mat with Cheerios given as treats.At the end of the study period, we found that rats in both handling groups urinated and defecated less during cage change than control rats, were quicker to voluntarily approach the hand of an unknown person, suggesting lower fear of humans, and also eliminated less when restrained for blood collection. However there were no differences between groups in glucose levels or in behaviour when the rats were tested in an elevated plus maze, suggesting that while fear of humans had been reduced, there was still some level of handling stress. We also noted that there were no differences between the low-handled and moderately-handled groups, suggesting that only 15 seconds of handling three times a week is sufﬁcient to reduce negative responses.The results of this initial study are promising, as they suggest that relatively little time needs to be invested to improve the experience of laboratory rats which has important implications for the overall cumulative experience of these animals. We plan to further investigate these effects, ﬁrstly by seeing if our result can be replicated and then by carrying out this study with female rats to see if their responses differ from males. We also hope to carry out a longer study to examine how long the effects of this simple handling protocol will last.A good life for laboratory rodents?I Joanna Makowska, University of British ColumbiaA ‘good life’ requires that animals be able to express a rich behavioural repertoire, use their abilities and fulﬁl their potential through active engagement with their environment. Although some types of research may not always be compatible with providing laboratory animals with a good life, it is possible to consider what the minimum day-to-day living conditions would be that contribute to a good life for laboratory rodents. There are three major aspects of animals’ lives which play a major role in having a good life: the animal’s life outside the research context, the interactions that animal has with humans and the animal’s physical environment. Here we focus on the physical environment but the importance of the animal’s life outside research and human-animal interactions are discussed in Makowska and Weary (2020).34A ‘standard’ cage for laboratory rodents has two main physical features – litter and shelter. The types of these features which are chosen can have a signiﬁcant impact on welfare – for example, in North America, corncob bedding is popular for its high absorbency, but has been found to be avoided in preference tests. An alternative is paper-based material which has fewer impacts on animal health but has lower absorbency. A simple way to improve welfare for rodents is therefore to provide paper material but use a deeper layer – this is preferred by mice, and also was found to lead to lower corticosterone levels, higher body temperature, lower food intake and lower ammonia levels in mouse cages, meaning that any higher cost of using more litter may be offset by lower food costs.35 With respect to shelters, open-ended PVC pipes are often used in rat cages, even though rats prefer hut-type shelters with Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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29August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfareonly one entrance. Mice tend to prefer nests as shelters and will choose soft paper towels or tissue paper as their main building material, even though they produce better-quality nests with crinkle paper. If both are provided, mice will use crinkle paper as the structural outer layer and paper towels as the inner layer, resulting in a comfortable and high-quality shelter.Another important aspect of the physical environment for laboratory rodents is the level of environmental complexity. Creating more complexity, either by adding structures which increase the amount of usable space, or by adding cage dividers, is preferred and leads to lower stress levels in both rats and mice. Increased complexity can also allow rodents to use separate areas for different activities – mice housed in three interconnected cages were found to build a nest in one cage and use another as a latrine,36 and – in general – mice provided with a demarcated area in their cage spontaneously use this area as a latrine.37 Cage designs should therefore promote this segregation of space, for example, a litter pan containing absorbent bedding can be placed near the food and water, as mice and rats prefer to eliminate close to food and water. Doing this would also allow the rest of the cage to be disturbed less frequently, as only the litter pan would need regular changing and would also allow more comfortable bedding to be used in the rest of the cage.Alternatives to the typical ‘shoebox’ cage, such as cages more similar to those used for pet rats, can provide an even greater level of environmental complexity and are associated with better welfare and a more complex behavioural repertoire. For example, rats housed in large cages containing soil were less stressed and performed behaviours not possible in a standard cage, such as burrowing, climbing and upright stretching, while mice housed in large, complex enclosures had less fat and stronger immune systems.38 When it is not possible to provide home cages with this level of complexity, animals will still beneﬁt from access to a ‘playpen’ – repurposed rabbit cages (for rats) or rat cages (for mice) that animals have regular access to can promote better welfare and a wider range of behaviours in rodents.The reﬁnements presented here, along with other reﬁnements such as less restrictive handling and good socialisation protocols, are simple ways to immediately improve the welfare of laboratory rodents and contribute to them having a better lifetime experience. Over a longer time-frame, there are even more potential avenues to explore, many of which should be the ultimate goal for how animals are used in future, for example, providing options which allow animals to free-range, training animals to voluntarily participate in procedures and using pets which naturally develop conditions for studying diseases rather than created models. Taking steps like these to give animals a good life is not only our duty but should be considered a prerequisite for their use, and a starting point around which we build our research programmes.Home Ofﬁce updateCharlotte Inman, Animals in Science Regulation Unit (ASRU)Under the Animals (Scientiﬁc Procedures) Act in the UK, any application to use animals in research is subject to a harm-beneﬁt analysis, to ensure that any harm that may be caused to the animals is justiﬁed by the expected beneﬁts for humans, animals or the environment. The experience of animals used under ASPA can be inﬂuenced by project-related effects (effects which are speciﬁc to the regulated procedures undertaken) and contingent effects (those which inherently arise from the experimental or scientiﬁc use of an animal). The net impact of these two groups of effects determine the cumulative severity of an animal’s experience over the course of its use. The use of severity classiﬁcation is required by law and qualiﬁes the likely (prospective), ongoing (during procedures) and actual nature of the experience of an animal. Understanding the cumulative nature of animal experiences presents multiple opportunities to inﬂuence the likely and actual severity experiences of animals in science.When considering cumulative experience, there is often a focus on the project-related effects, for example, in the case of administration of a substance, factors such as the route, the nature of the substance and the frequency can all have an impact, as will the application of good practices such as single-use needles and the use of anaesthesia and analgesia. However, contingent effects can also have a signiﬁcant impact, especially as they may affect animals prior to their use and between and after procedures. Contingent effects may be broader than project-related effects, such as provision of food and water, including reﬁnements such as the provision of wet mash post-surgery, handling, enrichment and housing conditions. Animal Technologists can have a major impact on how both project-related and contingent effects impact animals, as they can provide highly valuable expert input due to their speciﬁc qualiﬁcations and exposure to continuing professional development. Animal Technologists will also see a broad range of studies involving a range of species and so are well-placed to identify opportunities for translational reﬁnements or changes in practice across different studies or species. Finally, they are involved across the lifetime of an animal, not just when the animal is being used for an experiment and so are able to consider how to make incremental improvements to an animal’s lifetime experience.Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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30Animal Technology and Welfare August 2020Often, improvements to animals’ lives can be made that are not necessarily written into project licenses and Animal Technologists can be key in identifying these. For example, re-using needles for procedures can cause animals unnecessary pain and tissue damage, and can have a signiﬁcant impact on cumulative experience. A survey by ASRU in 2019 found that 73% of establishments were aware of this issue and that needle re-use was occurring in 35% of establishments and establishment culture was a major reason for this. Another example of the importance of the impact of animal care staff on cumulative experience is the use of reﬁned handling methods for mice. An ASRU themed inspection in 2019 found that 59% of establishments were only using these non-aversive methods of handling and that the primary factor for the success of these methods was engagement of Animal Technologists and agreement over the need for change. The best motivation for that change therefore came from within the technologist community. These examples demonstrate how important the role of the Animal Technologist is in helping to identify and implement positive change for animals.Animal Technologists have the relevant professional background, interact with animals across their lifetimes and are likely to interact with animals more frequently than researchers. They are also closely involved in the care of experimental animals before, during, between and after their use, and so have the opportunity to make a really positive impact on animal cumulative experiences through their input on both the project-speciﬁc and procedural effects. It is therefore important that animal technologists are empowered to make this positive contribution. Interactive discussionThe ﬁnal session of the day was an interactive discussion around the topic ‘how do we know if cumulative suffering is present in rodents cage-side?’. A brief survey of audience members at the start of the session showed that over 85% of the audience felt that cumulative severity was an issue for at least some, if not all their animals, but only 42% felt that they would be conﬁdent in identifying indicators of cumulative severity and just 38% said their establishment’s welfare assessment systems included indicators that detect cumulative effects. The discussion therefore focussed on indicators that can be used to identify cumulative effects.Some of the possible signs that may indicate issues with cumulative welfare which were suggested by participants included body condition, weight, posture and activity and it was agreed that activity levels, as well as particular activities or behaviours like nest building, can be used as indicators of cumulative welfare. This may especially apply to abnormal behaviours such as stereotypic behaviours, barbering or aggression. Another suggestion was that behavioural diversity and circadian rhythms, can be disrupted in response to stress or chronic stress, so noticing these changes can help identify poor welfare. Participants also discussed how an animal’s response to handling or other human interactions may change in response to a cumulative welfare issue, although these changes are difﬁcult to quantify and capture but are usually recognised by technologists who have the experience to recognise when an animal is ‘just not right’. Given the difﬁculty of quantifying some of these indicators, it was suggested that a way to help monitor some of them on welfare assessment score sheets would be free-text boxes, so that signs which do not appear on the lists of indicators but are recognised by technologists can be recorded. It was also suggested that score sheets should include a list of procedures done so that those interacting with an animal can see what the animal has previously experienced and that procedures which are not necessarily part of an experiment and may be thought of as ‘routine’, such as biopsies and marking for identiﬁcation, may still affect animals and therefore should be included on such a list. The discussion also covered how animals can be monitored to identify cumulative welfare indicators. For example, it was suggested that reﬁned handling methods can be a useful tool, as some indicators of poor welfare are likely to be easier to notice when using these low-stress techniques. Another point was the importance of the timing of monitoring: it was noted that animals are often looked at for only short periods and sometimes during the day when they are asleep. We therefore may need to think more about observing animals for longer periods of time or increasing the number of observations and using up-to-date home cage monitoring technologies and methods, as well as using reversed light-cycles if not already doing so (although this does not remove all issues (see Hawkins and Golledge39). It was agreed that it is generally more relevant to look at animals during their active phase and sometimes 100% of certain behaviours can be missed if animals are only monitored during the light phase. However, if there is some reason where animals cannot be monitored in the dark phase or if animals are being monitored in the light phase immediately after a procedure, placing animals in a playpen can be helpful. This is because animals are generally very active in the playpen even during the light phase, so lack of activity in a playpen can help to identify issues.One ﬁnal question that was raised by the participants was how the effects of ageing and cumulative welfare can be separated. On the one hand, it was felt that the ageing process is part of an animal’s lifetime experiences and so its effects cannot be separated Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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31August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfarefrom other aspects of cumulative welfare. On the other hand, it was considered important to compare amongst animals of the same age group to ensure that indicators which would not be accepted in a younger animal are not ignored simply because the animal is older. One participant also added that the beneﬁcial effects of exercise for older animals are under-estimated and that these may help limit some of the negative effects of ageing.The discussion session closed with a general agreement that of all the possible reﬁnements and interventions presented over the course of the day, the use of heat maps to assess cumulative severity was the one that most participants wished to try and implement in their own establishments.Action Points– Ask how your establishment keeps up to speed with new developments in animal welfare science. Does the Named Information Ofﬁcer (NIO) have the resources they need; how does the AWERB access information; are there any researchers who work in related ﬁelds?– Recognise the importance of considering how an animal’s cumulative experiences might affect that animal’s response to further procedures or experiences. You may like to raise the issue at your establishment, e.g, via the AWERB.– Consider whether an animal’s prior or cumulative experiences may cause some procedures or projects to exceed their severity limit.– Assessing the severity of procedures, consider: • how many clinical signs is the animal displaying? • how long has each sign been present? • how close is each sign to the humane endpoint? • what is the combined effect on the actual severity?– Review colony ‘all-cause’ morbidity and mortality data to see if there are any indicators that animals may be experiencing poor cumulative welfare which can be addressed but take mortality very seriously and prioritise preventing this.– Stereotypic behaviours and ‘inactive-but-awake’ behaviours indicate poor welfare and staff should keep an eye out for these indicators.– Keep a lookout for signs of sensitisation, such as exaggerated response to a ‘routine procedure’ or depression, such as inactivity or no longer using enrichment, which may suggest an animal is no longer coping with life in the laboratory.– Monitor group-housed male mice for signs of inter-male aggression and remember that aggression is frequently under-estimated. If aggressive male mice must be housed singly, provide individuals with extra bedding to help avoid cold stress.– Try and ﬁnd foods which your animals enjoy eating to help train animals to voluntarily ingest substances for experiments - or to use as treats.– Include enrichment items which provide cognitive stimulation and allow animals to exercise.– Try to incorporate low-stress handling into your interactions with laboratory rats and mice to improve human-animal interactions. Challenge assumptions that habituation and training (for both animals and humans) will take too long.– Provide preferred forms of litter, nesting material and enrichment to your animals, such as shredded paper over corncob bedding for mice and a mix of nest-building materials, and hut-type shelters for rats. You can research preferences in the literature or you should be supported to do your own trials.– Try giving rodents a dish or other demarcated area to use as a latrine to keep the cage clean and minimise how often animals have to be disturbed for cage change.– If animals cannot be housed in larger, more enriched cages, repurpose old cages and toys to create a playpen and give your animals regular access to this.Acknowledgements Thank you to all the speakers and online participants and to UFAW for providing the meeting platform. References1 Animals in Science Committee Harm-Beneﬁt Analysis Sub-Group (2017) Review of Harm-Beneﬁt Analysis in the Use of Animals in Research, Animals in Science Committee.2 Stevens C., Finnegan E., Clarkson J. et al. (2020) Report of the 2019 RSPCA/UFAW Rodent Welfare meeting. Animal Technology and Welfare, Vol. 19, 101–111.3 Gaskill B.N., Karas A.Z., Garner J.P. & Pritchett-Corning K.R. (2013) Nest building as an indicator of health and welfare in laboratory mice. Journal of Visualized Experiments, Vol. 82, e51012.4 Bateson M. & Poirier C. (2019) Can biomarkers of biological age be used to assess cumulative lifetime experience? Animal Welfare, Vol. 28, 41–56.5 Poirier C., Bateson M., Gualtieri F. et al. (2019) Validation of hippocampal biomarkers of cumulative affective experience. Neuroscience and Biobehavioral Reviews, Vol. 101, 113–121.6 Sierakowiak A., Mattsson A., Gómez-Galán M. et al. (2015) Hippocampal morphology in a rat model of depression: the effects of physical activity. Open Neuroimaging Journal, Vol. 9, 1–6.7 Rahman M.M., Callaghan C.K., Kerskens C.M., Chattarji S. & O’Mara S.M. (2016) Early hippocampal Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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32Animal Technology and Welfare August 2020volume loss as a marker of eventual memory deﬁcits caused by repeated stress. Scientiﬁc Reports, Vol. 6, 29127.8 Yun J., Koike H., Ibi D. et al. (2010) Chronic restraint stress impairs neurogenesis and hippocampus-dependent fear memory in mice: possible involvement of a brain-speciﬁc transcription factor Npas4. Journal of Neurochemistry, Vol. 114, 1840–1851.9 Malmkvist J., Brix B., Henningsen K. & Wiborg O. (2012) Hippocampal neurogenesis increase with stereotypic behavior in mink (Neovison vison). Behavioural Brain Research, Vol. 229, 359–364.10 Bechard A.R., Cacodcar N., King M.A. & Lewis M.H. (2016) How does environmental enrichment reduce repetitive motor behaviors? Neuronal activation and dendritic morphology in the indirect basal ganglia pathway of a mouse model. Behavioural Brain Research, Vol. 299, 122–131.11 Gottlieb D.H., Coleman K. & McCowan B. (2013) The Effects of Predictability in Daily Husbandry Routines on Captive Rhesus Macaques (Macaca mulatta). Applied Animal Behaviour Science, Vol. 143, 117–127.12 Greco B.J., Meehan C.L., Hogan J.N. et al. (2016) The Days and Nights of Zoo Elephants: Using Epidemiology to Better Understand Stereotypic Behavior of African Elephants (Loxodonta africana) and Asian Elephants (Elephas maximus<i/>) in North American Zoos. PLOS One, Vol. 11, e0144276.13 Fureix C., Walker M., Harper L. et al. (2016) Stereotypic behaviour in standard non-enriched cages is an alternative to depression-like responses in C57BL/6 mice. Behavioural Brain Research, Vol. 305, 186–190.14 Nip E., Adcock A., Nazal B. et al. (2019) Why are enriched mice nice? Investigating how environmental enrichment reduces agonism in female C57BL/6, DBA/2, and BALB/c mice. Applied Animal Behaviour Science, Vol. 217, 73–82.15 Walker M.D., Duggan G., Roulston N., Van Slack A. & Mason G. (2012) Negative affective states and their effects on morbidity, mortality and longevity. Animal Welfare, Vol. 21, 497–509.16 Leserman J. (2008) Role of depression, stress, and trauma in HIV disease progression. Psychosomatic Medicine, Vol. 70, 539–545.17 Satin J.R., Linden W. & Phillips M.J. (2009) Depression as a predictor of disease progression and mortality in cancer patients: a meta-analysis. Cancer, Vol. 115, 5349–5361.18 Cavigelli S.A. & McClintock M.K. (2003) Fear of novelty in infant rats predicts adult corticosterone dynamics and an early death. Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, 16131–16136.19 Diener E. & Chan M.Y. (2011) Happy people live longer: Subjective wellbeing contributes to health and longevity. Applied Psychology: Health and Wellbeing, Vol. 3, 1–43.20 Bice B.D., Stephens M.R., Georges S.J. et al. (2017) Environmental Enrichment Induces Pericyte and IgA-Dependent Wound Repair and Lifespan Extension in a Colon Tumor Model. Cell Reports, Vol. 19, 760–773.21 Lewejohann L., Schwabe K., Häger C. & Jirkof P. (2020) Impulse for animal welfare outside the experiment. Laboratory Animals, Vol. 54, 150–158.22 Habedank A., Urmersbach B., Kahnau P. & Lewejohann L. (2020) O mouse, where art thou? The Mouse Position Surveillance System (MoPSS) - an RFID based tracking system. bioRxiv, 379719.23 Kahnau P., Habedank A., Diederich K. & Lewejohann L. (2020) Behavioral Methods for Severity Assessment. Animals, Vol. 10, 1136.24 Gaskill B.N., Stottler A., Pritchett-Corning K.R. et al. (2016) He’s getting under my skin! Comparing the sensitivity and speciﬁcity of dermal vs subcuticular lesions as a measure of aggression in mice. Applied Animal Behaviour Science, Vol. 183, 77–85.25 Kappel S., Hawkins P. & Mendl M.T. (2017) To Group or Not to Group? Good Practice for Housing Male Laboratory Mice. Animals, Vol. 7, 88.26 Van Loo P.L.P., Van Zutphen L.F.M. & Baumans V. (2003) Male management: Coping with aggression problems in male laboratory mice. Laboratory Animals, Vol. 37, 300–313.27 Lidster K., Owen K., Browne W.J. & Prescott M.J. (2019) Cage aggression in group-housed laboratory male mice: an international data crowdsourcing project. Scientiﬁc Reports, Vol. 9, 15211.28 Jirkof P., Bratcher N., Medina L. et al. (2020) The effect of group size, age and handling frequency on inter-male aggression in CD 1 mice. Scientiﬁc Reports, Vol. 10, 2253.29 Scarborough J., Mueller F., Arban R. et al. (2020) Preclinical validation of the micropipette-guided drug administration (MDA) method in the maternal immune activation model of neurodevelopmental disorders. Brain, Behavior, and Immunity, Vol. 88, 461–470.30 Sauer M., Fleischmann T., Lipiski M., Arras M. & Jirkof P. (2016) Buprenorphine via drinking water and combined oral-injection protocols for pain relief in mice. Applied Animal Behaviour Science, Vol. 185, 103–112.31 Expert Working Group on Retrospective Severity Assessment (2012) National Competent Authorities for the implementation of Directive 2010/63/EU on the protection of animals used for scientiﬁc purposes: Working document on a severity assessment framework EU Commission.32 Gouveia K. & Hurst J.L. (2013) Reducing mouse anxiety during handling: effect of experience with handling tunnels. PLOS One, Vol. 8, e66401.33 Cloutier S., LaFollette M.R., Gaskill B.N., Panksepp J. & Newberry R.C. (2018) Tickling, a Technique for Inducing Positive Affect When Handling Rats. Journal of Visualized Experiments, Vol. 135, e57190.Report of the 2020 RSPCA/UFAW rodent and rabbit welfare meeting

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35August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2021 Animal Technology and WelfareEmotional challenges in our work with laboratory animals: tools that support caring for others and yourselfANGELA KERTON1 and JORDI L TREMOLEDA2,3 1 The Learning Curve (Development) Ltd, PO Box 140, Ware, Hertfordshire SG9 0ZN UK 2 The Blizard Institute, Barts and the London School of Medicine and Dentistry, 4 Newark St, London E1 2AT UK3 Biological Services, Barts and the London School of Medicine and Dentistry, 4 Newark St, London E1 2AT UKCorrespondence: angela@learningcurvedevelopment.co.uk Abstract Inevitably, most of us who work with laboratory animals will sometimes form bonds with the animals we are caring for. These relationships will positively enhance the care and wellbeing of the animals’ but they also pose important emotional challenges, as was clearly exposed with some contingency managements associated with the COVID-19 pandemic. It is important that the industry acknowledge the existence of these bonds and provide institutional support mechanisms to help Animal Technologists to deal with the emotional challenges of their profession. Current COVID-19 associated working logistics pose further challenges such as delegation of responsibilities, separation of working teams and contingency management of stock to name but a few, along with the individual health and social, economic and personal relationship challenges. This article provides some tools and ideas to support a more open, communicative and emotionally-supportive working environment. The importance of ‘self-care’ is also discussed. There is a growing commitment to nurture a Culture of Care, and supporting our colleagues by raising awareness of our emotional challenges may support this.IntroductionWorking within the animal research sector and dealing with a range of associated professional responsibilities can impact our emotions. It is important that staff recognise this and practice ‘self-care’ but equally know where and when to ask for additional support when it is needed. Emotional stressors in the work environment can inﬂuence our professional and personal integrity, directly affecting how we carry out our responsibilities and can change our attitude towards colleagues and the animals we are working with. Therefore, seeking a better acknowledgment and understanding of such emotional challenges and how to best manage them is crucial. We hope that by exploring supporting tools to promote openness and emotional resilience, like the use of mindfulness to reduce stress and supporting individual and team reﬂective practice will provide better coping mechanisms.Animal-human interactions directly inﬂuence the behaviour of the animals; this is in part associated with the dependence relationship between the animal and carer, as it is seen through habituation and positive reinforcement practices.1 This connection can affect positively the impact the research outcomes as such ‘positive’ animal and human bond have a ‘profound’ inﬂuence on the animals’ behaviour and physiology.2 Yet, continuing carrying of such bonds accounts important professional responsibilities. These expectations are to be maintained even in critical challenging personal and professional scenarios, like the current COVID-19 scenario (e.g. different working set ups, increase responsibilities and decision making, higher pressure and expectation, professional uncertainties, etc). Such animal-human bonds remain at the core of the empathetic challenge, as the stronger the bond the more robust the caring activity may be but also the more distressful the execution of experimental procedures or contingency decision (e.g. culling) can be.3 The COVID-19 pandemic has forced many institutions to scale back operations including animal research. Staff have been faced with difﬁcult decisions over what to do with research animals amid lockdowns, exacerbated by uncertainty on contingency provision for funding support and the unpredictability of the length of the lockdown.4,5

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36Animal Technology and Welfare August 2020Emotional challenges affect us personally and professionallyThe relationship between laboratory animals and professional staff who work with them has a overwhelming impact on the animals’ wellbeing, as well as on the emotional health of staff including veterinarian, caretakers, researchers and other support staff. Stressors like lack of support and communication, excessive workload, changes on working patterns, delivery of expectations4 need to be managed to promote an appropriate balance between expectations, motivation and excessive pressure to avoid leading to work-related stress. Indeed, working with other sentient beings’ lives and their wellbeing following the human-induced experimental procedures that can cause harm or distress remains a major emotional challenge. This is also pressured by existing regulatory frameworks and increasing social accountability.6,7 All this exposes the vulnerability of individual moral attitudes on care and compassion, along with the professional expectations and accountability. Emotional stressors, particularly those associated with critically harmful interventions and/or the need to humanely euthanise animals make a signiﬁcant contribution to the development of so called compassion fatigue.8,9 This associated ‘reduced awareness/ capacity in being empathetic’ may evolve in less caring attitudes and further physical and emotional distress and exhaustion.10,11 Importantly, such reduced empathy for others will diminish the quality of care towards the animals but also towards colleagues. Such emotional challenges can lead to what is associated with ‘compassion fatigue’, commonly recognised as a type of stress that results from helping or wanting to help those who are experiencing signiﬁcant pain, suffering and distress or are themselves under signiﬁcant emotional duress. Compassion fatigue symptoms can lead to lack of communication, excessive blaming, isolation from others, and excessive complaining attitudes that can easily progress towards bottling up emotions, and mental and physical tiredness and depression.12 Compassion fatigue is considered as form of burnout, associated with our social and professional interactions. Possibly it would be also more accurate to refer to empathy fatigue rather that compassion fatigue, as it is empathy that fatigues (tires), in care givers, not compassion.13Our aim is to provide guidance on supporting tools and avenues to help the community to speak out and build up resilience. We wish to raise awareness of speciﬁc mental health challenges in the current COVID-19 working environment and how discussion of these can be facilitated within an emotionally supportive workplace environment. For example, it is important that strategies to minimise euthanasia stress (that Emotional challenges in our work with laboratory animalsmay contribute to empathy/compassion fatigue) are embedded in workplace cultures, including an open atmosphere to encourage dialogue and expressions of grief, strong social support networks, explanations as to the necessity for the research and openness in the recruitment and training phase of the occupational requirements involved in animal-based research, including euthanasia.14-17 Using compassion skills for managing our emotional challengesHigh quality and conscientious animal care is good for the animals, science, Animal Technologists and public perception of research facilities. There are cross overs between different professions involved with providing a high degree of direct care during their day jobs and the potential for them to become ‘over attached’ or ‘emotionally vulnerable’ to speciﬁc work cases or situations they may encounter. Emotional burn-out and empathy fatigue are widely reported across a wide range of professions including care-workers, hospital workers, veterinary staff and medics.18,19 Such situations can challenge mental health including our emotional, psychological, and social wellbeing. This affects how people and animals think, feel, and act. It also inﬂuences how we handle stress, relate to others and make choices. In such situations it is important to promote a positive mental health approach allowing people to realise their full potential, cope with the stresses of life, work productively and make meaningful contributions to their communities. Figure 1. Compassion fatigue.

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37August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareEmotional challenges in our work with laboratory animalsSo, how can we make such working environment more ‘emotionally supportive’? Well recognised techniques for compassion skills are found in mindfulness meditation programmes which strengthen the need to provide personal space and tools to promote individual and group communication across all different professional layers. Even with short periods of compassion training, participants continue to feel empathy for the suffering of others but gain the capacity to feel positive emotions without feeling distress.20A practical insight into mindfulnessMindfulness can be described as being ‘fully aware of the present moment’, you are free from distraction and can be more focussed on even the simplest of tasks. Thoughts may still be ‘ﬂowing in the mind’ but you accept these thoughts without judgement or critical reﬂection. There are many resources which are freely available outlining the practice of mindfulness in more detail, these include books, online courses, apps and video tutorials. Many community groups or adult education centres (even organisational education CPD centres) offer courses on Mindfulness, many of them on a reduced budget or free of charge. The NHS also has a very useful resource centre on this topic https://www.nhs.uk/conditions/mental-health/self-help/tips-and-support/mindfulness/Mindfulness practiceStarting your day with a mindfulness practice can help charge your batteries with energy that you can draw on throughout the day. Spend time focusing inwards and connecting with yourself to charge yourself up. Choose from one of the suggested mindfulness practices listed below. This practice need not be long, even a few seconds to take some deep breaths can be helpful.(i) Three minute mindful breathing space– One minute for ACKNOWLEDGING what is happening and how you are feeling.– One minute for GATHERING your awareness around your breath.– One minute for EXPANDING awareness of your breath into your body – notice where you feel your breath most in your body.(ii) Mindful check-in– Whilst either sitting down or standing up, start to focus on your breath.– Once you feel comfortable focussing on your breath, move your focus into your body.– Notice any physical sensations you have.– If any of these sensations are overwhelming, move your focus to the feet – the feet are not generally affected by stress.– Go on to notice any thoughts, feelings or emotions you are having. There is no need to engage with them, just notice they are there.– Finally, return to your breath for the last few moments.Simple tips for boosting your mood1. Find the good stuffEach night, write down 3 things you are grateful for or enjoyed. Some people call this a ‘gratitude journal’, by writing down these thoughts it helps emphasise your feelings and focusses your thoughts. Take time to reﬂect on these thoughts and how they make you feel.2. Take a walkMoving our bodies and getting daylight (especially as the short days draw in during the winter months) helps to lift our mood and clears our mind. The simple pleasures of being more aware of our surroundings, interacting with the elements and exploring new places can have a positive effect on our mood.3. Ask for helpIf you have a problem or something is worrying you, then ask for support. Knowing when to ask for help is not a sign of defeat or failure, it is a sign of your inner strength than demonstrates that you aware of your mental health and are making decisions not to let it deteriorate. Asking others to help can build a connection, provides opportunity for discussion and gives them a wellbeing boost too! Think ‘Good Karma’ – when a person does something good and that individual’s positive actions seem to lead to positive consequences. Doing someone a favour activates the ‘feel-good reward centre’ in the brain, so not only does the person receiving the favour (or support) feel the beneﬁt, the person performing the action does too!Figure 2: Mindfulness

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38Animal Technology and Welfare August 2020The NHS website also contains some excellent ‘Stress Busting’ ideas and also links to ‘Stress busting apps’ https://www.nhs.uk/conditions/stress-anxiety-depression/reduce-stress/ The main author of this resource says the keys to good stress management are building emotional strength, being in control of your situation, having a good social network and adopting a positive outlook. Self-care – a mindful poem to reﬂect onDerek Walcott was a Caribbean poet and playwright who was awarded the Nobel Prize in Literature in 1992. His poem ‘Love after love’ is about being at ease with yourself, and loving who you are. It is also about owning all of your stories, experiences, strengths and weaknesses – and treating them with respect, compassion and love. It is often cited as a ‘Mindfulness poem’ and it serves as an anchor to reﬂect on.How to enhance workplace communication toolsAlongside your employer, you should recognise that you have a joint responsibility to look after your own wellbeing and that of your colleagues. Managers should be aware of triggers and risk factors for all team members and work to reduce them. Workplaces should foster and support a Culture of Care nurturing mental and physical safety, embracing OPENNESS and COMMUNICATION across all the team players.Such levels of empathetic openness can only proceed when staff feel emotionally/physically safe and valued.21 This caring professional attitude must be well supported by an organised institutional system and as such it translates into proactive management actions and good communication. Providing a safe space where the staff can share and reﬂect on any personal experiences at work, individually or in group and analysing them openly can inform learning towards an attitude of care. Such experiences can be, for example, monitoring, husbandry or clinical duties with the animals, reading a research article, managing order suppliers, attending a staff meeting, or a debrief with your manager. Team exploration and interactive reﬂection is crucial to improve care across the institution and build-up strength on staff expertise as individuals with different responsibilities will identify different issues and effects on their behaviour. Group reﬂection activities should be encouraged in the training schemes, along with protecting physical space and time for this, to support all staff categories, including senior management.Figure 3: Love after love, a mindful poem.Figure 4: Positive open communication is key to success.Communication across all staff levels including senior researchers is also crucial. Engaging communication across different levels of expertise will facilitate a broader perspective analysis e.g. understanding the best husbandry practice is as relevant as publishing a scientiﬁc paper.22 Hence any discussion or educational platforms must facilitate communication across the different roles and responsibilities in animal research. How to promote open discussions at workDisplaying emotions is complex in the working environment and possibly more complex within a laboratory animal set up which often feel formally focussed on compliance and biosafety regulations. “”Emotional challenges in our work with laboratory animals

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39August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareEmotional challenges in our work with laboratory animalsThis atmosphere would not seem to most to be an accommodating space for switching off and to openly reﬂect, sharing personal thoughts and concerns with colleagues. Thus, common areas need to be protected as generally this type of space in the units is prioritised for catering or bench administrative duties, resulting in limited social ‘relaxation’ space compounded with restricted natural lightning, (which can be beneﬁcial for mood enhancement). Outside of work there are often only limited spaces where staff feel comfortable discussing their work and its emotional impact. On a positive note, emotions can be contagious and as such, can be shared within the group; communication breaks for teams are crucial and must be encouraged and time protected. Promoting openness relies also on providing a tolerant, respectful and safe space!Laboratory animal professionals do not talk openly in public about their chosen career pathway for fear of disproval or personal security concerns. This may lead to feelings of ‘suppression’ and even ‘shame’ for working in a little understood professional sector being performed behind closed doors with little positive reports or career models openly highlighted. To help counteract this, many UK professional associations, such as The Institute of Animal Technology, have recently campaigned tirelessly for the recognition of animal care staff as ‘key workers’ in the biomedical ﬁeld in views of their instrumental work during the COVID-19 emergency.23,24Individual pro-action remains one of the key pillars of emotional balance. Techniques such as encouraging expression of emotion in a suitably controlled environment – openness without judgement, positive learning to personal experiences, changing towards optimist patterns, good judgement on speciﬁc situations, personal acceptance and avoid comparison with others (everyone is different!), build up pride in any small or large job, better involvement across your working routine, participative engagement with your colleagues and encourage emotional connections.Some organisations have chosen to display a voluntary ‘healthy workplace’ poster to help demonstrate their commitments to mental health and to encourage staff to talk openly about their feelings and any challenges they may face in the workplace. Such posters can be used to highlight the core principles and key resources every workplace should have.DiscussionA healthy workplace is vital to allow Animal Technologists to fulﬁl their professional obligations and continue to safeguard animal health and welfare and public health. It is vitally important to preserve the caring attitude displayed by animal staff and researchers. The Institute of Animal Technology has released supportive guidelines to support the mental health of Animal Technologists as they adapt to a range of new working measures but also highlighting the need to protect their work/life balance.25Figure 5: ‘Healthy workplace’ poster example.HEALTH AND WELLBEING – PROMOTING A POSITIVE CULTURE  Establish a good physical and mental wellbeing at the workplace, supportive of the needs of all team members. Personal safety always taking precedence, including over professional responsibilities- proactively manage workplace stress. Support, encourage and celebrate a good work-life balance - promote a culture of respecting personal health. Establish regular meetings (best one-to-one) to discuss current workload, agree priorities and raise other concerns in an informal environment; considering the value of resilience training, for individuals or the entire team. A culture of emotional safety, in which all team members’ mental health and wellbeing are supported. No tolerance of prejudicial, discriminatory or offensive language - ensure a fair, respectful and equal treatment of all team members. Mitigate the potential risks posed by emotional burden and empathetic fatigue, signposting appropriate helplines and support groups and making sure that all team members are aware of the support available. A positive workplace where staff communicates and shared values and attitudes Embrace the emotional challenges of our work with laboratory animals, being aware of the impact on personal and professional lives , and also the impact on the wider community .

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40Animal Technology and Welfare August 2020In this article we have also highlighted the importance for providing better opportunities for frank discussions on this sensitive topic, promoting better communication platforms. One of the UK’s leading mental health charities has the following quotation on its website: “In the past six years I have had counselling, a brief attempt at CBT [cognitive behavioural therapy] and routine meetings with mental health doctors but the thing I have found most helpful is open online forums full of people like me”. It is very normal to feel fearful, anxious, low or irritable. If you are having worrisome feelings, give yourself time to reﬂect on them and accept that they are normal reactions. Humans are programmed to respond to things that are threatening. If you can give yourself time to stay with your feelings for 60 seconds it will help them dissipate. Instead of getting upset by your own emotions, try to be kind to yourself and accepting of your emotional reactions. Small acts of kindness not only to others but to yourself can boost your mood. Why not treat yourself? The ‘Buddy Box’ subscription box service (https://www.blurtitout.org/buddybox) can be used as a ‘small treat’ to yourself. The contents of each box arrive each month and are designed to counter the pressures we face in modern life. Packed full of thoughtful, mood-lifting treats, the Buddy Box comforts, delights and gives you that warm, ‘I’ve been cared for’ feeling inside. In other words – it’s a hug in a box. However there are many other ways to practise ‘self-care on a budget’ and there are several good ideas here for you to consider https://www.blurtitout.org/2016/03/30/self-care-budget-10-things-try/The implementation of simple but effective approaches like secure discussion groups, virtual ‘coffee support groups’ and face to face training will represent an important step forward to assist with building emotional resilience. By learning that ‘It’s OK not to be OK’ and communicating personal emotions, it is still possible to be proud ambassadors for the care and welfare of the animals studied as vital research models. We feel it would be beneﬁcial to discuss how those closely working with animals can utilise some of the above-mentioned approaches to support managing emotion. To maintain good mental health and practise resilience techniques, it is likely to be necessary to improve staff’s self-conﬁdence at communicating any concerns on animal care and welfare and encourage greater openness, particularly across technical staff and researchers. The implementation of discussion platforms and resilience training opportunities that we have identiﬁed will improve not only animal welfare, staff wellbeing, but also the integrity of our research. References1 Bloomsmith, M. A., Perlman, J. E., Hutchinson, E. , Sharpless M. (2018) Behavioral Management Programs to Promote Laboratory Animal Welfare. In: Weichbrod RH, Thompson GA, Norton JN, eds. Management of Animal Care and Use Programs in Research, Education, and Testing. 2nd ed. Boca Raton (FL): CRC Press/Taylor & Francis; 63–82.2 Davis, H. (1996) How human/animal bonding affects the animals. In: Kru-lisch L Mayer S Simmonds RC eds. The Human\Research Animal Relationship. Greenbelt MD : Scientists Center for Animal Welfare, 67 – 75.3 Bayne, K. (2002) Development of the Human-Research Animal Bond and Its Impact on Animal Wellbeing, ILAR Journal, 43, 1, 4–9. https://doi.org/10.1093/ilar.43.1.4 4 Grimm, D. (2020) It’s heartbreaking.’ Labs are euthanizing thousands of mice in response to coronavirus pandemic. American Association for the Advancement of Science in Science. https://doi.org/10.1126/science.abb8633 5 Nowogrodzki, A. (2020) Cull, release or bring them home: Coronavirus crisis forces hard decisions for labs with animals. Nature. 580, 7801, 19. doi:10.1038/d41586-020-00964-y 6 Health & Safety Executive. (2019) Work-related stress, anxiety or depression statistics in Great Britain, 2019 https://www.hse.gov.uk/statistics/causdis/stress.pdf [Accessed October 2020]Figure 6: It’s okay, not to be okay. (image courtesy of A. Kerton)Emotional challenges in our work with laboratory animals

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41August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareEmotional challenges in our work with laboratory animals7 Home Ofﬁce. (2014) Guidance on the operation of the Animals (Scientiﬁc Procedures) Act 1986. Available at: https://www.gov.uk/guidance/guidance-on-the-operation-of-the-animals-scientiﬁc-procedures-act-1986 [Accessed October 2020]8 Ipsos MORI. (2016) Attitudes to animal research in 2016. Available at: https://www.ipsos.com/ipsos-mori/en-uk/attitudes-animal-research-2016 [Accessed October 2020]9 Cocker, F. & Joss, N. (2016) Compassion fatigue among healthcare, emergency and community service workers: a systematic review. Int J Environ Res Public Health 13, 1–18.10 Compassion Fatigue Awareness Project. (2017) Recognising compassion fatigue. Available at: http://www.compassionfatigue.org/pages/symptoms.html [Accessed October 2020]11 Scotney, R. L., McLaughlin, D., Keates, H. L. (2015) A systematic review of the effects of euthanasia and occupational stress in personnel working with animals in animal shelters, veterinary clinics, and biomedical research facilities. J Am Vet Med Assoc 247, 1121–1130. https://doi.org/10.2460/javma.247.10.1121. 12 Mathieu, F. (2007) Running on empty: compassion fatigue in health professionals. Available at: http://www.compassionfatigue.org/pages/RunningOnEmpty.pdf [Accessed October 2020]13 Klimecki, O., Singer, T. (2011) Empathic distress fatigue rather than compassion fatigue? Integrating ﬁndings from empathy research in psychology and social neuroscience. In: Oakley B, Knafo A, Madhavan G, et al. eds. Pathological Altruism. New York, New York: Oxford University Press, 1–23.14 Institute of Animal Technology (IAT) (2020) Let’s talk Euthanasia. Available at: https://6a18642f-f788-41d5-acd0-7bcff8782988.ﬁlesusr.com/ugd/a30180_a958a0feaf4b4904ba829846d330a9a9.pdf [Accessed October 2020]15 Davies, K., Lewis, D. (2010) Can caring for laboratory animals be classiﬁed as emotional labour? Animal Technology and Welfare 9, 1–6.16 Overhulse, K. A. (2002) Coping with lab animal morbidity and mortality: a trainer’s role. Lab Anim (NY) 31, 39–42.17 Rohlf, V., Bennett, P. (2005) Perpetration-induced traumatic stress in persons who euthanize nonhuman animals in surgeries, animal shelters, and laboratories. Soc Anim 13, 201-219. https://doi.org/10.1163/1568530054927753 18 Showalter, S. E. (2010) Compassion fatigue: what is it? Why does it matter? Recognizing the symptoms, acknowledging the impact, developing the tools to prevent compassion fatigue, and strengthen the professional already suffering from the effects. Am J Hosp Palliat Care 27, 239–242. https://doi.org/10.1177/1049909109354096 19 AALAS. Compassion Fatigue: The Cost of Caring. Human emotions in the care of laboratory animals. https://www.aalas.org/education/educational-resources/cost-of-caring [Accessed October 2020]20 Klimecki, O. M., Leiberg, S., Lamm, C., & Singer, T. (2013) Functional neural plasticity and associated changes in positive affect after compassion training. Cereb Cortex 23, 1552–1561.21 Heyhoe, J., Birks, Y., Harrison, R., O’Hara, J.K., Cracknell A. & Lawton R. (2016) The role of emotion in patient safety: Are we brave enough to scratch beneath the surface?. J R Soc Med.;109, 2, 52–58. doi:10.1177/014107681562061422 Reardon, S. (2016) A mouse’s house may ruin experiments. Nature, 530 7590, 1. https://doi.org/10.1038/nature.2016.19335 23 UK Research and Innovation.(2020) Guidance for the research and innovation communities. Available at: https://www.ukri.org/research/coronavirus/guidance-for-the-research-and-innovation-communities1/ [Accessed October 2020].24 Department for Education. (2020) Key workers in education letter. Available at: http://independenthe.com/wp-content/uploads/2020/04/200421-COVID-19-Keyworkers-Letter.pdf [Accessed October 2020].25 Institute of Animals Technology. (2020) It’s OK … not to be Okay … Let’s Talk Coping with Change … A Future of Hope …? Available at: https://www.iat. org.uk/;https://6a18642f-f788-41d5-acd0- 7bcff8782988.filesusr.com/ugd/a30180_ 84e91129693c42a79eca78dcfdd9acd7.pdf [Accessed October 2020]

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42Animal Technology and Welfare August 2020It’s OK … NOT TO BE OKAY …Let’s Talk Coping with Change … A Future of Hope …?Since the 11th of March, when WHO declared Covid-19 had become a pandemic and government advice led to a huge impact on businesses and personal lives. Businesses commenced lockdown measures and implemented pandemic contingency plans. Animal Techs and others associated with the industry were on the frontline continuing to provide excellent standards of care and welfare for our animals used in scientiﬁc research.For many Animal Technicians the implications of the contingency plans meant splitting into smaller teams with extra workloads, working on a rota basis and working more irregular hours than usual. Further pressures, such as being isolated from team members, scientists, managers and other colleagues who would usually encourage us and support us with day-to-day issues, have also had an impact on mental health and wellbeing. Some employees are trying to combine home schooling and other family responsibilities with work responsibilities, leading to a poor work/life balance.For many, factors such as social distancing and self-isolation, protecting vulnerable family members and anxiety over family and friends also causes extra pressure and uncertainty. Many employees are fearful about contracting the virus and many may have suffered bereavements during this time.Organisations are stating that they do not yet know exactly what the long term mental health impacts of Covid-19 will be. Early research (CIPD, May 2020 ) into the health impacts of lockdown found increased levels of fatigue, musculoskeletal conditions, poor work/life balance, reduced exercise and increased alcohol consumption. In addition to this, employees reported reduced motivation, loss of purpose, anxiety and isolation. Evidence from previous quarantine situations prior to the pandemic suggests that there are long lasting effects on mental health. Symptoms ranged from irritability and anger, to depression and post-traumatic stress symptoms.As many countries (including the UK) start to ease Coronavirus restrictions, BBC News reported (Butterly 2020) that mental health experts are starting to see increased anxiety about what life will be like after lockdown. While many people have developed their own routine of safety in lockdown, creating a bubble around themselves, physically distancing themselves from family and friends, there are bound to be uncertain feelings regarding what the future holds. What can Employers do?It is well known that many employees do not feel comfortable in speaking up about poor mental health; this is unlikely to change following the pandemic.Employers need to adapt a range of measures to support employees experiencing poor mental health as a result of COVID-19. These will range from supporting employees to regain an effective work/life balance and addressing fears about returning to work, right through to support for severe mental health conditions.Factors to consider as an Employer:l offering long-term mental health support for those continuing to work in this essential role as an Animal Technicianl continue to offer support for those who are currently working from home and may start to return to work on a phased basis over the coming weeks and monthsl provide an open culture where people feel able to talkl provide safe working environments for employees in order to comply with Covid-19 guidelines in the workplace It’s OK … NOT TO BE OKAY … Let’s Talk Coping with Change … A Future of Hope …?Mental Health Awarenesswww.iat.org.ukWhat can Employees do?l stay safe and follow government guidelinesl understand that it will be difﬁcult to get back into a routinel keep it simple, don’t rush back and overwhelm yourself with extra anxietyl be open and talk to friends, family and your employer if you are ﬁnding easing back into life after lockdown hardl continue to report symptoms and self-isolate if any symptoms are developedl be proud to have been an Animal Technician or in a supporting role during this time ensuring continued levels of excellent animal welfare standards while supporting vital researchFuture of Hope …There are many who identify with the theory that in all of the sadness, anxiety and struggles that Covid-19 has brought with it, there is a light at the end of the tunnel. Communities have come together in a way they have not before, caring, shopping, helping and looking after neighbours and friends who have needed it. People have reached out to the lonely and isolated though social media, window visits and doorstep chats, when required. Families have been able to spend more time together and there is an increased appreciation for all front line workers from lorry drivers, postmen, carers, etc ... to NHS staff . When you do start to develop your own strategy to ease out of lockdown and ﬁnd your own way forward through the Covid-19 blur, please remember that YOU are a key worker and you should be extremely proud to be an Animal Technician.Useful links:https://www.gov.uk/government/publications/support-for-those-affected-by-covid-19https://www.actionforhappiness.org/news/covid-19-how-to-respondhttps://www.mind.org.uk/information-support/coronavirus/coronavirus-and-your-wellbeing/https://www.nhs.uk/conditions/stress-anxiety-depression/References:Chartered Institute of Personnel and Development (CIPD 2020)Mind.org.ukhttps://www.bbc.co.uk/news/health-52443108Institute of Animal TechnologyCOUNCIL

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43August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareIt’s OK … NOT TO BE OKAY …Let’s Talk Coping with Change … A Future of Hope …?Since the 11th of March, when WHO declared Covid-19 had become a pandemic and government advice led to a huge impact on businesses and personal lives. Businesses commenced lockdown measures and implemented pandemic contingency plans. Animal Techs and others associated with the industry were on the frontline continuing to provide excellent standards of care and welfare for our animals used in scientiﬁc research.For many Animal Technicians the implications of the contingency plans meant splitting into smaller teams with extra workloads, working on a rota basis and working more irregular hours than usual. Further pressures, such as being isolated from team members, scientists, managers and other colleagues who would usually encourage us and support us with day-to-day issues, have also had an impact on mental health and wellbeing. Some employees are trying to combine home schooling and other family responsibilities with work responsibilities, leading to a poor work/life balance.For many, factors such as social distancing and self-isolation, protecting vulnerable family members and anxiety over family and friends also causes extra pressure and uncertainty. Many employees are fearful about contracting the virus and many may have suffered bereavements during this time.Organisations are stating that they do not yet know exactly what the long term mental health impacts of Covid-19 will be. Early research (CIPD, May 2020 ) into the health impacts of lockdown found increased levels of fatigue, musculoskeletal conditions, poor work/life balance, reduced exercise and increased alcohol consumption. In addition to this, employees reported reduced motivation, loss of purpose, anxiety and isolation. Evidence from previous quarantine situations prior to the pandemic suggests that there are long lasting effects on mental health. Symptoms ranged from irritability and anger, to depression and post-traumatic stress symptoms.As many countries (including the UK) start to ease Coronavirus restrictions, BBC News reported (Butterly 2020) that mental health experts are starting to see increased anxiety about what life will be like after lockdown. While many people have developed their own routine of safety in lockdown, creating a bubble around themselves, physically distancing themselves from family and friends, there are bound to be uncertain feelings regarding what the future holds. What can Employers do?It is well known that many employees do not feel comfortable in speaking up about poor mental health; this is unlikely to change following the pandemic.Employers need to adapt a range of measures to support employees experiencing poor mental health as a result of COVID-19. These will range from supporting employees to regain an effective work/life balance and addressing fears about returning to work, right through to support for severe mental health conditions.Factors to consider as an Employer:l offering long-term mental health support for those continuing to work in this essential role as an Animal Technicianl continue to offer support for those who are currently working from home and may start to return to work on a phased basis over the coming weeks and monthsl provide an open culture where people feel able to talkl provide safe working environments for employees in order to comply with Covid-19 guidelines in the workplace It’s OK … NOT TO BE OKAY … Let’s Talk Coping with Change … A Future of Hope …?Mental Health Awarenesswww.iat.org.ukWhat can Employees do?l stay safe and follow government guidelinesl understand that it will be difﬁcult to get back into a routinel keep it simple, don’t rush back and overwhelm yourself with extra anxietyl be open and talk to friends, family and your employer if you are ﬁnding easing back into life after lockdown hardl continue to report symptoms and self-isolate if any symptoms are developedl be proud to have been an Animal Technician or in a supporting role during this time ensuring continued levels of excellent animal welfare standards while supporting vital researchFuture of Hope …There are many who identify with the theory that in all of the sadness, anxiety and struggles that Covid-19 has brought with it, there is a light at the end of the tunnel. Communities have come together in a way they have not before, caring, shopping, helping and looking after neighbours and friends who have needed it. People have reached out to the lonely and isolated though social media, window visits and doorstep chats, when required. Families have been able to spend more time together and there is an increased appreciation for all front line workers from lorry drivers, postmen, carers, etc ... to NHS staff . When you do start to develop your own strategy to ease out of lockdown and ﬁnd your own way forward through the Covid-19 blur, please remember that YOU are a key worker and you should be extremely proud to be an Animal Technician.Useful links:https://www.gov.uk/government/publications/support-for-those-affected-by-covid-19https://www.actionforhappiness.org/news/covid-19-how-to-respondhttps://www.mind.org.uk/information-support/coronavirus/coronavirus-and-your-wellbeing/https://www.nhs.uk/conditions/stress-anxiety-depression/References:Chartered Institute of Personnel and Development (CIPD 2020)Mind.org.ukhttps://www.bbc.co.uk/news/health-52443108Institute of Animal TechnologyCOUNCIL

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44Animal Technology and Welfare August 2020AbstractThe article describes the care of the Olive python (Liasis olivaceus; including information on housing, feeding, handling, as well as the general health care of these animals. Some of the information presented is derived from Standard Operating Procedures which have been written by the author. It is hoped that the information presented may assist novice Animal Technicians and zookeepers who may be responsible for the care of these species in an animal research facility or a zoological setting. This work has been made possible under Scientiﬁc Licence number SL100342, held by Gary Martinic and issued by the National Parks and Wildlife Service of New South Wales. Background The Olive python is one of Australia’s largest snakes. Fully grown adults can vary in size from 2-5 metres and weigh between 10-20 kg. The body is thick and muscular and the tail tapers to a thin point. Their colouring ranges from a single colour of olive green to greenish-brown, reddish-brown or ‘off-white’. They generally have a whitish belly and pale lips. The distribution of Olive pythons occurs from the Pilbarra region of Western Australia through to Northern Queensland. They inhabit a variety of habitats from arid plains to sub-humid areas and are usually found in rocky areas and gorges especially those associated with water courses. These ground-dwelling snakes often inhabit rocks and caves and may be found in hollow logs. Like other pythons, these snakes are carnivores and kill their prey, usually birds, mammals and reptiles, by coiling around the prey and suffocating them before ingesting them whole. They are not venomous. Large, adult Olive pythons often consume mammals as large as rock wallabies. Noting that rock wallabies range in weight from 1-12 kg depending on the species. For instance the Monjon is Husbandry and healthcare of the Olive python (Liasis olivaceus)GARY MARTINIC Reptile House, K1 Animal Facility, Research Laboratories and Animal Facilities, Western Sydney University, Hawkesbury Campus, Richmond NSW AustraliaCorrespondence: G.Martinic@westernsydney.edu.au3 Figure 1. The inquisitive stare of an adult Olive python, in shed, as seen through the glass panel doors of its enclosure. Some grains of sand can be seen falling from the underside of its lower jaw. (Courtesy: G.Martinic) Background The Olive python is one of Australia’s largest snakes. Fully grown adults can vary in size from 2-5 metres and weigh between 10-20 kg. The body is thick and muscular and the tail tapers to a thin point. Their colouring ranges from a single colour of olive green to Figure 1. The inquisitive stare of an adult Olive python, in shed, as seen through the glass panel doors of its enclosure. Some grains of sand can be seen falling from the underside of its lower jaw. (Courtesy: G. Martinic) the smallest rock wallaby weighing an average of 1.3kg, whereas some males of the Yellow-footed rock wallaby species can weigh between 11-12kg.Clutches of up to 19 eggs are laid, the incubation period being approximately 7 weeks. The sex of individuals is determined by probing the inside of the cloaca (see Figure 2; how to sex snakes using probes and obtain Animal Technology and Welfare April 2021

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45August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfarethe training video link cited under references). The type of probe used is usually a 12-15 cm long surgical grade, stainless steel instrument with a round polished 5 mm ball tip (smaller ball tips are used for smaller snakes). When inserting the probe into the cloaca, the depth that the probe can be inserted indicates the presence or absence of hemipenes. (Hemipenes are a pair of intromittent organs in male snakes designed to deliver sperm when erect during mating. They are usually held inverted within the body of the snake but are everted outside of the body for mating). If the probe only enters 1-3 scales deep, then there are no hemipenes and the snake is female. If the probe can be inserted much further (9-15 scales deep), then this indicates the presence of hemipenes and the snake is male. The K1 reptile facility specimens are all captive bred and have an expected captive lifespan of approximately 30 years. The conservation status of the Olive python is currently classiﬁed as ‘vulnerable’. Loss of habitat and the depletion of food sources due to foxes and Australia’s feral cat population are the main threats to this species in the wild. Also, commonly mistaken for the very venomous King brown snake (Pseudechis australis), the Olive python is often killed by uninformed humans due to mistaken identity.Figure 2. Sexing of snakes using a probe. (Image Courtesy: Zoological Society London)1. Housing and environmental conditionsFor one adult Olive python, a 2.4m x 600 x 600 mm enclosure is recommended (Figure 3). The one used at the Reptile Facility, Western Sydney University has these dimensions and was custom-built of large solid wooden panels forming the sides, top and bottom of the enclosure with solid shelving providing three levels and it has two lockable glass panel doors at the front of the enclosure. This is large enough for the animal to uncoil and stretch out. A purpose-built solid wood hide box is also provided with an entrance opening large enough for the animal to enter without scratching its scales. The water dish should be sufﬁciently heavy to prevent it being tipped over and should be positioned at the cooler end of the enclosure away from the heat 6 Figure 2. Sexing of snakes using a probe. (Image Courtesy: Zoological Society London)1. Housing and Environmental Conditions For one adult Olive python, a 2.4m x 600 x 600 mm enclosure is recommended (Figure 3). The one used at the Reptile Facility, Western Sydney University, has these dimensions and was custom-built of large solid wooden panels forming the sides, top and bottom of the enclosure with solid shelving providing three levels and it has two lockable glass panel doors at the front of the enclosure. This is large enough for the animal to uncoil and stretch out. A purpose-built solid wood hide box is also provided with an entrance opening large enough for the animal to enter without scratching its scales. The water dish should be sufficiently heavy to prevent it being tipped over globes. The water container must be large enough for the animal to soak itself in, if it chooses. Washed sand is used as the substrate on each of the three levels of the enclosure. Prior to using washed sand, we used commercially-available Butcher’s paper, when this was used we placed the paper 2-3 layers thick into each level of the enclosure all of which was replaced weekly. In the case of washed sand, a 1 inch (25 mm) layer across the entire ﬂoor is provided and is spot cleaned daily and replaced completely every 3 months. The reptile rooms are maintained at constant temperature of 22oC (+/- 2oC) and a relative humidity of between 40-60%. The air conditioning system provides ventilation at between 10-12 changes/hour, a positive pressure is maintained. Figure 3. Olive python’s three-level enclosure showing wide opening glass panel doors at the front. The snake can be seen through the glass doors coiled up at the front of its enclosure. Note the sand substrate, water dish, ‘soaking’ dish, a custom-made wooden hide box (right), a cardboard hide and enclosed infra-red and lighting globes installed just below the top shelf. (Courtesy: G. Martinic) Enclosure heatingProviding adequate temperature gradients within the Olive python’s enclosure is essential for its health and wellbeing. They require a temperature gradient so they can move around the enclosure to where the temperature that suits them. Olive pythons require a hot basking spot maintained at approximately 34˚C. The cool end of the enclosure should be maintained between 24-26˚C. The enclosure should not drop below 18-21˚C at night. Temperatures should be regulated by a thermostat and checked daily to ensure the thermostat and globes are functioning correctly. 8 Figure 3. Olive python’s three-level enclosure showing wide opening glass panel doors at the front. The snake can be seen through the glass doors coiled up at the front of its enclosure. Note the sand substrate, water dish, ‘soaking’ dish, a custom-made wooden hide box (right), a cardboard hide and enclosed infra-red and lighting globes installed just below the top shelf. (Courtesy: G.Martinic) Husbandry and healthcare of the Olive python (Liasis olivaceus)

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46Animal Technology and Welfare August 2020Enclosure lightingAdequate lighting in the artiﬁcial environment of an enclosure, is critical to for most pythons as it is for other reptiles. A UVB 25-Watt globe provides suitable artiﬁcial UVB light and is connected to a timer to create a day and night cycle with 10 hours of UVB light (7 am – 5 pm). Higher intensity UVB globes should not be used in Olive python enclosures. Lights must be covered with a wire mesh shield to prevent the pythons from coming into contact with the globe or breaking them. In the unlikely occurrence of a bulb spontaneously bursting the wire shield will minimise the risk of broken glass being distributed into the enclosure. Olive pythons also beneﬁt from short periods of unﬁltered, natural light and are taken outside 1 – 2 times a fortnight for supervised sunning.Room lightingRoom lighting within the separate reptile holding rooms is maintained on an artiﬁcial photoperiod set at 12:12 hrs light/dark, provided by individual electronic light timer switches (with manual override features) to each room (Figure 4).10 periods of unfiltered, natural light and are taken outside 1 – 2 times a fortnight for supervised sunning. Room Lighting Room lighting within the separate reptile holding rooms is maintained on an artificial photoperiod set at 12:12 hrs light/dark, provided by individual electronic light timer switches (with manual override features) to each room (Figure 4). Figure 4. Electronic Light Timer Switch as used in each reptile holding room to regulate room lighting Figure 4. Electronic Light Timer Switch as used in each reptile holding room to regulate room lighting.HandlingAs they mature in captivity, they can become quite docile snakes especially when handled appropriately. The female Olive python (Figure 5) that we have at the Reptile House, which we have affectionately named ‘Scarf’ has been cared for by us for over 15 years. Scarf, at one point reached a bodyweight of 25kg, so action was taken to reduce her bodyweight to a more reasonable weight for her species. At the time of writing this article, Scarf weighed approximately 16kg and had a body length of 3.5m. Handlers must be aware that bites can occur during feeding and handle pythons accordingly to minimise risk. However, there has been no recorded instance of Scarf biting any of our Animal Technologists during the last 15 years. Olive pythons can be difﬁcult to remove while biting and once the snake has bitten, the sustained injury is not likely to worsen while the animal is attached but improper or hasty attempts to remove the snake may cause a larger laceration. Running cold water over their body or into the mouth can result in the python releasing its hold. Alcohol from an Alcohol wipe is also highly effective. Serious injury is unlikely but bites from larger specimens can be very painful.Some pythons, such as the much smaller Children’s pythons, which we also maintain, can often strike out as soon as the enclosure door is opened, especially in the days immediately preceding scheduled feeding days when the snake is hungry. It is good practice to remove the snake from its normal enclosure before you feed them, this way they do not associate the enclosure 13 Figure 5. Photograph of ‘Scarf’ our Olive python shown on the grassed area outside of the K1 Animal Facility, sunning itself alongside the author. We try and provide regular periods of ‘sunning’ for each of our pythons on a weekly basis. The photo also gives a reasonable indication of the size and length that these species can grow to when compared to an average-sized human (Courtesy: G.Martinic) Figure 5. Photograph of ‘Scarf’ our Olive python shown on the grassed area outside of the K1 Animal Facility, sunning itself alongside the author. We try and provide regular periods of ‘sunning’ for each of our pythons on a weekly basis. The photo also gives a reasonable indication of the size and length that these species can grow to when compared to an average-sized human. (Courtesy: G. Martinic)Husbandry and healthcare of the Olive python (Liasis olivaceus)

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47August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareHusbandry and healthcare of the Olive python (Liasis olivaceus)opening with feeding. When opening the enclosure to handle or remove a python, the handler should ﬁrst gently tap the snakes head with a thick gardening style glove. The glove cannot be mistaken for food and the snakes that we have in captivity have been conditioned such that this tap indicates handling rather than feeding. During periods of ‘sunning’ when the python is outside the enclosure (Figure 6), if it begins to coil around an arm or hand, the handler should keep the snake moving and support the python’s whole bodyweight; otherwise the python may adopt feeding behaviours and bite once the arm or hand is restricted. It is not recommended to handle pythons if they have been fed in the previous 24 hours. It is also not recommended to handle pythons if they are ‘blue’ and about to commence shedding. (See Skin Shedding section for a description of ‘blue). Packing and transportOn a few occasions throughout each year, our reptiles are used in static and live handling displays by our academic and technical staff in events such as University Open days, for teaching purposes and for public engagement events. When this occurs the reptiles need to be safely packed and transported between locations. Smaller lizards are normally placed into and transported in breathable cotton bags (one animal per bag) which are placed inside rigid plastic containers. For pythons that are too large for a cotton bag, they need to be carefully 14 Figure 6. The distinct almost glassy ‘bluish’ hue can be seen on this Olive python in the bright sunlight, making the skin look smoother than other species. The Olive python has a very high mid-body scale count of 61-72 scales. (Courtesy: G.Martinic) Packing and Transport On a few occasions throughout each year, our reptiles are used in static and live handling displays by our academic and technical staff in events such as University Open days, for teaching purposes and for public engagement events. When this occurs the reptiles need to Figure 6. The distinct almost glassy ‘bluish’ hue can be seen on this Olive python in the bright sunlight, making the skin look smoother than other species. The Olive python has a very high mid-body scale count of 61-72 scales. (Courtesy: G. Martinic)Figure 7. A large, mobile purpose-designed and built crate with a secure lid and ventilation screen used for transporting our large Olive python. 16 Figure 7. A large, mobile purpose-designed and built crate with a secure lid and ventilation screen used for transporting our large Olive python. Hygiene Infection control is one of the basic principles of good animal care. Staff should wash their hands and change gloves between enclosures. They should use a quality disinfectant-cleaner like F10sc (a broad-spectrum veterinary disinfect containing a combination of benzalkonium chloride and polyhexamethyline biguanide) to clean cages and furniture regularly. This will reduce the risk of bacterial transported in a large, mobile purpose-designed crates (Figure 7) that have a secure lid and a ventilation screen. They are always transported in air-conditioned vehicles by staff who have been appropriately trained to perform this task.

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48Animal Technology and Welfare August 2020HygieneInfection control is one of the basic principles of good animal care. Staff should wash their hands and change gloves between enclosures. They should use a quality disinfectant-cleaner like F10sc (a broad-spectrum veterinary disinfect containing a combination of benzalkonium chloride and polyhexamethyline biguanide) to clean cages and furniture regularly. This will reduce the risk of bacterial build up and the risk of Salmonella in particular. Staff should remove shed skins, uneaten foods and droppings daily and change water every two days. Food and water dishes should be washed thoroughly and enclosure substrates changed weekly. Staff should wash their hands thoroughly before and after handling reptiles.Skin sheddingOlive pythons have a very high mid-body scale count often making their skin look smoother than other python species (Figure 5) and sometimes providing a somewhat ‘glassy’ looking appearance when being sunned. It should be remembered that healthy Olive pythons will shed regularly, particularly when they are growing. Pythons, especially the larger ones, should not be handled during the shed as they can be defensive during this time. For this reason, the python’s shedding is recorded and enclosures of snakes about to shed are labelled ‘blue’. Snakes are identiﬁed as ‘blue’ when the brille (the scale covering the eye) becomes a milky blue and/or their scales generally become dull indicating they are about to shed. After three to four days, the eyes become clear again and the snake begins seeking out rough surfaces in its enclosure such as branches and rocks (which should be relatively smooth – not pumice) and should be readily accessible, this may include course or naturally rock-shaped hides. The shedding will progress from nose to tail and takes between 7 to 14 days.Olive pythons should not be handled if they are showing signs of an impending shed or are actively shedding. Snakes will generally not eat during a shed. Force-feeding during this time is not necessary and in fact can be harmful. Once complete, the shed skin should be removed and the snake checked for a complete shed, including the brille (eye scales).Incomplete shedsThere are many reasons for the shedding process to be incomplete or improper, referred to as dysecdysis. The most common causes are related to poor husbandry and/or nutrition. Dysecdysis is a symptom of another problem and not a primary problem in itself. Persistent or incomplete sheds should be inspected by an experienced reptile veterinarian or experienced herpetologist. This will help to rule out medically treatable causes such as mites or bacterial infections of the skin. Other causes of dysecdysis include: trauma, dermatitis, malnutrition, and over-handling. The veterinarian will advise the appropriate treatments once the underlying cause has been determined.Husbandry Techniques used to assist with incomplete shedsHumidity is very important for reptiles with requirements varying from species to species. Most snakes require an environment of 50% to 70% humidity. Incomplete sheds can often be managed by increasing humidity. Spraying with luke-warm water (approx. 35-40˚C) from head to tail on a daily basis may be beneﬁcial. Butchers paper can also be soaked to increase the humidity of a speciﬁc enclosure. Snakes which retain their shedding for an extended period can be lightly sprayed with ‘Shed-Ezi’ spray or similar product on the residual scale which can then be gently rubbed off.Sometimes the snake may be placed in a large container with warm water (up to 43˚C) deeper than the girth of the snake to allow submersion but shallow enough such that it can easily keep its head above water while resting on the bottom of the container. Never leave a soaking snake unattended. After 10 – 15 minutes soaking in the tub the residual shed should be easily removed with gentle rubbing. Retained eye caps (or spectacles/brille) can be very dangerous for snakes. They can harbour pathogenic (potentially disease-causing) bacteria as well as making it difﬁcult for the snake to see. Removing them is not difﬁcult but can permanently damage the cornea of the snake if not done correctly. Staff must ﬁrst be trained in this procedure before attempting to remove retained eye-caps, or otherwise organise a consultation with an experienced herpetologist or reptile veterinarian to remedy the condition.FeedingIn the wild, the Olive python’s diet consists of birds, mammals and other reptiles but in captivity they can be fed exclusively on large rodents (mainly rats). As they grow, they are fed progressively larger feeds starting with adult mice, then progressing eventually to large 300-400g adult rats. The adult Olive python in the K1 Reptile House is fed primarily adult rats, although on occasion they may be fed adult Guinea pigs or even rabbits, when available. Adults are generally fed 5% of their bodyweight weekly (or 10% fortnightly) unless ‘blue’.The food (rats) is always frozen for at least 48 hours before use and thawed freshly in a tub of hot water for over an hour, then dried (Figure 8) Ensure the prey is fully thawed and dry before feeding. Dry the prey with paper towel and allow it to cool for 5-10 minutes before offering it to ensure the food is not too hot. The prey Husbandry and healthcare of the Olive python (Liasis olivaceus)

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49August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareHusbandry and healthcare of the Olive python (Liasis olivaceus)should be presented to the snake using forceps, tongs or a gloved hand. They will generally strike at it very quickly and coil around it. If the snake does not strike, the food item can be left in the enclosure while the other snakes are feeding. If the snake does not constrict or consume the food within 30 minutes the food should be removed and disposed of. Feeds and refusals should be recorded on a room activity log sheet for that animal room. HealthcareEven with proper care Olive pythons may become sick. It is important to seek specialist reptile veterinary care before an illness progresses too far. Observation is the best way to recognise ill health, watch for changes in behaviour, weight loss, loss of appetite, changes in colour, changes in faeces, changes in muscle tone or coordination, incomplete or absence of ecdysis (shedding) and persistent aggression. A range of conditions and illnesses that can afﬂict Olive pythons, have been presented below for the beneﬁt of the Animal Technologists and Zookeepers:InfectionsSalmonellosis – Salmonella bacteria are readily found in the environment and can cause disease when present in large volumes in food. It is often associated with poultry. Although most pythons asymptomatically carry and shed some serotypes of salmonella, large gut loads of serotypes that they do not usually harbour, can cause disease which is often fatal. Salmonella causes gastrointestinal infections. Signs of gastrointestinal infection are weight loss, lack of appetite, and foul smelling diarrhoea, and vomiting. This type of illness must be treated by a veterinarian.Stomatitis – Commonly referred to as mouth rot or canker, usually results from poor husbandry and sanitation practices. It may appear when a snake’s immune system has been weakened by a concurrent disease (e.g., pneumonia), low temperatures, or excessive humidity. Symptoms of stomatitis include accumulation of cheesy material along the gums and around the tongue sheath, blood spots and bruising in the gums, an inability to shed skin, dribbling saliva, in severe cases swelling of the gums and subcutaneous tissue along the jaw may be seen, possibly spreading to much of the head and neck. Specialist veterinary advice should be sought if stomatitis is suspected.Necrotising Dermatitis – Often referred to as Scale Rot and Blister Disease, often results from unhygienic conditions and excessive dampness, most commonly affecting reptiles that come from dry, arid regions of Australia. Occasionally, a deﬁciency of vitamins A and C can be a contributing factor. Symptoms of scale rot include: yellow, red or greenish-black discolouration of 21 of. Feeds and refusals should be recorded on a room activity log sheet for that animal room. Figure 8. Preparation of food for the snakes - photograph showing four adult rats (top) and three adult mice (bottom; used for smaller pythons) drying in a tub after having been soaked in warm water (in the tub at left) then being thoroughly damped dry with paper towel and left to air dry for up to 10 mins before feeding. The long stainless steel forceps (used to present the food to the snake) can be seen sitting over the tubs (Courtesy: G.Martinic) Figure 8. Preparation of food for the snakes – photograph showing four adult rats (top) and three adult mice (bottom; used for smaller pythons) drying in a tub after having been soaked in warm water (in the tub at left) then being thoroughly damped dry with paper towel and left to air dry for up to 10 mins before feeding. The long stainless steel forceps (used to present the food to the snake) can be seen sitting over the tubs. (Courtesy: G. Martinic)

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50Animal Technology and Welfare August 2020the scales, particularly along the underside, softening or swelling of the skin surface caused by serum seeping through, sloughing of affected skin exposing subcutaneous tissue, ﬂuid-ﬁlled blisters in the scales, again primarily on the ventral surfaces, and bruising due to blood in the tissue beneath the scales in advanced cases. Seek veterinary advice if dermatitis is suspected.Respiratory infectionsPneumonia – respiratory disease or pneumonia is quite common when conditions are too cold or damp for reptiles. While the condition is normally the result of bacterial infection, lungworms, fungal disease and tumours can also cause similar signs. Diagnosis may require a lung wash or radiology. Symptoms of pneumonia include: open-mouthed breathing, resting of the head in elevated positions, tongue tips stick together or snake is unable to ﬂick its tongue, gurgling sound while breathing, accumulation of frothy mucus at the back of the throat. Specialist veterinary advice should be sought if pneumonia is suspected. Fungal infections can be exacerbated by a warm and damp environment. These infections can occur in a cut or scrape and should be treated with an antifungal ointment under the direction of a veterinarian.Parasite infestationsHelminthic (internal parasite) Infestations – Olive pythons can become infected with internal parasites, notably nematodes (roundworms) and cestodes (tapeworms). These infections can be treated with Panacur™ (fenbendazole) for the nematodes and Drontal™ (Praziquantel, Pyrantel Embonate and Febantel). dog worming tablets for the tapeworms. These medications can be inserted into food and fed to the adults. This should only be done under veterinary or experienced herpetologist instruction.Ectoparasitic (external parasite) Infestations – Olive pythons can become infested with mites although this is unlikely in the reptile house unless new animals are introduced. However, mites could be inadvertently brought in by a student or staff member if proper hygiene controls are not followed. Symptoms of ectoparasite infestation include: lying in the water bowl, excessive rubbing against terrarium furnishings, skin shedding which is slow, uneven or does not occur, the presence of very small red-brown insects on the skin. In the case of mites, when a white pillowcase is placed in terrarium overnight, mites will be visible moving slowly on the bottom the following morning. Mites are extremely difﬁcult to eliminate and require a two-stage response, which is to treat the animal, as well as the habitat, under veterinary direction.AcknowledgementsI gratefully acknowledge the dedication and skilful assistance of our Animal Technologists from the Animal Facilities team. Technical Support Services including Nikola Mills, Lauren Hughes, Alex Hosking, Kieran Burns, Vamsi Inampudi, Amy Woodley, Sophie Ball and Lauren Grote. I thank Gavin McKenzie, Cluster Manager – Institutes, at the University, for reviewing the manuscript and for supporting our work. References1 Laszlo, J. (1975) Probing as a practical method of sex recognition in snakes. International Zoo Yearbook 15:178-179.Also See: ‘How to Probe Sex Snakes’ on YouTube by Ultimate Exotics: https://www.youtube.com/watch?v=uKlW6DXiElwBibliographyEhmann, H. (1992). Encyclopedia of Australian Animals. Reptiles. Australian Museum, Angus & Roberston, Sydney.Greer, A.E. (1989). The Biology and Evolution of Australian Lizards. Surrey Beatty & Sons, Sydney.NSW Department of Environment and Climate Change, (2008). ‘Hygiene protocol for the control of disease in captive snakes’, NSW Department of Environment and Climate Change, Sydney.OEH (2013). Code of Practice for the Private Keeping of Reptiles, State of NSW and Ofﬁce of Environment and Heritage, Sydney, ISBN: 978 1 74293 323 8.Weigel, J. (1988). Care of Australian Reptiles in Captivity, Reptile Keepers’ Association, Gosford.Husbandry and healthcare of the Olive python (Liasis olivaceus)

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51August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2021 Animal Technology and WelfarePAPER SUMMARY TRANSLATIONSCONTENU DE LA REVUEManipulation de souris à l’aide de gants pulvérisés avec un désinfectant pour les mains à base d’alcool: effets aigus sur le comportement de la souris NOELIA LOPEZ-SALESANSKY, DOMINIC J WELLS, NATALIE CHANCELLOR, LUCY WHITFIELD, CHARLOTTE C BURN Correspondence: cburn@rvc.ac.uk Résumé Les alcools sont couramment utilisés dans les installations de laboratoire pour désinfecter les mains, l’équipement et les environnements de laboratoire. L’effet sur les souris est inconnu. Nous avons donc observé des souris mâles et femelles C57BL/6J et BALB/c pendant et après leur manipulation avec des gants en nitrile qui ont été vaporisés avec un désinfectant à 70 % d’alcool (environ 67 % d’éthanol, ~3 % de méthanol et 30 % d’eau), ou qui n’ont pas été pulvérisés. Nous avons émis l’hypothèse que, si les souris considéraient ce désinfectant pour les mains comme un désinfectant aversif, son application sur les gants avant de les manipuler augmenterait les indicateurs comportementaux de peur ou de défense ; cela pourrait également affecter les interactions sociales et le toilettage dans les deux sens. Les souris manipulées avec des gants aseptisés étaient plus nombreuses à se relever le long de la paroi de la cage et à adopter un comportement d’auto-toilettage, de toilettage social, de reniﬂage des autres souris de la même cage et de consommation de nourriture/d’eau parmi une ou les deux souches de souris. Chez les mâles, la manipulation avec des gants aseptisés réduisait également l’agression initiale dans la cage, qui était remplacée par le toilettage, mais le fait de savoir si l’agression avait vraiment diminuée ou était simplement retardée n’a pas été établi. Aucun effet statistiquement signiﬁcatif du traitement sur les comportements d’évitement n’a été démontré lors d’un test d’interaction avec la main. L’enfouissement défensif se produisait avec les gants aseptisés et les gants de contrôle au cours du premier test d’interaction avec la main et a considérablement diminué au cours des 4 semaines de l’étude, ce qui suggère un effet de nouveauté. Les résultats indiquent que la manipulation de souris avec un désinfectant pour les mains à base d’alcool affecte leur comportement , notamment en ce qui concerne les interactions sociales, bien que la réplication soit nécessaire car il n’a pas été possible que l’observateur ait connaissance du traitement. D’autres recherches sont nécessaires pour évaluer les effets à long terme de l’utilisation de désinfectant pour les mains à base d’alcool et de désinfectants de rechange lors de la manipulation de souris de laboratoire aﬁn de formuler des recommandations de rafﬁnement. Mots-clés. Comportement des animaux; bien-être animal; désinfectant; manipulation; hygiène; souris

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52Animal Technology and Welfare August 2020Rapport de la réunion de 2020 du Groupe de protection des rongeurs de la RSPCA/UFAWCHLOE STEVEMS, PENNY HAWKINS, TOM V SMULDERS, AILEEN MACLELAN, LARS LEWEJOHANN, PAULIN JIRKOF, JACKIE BOXALL, HELEN MURPHY, CARLEY MOODY, PATRICIA V TURNER, I J MAKOWSKA, CHARLOTTE INMAN Résumé Le Groupe de protection des rongeurs et des lapins de la RSPCA/UFAW tient chaque automne depuis 27 ans une réunion d’une journée aﬁn de permettre à ses membres de discuter de la recherche actuelle sur le bien-être, d’échanger des points de vue sur les questions de bien-être et de partager des expériences de la mise en œuvre des 3R de remplacement, réduction et rafﬁnement en ce qui concerne l’utilisation de rongeurs. L’un des principaux objectifs du groupe est d’encourager la réﬂexion concernant l’expérience de vie entière des rongeurs de laboratoire, en veillant à ce que chaque impact négatif potentiel sur leur bien-être soit examiné et minimisé.Cette année, la réunion s’est déroulée en ligne pour la première fois et a réuni plus de 400 délégués de près de 40 pays. Elle avait pour thème les « expériences cumulées », avec des sessions sur « la science de la gravité cumulative » et « les rafﬁnements pratiques permettant de réduire la gravité et de promouvoir le bien-être ». Les présentations comprenaient un discours d’introduction qui expliquait la raison pour laquelle les expériences cumulatives sont importantes et la manière dont les expériences positives et négatives qui s’accumulent au cours de la vie d’un animal sont susceptibles d’avoir des répercussions à long terme sur son bien-être. D’autres discussions ont porté sur les différentes façons de reconnaître et d’évaluer la gravité cumulative, les effets cumulatifs des petits rafﬁnements et le concept de « belle vie » ainsi que sa signiﬁcation pour les rongeurs de laboratoire. L’unité de réglementation des animaux utilisés à des ﬁns scientiﬁques (ASRU) du Home Ofﬁce a également fourni une mise à jour concernant la façon dont les expériences cumulatives inﬂuencent la gravité vécue par les animaux utilisés à des ﬁns scientiﬁques. La journée s’est terminée par une séance de discussion interactive sur les moyens d’identiﬁer les souffrances cumulées chez les rongeurs encagés. Ce rapport résume la réunion et se termine par une liste de points d’action que les lecteurs peuvent envisager de soulever au sein de leurs propres établissements.Mots clés. Les 3R, rongeurs, lapins, expérience cumulative, sévérité, rafﬁnement★ ★ ★Déﬁs émotionnels dans le cadre du travail réalisé avec les animaux de laboratoire: outils permettant de prendre soin des autres et de vous-même ANGELA KERTON ET JORDI L TREMOLEDA Correspondence: angela@learningcurvedevelopment.co.uk Résumé Il arrivera inévitablement à la plupart des personnes qui travaillent avec des animaux de laboratoire de former parfois des liens avec les animaux dont elles s’occupent. Ces relations renforceront positivement les soins et le bien-être animal, mais elles posent également des déﬁs émotionnels importants qui ont été clairement exposés par certaines règles de gestion associées à la pandémie de COVID-19. Il est essentiel que le secteur reconnaisse l’existence de ces liens et fournisse des mécanismes de soutien institutionnel visant à aider les techniciens animaliers à faire face aux déﬁs émotionnels inhérents à leur profession. La logistique de travail actuellement associée à la pandémie de COVID-19 pose d’autres déﬁs, dont notamment la délégation des responsabilités, la séparation des équipes de travail et la gestion des stocks en cas d’urgence, pour n’en nommer que quelques-uns, ainsi que les déﬁs individuels en matière de santé et de relations sociales, économiques et personnelles. Cet article fournit des outils et des idées visant à favoriser un environnement de travail plus ouvert, communicatif et fournissant un soutien émotionnel. L’importance de « l’auto-soin » est également discutée. Il existe un engagement croissant en faveur d’une culture de soins et de soutien pour soutenir nos collègues en les sensibilisant à nos déﬁs émotionnels. Mots clés. Animaux, déﬁ émotionnel, COVID-19, techniciens animaliers, outils de soutien.Paper Summary Translations

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54Animal Technology and Welfare August 2020INHALTVERZEICHNISAkute Verhaltensauswirkungen auf Mäuse durch Handhabung mit Handschuhen, die mit Handdesinfektionsmittel auf Alkoholbasis besprüht wurden NOELIA LOPEZ-SALESANSKY, DOMINIC J WELLS, NATALIE CHANCELLOR, LUCY WHITFIELD, CHARLOTTE C BURN Korrespondenz: cburn@rvc.ac.uk Abstract Alkohole ﬁnden in Versuchstiereinrichtungen zur Desinfektion von Händen, Geräten und Laborumgebungen breite Anwendung. Da die Auswirkung auf Mäuse bisher nicht bekannt war, haben wir männliche und weibliche C57BL/6J- und BALB/c-Mäuse während und nach ihrer Handhabung mit Nitrilhandschuhen beobachtet, die mit 70%igem Alkohol-Desinfektionsmittel (~ 67 % Ethanol, ~ 3 % Methanol und 30 % Wasser) besprüht bzw. nicht besprüht worden waren. Wir gingen von der Annahme aus, dass, wenn Mäuse dieses Handdesinfektionsmittel als aversiv empﬁnden, vor der Handhabung damit behandelte Handschuhe Verhaltensindikatoren für Angst oder Abwehr verstärken würden und auch soziale Interaktionen und Grooming in beiden Richtungen beeinﬂussen könnten. Die Handhabung von Mäusen mit desinﬁzierten Handschuhen führte bei einem bzw. beiden Mäusestämmen zu vermehrtem Aufrichten an der Käﬁgwand, Grooming, Allogrooming, Beschnüffeln von Käﬁgnachbarn sowie Fressen/Trinken. Bei den männlichen Tieren verringerte sich anfänglich auch die Aggression im Heimkäﬁg, an deren Stelle Grooming trat, wobei jedoch unklar ist, ob aggressives Verhalten tatsächlich abnahm oder nur verzögert wurde. Es gab keine statistisch signiﬁkanten Auswirkungen der Behandlung auf in einem Hand-Interaktionstest gezeigtes Vermeidungsverhalten. Defensives Graben trat sowohl mit desinﬁzierten als auch mit Kontrollhandschuhen während des Ersthand-Interaktionstests auf und nahm im Laufe der 4-wöchigen Studie signiﬁkant ab, was einen Neuheitseffekt nahelegt. Die Ergebnisse deuten darauf hin, dass die Handhabung von Mäusen mit alkoholbasiertem Handdesinfektionsmittel das Verhalten der Tiere, einschließlich sozialer Interaktionen, beeinﬂusst, wenngleich eine Replikation erforderlich ist, da wir den Beobachter der Behandlung nicht verblinden konnten. Weitere Untersuchungen sind erforderlich, um die langfristigen Auswirkungen der Verwendung von alkoholbasierten Handdesinfektionsmitteln und alternativen Desinfektionsmitteln beim Umgang mit Labormäusen bewerten und entsprechende Empfehlungen für eine Verbesserung aussprechen zu können. Schlagwörter: Tierverhalten, Tierschutz, Desinfektionsmittel, Handhabung, Hygiene, Mäuse Paper Summary Translations

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55August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsBericht über die Tagung der RSPCA/UFAW-Tierschutzgruppe für Nager 2020CHLOE STEVEMS, PENNY HAWKINS, TOM V SMULDERS, AILEEN MACLELAN, LARS LEWEJOHANN, PAULIN JIRKOF, JACKIE BOXALL, HELEN MURPHY, CARLEY MOODY, PATRICIA V TURNER, I J MAKOWSKA, CHARLOTTE INMAN AbstractDie RSPCA/UFAW-Tierschutzgruppe für Nager und Kaninchen veranstaltet seit 27 Jahren jeden Herbst ein eintägiges Treffen, bei dem die Mitglieder über Themen aktueller Tierschutz-Forschung diskutieren und Erfahrungen und Meinungen über Tierschutzfragen sowie über die Umsetzung des 3R-Prinzips (Vermeidung, Reduktion und Verbesserung) beim Einsatz von Nagern austauschen können. Ein Hauptziel der Gruppe ist es, zum Nachdenken über die Gesamtheit der Lebenserfahrungen von Nagern als Laborversuchstiere anzuregen und sicherzustellen, dass jede potenzielle Beeinträchtigung ihres Wohlergehens geprüft und auf ein Minimum reduziert wird.Am diesjährigen Treffen, das zum ersten Mal online stattfand, nahmen über 400 Delegierte aus fast 40 Ländern teil. Es stand unter dem Motto „kumulative Erfahrungen“, zu dem Sitzungen über „die Wissenschaft kumulativer Belastung“ und „praktische Verbesserungen zur Verringerung der Belastung und Förderung des Wohlbeﬁndens“ veranstaltet wurden. Die Präsentationen umfassten einen Einführungsvortrag, in dem erläutert wurde, warum kumulative Erfahrungen wichtig sind und wie sich sowohl positive als auch negative Erfahrungen im Laufe des Lebens eines Tieres akkumulieren und langfristig auf sein Wohlbeﬁnden auswirken können. Weitere Beiträge befassten sich mit verschiedenen Möglichkeiten zur Erkennung und Bewertung von kumulativen Belastungen, kumulativen Auswirkungen kleiner Verbesserungen sowie mit dem Konzept eines „guten Lebens“ und was dies für Labornager bedeutet. Zudem gab es ein Update der Home Ofﬁce Animals in Science Regulation Unit (ASRU, Regulierungsstelle des britischen Innenministeriums für in der Wissenschaft verwendete Versuchstiere), das untersuchte, wie kumulative Erfahrungen die von Versuchstieren erlebten Belastungen beeinﬂussen. Der Tag endete mit einer interaktiven Diskussionsrunde über Möglichkeiten der Erkennung kumulativen Leidens bei Nagetieren in Käﬁgen. Dieser Bericht fasst die Tagung zusammen und endet mit einer Liste von Aktionspunkten, die Leser für ihre eigenen Einrichtungen in Betracht ziehen können.Schlagwörter: 3R-Prinzip, Nager, Kaninchen, kumulative Erfahrung, Belastung, Verbesserung★ ★ ★Emotionale Herausforderungen bei der Arbeit mit Labortieren: Instrumente zur Unterstützung von Fürsorge und Selbstfürsorge ANGELA KERTON UND JORDI L TREMOLEDA Korrespondenz: angela@learningcurvedevelopment.co.uk Abstract Unweigerlich gehen die meisten von uns, die mit Labortieren arbeiten, zuweilen Bindungen mit den Tieren ein, für die wir sorgen. Diese Beziehungen beeinﬂussen die Pﬂege und das Wohlbeﬁnden der Tiere positiv, stellen aber auch eine große emotionale Herausforderung dar, wie sich bei einigen Maßnahmen des Kontingenzmanagements im Zusammenhang mit der COVID-19-Pandemie deutlich gezeigt hat. Es ist wichtig, dass die Branche das Bestehen dieser Bindungen zur Kenntnis nimmt und institutionelle Unterstützungsmechanismen bereitstellt, um Tiertechnikern zu helfen, mit den emotionalen Herausforderungen ihres Berufs umzugehen. Die derzeitige mit COVID-19 verbundene Arbeitslogistik bringt weitere Schwierigkeiten mit sich, wie z. B. die Delegierung von Aufgaben, die Trennung von Arbeitsteams und das Kontingenzmanagement von Tierbeständen, um nur einige zu nennen. Hinzu kommen die individuellen Herausforderungen in gesundheitlicher, sozialer und wirtschaftlicher Hinsicht ebenso wie das mit den persönlichen Bindungen verbundene Dilemma. Dieser Artikel bietet einige Instrumente und Ideen zur Unterstützung eines offeneren, kommunikativeren und emotional unterstützenden Arbeitsumfelds. Ebenso wird die Bedeutung der „Selbstfürsorge“ erörtert. Es gibt zunehmend Bestrebungen hin zu einer Kultur der Fürsorge, und die Unterstützung unserer Kollegen durch eine Sensibilisierung für unsere emotionalen Herausforderungen kann dieser förderlich sein. Schlagwörter: Tiere, emotionale Herausforderung, COVID-19, Tiertechniker, unterstützende Instrumente

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57August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsINDICE DELLA REVISTAManipolazione dei topi con l’uso di guanti spruzzati con igienizzante per le mani a base di alcol: effetti acuti sul comportamento del topo NOELIA LOPEZ-SALESANSKY, DOMINIC J WELLS, NATALIE CHANCELLOR, LUCY WHITFIELD, CHARLOTTE C BURN Corrispondenza: cburn@rvc.ac.uk Abstract L’alcol è comunemente utilizzato nei laboratori animali per la disinfezione di mani, apparecchiature e ambiente in genere. L’effetto sui topi non è noto, per cui abbiamo osservato topi C57BL/6J e BALB/c di sesso maschile e femminile durante e dopo la loro manipolazione con guanti in nitrile spruzzati con un igienizzante contenente 70% di alcol (~67% etanolo, ~3% metanolo e 30% acqua) oppure non ricoperti di igienizzante. Abbiamo ipotizzato che, se i topi percepivano questo igienizzante per le mani come avverso, la sua applicazione sui guanti prima della manipolazione avrebbe esacerbato gli indicatori comportamentali di paura o difesa; inoltre, avrebbe potuto incidere sulle interazioni sociali e sulla toelettatura in entrambe le direzioni. La manipolazione dei topi con guanti disinfettati ha accresciuto i casi di posizionamento eretto contro la parete della gabbia, l’auto-toelettatura, il social grooming, gli annusamenti di altri animali presenti nella gabbia e il consumo di cibo/liquidi in uno o entrambi i ceppi di topi. Nei maschi, ha ridotto anche l’aggressione iniziale nella gabbia di stabulazione, sostituendola con la toelettatura, ma non è chiaro se l’aggressione sia effettivamente diminuita o fosse semplicemente tardiva. Non si sono notati effetti statisticamente signiﬁcativi del trattamento sui comportamenti di annullamento mostrati durante un test di interazione manuale. Un comportamento di seppellimento difensivo si è veriﬁcato con guanti disinfettati e di controllo durante il primo test di interazione manuale ed è diminuito notevolmente nel corso delle 4 settimane dello studio, suggerendo un possibile effetto di novità. Le conclusioni indicano che la manipolazione di topi con igienizzante per le mani a base di alcol incide sul loro comportamento, comprese le interazioni sociali, anche se è necessario eseguire ulteriori repliche, dal momento che non abbiamo potuto rendere ‘ciechi’ gli osservatori al trattamento. Sono necessarie ulteriori ricerche per valutare gli effetti a lungo termine legati all’uso di igienizzanti per le mani a base di alcol e disinfettanti alternativi durante la manipolazione di topi da laboratorio prima di poter offrire raccomandazioni di perfezionamento. Parole chiave: comportamento animale; benessere animale; disinfettante; manipolazione; igiene; topi

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58Animal Technology and Welfare August 2020Resoconto dell’incontro del RSPCA/UFAW rodent and rabbit welfare group del 2020CHLOE STEVEMS, PENNY HAWKINS, TOM V SMULDERS, AILEEN MACLELAN, LARS LEWEJOHANN, PAULIN JIRKOF, JACKIE BOXALL, HELEN MURPHY, CARLEY MOODY, PATRICIA V TURNER, I J MAKOWSKA, CHARLOTTE INMAN AbstractOgni autunno, da ormai 27 anni, il RSPCA/UFAW Rodent and Rabbit Welfare Group (Gruppo sul benessere dei roditori e dei conigli) organizza un incontro di un giorno per consentire ai suoi membri di discutere degli studi attuali di ricerca sul benessere, di scambiarsi opinioni sulle questioni legate al benessere e di condividere esperienze di applicazione del principio delle 3 R, sostituzione (replacement), riduzione (reduction) e perfezionamento (reﬁnement), in relazione all’uso di roditori. Uno degli obiettivi primari del Gruppo è quello di invitare a prendere in considerazione l’intero percorso di vita dei roditori da laboratorio, accertandosi che venga valutato e minimizzato ogni possibile impatto negativo sul loro benessere.L’incontro di quest’anno si è tenuto online per la prima volta e ha visto la partecipazione di oltre 400 delegati di quasi 40 Paesi. È stato affrontato il tema delle ‘esperienze cumulative’, con sessioni dedicate alla ‘scienza della gravità cumulativa’ e a ‘perfezionamenti pratici volti a ridurre la gravità e a promuovere il benessere’. Le presentazioni hanno incluso una relazione introduttiva che ha spiegato l’importanza delle esperienze cumulative e il fatto che le esperienze positive e negative possono accumularsi durante il percorso di vita di un animale e avere impatti a lungo termine sul benessere. Ulteriori discussioni hanno esaminato i vari metodi per riconoscere e valutare la gravità cumulativa, gli impatti cumulativi di piccoli perfezionamenti e il concetto di una ‘buona vita’ e di cosa signiﬁchi per i roditori da laboratorio. L’Animals in Science Regulation Unit (ASRU) del Ministero dell’Interno britannico ha presentato, inoltre, un aggiornamento su come le esperienze cumulative inﬂuenzano la gravità vissuta dagli animali in campo scientiﬁco. La giornata si è conclusa con una sessione interattiva su come identiﬁcare la sofferenza cumulativa nei roditori in gabbia. Il presente resoconto riassume l’incontro e termina con un elenco di punti di intervento che i lettori possono portare all’attenzione delle loro strutture.Parole chiave: 3 R, roditori, conigli, esperienza cumulativa, gravità, perfezionamento★ ★ ★Sﬁde emotive sul lavoro con gli animali da laboratorio: strumenti che aiutano a prendersi cura di se stessi e degli altri ANGELA KERTON E JORDI L TREMOLEDA Corrispondenza: angela@learningcurvedevelopment.co.uk Abstract Inevitabilmente, la maggior parte di noi a contatto con animali da laboratorio forma talvolta un legame con gli animali di cui si prende cura. Queste relazioni avranno un impatto positivo sulla cura e sul benessere degli animali, ma comportano anche importanti sﬁde emotive, come è emerso chiaramente da alcuni casi di gestione della contingenza associati alla pandemia da COVID-19. È importante che l’industria riconosca l’esistenza di questi legami e fornisca meccanismi di supporto istituzionale per aiutare gli stabularisti a far fronte alle sﬁde emotive emergenti dalla loro professione. Le attuali modalità operative dettate da COVID-19 presentano ulteriori sﬁde, come la delega delle responsabilità, la separazione dei team di lavoro e la gestione della contingenza della popolazione animale, per citarne solo alcune, unitamente ai problemi sanitari e sociali, economici e interpersonali dei singoli individui. Questo articolo condivide strumenti e idee a sostegno di un ambiente di lavoro più aperto, comunicativo e improntato sul supporto emotivo. Pone l’accento anche sull’importanza della “cura di sé”. Si è sempre più dediti a promuovere una cultura di cura e il sostegno nei confronti dei colleghi attraverso una maggiore consapevolezza delle nostre sﬁde emotive potrebbe aiutare in tal senso.Parole chiave: Animali, sﬁda emotiva, COVID-19, stabularisti, strumenti di supporto. Paper Summary Translations

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60Animal Technology and Welfare August 2020INDICE DE LA REVISTAManipulación de roedores utilizando guantes a los que se les ha aplicado previamente desinfectante para manos basado en alcohol NOELIA LOPEZ-SALESANSKY, DOMINIC J WELLS, NATALIE CHANCELLOR, LUCY WHITFIELD, CHARLOTTE C BURN Correspondencia: cburn@rvc.ac.uk Resumen El alcohol suele utilizarse en las instalaciones de animales de laboratorio para desinfectar manos, equipos y entornos del laboratorio. Los efectos en los roedores se desconocen, por lo que observamos a los roedores C57BL/6J y BALB/c macho y hembra durante y después de la manipulación con guantes de nitrilo a los que se les administró un desinfectante con 70 % de alcohol (~67 % etanol, ~3 % metanol y 30 % agua) o con guantes sin ningún tipo de desinfectante. Especulamos que, si los roedores percibían este desinfectante de manos como algo desagradable, su aplicación en guantes antes de la manipulación aumentaría los indicadores de comportamiento de miedo o defensa; asimismo podría afectar las interacciones sociales y el acicalamiento en cualquier dirección. La manipulación de roedores con guantes desinfectados hacía que estos se pusieran de pie más de lo habitual en los bordes de la jaula y aumentaba también el autoacicalamiento, el acicalamiento social entre ellos, el olfateo de otros miembros de la jaula y la actividad de comer/beber en una o ambas cepas de roedores. En los machos, también redujo las agresiones iniciales en jaulas, siendo reemplazadas por un acicalamiento, pero no está claro si las agresiones disminuyeron realmente o simplemente se demoraron. No hubo efectos estadísticamente signiﬁcativos del tratamiento respecto a comportamientos evasivos observados en una prueba de interacción manual. Se observó una excavación defensiva tanto con guantes desinfectados como con guantes de control durante la primera prueba de interacción manual y una disminución signiﬁcativa durante el estudio de 4 semanas, lo que sugiere un efecto novedoso. Las conclusiones indican que la manipulación de roedores con desinfectante para manos basado en alcohol afecta al comportamiento de los ratones, lo que incluye las interacciones sociales, aunque es necesario realizar una replicación ya que no pudimos cegar al observador del tratamiento. Es necesario realizar más estudios de investigación para evaluar los efectos a largo plazo del uso de desinfectante para manos basado en alcohol y desinfectantes alternativos al manipular roedores de laboratorio para poder realizar recomendaciones para su reﬁnamiento. Palabras clave: Comportamiento animal, bienestar animal, desinfectante, manipulación, higiene, roedoresPaper Summary Translations

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61August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsInforme sobre la reunión del Grupo para el bienestar de roedores RSPCA/UFAW 2020CHLOE STEVEMS, PENNY HAWKINS, TOM V SMULDERS, AILEEN MACLELAN, LARS LEWEJOHANN, PAULIN JIRKOF, JACKIE BOXALL, HELEN MURPHY, CARLEY MOODY, PATRICIA V TURNER, I J MAKOWSKA, CHARLOTTE INMAN ResumenEl Grupo para el bienestar de conejos y roedores RSPCA/UFAW ha celebrado una reunión de un día cada otoño durante los últimos 27 años para que sus miembros puedan debatir sobre la investigación actual sobre bienestar, intercambiar opiniones sobre temas relacionados con el bienestar y compartir su experiencia respecto a la implementación de las 3 R (reemplazo, reducción y reﬁnamiento) en relación al uso de roedores. Uno de los objetivos primordiales del Grupo es fomentar que las personas piensen sobre la experiencia vital de los roedores de laboratorio garantizando a su vez que cualquier impacto negativo posible en su bienestar sea revisado y reducido.La reunión de este año se celebró en línea por primera vez y asistieron más de 400 delegados de casi 40 países. El tema era «experiencias acumulativas», con sesiones sobre «la ciencia de la gravedad acumulativa» y los «reﬁnamientos prácticos para reducir la gravedad y fomentar el bienestar». Las presentaciones incluyeron una charla de presentación en la que se explicaba el motivo por el que las experiencias acumulativas son importantes y cómo tanto las experiencias positivas como las negativas pueden ir acumulándose durante la vida de un animal y tener un impacto a largo plazo sobre su bienestar. Otras charlas trataron distintas formas de reconocer y evaluar la gravedad acumulativa, los impactos acumulativos de pequeños reﬁnamientos y el concepto de una «buena vida» y lo que esto implica para los roedores de laboratorio. También se asistió a una actualización de la Home Ofﬁce Animals in Science Regulation Unit (ASRU) que se centró en cómo las experiencias acumulativas inﬂuyen en la gravedad experimentada por los animales utilizados para ﬁnes cientíﬁcos. El día se clausuró con un debate interactivo sobre las distintas formas de identiﬁcar el sufrimiento acumulativo en roedores observándolos desde fuera de la jaula. Este informe resume la reunión y ﬁnaliza con una lista de puntos de acción para que los lectores consideren presentar en sus propias instalaciones.Palabras clave: Las 3 R, roedores, conejos, experiencia acumulativa, gravedad, reﬁnamiento★ ★ ★Desafíos emocionales en nuestro trabajo con animales de laboratorio: herramientas que apoyan el cuidado de los demás y de uno mismoANGELA KERTONY JORDI L TREMOLEDACorrespondencia: angela@learningcurvedevelopment.co.uk Resumen Inevitablemente, la mayoría de los que trabajamos con animales de laboratorio creamos a veces un vínculo con los animales a los que cuidamos. Estas relaciones mejorarán positivamente el cuidado y el bienestar de los animales, pero también suponen unos retos emocionales, tal y como se pudo observar con algunos tratamientos de contingencias en relación con la pandemia de la COVID-19. Es importante que el sector admita la existencia de estos vínculos y ofrezca mecanismos de soporte institucional para ayudar a los tecnólogos de animales a gestionar los retos emocionales de su profesión. La logística actual de trabajo relacionada con la COVID-19 supone más retos como la delegación de responsabilidades, la separación de equipos de trabajo y el tratamiento de contingencias de existencias de ejemplares, por nombrar algunos, junto con los retos individuales de relaciones personales, económicos, sociales y sanitarios. Este artículo presenta algunas herramientas e ideas para respaldar un entorno de trabajo más abierto, más comunicativo y con más apoyo emocional. También se trata la importancia del «autocuidado». Existe un compromiso creciente para fomentar una cultura de cuidado y,apoyando a nuestros compañeros mediante la creación de concienciación sobre nuestros retos emocionales puede ayudar a conseguir este objetivo.Palabras clave: Animales, reto emocional, COVID-19, tecnólogos de animales, herramientas de apoyo.

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63August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareGuiding principles to help deliver the ethics learning outcomes of module 2for personal licenseesM JENNINGS1,2 and M BERDOY1,3 (editors), A-M FARMER,1,4 P HAWKINS,2 E LILLEY,1,2 A KERTON,1,5 B LAW,1,6 JORDI L TREMOLEDA,1,7 C STANFORD,1,8 L WHITFIELD1,9 and K RYDER101 LASA Education, Training and Ethics Section2 Research Animals Department, RSPCA3 Oxford University4 Cambridge University5 The Learning Curve (Development) Ltd6 Leeds University7 Queen Mary University of London8 University College London 9 Agenda Veterinary Services 10 Home Ofﬁce observerCorrespondence: info@lasa.co.ukReprinted at the request of the Laboratory Animal Science Association (LASA)IntroductionDelivering – and assessing – the ethics learning outcomes in personal licensee (PIL) training courses (EU functions A and B1, Home Ofﬁce PIL courses) is an important component of licensee training, yet experience shows that this is not easy. There is often limited time and resource available for the course as a whole. As a consequence, current licensee training may not be able to deliver the intended long-term effects on attendees’ knowledge, understanding, attitudes and behaviours. Therefore, it is clearly important to ﬁnd ways of maximising the impact of what can be delivered in the time available. Trainers, particularly those delivering module 2 ‘Ethics, Animal Welfare and the Three Rs’, have reported that they would welcome guidance on how to achieve the ethics learning outcomes. Access toup-to-date training resources would also be helpful. This document aims to address these needs by focussing on the relevant ethics learning outcomes of EU module 2 for personal licensees (see appendix 1; module 2 is also a ‘core module’ for project licensees although, in addition, they are required to do module 9 which addresses the same issues in greater depth).The ﬁrst part of the document deals with general principles and addresses the following points:1. The overall aim of ethics training – what each learning outcome should cover and what the overall outcome for personal licensees should be.2. What ethics is, why it is important and how it relates to the use of animals in research.3. The principles underlying good practice in teaching this topic, such as the need for a relevant and practical approach that integrates ethics throughout the whole training course.The second part of the document focusses on the practical aspects of delivering the learning outcomes (LOs) of module 2 that speciﬁcally relate to ethics. Not all the fourteen LOs directly address this topic; those that relate to the 3Rs, animal welfare or legislation (see those in italics in appendix 1) are practical issues that are generally easier to address, and many can also be dealt with as part of other modules. For example, LO 2.9 on severity classiﬁcation is also dealt with in module 5; LO 2.11 on ‘the importance of good animal April 2021 Animal Technology and Welfare

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64Animal Technology and Welfare August 2020welfare for good science’ also appears in module 3. They are, however, based on ethical principles and it is important to make this clear when delivering them.A number of learning outcomes however (i.e. LOs 2.1 to 2.4 and 2.12), are directly linked to ethics and part 2 of this document focusses on these. It highlights how they could be addressed and delivered and the key points to get across, as well as suggesting some useful resources and opportunities for CPD (e.g. via the Animal Welfare and Ethical Review Body). Part 1: General principles1. Overall aim of the ethics learning outcomesOn completion of the ethics aspects of EU module 2 trainees should understand what ethics is (see deﬁnition in section 2 below) and be clear about the practical application of ethical values to animal research. They should be able to identify the ethical issues within their own work and that of the establishment as a whole, and see how ethics is integral to establishing and maintaining a Culture of Care. They need to understand the importance of maintaining an open mind and respect for other people’s opinions, perspectives and beliefs, of acting responsibly at all times, and of accepting the consequences of their actions for animals and other people.The aim, together with the other modules in the course, is to create thoughtful and reﬂective licensees, who are prepared to challenge themselves and strive for continuous improvement in the work that they do. They should be better able to think through the harms, beneﬁts and justiﬁcation for their work whatever their role or level of input. They should feel able and comfortable to question and, where necessary, challenge practices based on the ‘I’ve always done it like this’ way of thinking and operating, thus helping to ensure good science with minimal animal use and suffering.2. Deﬁning ethics and why it is importantTrainees need to be clear that attitudes, decisions and laws regarding the use of animals in science are based on an ethical framework. They need to understand the ethical components of such frameworks and how they are identiﬁed, developed and applied in practice.The following points in this section list the main issues to address. These could be developed into a pre-course handout with references or links to further reading.i) What is ethics?Arguably, everything starts with ethics. Ethics is a system of moral principles that includes ideas about right and wrong and how people should, or should not, behave in general and speciﬁc instances. The term is used in several ways including:– To describe ways of life (for example, Buddhist or Christian ethics).– To help deﬁne practitioners’ rights and responsibilities within professional codes of conduct and provide guidance on what are good or bad moral decisions. Examples are the World Medical Association Declaration of Helsinki for medical ethics2 and the UK Code of Conduct for Veterinary Surgeons.3 In science, both funders and journals have ethical guidelines deﬁning the research they will fund and publish.4,5– Interchangeably with the term ‘moral’ in public life to describe ‘desirable’ and ‘undesirable’ behaviour - what ought and ought not to be done.Societal views evolve over time. Activities or actions that are considered acceptable today may be considered unacceptable in the future. This has been very noticeable in animal research as advances in our understanding of animal suffering and animal sentience has led to a greater commitment within the research community and wider society to reduce animal use, reduce suffering and improve the welfare of those animals that are used.ii) Ethics and the lawBehaving ethically and legally are not always synonymous. Laws take time to create or amend and, in our rapidly changing world, some may not reﬂect current ethical values or thinking. Moreover behaving ethically involves more than just following the letter of the law.6 Laws such as the Animals (Scientiﬁc Procedures) Act 1986 (ASPA)7 lay down certain baseline rules and boundaries but within these there is room for judgement about what is and is not ethically acceptable.iii) Ethics and philosophyEthics is part of the academic ﬁeld of moral philosophy. This has an extensive theoretical base commonly divided into three areas.8 Meta-ethics deals with the nature of moral judgement, exploring the origins and meaning of ethical principles. Normative ethics is concerned with the content of moral judgement and the criteria for what is right or wrong. Applied ethics deals with the practical application of moral considerations. Understanding how to make sound choices and providing a framework for deciding how to make them, is particularly relevant to many societal concerns such as abortion, biotechnology, gene therapy and artiﬁcial intelligence.Applying ethics in practice to the use of animals in science under the ASPA is most relevant to licensees and should form the basis for ethics training.iv) Ethics and the use of animals in scienceThe most obvious application of an ethical framework is that of the harm/beneﬁt analysis that is at the centre of drafting and reviewing a project licence. However day-to-day decisions have ethical aspects that require recognition and careful consideration of competing Guiding principles to help deliver the ethics learning outcomes of module 2 for personal licensees

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65August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareGuiding principles to help deliver the ethics learning outcomes of module 2 for personal licenseeshuman and animal interests, and it is important to get this message across. For example it may be thought that the use of analgesia or an enriched environment will interfere with aspects of the science whereas the alternative view is that these are essential for the wellbeing of the animals. Practical ethics involves exploring the different opinions and perspectives behind such dilemmas in order to determine what is regarded as the best solution in each case.v) Common misconceptionsIt is also important to understand the broad remit of ethics and to avoid misconceptions. For example, in the ﬁeld of animal research, ethics is often equated just with implementing the 3Rs and improving animal welfare. However although both these issues concern the reduction of overall animal suffering and are thus a factor in ethical decision-making, ethics encompasses a much wider set of considerations and reﬂection about what it is justiﬁed to do to animals in the name of science.A second misconception is that debate over animal experiments exists solely in terms of the polarised extremes. However public opinion is much more nuanced than this, encompassing a wide diversity of perspectives. On balance, the public are ‘conditional acceptors’ of animal experiments but only provided that these are carried out to high legal, animal welfare, scientiﬁc and ethical standards.93. Good practice teaching principlesIt is important for trainees to recognise that licensee training is not just about acquiring a personal licence. The modules have been carefully developed by laboratory animal scientists across Europe and are designed to give licensees a good foundation for their future work.Training content and delivery therefore needs to be relevant to the participants. In the case of ethics, this means that a didactic lecture on moral philosophy is unlikely to be useful. Nevertheless as stated above, trainees need to understand the ethical issues around the use of animals and why this is important in their work. It is therefore a good idea to deﬁne these at the start of the course since all other aspects of the course including legislation, welfare, husbandry, 3Rs and harm/beneﬁt analysis are based on ethical principles. This could be done as a short talk or discussion session using the information in section 2, with additional information and references (e.g. on different ethical perspectives) provided as a pre-course handout for those who want more detail.Such face to face interaction needs to be related to any pre-course learning to ensure that this is consolidated. Some (simple) ethical principles using local examples or examples otherwise relevant to the trainees, could be described and these could then be built on by ‘spiralling’ the ethical discussion throughout other elements of the course (see box below). This means explaining how In summary– Ethics is a system of moral principles that encompasses ideas about right and wrong, and how people should, or should not, behave: a ‘we can – but should we?’ approach.– Willingness to listen open-mindedly to differing opinions, perspectives, beliefs and values, whether in the context of culture, religion, experience or society, is key to taking part in ethical discussions with integrity.– Although it may be helpful to understand the philosophical background, the key aspect for licensees is practical ethics and how this relates to science and their own work.– Ethics provides an approach for decision-making when faced with disagreements and dilemmas.– Such dilemmas require careful identiﬁcation and consideration of all the relevant issues, competing interests and perspectives; ethics helps reconcile disagreement and contributes to sound and consistent decisions.– Ethics applies throughout a licensee’s day-to-day work and is not conﬁned to the ethical review within project evaluation.– Applying the 3Rs alone does not equate to ‘doing ethics’ – practical ethics encompasses a much wider set of considerations about what is, and is not, acceptable to do to animals and for what purpose.Spiral curriculumA spiral curriculum is an educational approach that involves the student re-visiting the same topics over the course of their education. This iterative approach helps to reinforce learning over time, using prior information to deepen understanding of the subject, consolidate learning and inform the students’ approach to future learning. It stems from a behaviourist theory that spaced repetition of learning is the most helpful way to deepen understanding and promote long term retention of information.According to Bruner (p. 141), there are three components to this approach:1. Cyclical: the student returns to the same topic several times during the course of their study.2. Complexity: each time the topic returns, it is explored in more depth/ complexity.3. Prior knowledge: the knowledge gained previously is used, so that students explore the topic from a certain level of understanding, rather than starting from fresh each time.10 Developing a spiral system of learning (e.g. for ethics) requires an integrated approach between tutors for different parts of the course, such that the whole course hangs together and is delivered in a connected way.11

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66Animal Technology and Welfare August 2020issues in other modules, such as husbandry, pain and distress, handling and breeding and colony management, all have ethical elements.It can help trainees ‘normalise’ the application of ethics if they see it as an issue that pervades not only science in many contexts, but also day-to-day life whenever decisions have to be made about what should or should not be done in any given situation. It is also helpful to note the application of ethics in ensuring research integrity12 and to explain that applied ethics is also important in other contexts such as professional medical and veterinary ethics, research funding and publishing. Explaining that ethical principles permeate the culture of care,13 which encompasses the way both animals and staff are treated, helps to emphasise further its importance and practical application. A good discussion example to use here would be the ethical issues around humane killing of animals – whether loss of a life is in itself a harm and how killing animals affects those who have to carry this out.Aside from setting out the deﬁnitions and principles relating to ethics, the topic is best addressed by guided discussions, preferably in small groups. A discussion session, that brings together all elements of the learning outcomes, at the end of the whole training course, when students are likely to be more communicative, can work well and is recommended.There are now a wide range of polling devices which can be used to gather responses anonymously and stimulate discussion of a variety of ethical dilemmas or statements if participants are reticent about speaking up. Ideally when courses are run in-house it is helpful to bring in the named people and/or existing personal or project licensees to contribute to discussions and provide their personal views on ethical issues, other aspects of the module and the course as a whole. It may also be possible to make use of the range of research interests and ethical viewpoints within different research groups. These can range from basic to translational research to animal welfare and conservation studies. This will help trainees see the diversity of views and perspectives that can be present even within an establishment.The AWERB and the Home Ofﬁce Liaison Contact (HOLC) could both be a helpful source of in-house examples for discussion (assuming this is permitted), for example where a project proposal has generated a lot of debate during a committee meeting. This could provide a local resource showing: what made the AWERB think; why the proposal raised ethical concerns; whether everyone agreed; and how the issue was resolved. It should be possible to develop some case studies to discuss in the course and then put the ‘real life solution or agreement’ up at the end of the discussion. Asking the AWERB chair to introduce the issue would have the additional function of raising trainees’ awareness of the AWERB and its role.3.1 Pre-course workPre-course work helping trainees understand the ethics learning outcomes and what will be expected of them will facilitate better use of the teaching time available (see The Flipped Classroom box). This can also help provide a more consistent starting position when trainees come from different educational backgrounds and/or cultures.Pre-course work should not only provide information (e.g. on what ethics is, as mentioned above and see reference list) but also stimulate thinking. Some options are:– An activity, such as providing a scenario for students to think about and discuss later in group work. Or asking students to set out their own ideas on ethical dilemmas in life generally or speciﬁc to their work.– Delegates could also be presented in advance with a range of scenarios and score these for whether or not they are ‘acceptable’ or ‘not acceptable’. These could be collated and used as a discussion prompt in class: Why did you make this response? Did you consider this? Is there is a split of opinion in class or a consensus? Following the ethics section of the course, the same questions could be asked again to see if trainees’ opinions change.– Online facilities also offer the possibility of increasing the interactivity of pre-course work. For example, participants can be faced with iterative choices of dilemmas based on their previous choices.Further examples are given in the tables in Part 2.3.2 The importance of feedbackAs with all forms of training, it is important to ask course attendees to evaluate the ethics component. Has it been helpful and worthwhile? Has it allowed them to reﬂect on the issues? Has it been successful in addressing the learning outcomes? The feedback must then be used to inform and improve the course.The Flipped ClassroomTraditional lecture-based teaching can be a passive form of education. The “Flipped classroom” is a learning methodology that aims to facilitate deep learning, better retention and critical thinking by moving some information- transmission teaching out of the class. Students are required to complete pre (and/or post) classroom activities or assignments (some taking advantage of technological innovations) which are then used for active learning (e.g. problem solving, case studies, discussion groups) during the crucial class time with a teacher.14-16Guiding principles to help deliver the ethics learning outcomes of module 2 for personal licensees

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67August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareGuiding principles to help deliver the ethics learning outcomes of module 2 for personal licensees3.3 The importance of CPDA single session on ethics during licensee training will not be sufﬁcient to encourage trainees to think routinely about the ethical implications of their own work and that of others. It is therefore important to use other opportunities such as lectures, workshops or dedicated training sessions as CPD to reinforce the initial messages and stimulate further thought and reﬂection (see spiral curriculum and ﬂipped classroom boxes above). The AWERB in its role as providing a forum for discussion and development of ethical advice could be a useful resource in this respect17 (see appendix 2). Explain what ethics is, why it is important and how it relates to animal research. Use local examples as illustrations of where ethical debate has occurred. ‘Normalise’ ethics e.g. by reference to ethics in daily life, other professional codes of conduct and with respect to research integrity. ‘Spiral’ (i.e. refer to) ethics throughout the whole course and pick out examples of how it relates to other topics such as legislation, welfare, humane science. Show how ethics relates to licensees’ own work and encourage them to reﬂect on this. Allow as much time as possible for the course including time for discussion. Bring in named persons, scientists or staff from other disciplines (e.g. medical ethics) to participate in discussions. Re-enforce the ethics learning outcomes in the rest of the module and in other modules where relevant. Make the course interactive – and ensure trainees are ‘engaged’. Ask the trainees for feedback and use it to tailor and improve the course. Ask the AWERB to organise ethics-related activities. Covering ethics with a half hour lecture at the start and then putting it aside for the rest of the course.Delivering theoretical lectures on utilitarianism, deontology, virtue ethics, etc.Quoting historical philosophers and expecting trainees to know the relevance of their names.Confusing ethics with the 3Rs and animal welfare. Interpreting it as the polarised extremes of animal rights versus legitimate science.Imposing your views on others; stating there is ‘only one right way’.Implying that ethical beliefs, perspectives and dilemmas are static and cannot change over time.Dumbing down the importance of ethics.Making it boring!Things to avoid...Good practice points…Summary

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68Animal Technology and Welfare August 2020Part 2: Some practical examples for use in teachingThe following tables present the key points to cover under each of the ethics learning outcomes (LOs), together with ideas in the right-hand column of how these points could be addressed. Learning outcomes addressing similar issues are coloured similarly since there may be overlap between them and how they are delivered.Key points to get across:– There is a wide range of opinions on the use of animals in science and views are now quite nuanced.– The polarised animal experimentation debate (pro- vs anti-vivisection) is outdated and trainees need to recognise this and that it is unhelpful to present society’s views in this way.– Trainees need to think beyond the polarised extremes and recognise that focussing on this type of debate prevents people addressing the real ethical issues in their work.– It is important to be open-minded, listen to and respect other peoples’ views and try to understand the basis for these.– People’s views – and hence ethical values – are constantly evolving. Their views will also depend on their country of origin, culture, individual background and experience, circumstances, age and gender. Views are also affected by the type of use, species and age of animals.How delivery could be achieved:– Start by presenting a series of viewpoints on the use of animals and ask trainees to identify their position in the spectrum of views with their reasons. Repeat at the end of the course/module.– Ask trainees to deﬁne what ‘respecting other people’s views means to them. Collate their thoughts and use these as a platform to promote discussion.– Use examples of research that was once considered acceptable (either its purpose or how it was done) but is not thought so now. Examples include using death as an end-point and tail tipping for genotyping. Where training is in-house, try to use local examples to make it directly relevant e.g. a Contract Research Organisation (CRO) may no longer be prepared to test certain types of product or carry out particular tests requested by a client; a research institute may decide not to carry out any research that causes severe suffering or use certain species.Learning outcome 2.1 Describe the differing views, within society, relating to the scientiﬁc uses of animals and recognise the need to respect these.Key points to get across:– This LO links to module 1 on national legislation, so it is helpful to refer to details of the legislation to re-enforce the responsibility message.– Everyone coming into contact with animals has a responsibility to treat them with respect and consideration, minimise use and suffering and raise issues of concern so that safeguards can be implemented or maintained. Good welfare and thoughtful use of animals in experiments is essential for good science.– The use of animals is a privilege not a right. The ASPA sets minimum standards for deciding whether and how animals are used (see module 1: national legislation) but trainees have a responsibility to try to improve on these: i.e. to implement the spirit of the ASPA not just the letter of the law.– PIL standard conditions set out the speciﬁc responsibilities of personal licensees for the welfare of the animals they perform procedures on.How delivery could be achieved:– Ask trainees to describe their impression of a day in the life of laboratory animals and of the impact on animals of a procedure they have seen used. Then discuss within the class whether they have accurately represented the harms/welfare issues including lifelong harms, or underplayed these.– Introduce a discussion on transparency and reporting and ask trainees under what circumstances they would feel able to raise concerns about a colleague.– Introduce concept of professionalism and integrity in research and ask trainees to discuss their thoughts on appropriate professional behaviours with regard to both animals and people.– Ask trainees to read the PIL standard conditions and identify any that they think are not relevant to their area of work.Learning outcome 2.2 Describe the responsibility of humans when working with research animals and recognise the importance of having a respectful and humane attitude towards working with animals in research.Guiding principles to help deliver the ethics learning outcomes of module 2 for personal licensees

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69August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareGuiding principles to help deliver the ethics learning outcomes of module 2 for personal licenseesKey points to get across:– Ethics encompasses not only how an animal is used, but also whether an animal is used i.e. the justiﬁcation for the research. Too often ethics is interpreted only in terms of animal welfare and application of the 3Rs. Both of these issues are important as they concern the reduction of animal suffering and are thus a factor in ethical decision-making. However ethics encompasses a much wider set of considerations about what it is justiﬁed to do to animals in the name of science.– Application of the 3Rs to reduce animal use and suffering and enhance animal welfare must be considered throughout the lifetime of the animal up to and including euthanasia.– The importance of constructive criticism (by and to themselves), of being able to challenge assumptions and of thinking more broadly about the wider implications of their work for society at large.– The sanctions that can be applied if PILs do not consider the consequences of their actions.– Methods are updated and reﬁned and what is considered acceptable changes over time. Use of ‘we’ve always done it this way’ as an argument for maintaining unreﬁned and poor practice does not advance the quality and utility of science.How delivery could be achieved:– Use examples (preferably local and targeted to the work trainees will be doing) to illustrate how to identify ethical issues, to start them thinking. Seek peer support from existing thoughtful licensees or AWERB members.– Describe a surgical procedure (and potential harms) in a rat (e.g. an IV cannula with tether) without context and then for a range of research purposes. Ask whether the research context changes how the trainees feel about the harms caused to the animal.– Provide a section of a project licence or grant proposal and ask trainees to identify the ethical issues within it.– Ask trainees to give an example of (i) an ethical and (ii) a welfare issue. Review their suggestions making sure they recognise that ethics is not just about welfare, and discuss.– Show how to design a procedure that is appropriate to the experimental aims and causes the least harm to the animals and visit the facility to illustrate this; use simple examples of common procedures that people can relate to when putting experiments together.– Ask them to think through:– What questions would I ask of my study before I start?– What questions should I ask at the end?– How would I describe the beneﬁts of the work and how/why do they outweigh the harms?– How would I justify my work to myself and my colleagues, the Home Ofﬁce Inspector, or a stranger?– Use discussion topics to consider the ethical issues around animal use for studies/procedures such as:– Research into diseases caused / worsened by human behaviour e.g. pollution from vehicles and the effects this has on COPD; safety assessment of vaping; effects of night club noise on hearing; drug addiction studies.– Development of brain organoids and their implantation into animals.– Treating or triaging patients based on survival predictions (casualty, emergency situations, humanitarian aid).– Cloning of agricultural or competition animals.– Long term housing of animals; removal of enrichment for an experimental purpose.– Provide an example of why a project was turned down.– Turn trainees into an AWERB getting them to role-play different categories of staff with an example project licence to review.– Use a dilemma website tool or ethical reasoning tool.Learning outcome 2.3 Identify ethical and animal welfare issues in their own work and be aware and able to reﬂect on the consequences of their own actions.

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70Animal Technology and Welfare August 2020Key points to get across:– Scientiﬁc research uses public money and is done ‘for public beneﬁt’. Therefore information should be available to the public on what is done.– Openness and transparency contribute to ensuring public trust and conditional acceptance of animal use. Explain the role and content of the non-technical summary (NTS) and the Concordat on Openness in this respect.18,19– Accurate information is critical to informing opinion. A decision based on inadequate information on all the relevant factors is a prejudiced decision not an ethical one.– It is obviously acceptable to hold differing views but opinions need to be based on correct facts, not rhetoric (from any source) and it is essential to be able to recognise and acknowledge the difference.– Everyone can have their own beliefs but all must work within the ethical framework of the ASPA.How delivery could be achieved:– Use good and poor examples from the NTS as discussion points.– Use recent Mori poll information9 – discuss the questions and responses and how trainees would have responded.– As a thought-starter, ask trainees what would make them trust people working in a different ﬁeld such as climate change or human cloning. What expectations would they have? Then relate this back to their own ﬁeld and ethical values.– Provide links to the Concordat on Openness and discuss what could be done to deliver it. Ask trainees what their own establishments do about openness and suggest they ﬁnd out if they do not already know.Learning outcome 2.4 Recognise that compliance with ethical principles may contribute to the long-term trust and acceptance in scientiﬁc research from the general public.Key points to get across:– All establishments must have a Culture of Care and trainees have a responsibility to ﬁnd out about the elements of this.– The deﬁnition and expectations of a culture of care and what this looks like in practice, emphasising it is for people (treating them thoughtfully and with consideration and respect) as well as for animals.– The Culture of Care should deﬁne the local standards expected but as individuals, trainees should always look for ways to improve current practice. They should lead by example and be aware of the effect their behaviour has on others around them.How delivery could be achieved:– The deﬁnition of a Culture of Care can be found in the RSPCA/LASA Guiding Principles on good practice for AWERBs20 so this section of the document could be provided as a handout and talked through. The Norecopa International Culture of Care website13 also has excellent information on the key factors which foster a culture of care with details of how this can be developed and promoted.– Ask trainees to think about what the term means to them and what they think are the key components for their research group.– Some trainers teach this later in the course bringing in the named people, AWERB members or project licensees to talk about their own role, expectations and experiences.Learning outcome 2.12 Describe the need for a culture of care and the individual’s role in contributing to this.Key points to get across:– This LO is relevant to all other LOs and all modules.How delivery could be achieved:– Provide a handout with up to date references and links to useful websites.– Provide information on the roles of all the Named Persons and the Home Ofﬁce Liaison Contact (HOLC) and advise trainees to ﬁnd out who they are.– Advise trainees to sign up to key newsletters and to stay up to date.Learning outcome 2.13 Describe relevant sources of information relating to ethics, animal welfare and the implementation of the Three Rs.Guiding principles to help deliver the ethics learning outcomes of module 2 for personal licensees

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71August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareGuiding principles to help deliver the ethics learning outcomes of module 2 for personal licenseesAppendix 1 EU Module 2: Ethics, animal welfare and the Three Rs (level 1) [Core]This module provides guidance and information to enable individuals working with animals to identify, understand and respond appropriately, to the ethical and welfare issues raised by the use of animals in scientiﬁc procedures generally and, where appropriate, within their own programme of work. It provides information to enable individuals to understand and to apply the basic principles of the Three Rs.Learning Outcomes Trainees should be able to:2.1 Describe the differing views, within society, relating to the scientiﬁc uses of animals and recognise the need to respect these.2.2 Describe the responsibility of humans when working with research animals and recognise the importance of having a respectful and humane attitude towards working with animals in research.2.3 Identify ethical and animal welfare issues in their own work and be aware and able to reﬂect on the consequences of their own actions.2.4 Recognise that compliance with ethical principles may contribute to the long-term trust and acceptance in scientiﬁc research from the general public.2.5 Describe how the law is based on an ethical framework which requires 1) weighing the harms and beneﬁts of projects (the harm/beneﬁt assessment) 2) applying the Three Rs to minimise the harm, maximise beneﬁts and 3) promote good animal welfare practices.2.6 Describe and discuss the importance of the Three Rs as a guiding principle in the use of animals in scientiﬁc procedures.2.7 Explain the Five Freedoms and how these apply to laboratory species.2.8 Describe the concept of harms to animals including avoidable and unavoidable suffering, direct, contingent and cumulative suffering.2.9 Describe the severity classiﬁcation system, and give examples of each category. Describe cumulative severity and the effect this may have on the severity classiﬁcation.2.10 Describe the regulations regarding re-use of animals.2.11 Describe the importance of good animal welfare including its effect on scientiﬁc outcomes as well as for societal and moral reasons.2.12 Describe the need for a Culture of Care and the individual’s role in contributing to this.2.13 Describe relevant sources of information relating to ethics, animal welfare and the implementation of the Three Rs.2.14 Be aware of different search tools (e.g. EURL ECVAM Search Guide) and methods of search (e.g. systematic reviews,21 meta analysis).22Appendix 2 Ethics and the AWERBEthics is integral to the functions and tasks of the AWERB as would be expected by its title. Ethics is speciﬁcally mentioned in two of the four key AWERB functions, including with respect to training i.e. –• to provide a forum for discussion and development of ethical advice to the establishment licence holder on matters relating to animal welfare, care and use and• to support named persons and other staff dealing with animals, on animal welfare, ethical issues and provision of appropriate training.To meet the need for a forum for discussion, AWERBs could organise ethics related activities as suggested in both the ‘Guiding Principles on good practice for Animal Welfare and Ethical Review Bodies’ and the ‘AWERB as a Forum for Discussion’ documents.17,20 For example:• considering whether an establishment wants to rule out certain types of work, or use of certain species or techniques; or how it deals with severe procedures• organising informal lunchtime discussions enabling people to bring novel ideas or consider controversial issues, or discuss their own work and its ethical implications• recruiting (to the above) someone from a related but different ﬁeld to compare viewpoints e.g. a zoologist vs a laboratory animal viewpoint• appointing ‘ethics champions’ who can raise difﬁcult underlying ethical questions• considering the emotional wellbeing of staff involved in killing animals• for people who have worked abroad, discuss what they are permitted to do in their country and whether and how this differs from the UK.Making such discussions widely open to staff encourages them to understand and be more aware of ethical issues and consider the implications for their own work, so contributing to the culture of care. It also has the additional advantage of exposing AWERB members themselves to ethical discussions since many will probably not have had any ethics training unless they have attended the relevant modules.AcknowledgementsThe authors would like to thank trainers from the three UK training bodies for their valuable input and comments during the development of this document.Suggested background reading and resourcesBioethicsNufﬁeld Council on Bioethics animal research web page: nufﬁeldbioethics.org/topics/animals-food-and-environment/ animal-research

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72Animal Technology and Welfare August 2020Susan Gilbert, Gregory E. Kaebnick, and Thomas H. Murray, eds., Animal Research Ethics: Evolving Views and Practices, Hastings Center Special Report 42, no. 6 (2012): S1–S40. animalresearch.thehastingscenter.org/special-report/RC Simmons et al. (2018) Bioethics and Animal Use in Programs of Research, Teaching, and Testing. Boca Raton (FL): CRC Press/Taylor & Francis. pubmed.ncbi.nlm.nih.gov/29787201/Kraus, A. Lanny, and Renquist, David, eds. (2000) Bioethics and the Use of Laboratory Animals: Ethics in Theory and Practice. ACLAM. aclam.org/media/d9ecd55d-5edc-4fce-b0a4-9ad6e7705da0/P-RsCw/ ACLAM/Publications/Bioethics_Kraus.pdfAWERBs and ethical reviewRSPCA/LASA Guiding principles on good practice for AWERBs: lasa.co.uk/PDF/AWERB_Guiding_Principles_2015 _ﬁnal.pdfRSPCA Lay Members’ Resource Book, Appendix A: What is Ethics: tinyurl.com/RSPCALMRBRSPCA/LASA/LAVA/IAT meeting report: Putting Ethics into the AWERB: tinyurl.com/AWERB-UK2017RSPCA Ethical Review webpages. science.rspca.org.uk/ sciencegroup/researchanimals/ethicalreviewRöcklinsberg H, Gamborg C, Gjerris M. (2014) A case for integrity: gains from including more than animal welfare in animal ethics committee deliberations. Laboratory Animals 48(1):61- 71. doi: 10.1177/ 0023677213514220References1 European Commission (2013) A working document on the development of a common education and training framework to fulﬁl the requirements under the Directive. ec.europa.eu/environment/chemicals/lab_animals/pdf/guidance/education_training/en.pdf2 World Medical Association (2013) Declaration of Helsinki. wma.net/what-we-do/medical-ethics/declaration-of-helsinki/3 Royal College of Veterinary Surgeons UK Code of Conduct for Veterinar y Surgeons. rcvs.org.uk/setting-standards/advice-and-guidance/code-of-professional-conduct-for- veterinary-surgeons/4 NC3Rs/BBSRC/Defra/MRC/NERC/Royal Society/Wellcome Trust (2019) Responsibility in the use of animals in bioscience research: expectations of the major research councils and charitable funding bodies, 3rd edition. London: NC3Rs. nc3rs.org.uk/responsibility-use- animals-bioscience-research5 S Jarvis, JEL Day, B Reed (undated) Ethical guidelines for research in animal science (for publication in Animal) animal-journal.eu/instructions-and-policies/6 Boyd KM, Higgs R, Pinching AJ (eds) (1997) A new dictionary of medical ethics. BMJ Publishing group: London.7 Home Ofﬁce (2012) Animals (Scientiﬁc Procedures) Act 1986 (ASPA). gov.uk/government/publications/consolidated-version-of-aspa-19868 Ethics from The Internet Encyclopedia of Philosophy (IEP) (ISSN 2161- 0002) iep.utm.edu/ethics/9 Ipsos MORI (2018) Public attitudes to animal research in 2018. ipsos.com/ipsos-mori/en- uk/public-attitudes-animal-research-201810 Bruner, J. S. (1960). The Process of Education. Cambridge, Mass: Harvard University Press.11 Education Partnerships, Inc. (2012) The Spiral Curriculum. ﬁles.eric.ed.gov/fulltext/ED538282.pdf12 UK Research Integrity Ofﬁce (2019) Research Integrity: A Primer on Research Involving Animals. UKRIO, Croydon, UK. doi.org/10.37672/UKRIO.2019.01.animals13 The International Culture of Care Network: norecopa.no/coc14 Lakmal Abeysekera & Phillip Dawson (2015) Motivation and cognitive load in the ﬂipped classroom: deﬁnition, rationale and a call for research, Higher Education Research & Development, 34:1, 1-14, DOI: 10.1080/07294360.2014.93433615 Bouwmeester Rianne A.M. [et al.]. (2019) Flipping the medical classroom: Effect on workload, interactivity, motivation and retention of knowledge. Computers and Education 139: 118-128. doi.org/10.1016/j.compedu.2019.05.00216 Anna Therese Steen-Utheim & Njål Foldnes (2018) A qualitative investigation of student engagement in a ﬂipped classroom, Teaching in Higher Education, 23: 3, 307-324, DOI: 10.1080/13562517.2017.137948117 RSPCA, LASA, LAVA, IAT, University of Nottingham & ESRC (2017) Delivering Effective Ethical Review: The AWERB as a ‘Forum for Discussion’. view.pagetiger.com/AWERB/AWERB18 Understanding Animal Research (undated) Writing Non-Technical Summaries: A Researcher’s Guide. understandinganimalresearch.org.uk/ﬁles/5115/ 3235/6558/Writing_NTS_summaries.pdf19 Concordat on Openness on Animal Research in the UK: concordatopenness.org.uk/20 RSPCA/LASA Guiding Principles on Good Practice for AWERBs (2015) lasa.co.uk/PDF/AWERB_Guiding_Principles_2015_ﬁnal.pdf21 Systematic Review Center for Laboratory animal Experimentation (SYRCLE) radboudumc.nl/en/research/ departments/health-evidence/systematic-review-center-for-laboratory-animal-experimentation22 Carlijn R. Hooijmans, Joanna IntHout, Merel Ritskes-Hoitinga, Maroeska M. Rovers, Meta-Analyses of Animal Studies: An Introduction of a Valuable Instrument to Further Improve Healthcare, ILAR Journal, Volume 55, Issue 3, 2014, Pages 418-426, https://doi.org/10.1093/ilar/ilu042Guiding principles to help deliver the ethics learning outcomes of module 2 for personal licensees49Haven’t the time to write a paper but want to have something published? Then read on!This section offers readers the opportunity to submit infor mal contributions about anyaspects of Animal Technology. Comments, observations, descriptions of new or refinedtechniques, new products or equipment, old products or equipment adapted to new use,any subject that may be useful to technicians in other institutions. Submissions can bepresented as technical notes and do not need to be structured and can be as short or aslong as is necessary. Accompanying illustrations and/or photos should be high resolution.NB. Descriptions of new products or equipment submitted by manufacturers are welcomebut should be a factual account of the product. However, the Editorial Board gives nowarranty as to the accuracy or fitness for purpose of the product.What 3Rs idea have you developed?EMMA FILBYMira Building, University of Cambridge, University Biomedical Services,Charles Babbage Road, Cambridge CB3 0FSCorrespondence: emma.filby@admin.cam.ac.ukBased on an article written for the National Centre for the 3RsApril 2020 Animal Technology and WelfareTECH-2-TECHBackgroundEmma was invited to write an article as a 3Rschampion in NC3Rs ‘Tech 3Rs’ Issue 5, November2019.Here is her response describing how she has used anautomated system to reduce how frequently mousecage bedding ischanged without compromisingcleanliness.IntroductionOur unit opened in 2017, during the procurement ofnew equipment we had the opportunity to purchase adigital ventilated rack system from Tecniplast UK. Thecages are referred to as the Digitally Ventilated Cage orDVC. This system uses the data collected by sensorsbelow the cage to flag when to clean out based on thechange in an electromagnetic signal. To have thisfunctionality we first needed to create an algorithmduring a learning phase.The learning phase: devising analgorithmWe held a meeting to agree what warranted a cage basechange based on pictures to avoid being subjective. Wereferred to the Home Office Codes of Practice for thehousing and care of animals bred, supplied or used forscientific purposes (HOCoP) for advice on husbandrypractices to set our criteria, balancing hygiene and theimportance of olfactory cues to rodents and their needfor control over their environment.1We started the trial, noting when the cage reached thepoint it required a base change. We assessed airquality, what proportion of the cage base was wet andwhether the animals still had choice over theirenvironment and their ability to show spatial separationof different behaviours such as nesting and excretion,for example their nest was free of faeces. During the‘learning phase’ we asked our Named VeterinarySurgeon (NVS) and Home Office inspector (HOI) tocheck that they agreed with our assessment.APRIL_1-628207435_4-628196990.e$S:Animal Technology and Welfare 24/9/20 06:51 Page 49

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73August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2021 Animal Technology and WelfareSummaryThis article is based on a workshop presented at the Royal Society of Biology (RSB) Animal Science Group and the Home Ofﬁce Animals in Science Regulation Unit annual Animal Science Meeting on the 11th December 2020. At the workshop we aimed to:• share and summarise the results of the pre-workshop survey• concentrate on common themes• produce 3 ideas to improve training, for ASRU and RSB to take forwardBackground The RSB Animal Science Group and the Home Ofﬁce Animals in Science Regulation Unit (ASRU) annual Animal Science Meeting is for researchers, support staff and administrators in animal research. In 2020, the meeting was held online over two afternoons, with talks on the ﬁrst day and a series of workshops on the second. Pre-registration was required for the meeting with the number of participants limited and within those accepted to attend, fourteen allocated for each of the workshops. Attendees chose the workshop they wished to attend. The audience for this workshop ranged from 49Haven’t the time to write a paper but want to have something published? Then read on!This section offers readers the opportunity to submit infor mal contributions about anyaspects of Animal Technology. Comments, observations, descriptions of new or refinedtechniques, new products or equipment, old products or equipment adapted to new use,any subject that may be useful to technicians in other institutions. Submissions can bepresented as technical notes and do not need to be structured and can be as short or aslong as is necessary. Accompanying illustrations and/or photos should be high resolution.NB. Descriptions of new products or equipment submitted by manufacturers are welcomebut should be a factual account of the product. However, the Editorial Board gives nowarranty as to the accuracy or fitness for purpose of the product.What 3Rs idea have you developed?EMMA FILBYMira Building, University of Cambridge, University Biomedical Services,Charles Babbage Road, Cambridge CB3 0FSCorrespondence: emma.filby@admin.cam.ac.ukBased on an article written for the National Centre for the 3RsApril 2020 Animal Technology and WelfareTECH-2-TECHBackgroundEmma was invited to write an article as a 3Rschampion in NC3Rs ‘Tech 3Rs’ Issue 5, November2019.Here is her response describing how she has used anautomated system to reduce how frequently mousecage bedding is changed without compromisingcleanliness.IntroductionOur unit opened in 2017, during the procurement ofnew equipment we had the opportunity to purchase adigital ventilated rack system from Tecniplast UK. Thecages are referred to as the Digitally Ventilated Cage orDVC. This system uses the data collected by sensorsbelow the cage to flag when to clean out based on thechange in an electromagnetic signal. To have thisfunctionality we first needed to create an algorithmduring a learning phase.The learning phase: devising analgorithmWe held a meeting to agree what warranted a cage basechange based on pictures to avoid being subjective. Wereferred to the Home Office Codes of Practice for thehousing and care of animals bred, supplied or used forscientific purposes (HOCoP) for advice on husbandrypractices to set our criteria, balancing hygiene and theimportance of olfactory cues to rodents and their needfor control over their environment.1We started the trial, noting when the cage reached thepoint it required a base change. We assessed airquality, what proportion of the cage base was wet andwhether the animals still had choice over theirenvironment and their ability to show spatial separationof different behaviours such as nesting and excretion,for example their nest was free of faeces. During the‘learning phase’ we asked o ur Named VeterinarySurgeon (NVS) and Home Office inspector (HOI) tocheck that they agreed with our assessment.APRIL_1-628207435_4-628196990.e$S:Animal Technology and Welfare 24/9/20 06:51 Page 49ASRU-RSB workshop: impact of COVID-19 on present and future training for animal research – issues and opportunities for training and CPD in light of COVID-19LINDA HORAN1 and NGAIRE DENNISON2 1 University of Strathclyde, John Arbuthnott Building, SIPBS BPU, 161 Cathedral Street, Glasgow G4 0RE2 University of Dundee, Nethergate, Dundee DD1 4HNCorrespondence: linda.horan@strath.ac.uk; n.dennison@dundee.ac.ukTECH-2-TECH

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74Animal Technology and Welfare August 2020a lay person to a Director of Scholarship from the Open University. The small and varied cohort means that the responses collected are possibly not representative across establishments. We asked participants to complete a pre-workshop questionnaire and collated the results which are set out below and then during the session we addressed the issues raised and shared tips to assist with problems. The low numbers of responses are likely to be a reﬂ ection of the fact that the participant roles means that they probably did not have the knowledge needed to answer the questions.3 they wished to attend. The audience for this workshop ranged from a lay person to a Director of Scholarship from the Open University. The small and varied cohort means that the responses collected are possibly not representative across establishments. We asked that participants completed a pre-workshop questionnaire and collated the results which are shared below and then during the session we addressed the issues raised and shared tips to assist with problems. The low numbers of responses are likely be a reflection of the fact that the participants roles means that they probably didn’t have the information needed to answer the questions Figure 1. Concerns about Named Person Training: Of the respondents’ the majority said that they did not have concerns about training for Named People. We asked this question to see if there was a lack of a particular training courses or if people were Of the respondents, the majority commented that they did not have concerns about training for Named People. We asked this question to see if there was a lack of a particular training course or if people were concerned about the quality of online courses. The responses we received would suggest not. However, there were only 6 respondents.Figure 2. Appropriate training for Personal Licensees and techniques?We have highlighted two areas in yellow – training for physical methods of euthanasia and for methods new to the establishment, as these became a recurring theme. The main concerns revolved around issues caused by the need for social distancing between trainer and new trainees related to the COVID-19 environment. The Schedule 1 issue resurfaced several times, so we wondered whether this indicated that training in physical methods of Schedule 1 killing is a more general issue. Therefore in response we added some Schedule 1 advice into the end of the workshop.Figure 1. Concerns about Named Person training. 4 concerned about the quality of on-line courses, the responses we received would suggest not, however, there were only 6 respondents’. Figure 2. Appropriate training for Personal Licensees and techniques? We have highlighted two areas in yellow; training for physical methods of euthanasia and for methods new to the establishment as these became a recurring theme. The main concerns revolved around issues caused by the need for social distancing between trainer and new trainees related to the Covid-19 environment. The Schedule 1 issue resurfaced several times, so we wondered whether We wondered if there were particular issues with large animals however it was obvious from responses that the audience was too small to gain this information. Although there is a comment that there are difﬁ culties for less usual species. Schedule 1 was again highlighted as a problem.Figure 3. Species related problems.5 this indicated that training in physical methods of Schedule 1 killing is a more general issue, therefore in response we added some Schedule 1 advice into the end of the workshop. Figure 3. Species related Problems. We wondered if there were particular issues with large animals, however, it was obvious from responses that the audience was too small to gain this information, although there is a comment that there are difficulties for less usual species. Schedule 1 was again highlighted as a problem. Figure 4. Asepsis / surgical training.6 Figure 4. Asepsis / surgical trainingThe point behind this question was that we had assumed that methods of training for surgery would have had to have changed in the current environment and we hoped to share that information with the group. It became clear during discussion that some establishments had taken the view that the risk of Covid-19 was too great and no recommencement of training has started.The point behind this question was that we had assumed that methods of training for surgery would have had to have changed in the current environment and we hoped to share that information with the group. It became clear during discussion that some establishments had taken the view that the risk of COVID-19 was too great and no recommencement of training had started.Tech-2-Tech

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75August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareTech-2-TechThe responses provided here reinforced our initial premise that practical training was a problem.Figure 5. Areas of concern.7 Figure 5. Areas of ConcernThe responses provided here reinforced our initial premise that practical training was a problem. Figure 6. New practises implemented.8 Figure 6. New Practises Implemented. A nice practical solution to inconsistencies in training was suggested in response 2. While the problem of setting up a new surgical model was again highlighted in discussions around response 5. The possible need to establish a training network was identified. There was a positive comment on the accessibility of on-line training courses. An acceptable practical solution to inconsistencies in training was suggested in response 2. While the problem of setting up a new surgical model was again highlighted in discussions around response 5. The possible need to establish a training network was identiﬁ ed. There was a positive comment on the accessibility of online training courses.By asking this question we hoped to learn from other mistakes. The response rate was very low but response 2 highlights a concern around a lack of control on the quality of on-line training.Highlighted issuesIssues primarily due to/driven by COVID-19 –• issues with practical training• establishment of new models• online training and external course QAGeneral issues but highlighted by COVID-19 related difﬁ culties.• physical methods of killing (Schedule 1)Practical solutionsSmall rooms Figure 7. New practises that did not work.9 Figure 7. New Practises that did not work. By asking this question we hoped to learn from other mistakes, the response rate was very low but response 2 highlights a concern around a lack of control on the quality of on-line training. Highlighted IssuesIssues primarily due to / driven by COVID-19 - Issues with practical training. - Establishment of new models. - Online training and external course QA. 10 General issues but highlighted by COVID-19 related difficulties. - Physical methods of killing (schedule 1). Practical solutions Small rooms Figure 8a & 8b. Pop-up stand room dividers. Purchasing a clear, not too expensive, pop-up poster stand allows the division of a space or small room so that where a 2 metre distance cannot be maintained, training on bespoke equipment can 10 General issues but highlighted by COVID-19 related difficulties. - Physical methods of killing (schedule 1). Practical solutions Small rooms Figure 8a & 8b. Pop-up stand room dividers. Purchasing a clear, not too expensive, pop-up poster stand allows the division of a space or small room so that where a 2 metre distance cannot be maintained, training on bespoke equipment can Figure 8a and 8b. Pop-up stand room dividers.

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77August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareTech-2-Techthe hamster. This unit has a height adjustment which makes the task easier. It is still a little awkward but both people are protected by the barrier and airﬂow.For basic handling training for mice using tube and cupping method, parallel ‘side by side’ training with a screen between can be very effective as the trainee can ‘mirror’ the handler. Figure 10a and 10b. Shows use of double-sided cage changing station in use for procedures. 13 An empty room was transformed into makeshift training space for PILC training. Each attendee had a trolley work-station which was set up in advance and the clear pop up screens were utilised. The visualiser (a webcam connected to a projector to allow demonstration of techniques to a whole class) was taken from a meeting room (which are no longer in use) and was used to demonstrate the suturing. Procedures Figure 10a & 10b. Shows use of double-sided cage changing station in use for procedures. 13 An empty room was transformed into makeshift training space for PILC training. Each attendee had a trolley work-station which was set up in advance and the clear pop up screens were utilised. The visualiser (a webcam connected to a projector to allow demonstration of techniques to a whole class) was taken from a meeting room (which are no longer in use) and was used to demonstrate the suturing. Procedures Figure 10a & 10b. Shows use of double-sided cage changing station in use for procedures. Figure 11 c and 11 d. Conﬁguration of wheeled ﬁxed screens stands for animal handling training. 14 The double-sided cage changing station was utilised for injections where the animal must be restrained by a technician due to the parasites being injected into the hamster. This unit has a height adjustment which makes the task easier, it is still a little awkward but both people are protected by the barrier and airflow. For basic handling training for mice using tube and cupping method, parallel “side by side” training with a screen between can be very effective as the trainee can “mirror” the handler. Figure 11 c & 11 d: Configuration of wheeled fixed screens stands for animal handling training. 14 The double-sided cage changing station was utilised for injections where the animal must be restrained by a technician due to the parasites being injected into the hamster. This unit has a height adjustment which makes the task easier, it is still a little awkward but both people are protected by the barrier and airflow. For basic handling training for mice using tube and cupping method, parallel “side by side” training with a screen between can be very effective as the trainee can “mirror” the handler. Figure 11 c & 11 d: Configuration of wheeled fixed screens stands for animal handling training.

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78Animal Technology and Welfare August 2020Use of digital media to help with training and assessment of competenceAs well as training being difﬁcult due to social distancing measures, assessment of the competence of those performing procedures has offered challenges. Several methods have been developed to allow remote training and assessment (or re-assessment) of those performing procedures. – The most effective idea tried so far was pairing of Apple devices such as iPhone, iPads or a Mac paired with a second Apple mobile device and connecting via Facetime. This provides encrypted video calls between devices, thus minimising any security risks. The person being assessed may need to ﬁnd a method to ‘prop’ up their device (tripods can be purchased for around £20) to allow visualisation of what they are doing. The person assessing can be socially distanced in the room, next door or in another part of the unit/building. Something similar could be set up via other Apps such as WhatsApp.– A webcam is mounted on a tripod (or worn on the head) and connected to a laptop or PC either wirelessly or by a long USB cable. This was used to give a 2m separation for demonstrating stereotaxic cranial surgery. The surgeon performed their normal surgery whilst the trainee was able to observe what was happening at the surgical site and ask questions or discuss the procedure with the surgeon, all with social distancing. It is possible to get USB cables in longer lengths and direct connection avoids potential security risks associated with Wi-Fi.– Wi-Fi go-pro type camera streaming to a phone has also been tried for assessment of competencies but it was not straightforward in the unit it was tried in due to Wi-Fi problems, – Remote NVS visits can be achieved using similar methods, the purchase of a Gimbal (a pivoted support that allows the rotation of an object about a single axis) to steady the camera can be a useful tool to utilise.Highlighted issues New modelsWe were already aware from our own establishments that setting up new models, such as the complex ones displayed in Figures 12 a-d is a complicated process especially when there is no expertise already on-site, but the COVID-19 environment has made it even more difﬁcult. One solution considered by the group that would help in this regard would be to set-up a network of establishments and individuals within them, with skills in speciﬁed techniques/models that they are willing to help train other people to do. This might be something that IAT/LAVA or other laboratory science organisations might be able to work together to set up. Figure 12a – 12d. Complex new models such as those seen above, can be difﬁcult to set-up in the current environment.17 17 17 17 Tech-2-Tech

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79August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareTech-2-TechOnline training and external course QAHow do we assess the quality of the courses currently provided online? Many courses had to rapidly change delivery from face to face to online. Has this affected the quality? We feel this is highly likely for at least some courses, as formats and approaches need to be tailored to the online platform and the subject of the training. There are a number of limitations for the online platforms that give different challenges for training than those experienced face to face. At least one person at the workshop was aware of online training for Named Persons that had been very unsuccessful as the structure, previously successful for face to face did not encourage sufﬁ cient discussion in an online meeting. Whilst accreditation should create consistent quality, it was felt that there was still signiﬁ cant variation in course quality within and between different accrediting organisations and many of the courses moved online were accredited for face-to-face delivery rather than the current online content. Ultimately, it was felt that the only way for an NTCO to be conﬁ dent in the quality of a course at this time would be to attend it – this obviously has signiﬁ cant cost and time implications. As more trainees attend new online courses and provide feedback to the organisers and NTCOs, a better idea of what is and is not working well will hopefully be achieved. Physical methods of Schedule 1 e.g. Cervical dislocationCervical dislocation is the manual, rapid separation of the cervical vertebrae with accompanying lethal trauma to the spinal cord. When performed correctly, cervical dislocation appears to be a humane method of euthanasia. The discussion of this topic was more general and not limited to the issues seen due to social distancing requirements. For all practical methods, there comesa point where the trainee needs to ‘have a go’ themselves. The issue for practical euthanasia methods is the concern around the consequences to the animal if the technique is not immediately successful. Currently other than cadavers and videos, there are no training aids that are helpful in training for physical methods of euthanasia Cervical dislocation being a method highlighted as creating particular difﬁ culties in moving the trainee from practicing on dead animals to performing the method on live ones. Whilst not speciﬁ c to this method, the use of the Direct Observation of Practical Skills (DOPS) system was discussed. Developed from similar systems used in clinical settings, this is becoming a widely used method for training of practical skills relating to laboratory animals and is useful in setting expectations for trainer and trainee as to what the trainee must achieve in order to be considered competent. An example DOPS assessment sheet was shown (Figure 13a). BIOLOGICAL SERVICES – ASSESSMENT OFPRACTICAL SKILLSTECHNIQUE – CERVICAL DISLOCATIONADULT MOUSEPrior to attempting this you need to completethe following e-learning moduleHumane Methods of Killing Laboratory AnimalsYou will be sent an online invitation for thisYou will be assessed in handling at least 5 animals Assessment criteria/Components of the task:a) Understand the legal and ethical requirements surrounding euthanasiab) Personal Training and Schedule 1 Registerc) Choose Suitable PPE (mask or ﬂ ow hood, gloves)d) Conﬁ rm animal’s ide) Handle and restrain mouse correctly and sympatheticallyf) Dislocate the head and neck at the cranial vertebral level on the ﬁ rst attempt using ﬁ ngers or a suitable sized metal bar g) Conﬁ rm death by a suitable method e.g severing of neck or femoral vesselsh) Maximum of 3 attempts to restrain and dislocatei) Know the correct procedure for disposal of the carcass/tissuesj) Correctly amend cage label/room recordsk) Demonstrate a professional attitude towards performing the procedureGrading/Threshold statements‘Below expectations’• Failure to check legal authorities, animal details• Failure to choose suitable PPE• Poor animal handling; animal showing signs of distress• Euthanasia proposed when other animals are within sight/hearing distance• Death not conﬁ rmed• Candidate unwilling or nervous about procedureRequirements for ‘Meets Expectations’• Correctly states legal authorities and animal’s id• Correctly removes mouse from cage, handle and restrain empathetically• Conﬁ dently and rapidly applies dislocation technique at upper cervical level• Death conﬁ rmed as set out in Schedule 1• Cage label/record updated• Correct carcass/sharps disposalRequirements for ‘Exceeds Expectations’As ‘meets’ and• Explain reasons for choice of technique, can suggest alternatives• Excellent manual dexterity and sympathetic animal handling• Understands possible psychological effects on staff of performing euthanasia21BIOLOGICAL SERVICES - ASSESSMENT OF PRACTICAL SKILLSTECHNIQUE – CERVICAL DISLOCATION ADULT MOUSEPrior to attempting this you need to complete the following e learning moduleHumane Methods of Killing Laboratory AnimalsYou will be sent an online invitation for thisYou will be assessed in handling at least 5 animals Assessment criteria/ Components of the task:a) Understand the legal & ethical requirements surrounding euthanasiab) Personal Training & Schedule 1 Registerc) Choose Suitable PPE (mask or flow hood, gloves)d) Confirm animal’s ide) Handle & restrain mouse correctly and sympatheticallyf) Dislocate the head and neck at the cranial vertebral level on the first attempt using fingers or a suitable sized metal bar g) Confirm death by a suitable method e.g severing of neck or femoral vessels h) Maximum of 3 attempts to restrain & dislocatei) Know the correct procedure for disposal of the carcase/ tissuesj) Correctly amend cage label / room recordsk) Demonstrate a professional attitude towards performing the procedureGrading/ Threshold statements“Below expectations”• Failure to check legal authorities, animal details• Failure to choose suitable PPE• Poor animal handling; animal showing signs of distress• Euthanasia proposed when other animals are within sight/ hearing distance• Death not confirmed• Candidate unwilling or nervous about procedure Figure 13a.DOPS Assessment Criteria and Grading/Threshold statements for Cervical Dislocation.

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80Animal Technology and Welfare August 2020DOPS assessment should be provided to the trainee from the outset of training so they understand the expectations. A DOPS assessment should have grading/threshold statements, to clarify what would be considered below expectations, what needs to be achieved to meet expectations and may also have requirements for exceeding expectations. The assessment can be used to re-assess competence and this last category of requirements can help demonstrate improvement in conﬁdence or in the technique. The workshop did not come up with a solution to the issue of training in physical methods of killing but it was considered improved training aids would be extremely helpful.SummaryThe disparate people within the workshop allowed some good sharing of ideas but the limitation on numbers also meant that the discussions were limited to areas for which those people had knowledge.Several possible solutions to speciﬁc problems were presented, with more general solutions suggested in relation to: – Networking to improve knowledge transfer. This might be best done by Mentoring and/or having a list of those skilled in speciﬁc models and prepared to share their experience.– A need for new training aids, particularly for speciﬁc techniques where current training is difﬁcult such as physical methods of Schedule 1 killing.– Continued sharing and updating of DOPS assessments.DOPS marking sheet for: Euthanasia of mouse by Cervical Dislocation Level Feedback/ (eg ME) commentsUnit Induction Registered UserLegal and compliance:Checks PIL and PPL and/or Humane Killing RegisterCorrect PPE/ﬂow hood operationConﬁrm animal’s idAmend cage label/recordCorrect carcase/sharps disposalNumbers of mice?Condition/age/sex (as appropriate/relevant to handling needs?)Animal Welfare:Empathetic, safe animal handling and restraint Euthanasia out of sight/sound/ smell of conscious animals Knows backup euthanasia method in case of technique failureProcedural:State method of dislocation (ﬁngers or bar)Have suitably sized implement to handCervical dislocation per formed rapidly, at ﬁrst attemptPalpate gap between head/spineConﬁrm death by second method severing of either the femoral vessels or the jugular/carotid vessels of the neck.Cadaver/Tissue disposalProfessionalism: Procedure carried out humanely, in timely mannerRespect for animals and colleaguesWorkspace left tidyCommunicates/ Knows own limits 3Rs: Demonstrates understanding of reﬁnements; possible effects on staff moraleRequirement to be assessed as COMPETENT: Meets Expectations or better.Candidates scoring ‘Below Expectations’ on any point are recommended to ‘continue supervision’Candidate’s name:................................... Date: ................... Assessor’s name: ................................................................. Assessor’s signature: ............................................................GLOBAL RATING: COMPETENT CONTINUE (Circle) SUPERVISION Tech-2-Tech

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81August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2021 Animal Technology and WelfareConsisting of 13 chapters over 190 pages, with references given at the end of each chapter. Contributors are distributed over the USA, UK and Europe so a balanced view is achieved. The chapters cover general issues such as human-animal interactions, Culture of Care, animal emotions, abnormal behaviour, animal learning and animal training. The remaining chapters deal with species speciﬁc topics of the more common laboratory species namely Zebraﬁsh, mouse, rat, rabbit, dog, non-human primates (NHP) and the pig. It should be noted that the use of NHPs is probably more common in the USA and the Far East than it is in the UK and Europe. Although people frequently skip reading the preface, it is important that you do not in this case, as it gives the background to animal centric management and explains what the book is trying to accomplish. The editors acknowledge the inﬂuence of Russell and Burch’s book and their concept of the 3Rs, namely Replacement, Reduction and Reﬁnement. They believe that currently most applications of the 3RS are scientiﬁcally centred and that we need to change to an animal centred approach. They use the example of planning the procedures for modelling E coli induced diarrhoea in young pigs where study planning will typically focus on inoculation and frequency of faecal sampling, ignoring how oral gavage and repeated restraint may add stress to the general experience for the animals. Selection of measures used to ensure welfare within the experimental setting is determined once the experimental procedures have been set. Perhaps the pigs will be given additional enrichment such as an empty cardboard box to manipulate and extra straw for rooting, in other words, the welfare of the pigs throughout the study is considered only within the limits and the framework set by the study design. What the book is trying to accomplish is to show that instead of ﬁtting the animals into experimental conditions one should ﬁrst strive to adjust the baseline conditions (husbandry and housing) to better meet the needs and preferences of the animals and then ﬁt the experimental conditions within those.The initial chapter of the book deals with human-animal interactions and points out that these are daily, signiﬁcant and often unavoidable components that can impact not only on the research animals but also on the people who work with them and the eventual research outcomes. The interactions, unless deliberate effort is made, can negatively affect both animals and humans. The authors deﬁne human-animal interactions and provide a useful graphic of the associated terminology. Impacts are divided into several frameworks and in some cases further divided into harmful and beneﬁcial and for animals and humans. They use the term human-animal interactions rather than human-animal relationships as they consider this term restrictive. Suggestions for changing human behaviour to improve animal welfare are also made, as are proposals for further work. The chapter on a Culture of Care suggests a working concept in relation to animal research and testing that includes elements that relate to appropriate attitudes, behaviour, mindsets and mutual respect between staff with different roles. The authors consider that regardless of the context the term should be used thoughtfully rather than it becoming a watch-word that is used without thought or what is meant by the term. They discuss implementation of a Culture of Care from BOOK REVIEWSAnimal-centric care and management: Enhancing Reﬁnement in Biomedical ResearchEdited by Dorte Bratbo Sørensen, Sylvie Cloutier and Brianna N GaskillJas Barley

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82Animal Technology and Welfare August 2020various perspective, the tools used to shape the Culture of Care, role of local ethics committees on animal care and use, the challenge involved and assessment of the culture. Opportunities for promoting the culture are also considered. Further chapters go on to discuss Animal Emotions and Moods and why they are important, together with abnormal behaviour A very useful table is provided of what are considered as abnormal behaviours in the species covered in the book. Why those involved with animals should care about abnormal behaviour is also covered. The chapters on Animal Learning and Animal Training should be of great interest to all readers as the successful application of training an animal and dare we also suggest, the humans, to act in a particular way during husbandry and research activities can greatly reduce the associated stress of such activities for all concerned. Obviously, it is important that both aspects should be considered as inappropriate methods could lead to more harm being inﬂicted on the animals and trainers. The chapters on the various species all take a slightly different approach which is probably due to the limits of available information. The chapter on Zebraﬁsh includes biological health, environmental conditions and natural behaviour, psychological and emotional wellbeing (possibly not an aspect of a ﬁsh care normally considered), special considerations including strains of Zebraﬁsh, human animal interaction and restrictive types of research and special considerations. Mice are considered under sections that discuss the cognitive abilities and emotions of the species, the environment and how to enrich it and what is practical and then concentrates on handling both from the animal’s and the handler’s perspective including the use of non-aversive handling and standardising this in research. The authors of the rat chapter talk about free ranging rats and who they are, housing for rats in laboratories and covers the historical perspective and current knowledge including shelters exercise and mobility and complexity with recommendations for implementation. Relationships with rats in laboratories are also discussed including the historical perspective, current knowledge and recommendations for implementation.Information on rabbits looks at general background, environment, social needs, social housing, handling, the human animal bond and training. Whereas for dogs the authors consider dog senses and cognition, housing environment, enrichment in the dog laboratory facility, handling and management, training and treating dogs as individuals. Strangely the information on non-human primates feels relatively limited, perhaps due to the authors considering that more consideration to animal-centric managements has already been given to the species concerned than it has to rodents and other species. Covered in this section is basic biology and behaviour, social dynamics and group constellations relating to macaques, marmosets and tamarins including information on social housing of primates in captivity, the environment, foraging and food rich enrichment and socialisation and training. The pig chapter authors provide information under the headings of characterisation: morphology, domestication and breeds, cognitive and emotional abilities, the environment including design for comfort and enrichment as well as social environments, behavioural management including: handling, habituation and training are included as is consideration of how to proceed. The book is comprehensive, well- written and exceedingly well laid out. The principles of Animal-centric care and management may sound a new idea but for many animal care staff it is what we have been doing for many years, although there is always room for new ideas. However, the concept of designing experiments studies to consider the animals needs ﬁrst rather than the scientiﬁc needs is something is entirely new. Although only the common animals are covered, the principles are applicable to all species used in research, although some in depth knowledge of some of the more unusual species needs to be acquired. I commend this book to you and would recommend that it is made available to all staff from the most junior technician to the head of teams using animals in research. Book Reviews

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83August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareBook ReviewsAAALAC Fellowship – Coming soon!The AAALAC Committee have made the very difﬁcult decision to postpone applications for the AAALAC Fellowship for 2021. Uncertainty about international travel restrictions and whether facilities will be able to host visitors means that the Fellowship programme has been paused for 2021.However we are very excited to announce that when we return to the programme in 2022, we will be launching our new application process. This will be a simpliﬁed, online application and will be accessed via the AAALAC website. Details on how to apply can be found on the AAALAC website (www.aaalac.org/awards/fellowship-award), as well as via links from the AALAS (www.aalas.org) and IAT (www.iat.org.uk) websites. PET PORTRAITS Watercolour painting from your chosen photograph, provided framed Dimensions approximately 35cm x 25cm £120 All profits going to AS-ET For more details contact wendy.steel1@outlook.com Registered Charity Number 1133119 Bulletin16 •July 2020AS-ET NewsThe Trustees of AS-ET would like to send our best wishes to all those of you whoare having to work in difficult circumstances, ensuring your animals are cared forand essential science carries on. It is an amazing achievement to have continuedto get to work while public transport has been lacking and while avoiding beinginfected yourselves. Congratulations to all of you for demonstrating your cultureof care in a really practical way in these dangerous times. Please make sure tostill stay safe now that the restrictions have been lifted slightly – the incidence ofinfections has slowed down but the virus is still around.As you can imagine this has been a quiet time for AS-ET but we are ready forwhen circumstances allow people to get back to enrolling on courses. Our plansfor events to mark our tenth anniversary will remain on hold until the countrybecomes more settled, however remember the Congress Bursary Competition isstill running so start writing. Congress Bursary CompetitionIf you are studying for the IAT level 2 qualification or you passed it in 2018, 2019or 2020 and you are employed as a laboratory animal technician in the UK or theRepublic of Ireland you can enter the competition. This year’s topic is —‘What challenges and rewards have you experienced as an animal technician?’ Your 1,000 word essay should be submitted by 20th October 2020. Full details areon our website (www.as-et.org.uk).Patron: Professor Lord Naren Patel KTChair of Trustees: Professor Sir Richard Gardner FIAT (Hon) FRSSecretary to the Trust: Ken Applebee OBE FIAT FRSBTrustees: Stephen Barnett MSc (Hon) FRSB, Jasmine Barley MSc FIAT,Karen J Gardner, Wendy Steel BSc (Hons) FIATRegistered Charity Number 113319Registered Office: 5 South Parade, Summertown, Oxford OX2 7JLJul20:IATB NEW 10/7/20 10:02 Page 16

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84Animal Technology and Welfare August 2020This revised 3rd edition of the Introduction to Laboratory Animal Science, Technology and Welfare is a comprehensive, well organised, and beautifully illustrated manual for the education and training of people involved with laboratory animal technology and working in the ﬁeld of animal care and use. The edition is comprised of over 200 pages of text and almost 200 photographs, illustrations and tables organised into 16 chapters with a glossary and index. Although the targeted audience is for the Institute of Animal Technology (IAT) levels 1 and 2 laboratory animal technician certiﬁcations in the United Kingdom, the book also can be used as a training tool for educators, research technicians and principal investigators around the world.The book begins with an opening chapter on ‘Animal Health’, thus emphasising a most important topic for animal care and wellbeing. It continues with practical chapters on caging, housing, the facility, facility environment, nutrition, husbandry, hygiene, breeding, care of young animals, identiﬁcation methods, experimental procedures, euthanasia, handling, sexing and safety. Also included are chapters on laws, regulations, recommended guidelines and ethics and animal welfare. The chapters cover the breadth of common laboratory animal mammals and Zebraﬁsh used in research. Most likely because of the limited use of non-human primates in the UK, this species is not included in this edition. While the information on euthanasia and the governing laws, guidance and recommendations (Animals Scientiﬁc Procedures Act, 1986, amended 2012), is a requirement to workers in the UK, readers outside of the UK can easily utilise the information presented as an education tool, with practical comparisons to their own governance applications and guidance in programmes of laboratory animal care and use. The other chapters relating to aspects of laboratory animal technology are very informative and help guide the reader in identifying current ‘best practices’.The ﬁnal chapter is an extensive review of the common discussions involving ethics, philosophical theories and the impact of Russell and Burch’s 3Rs. This edition concludes with an excellent glossary and index.The book is relatively small and compact in size, thus enhancing its use as a teaching tool and quick reference manual. It is also very modestly priced at £20 ($25-30 US). The editors are commended for the information selected to present in this edition, the use of beautiful graphics and putting forward such a practical approach towards reﬁnement of laboratory animal care and use techniques. We highly recommend Introduction to Laboratory Animal Science, Technology and Welfare, 3rd Edition as an education and training reference in this ﬁeld. We also applaud the IAT for recognising the need, supporting the development and production of this important revised 3rd edition reference book.A personal note from Gail A. Thompson (IAT Vice-president):I wish to thank the IAT for the strong support the Institute has provided since its organisation in 1950, for producing training programmes and materials. As a young, enthusiastic technician in the late 60s and early 70s, it was difﬁcult for me to ﬁnd study materials for preparation for the AALAS certiﬁcation exams. Although AALAS was producing ‘Syllabus’ materials, the exams were not restricted to material in those recently produced documents. The ‘Bible’ for study material at that time was ‘The IAT Manual of Laboratory Animal Practice & Techniques’, edited by Douglas J. Short and Dorothy P. Woodnott, ﬁrst published in 1963. My copy was from the printing in 1971. My copy is highlighted and underlined on almost every page. What a valuable tool for our scientiﬁc ﬁeld it was when, so few resources were available. A few years ago, I was privileged to meet Dorothy Woodnott, a hero in our industry. This current manual produced by the IAT and revised by Stephen W. Barnett is but another sterling example of how the IAT has continued to produce and support training and reference materials for the proper care and humane use of laboratory animals, worldwide.Book ReviewsIntroduction to Laboratory Animal Science, Technology and Welfare, 3rd EditionRevised by Stephen W BarnettBruce W Kennedy, Heather L Narver, Gail A Thompson and Robert H Weichbrod