ATW August 2019 : simplebooklet.com

Editorial ixJas Barley, Chair of the Editorial BoardReport of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting 81Chloe Stevens, Penny Hawkins (Secretary), Robin Lovell-Badge, Robert Hubrecht, Huw Golledge, Anna Slaviero,Clare Ellis, Demi Minhinnett, Rebecca Terry, Katharina Hohlnaum, Dominic Wells, Thomas Snoeks and John MarshallA comparison of enrichment items for the promotion of natural gnawing behaviour in laboratory mice 93Irene Lopez JuaristiLegal and ethical aspects of using animals in research: A NACWO’s guide 99Matthew BiltonPAPER SUMMARY TRANSLATIONS 103OPINION ARTICLEThe 3Rs and optimisation in a decentralised research world: new perspectives on an established 117paradigmDarrell HoskinsTECH-2-TECHPro’s and pro’s of selective cleaning 121Mark WhiteTeam awesome: Why we can be proud 127Joanna KingNACWO my first year – dream job or nightmare? 133Rhys PerryPOSTER PRESENTATIONSRefinement: promoting Gallus Gallus welfare in an experimental poultry unit 137Katie Harris, Rachel New, Paul Smith, Billy Matthews and Jess WantAnimal handling in containment 140Christine Zverev, Shaun Baker and Dhruti PatelIn-vivo imaging at NIBSC and how we use it to promote the 3Rs 143Luke GurneyAn investigation of how a photocatalytic oxidation system can be used to reduce airborne allergens 145Joseph Pennock and Hayley BuckleFailure to provide food and water: implications and prevention 147Murad MiahThe effect of relative humidity on water intake in C57BL/6J mice 150Rebecca Towns and Karen Ekkelund PetersonBottle or bowl? Feeding and dosing in juvenile Göttingen minipigs 153Alice McNamara and Lucy AllenRefining oestrus selection in the animal unit to optimise transgenic production 155Christopher Wilson, Mike Walls, Davie Black and Julie ThomsonInstructions to Authors 157CONTENTSVol 18 No 2 August 2019iLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:18 Page i

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iiIAT REPRESENTATIVESOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richard Gardner MA PhD FRSB FIAT (Hon) FRSVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS,Stephen Barnett BA MSc FIAT (Hon) CBiol FRSB RAnTech,Miles Carroll PhD, Brian Cass CBE, Paul Flecknell MA Vet MB PhDDLAS DipLECVA MRCVS, FIAT (Hon), Penny Hawkins PhD BSc,Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSBDVMS (h.c.), DipECLAM FRAgS DipACLAM MRCVS,Fiona McEwen BSc BVM&S MSc MRCVS, Tim Morris BVetMed PhDDipACLAM DipECLAM CBiol FRSB CertLAS MRCVS,José Orellana BVSc MSc, Clive Page OBE PhD BSc,Jan-Bas Prins PhD MSc, Vicky Robinson CBE BSc PhD,Paul Sanders MIAT RAnTech, David Spillane FIAT,Gail Thompson RLATG, Robert Weichbrod PhD RLATGLife MembersCharlie Chambers MIAT RAnTech, Roger Francis MSC FIAT RAnTech,Pete Gerson MSc FIAT RAnTech, Cathy Godfrey FIAT RAnTech,John Gregory BSc (Hons) FIAT CBiol FRSB RAnTech, Patrick HayesFIAT DipBA RAnTech, Rober t Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills HonFIAT RAnTech,Honorary MembersMark Gardiner MIAT RAnTech, Andy Jackson MIAT, Sarah Lane MScFIAT, Brian Lowe MSc FIAT RAnTech, Sue McHugh BSc FIAT,Norman Mortell BA (Hons) MIAT RAnTech, Terr y Priest MBE FIATRAnTech, Trevor Richards BEM MIAT, David Spillane FIAT,Wendy Steel BSc (Hons) FIAT, Pete Willan DMS FInstLM MIATMembers of CouncilKen Applebee, Matthew Bilton, Kally Booth, Charlie Chambers,Steven Cubitt, Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Alan Graham, Nathan Hill, Linda Horan, Sam Jameson,Elaine Kirkum, Adele Kitching, Theresa Langford, Sylvie Mehigan,Steve Owen, Alan Palmer, Allan Thornhill, John Waters,Lynda Westall, Carole Wilson, Adrian WoodhouseCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary: Simon Cumming BSc FIAT RAnTechTreasurer: Glyn Fisher FIAT RAnTechAssistant Treasurer: Charlie Chambers MIAT RAnTechChair of Board of Educational Policy: Steven Cubitt MSc FIATRAnTechChair of Board of Moderators: Haley Daniels MBA MSc MIATRAnTech CIPDChair Registration & Accreditation Board: Ken Applebee OBE FIATCBiol FRSB RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATBranch Liaison Officer: Lynda Westall BSc (Hons) FIAT DMS RAnTechEFAT Representative: Charlie Chambers MIAT RAnTechWebsite Coordinator: Allan Thornhill FIAT RAnTechAnimal Welfare Officers and LABA Representatives:Matthew Bilton, Kally Booth, Lois Byrom, Simon Cumming,Nicky Gent, Sylvie Mehigan, John WatersATW/Bulletin Editorial Board: Jas Barley (Chair), Matthew Bilton,Nicky Gent, Patrick Hayes, Elaine Kirkum, Carole Wilson,Lynda WestallBoard of Educational Policy: Steven Cubitt (Chair), Steven Cubitt(Secretary), Adele KitchingBoard of Moderators: Haley Daniels (Chair), Simon Cumming,Cathy GodfreyModerators: Anthony Iglesias, Theresa Langford, Jenny Parks,Sarah ReedCommunications Group: Adrian Woodhouse (Chair), Nathan Hill,Elaine Kirkum, Teresa Langford, Sylvie Mehigan, Allan Thornhill,Lynda WestallCPD Officer: Charlie ChambersRegistration and Accreditation Board: Ken Applebee (Chair),Glyn Fisher (Secretary), Charlie Chambers, John Gregory,Cathy Godfrey, Kathy Ryder (Home Office), Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee: Alan Graham (Chair), Haley Daniels,Linda Horan, Adele Kitching, Allan Thornhill, John WatersDiversity Officer: Haley Daniels MBA MSc MIAT RAnTech CIPDUK Biosciences ASG Representative/Home Office: Steve Owen,Charlie Chambers, Alan PalmerIAT OFFICERS MAY BE CONTACTED VIA:IAT Administrator: admin@iat.org.uk OR 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 the journal, the Institute of Animal Technologywish to expound that the data and opinions appearing in the articles, posterpresentations and advertisements in ATW are the responsibility of thecontributor and advertiser concerned. Accordingly the IAT, Editor and theiragents, accept no liability whatsoever for the consequences of any suchinaccurate or misleading data, opinion, statement or advertisement beingpublished. Furthermore the opinions expressed in the journal do notnecessarily reflect those of the Editor or the Institute of Animal Technology.© 2019 Institute of Animal TechnologyAll rights reserved. No part of this publication may be reproducedwithout permission from the publisher.BRANCH SECRETARIES 2019Cambridge: Sarah Shorne cambridgebranch@iat.org.ukEdinburgh: Janice Young edinburghbranch@iat.org.ukHertfordshire & Essex: Joanna Cruden hertsessexbranch@iat.org.ukHuntingdon, Suffolk & Nor folk: 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: Rachel Sandy and Joanne Bland northeastbranch@iat.org.ukNorth West: Nicky Windows cheshirebranch@iat.org.ukOxford: April Shipton 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: Linda Horan westscotlandbranch@iat.org.ukLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:18 Page ii

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August 2019 Animal Technology and WelfareEditorialJas BarleyChair of the Editorial BoardFood and water is a fundamental necessity for most living things. However, huge numbers of people, especially children and the elderly,die every year from lack of food and not just in developing countries. Of course, poor quality drinking water of the standard that I wouldhesitate to water the garden with, due to the levels of contamination, does not improve human or animal chance of survival.On the other side of the coin in other areas of the world people are killing themselves by eating too much food in general or too muchof the wrong type of food. It would be an unusual person that is totally unaware of the concern about obesity especially in childreneven if they are doing nothing about it. When I was commuting I often used to see children on the way to school accompanied by anadult, eating bags of crisps and being washed down by lurid coloured fizzy drinks – healthy diet, I do not think so.Conditions which affect people’s perception of body image such as anorexia nervosa or bulimia, thus leading to starvation diets orgross overeating, are also a feature of modern life and perhaps reflect the emphasis society puts of food.The way we view and our knowledge about food is also reflected in the amount food wasted in this country and, I suspect, in mostdeveloped countries. It is something we should be ashamed of especially when many people are struggling to provide food for theirfamilies. At some point in the last 50 years we seemed to have arrived at the idea that food needs to be sanitised and perfect, witheverything washed and wrapped in ‘nice’ plastic bags and we reject food that falls below this ideal. The fact that shops need to market‘wonky’ vegetables as a separate item amazes me – they should see the crops that I produce in my garden, all shapes and sizes andall delicious.Similarly, animals, although that is usually down to indulgent owners, seem to suffer from a feast or famine situation when it comesto food. Where I live there are a lot of dog walkers or to be more accurate dog carriers, one even gets to ride in its own trolley becauseit is too fat to walk more than a few feet. Others are not so fortunate, many of the animals start their stay at the local RSPCA rehomingcentre due to being undernourished, if not starved.This issue of the Journal also concerns itself with food and water, or perhaps I should say the lack of both. The Report of the 2018RSPCA/UFAW Rodent and Rabbit Welfare Group meeting contains an update from the Home Office Inspectorate on the failure toprovide food and water to animals. It represents a significant cause of non-compliance, accounting for approximately 20% of non-compliance cases annually. Everyone recognises that Animal Technologists care passionately about the animals in the care and thatno one leaves an animal without food or water intentionally. The IAT Animal Welfare Group, have produced guidance on providing foodand water and perhaps more importantly, ways to prevent animals being left without either or both. The guidance the group hasproduced is available from the IAT website at www.iat.org.uk– look under media and documents. The topic is also discussed in theposter from Murad Miah which suggests some interesting ideas as both cause and possible ideas for prevention.Food and water are also the concern of two further posters, Rebecca Towns and Karen Ekkelund Petersen look at the effect of relativehumidity on water intake in C57BL/6J mice and Alice McNamara and Lucy Allen discuss the feeding of young Göttingen minipigs.As well as the RSPCA report our formal papers offer a comparison of enrichment items for the promotion of natural gnawing behaviourin laboratory mice by Irene Lopez Juaristi and Matthew Bilton’s NACWO guide to legal and ethical aspects of animal research.Finally, we offer a new approach for the Journal in that we publish the first of hopefully a regular Opinion Article. This first article isfrom Darrell Hoskins and looks at a new way of considering the 3Rs, particularly replacement.THE INSTITUTE OF ANIMAL TECHNOLOGYETHICAL STATEMENT“In the conduct of their Professional duties, Animal Technologists have a moral andlegal obligation, at all times, to promote and safeguard the welfare of animals intheir care, recognising that good laboratory animal welfare is an essential componentof good laboratory 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.”ixLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:18 Page ix

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August 2019 Animal Technology and Welfare81Report of the 2018 RSPCA/UFAW Rodentand Rabbit Welfare Group meetingCHLOE STEVENS,1PENNY HAWKINS (SECRETARY),1ROBIN LOVELL-BADGE,2ROBERT HUBRECHT,3HUW GOLLEDGE,3ANNA SLAVIERO,4CLARE ELLIS,5DEMI MINHINNETT,6REBECCA TERRY,7KATHARINA HOHLNAUM,8,9DOMINIC WELLS,10THOMAS SNOEKS2and JOHN MARSHALL111Research Animals Department, Science Group, RSPCA, Wilberforce Way, Southwater,West Sussex RH13 9RS2The Francis Crick Institute, 1 Midland Road, London NW1 1AT3UFAW, The Old School, Brewhouse Hill, Wheathampstead, Hertfordshire AL4 8AN4University of Surrey, Guildford, Surrey GU2 7AL5University of Northampton, University Drive, Northampton, Northamptonshire NN1 5PH6Durham University, Stockton Rd, Durham, Northumberland DH1 3LE7University College London, Cruciform Building, Gower Street, London WC1E 6BT8Institute of Animal Welfare, Animal Behaviour and Laboratory Animal Science, Department ofVeterinary Medicine, Freie Universität Berlin, Germany9Institute of Pharmacology and Toxicology, Department of Veterinary Medicine, FreieUniversität Berlin, Germany10Department of Comparative Biomedical Sciences, Royal Veterinary College, Royal CollegeStreet, London NW1 0TU11Home Office Animals in Science Regulation Unit, 14th Floor, Lunar House, 40 WellesleyRoad, Croydon CR9 2BYIntroductionThe RSPCA/UFAW Rodent (and now Rabbit) WorkingGroup has held a one-day meeting every autumn for thelast 25 years, so that its members can discuss currentwelfare research, exchange views on welfare issuesand share experiences of the implementation of the3Rs of replacement, reduction and refinement withrespect to rodent and rabbit use. A key aim of theGroup is to encourage people to think about the wholelifetime experience of laboratory rodents and rabbitsensuring that every potential negative impact on theirwellbeing is reviewed and minimised.This year’s meeting was held at The Francis CrickInstitute in London on 30th October 2018 and wasattended by over 80 delegates from the UK andoverseas. To mark 25 years of Rodent and Rabbitmeetings, the day opened with a retrospective look athow animal technology has developed over the past 25years, and how these developments have impactedlaboratory rodent and rabbit welfare. This was followedby a look to the future, with a talk that discussed howanimal welfare science and practices might changeover the next 25 years. Other presentations coveredways to encourage laboratory rats to nest-build bygiving them appropriate building materials, tips ondesigning new rabbit facilities to best promote rabbitwelfare and a discussion of how imaging techniquescan be used to refine experimental procedures andreduce the number of animals used in studies. The dayended with a presentation from the Home OfficeAnimals in Science Regulation Unit and an interactivediscussion session, both on the topic of ensuring thatlaboratory rodents and rabbits never go without food orwater. This report summarises the meeting and endswith a list of action points for readers to considerraising at their own establishments.Advances in animal welfare andtechnology over the last 25 yearsRobin Lovell-Badge, The Francis CrickInstituteA lot has happened since 1993. There have beenLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 81

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting82significant, sometimes dramatic, advances intechnology covering aspects such as animalhusbandr y, surgical practice, methods ofcryopreservation, imaging and, of course, methods ofgenetic alteration. Many of these have led to betterwelfare, to more precise ways to test hypotheses andto reduced animal usage; others have led tosubstantial increases in the numbers of animals usedfor certain types of experiment, or to altered practicesthat may not always be beneficial to the animals or toscientific understanding.One of the most notable developments of the last 25years is the advance in methods of genome editing.Genome editing allows us to alter DNA sequences veryprecisely and probably gives us the ability to modify anyliving organism which, along with other techniques suchas directed differentiation of stem cells anddevelopment of organoids, may continue to helpreplace animals and yield better ‘models’ of humandisease.Another significant area of development has been inthe generation and archiving of mouse lines which hasgreatly improved. This means that fewer animals needto be maintained in the laboratory. For example, thereare now better methods of cryopreser vation forembryos, sperm, oocytes and ovaries. New methodshave arisen in reproductive biology, such as somaticcell nuclear transfer (SCNT), the method which wasused to produce Dolly the sheep in 1997.1,2A further area of development is in the use offluorescent protein markers,3or other types of reportersuch as luciferase. These allow us to follow cells in vitroor in vivo and give us the ability to do live imagingstudies. Imaging modalities such as MRI and ultrasoundhave also developed, allowing us to reduce laboratoryanimal numbers by carrying out longitudinal studies.The drivers of these advances in technology arewidespread. They include curiosity about how genes areexpressed during development, a greaterunderstanding of the roles of specific genes, increasingknowledge of stem cell biology and cancer biology andbetter ways to follow cell fate decisions duringdevelopment. Additionally, there has been interest inpractical applications of this technology in animals – forexample, how to make them more productive, growfaster, have disease resistance or how to use them as‘bioreactors’ to produce valuable human proteins.However, these possible applications come with theirown suite of ethical and animal welfare issues toaddress.The advances in Animal Technology over the last 25years have also impacted on animal welfare, husbandryand the 3Rs, though not always with positive results.Cages have largely switched from open-topped cages toindividually ventilated cages (IVCs) with in-built wateringsystems, which may help maintain the health status ofthe animals but may not be good for the animals’natural behaviour or normal physiology as they cannotsmell neighbours or interact with each other if singlyhoused. The increase in Specific-Pathogen-Free (SPF)facilities is good for the animals’ health but also leadsto animals having an underdeveloped immune systemand simpler gut microbiota which does not accuratelyreflect animals in the wild or humans. There has alsobeen an increase in the general understanding ofenvironmental enrichment needs and animal handlingtechniques which cause less stress and promote betterwelfare.Alongside the technological advances of the last 25years, we have seen better training programmes andcareer structures becoming available for AnimalTechnologists. This means that higher skilledindividuals are looking after the animals and are able tocarry out procedures but may also mean that scientistsare less likely to visit animal facilities and as a resultmay have unrealistic expectations or develop lessempathy for their animals. Similarly, improveddatabases and animal management systems mayappear to be beneficial as they offer more streamliningand centralised control but may further contribute tokeeping scientists out of the animal facility.Two encouraging trends underlie all the developmentsdiscussed above. The first trend is one of much greatertransparency and openness in science which can beseen in initiatives such as the Concordat on Opennessand the rise of open access publishing. The second isan increase in genuine concern about animal welfare byall those involved in research and, reflecting this, therehas been widespread adoption of the 3Rs, greatlysupported and assisted by Animal Technologists.Rodent and rabbit welfare – whatmight the next 25 years hold?Robert Hubrecht and Huw Golledge, UFAWLaboratory animal science has come a long way in thepast 25 years. The genetic revolution has transformedthe way mice are used in research, bringing bothchallenges and opportunities. We have also madegreat strides in the way we care for rodents in thelaboratory, both by better understanding their needsand by spreading that knowledge through training andeducation. What might the next 25 years bring?There are a variety of opportunities for replacing animalmodels with non-animal alternatives, such asorganoids, ‘organ-on-a-chip’ technology, tissue culture,imaging in humans, the use of data acquired fromanimals undergoing routine clinical veterinar yLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 82

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting83treatment, risk or hazard assessment and modelling.Some of these techniques are already established,although it is not easy to predict how they may develop.There are also oppor tunities which may lead toreductions in animal numbers, although it is difficult toestimate how many animals might be used in 25 years(see Table 1 for the number of procedures conductedon rodents and rabbits in 2017). For example,initiatives like ShARM (Shared Ageing ResearchModels) allow researchers to share resources such assurplus tissues, reducing the total number of animalsneeded to generate samples.4Sharing of data is alsolikely to provide a major opportunity, through creationof repositories of animal studies, or the use of ‘big’epidemiological data from companion animals to studydrug efficacy in naturally occurring disease models.Data sharing might also allow ‘read-across’ approaches– where similar chemical substances are grouped into‘families’ for chemical safety assessments, so thatinformation on the toxicology of a well-understoodsubstance can be used to make inferences aboutsimilar substances for which less information isavailable without having to repeat animal studies.A further way to reduce the numbers of researchanimals is through better experimental design.Evidence suggests that a large proportion of studiesthat use animals fail to adhere to several principles ofgood experimental design, such as randomisation orblinding.6Addressing this problem would also helpimprove the quality of science.There will also be oppor tunities to further applyrefinements in animal research. Progress has alreadybeen made, from the improved training of technologiststo better housing conditions (e.g. double-decker cagesfor rats). However, progress can be slow – for example,although providing nesting material for mice is nowstandard in the UK, this is not necessarily the caseelsewhere. To make further progress in refinement, anoverall better understanding of long-term welfareimpacts is needed, including insights into ‘cumulativesuffering’. By better understanding the welfare impactsof both scientific procedures and life in the laboratoryon animals, we can gain a better understanding ofwhere refinement or replacement is most important.The rise of automation in animal facilities offersopportunities to improve welfare, as monitoring orphenotyping of animals can be less invasive and yieldbetter data. Automation can also help avoid humanerror – for example, by avoiding the misidentification ofanimals. However, there is also the possibility thatlosing human input could lead to problems beingmissed – thus, a mixture of automation and humanattendance is necessary.The techniques used for euthanasia in laboratoryanimals offer yet another opportunity for refinement.Carbon dioxide is now widely understood to be aversive7but, subject to proper evaluation, new technologiescould offer the possibility of humane and practicalalternatives, such as Low Atmospheric PressureStunning (LAPS) or focussed microwave irradiation.As with the other areas of the 3Rs, there are threats tothe future of refinement. For example, new animalmodels developed in the future may be found to havesignificant or unpredictable welfare implications.Threats to refinement also exist where there is still alack of knowledge about animals’ needs – for example,relatively little is known about the social needs ofrabbits, or how best to keep male mice.There may in future, also be occasions where reductionand refinement conflict, for instance where feweranimals can be used in longitudinal studies whichrequire repeated imaging under anaesthesia as analternative to using more animals which must be killedto obtain samples. In such cases the cumulativeimpact on individual animals may be greater (sinceanaesthesia has an impact on welfare) but far feweranimals will need to be used. In such circumstancesmany prioritise the harm caused to individual animalsbut the harm-benefit analysis will need to carefullyweigh Reduction against Refinement.To conclude, it is possible that we will no longer beusing mammalian models in 25 years – although if weare, we can expect them to be more valid. However, thebest prediction we can make is that we will seesignificant fur ther advances in the 3Rs, although wemust not forget that threats and challenges are alsolikely to arise. Animal Technologists will continue to beessential advocates for animals in the future.Table 1. Numbers of rodents and rabbits used forexperimental procedures in the UK in 2017 accordingto UK Home Office annual statistics.5Species Experimentalprocedures––––––––––––––––––––––––––––––––––––––––––––––––––Mouse (Mus musculus) 1,094,867––––––––––––––––––––––––––––––––––––––––––––––––––Rat (Rattus norvegicus) 233,676––––––––––––––––––––––––––––––––––––––––––––––––––Guinea-pig (Cavia porcellus) 22,560––––––––––––––––––––––––––––––––––––––––––––––––––Hamster (Syrian) (Mesocricetus auratus) 1,126––––––––––––––––––––––––––––––––––––––––––––––––––Hamster (Chinese) (Cricetulus griseus)0––––––––––––––––––––––––––––––––––––––––––––––––––Mongolian Gerbil (Meriones unguiculatus) 311––––––––––––––––––––––––––––––––––––––––––––––––––Other rodent 2,105––––––––––––––––––––––––––––––––––––––––––––––––––Rabbit (Oryctolagus cuniculus) 10,362LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 83

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting84Building refinements for rabbitsinto a new facilityAnna Slaviero, University of SurreyRecent years have seen many new refinements ofvarious aspects of housing and husbandr y of rabbits.This has also been reflected in the new ‘Code ofPractice for the housing and care of animals bred,supplied or used for scientific purposes’ which sets outthe standard of care and accommodation for animalsunder the Animals (Scientific Procedures) Act 1986(ASPA).8At the University of Surrey we have aimed to improveevery aspect of our research work with rabbits over thepast three years. This has been beneficial for bothanimals and research, as it is widely recognised that ahigh standard of animal welfare makes sense fromethical, legal, economic and scientific points of view.The opening of our new animal facility was the chanceto apply and implement refinements on housing fromthe early stage of facility design. This consequentlyincreased our opportunities to apply refinements inother areas, such as husbandry and enrichment, toimprove animal welfare. It also allowed us to have astate-of-the-art facility to further develop our work tochallenge and improve standard practices in differentanimal procedures carried out at the University ofSurrey. This section tells the stor y of theimplementation of our rabbit housing refinements, andhow this affects refinements in husbandr y andenrichment.Prior to our redesign, our rabbit housing was typical ofmany facilities. Rabbits were housed in cages singly orin pairs, with the light, humidity and temperature of theroom controlled centrally. Although initial attempts toimprove rabbit welfare through socialisation, breedingand habituation programmes were successful, theopportunity to develop a new facility allowed us to takethese improvements a step fur ther.In designing the new facility, planning discussionsinvolved as many stakeholders as possible: theEstablishment Licence Holder, the Home Officeinspector, architects and builders, NACWOs,Veterinarians, Researchers and Animal Technologists.We also had to consider several factors: did we want torefurbish our old unit, or build a brand new unit? Whereshould it be located? What was our budget? And howcould we meet both animal welfare and researchneeds? The final design for the facility divided the unitinto three blocks. The first, the ‘noisy’ block, containsthe changing room, cage washers, autoclave, necropsysuite and so on, in order to contain all areas that wouldinvolve noise and minimise disturbance throughout therest of the facility. Animal housing and procedurerooms are contained in the second ‘quiet’ block. Thethird ‘super-quiet’ block, contains the surger y suite andsleep suite. Building materials were chosen tominimise noise transfer from one block to the next.In contrast to the central control in the previous unit,temperature and humidity are controlled separately foreach room. This allows us to have dif ferenttemperature and humidity for mouse or rabbit roomsbut also for the other rooms like the surgery suite andcage wash. Two lighting systems are in place – redlights and LED lights – to allow visual access to animalsout-of-hours without causing disturbance. The healthstatus of the animals is maintained through theexistence of negative pressure and air filters,Individually Ventilated Cages (IVCs), decontaminationprocedures using autoclaving or Vaporised HydrogenPeroxide (VHP).Building on the refinements to rabbit housing andhusbandry we introduced in our old facility, the rabbitrooms have various features to promote rabbit welfare.Rooms have anti-slip flooring so that rabbits can moveeasily when let out for play and exercise. A bespokemodular pen system allows the pens to be adapted tothe needs of different rabbits. For example, new rabbitscan be settled into the facility in phases, starting in asmaller, quiet space, with new areas opened as therabbits become more settled. Enrichment to encourageplay behaviour is available to the rabbits and theequipment provided can be restructured to maintainnovelty. Enrichment, social experience and exercise areall available to rabbits in our class II room as well asthe main rabbit room.Furthermore refining the design of the facility, we alsointroduced refinements relating to staff behaviour.Technologists spend 10-15 minutes with the rabbitseach day in order to allow the rabbits to habituate totheir presence and to being held and checked.Researchers are now required to visit the facility atleast three times over at least one week prior tostarting experiments to let rabbits habituate to them aswell. One outcome of this rule was that researchersshowed more concern for rabbit welfare when theyspent more time with the animals. We also introducedpositive reinforcement training procedures, such aspresenting rabbits with a basil leaf when they hop on ascale, or training rabbits to associate certain odourcues or music with positive experiences to promotebetter welfare when rabbits are moved to the class IIroom.Although our refinements have helped improve rabbitwelfare, new refinements are being planned all thetime. For example, we hope to introduce burrowing pitsto encourage more natural behaviours. Continuallychecking data, reviewing the results and puttinglessons learned into practice will continue to promoteand improve the welfare of our rabbits.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 84

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting85Exploring rabbit personalitiesClare Frances Ellis1, Wanda McCormick2,Ambrose Tinarwo3and Helen Clegg41University of Northampton,2Anglia Ruskin University,3Hadlow College and4University of BuckinghamshirePersonality in animals has been an area of growinginterest over the past 10-15 years.9-11However,attention from a variety of disciplines has led to a widerange of different methods being employed to test forand assess personality. While there have been anincreasing number of attempts to explore and describedog and cat personality in recent years, only limitedstudies have explored personality and individualbehavioural profiles for rabbits. Validated tools toassess personalities in rabbits may have applicationsacross a wide range of contexts, including at rehomingcentres, to select suitable animals for use in animal-assisted therapy or specific research paradigms and tounderstand the behaviour of rabbits in a laboratorysetting. Such tools may help in the selection of traitsthat indicate which rabbits might cope better in captivityor support the matching of individuals to particularsettings. In order to develop tools to explore possiblerabbit personalities and identify suitable assessmentmethods for this species, we aimed to answer severalquestions. Firstly, do rabbits show between-individualvariation and within-individual consistency in behaviour?If so, what traits are important to a captive setting? Andfinally, which tools are suitable to measure these?The first tool we developed to explore rabbit personalitywas based on a suite of behavioural tests. A sample of52 mixed-sex adult rabbits from four land-based collegetraining units were assessed in two trials, spaced threemonths apart. Trials consisted of an open field test,time taken to exit a carrier, a novel substrate test anda novel object test. Our results suggested that rabbitsshowed evidence of individual differences in boldness,activity and exploratory behaviour. Ten of the twentyvariables studied were consistent over time, indicatingthat individual rabbits do show consistent differencesin these personality traits.The second tool, the Rabbit Behaviour Rating Tool(RaBRT), was derived from a literature search of rabbitbehaviour articles. We identified 47 behaviouraldescriptors and each was rated on a 5-point scale bypet owners and people that work with rabbits, with1172 full responses received. Only 17 itemsdemonstrated fair to excellent inter-rater reliability.Statistical analysis identified three key behaviouralindicators, which related to social interactions withhumans, activity levels and antisocial interactions withhumans.In conclusion, we found that rabbits did show betweenindividual variation in behaviour which could bedetected using both tools, although the specific traitsthat could be measured depended on the tool beingused. Behavioural tests also indicated a low-moderatelevel of individual consistency over time. Furthervalidation studies are underway, including validation ofthese potential behaviour assessment tools andcomparisons to home cage behaviour observations.A natural approach – how toincrease rat nest buildingbehaviour in a laboratoryenvironmentDemi Minhinnett, Durham UniversityRats account for a large proportion of scientificprocedures on animals every year in the UK; in 2017rat use accounted for 6.3% of all procedures.5This isone reason why it is vital to focus on the welfare oflaboratory rats, including allowing them to exhibitnatural behaviours, such as nest building. However, itis not always easy to facilitate natural behaviours andnest building behaviour in rats (in the laborator y) is notas commonly observed as it is in mice. Whilst micehave been observed to spontaneously nest build whenprovided with nesting materials in laboratory settings,rats may not do so, suggesting that nest building in ratsis a learned, rather than innate, behaviour.12Weproposed to give rats the opportunity to learn to buildnests, trialling different nesting materials and notingthe effects on nest building behaviour.The nesting materials included in the trial wereselected by considering the kinds of materials ratswould naturally encounter in the wild, such as grasses.Rats were therefore provided with one of three differentkinds of material for this study: hay, paper wool, or amix of hay and paper wool. The rats used were allbreeding females from either Wistar or Lister Hoodedstrains, and none had exhibited nest-building behaviourbefore being included in the trial. The quality of thenests produced was also assessed based on a systemused to score mouse nests.13Several rats in the trial exhibited nest buildingbehaviour but only when provided with hay, or a mix ofhay and paper wool. This was thought to be due to thearchitectural properties of hay: providing hay allowednests to be built upwards and outwards to create ball-shaped nests, which score highly when assessing nestquality. The presence of hay was also beneficial forlitter production: when rats were provided with paperwool and no hay, pups tended to be found scatteredthroughout the nest (Figure 1) and mother rats showedmore signs of disturbance when the cage was opened– scurrying around the cage and attempting to movepups under the hopper. Two litters were alsoLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 85

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting86abandoned and two dead pups were found. In contrast,when hay was provided, pups tended to be found in acluster in a nest (Figure 1), mother rats showed lessdisturbed behaviour and there were no litterabandonments or pup deaths.This trial highlights the importance of looking at ananimal’s natural history to find ways to promote naturalbehaviours, which may have notable impacts on theanimal’s welfare. By attending to the natural historyand natural behaviour of laborator y animals, we canlearn more about how to best support them in alaboratory setting.Modifying laboratory rat housingfor improved welfareRebecca Terry, University College LondonThe importance of allowing rats space to stand on theirhind legs is largely recognised (e.g. by the NC3Rs andRSPCA)14–16but most current caging does not providethe height to allow for this. Not being able to stand uplimits rats’ ability to express natural behaviours and isthought to lead to muscle wastage in their hind legs.Conventional cages may also not provide enough roomfor rats to express play behaviour, and can exposealbino rats to too much light, causing retinaldegradation.17We therefore aimed to improve thewelfare of the rats kept at UCL Cruciform by modifyingthe cages to allow this additional height.Various factors had to be considered in order toproduce a feasible cage design. The room needed to beable to house a similar number of rats as could behoused in the conventional cages, but researchequipment stored in the room could not be moved,space was limited, racks could not be made larger, andcages which would require additional equipment, suchas air handling units for IVCs, were not feasible. Thenew cage designs had to meet several requirementswithin these limits: cages needed to provide adequateheight for rats to stand at full height, provide a higherlevel of enrichment and have space to provide hides tominimise potential retinal damage. Cages also had tobe a financially feasible option for other facilities andthe change to new cages had to be hassle-free in orderto encourage others to adopt the new design.In order to provide more space within these limits, thetotal number of cages in the room was reduced from 80to 64. This did not affect rat research as the room wasrarely operating at full capacity. This revision createdmore vertical space in each rack, allowing a total of 14cm extra height to be added to each cage. To add thisspace, a modified raised hopper based on theTecniplast -123 series was added to the conventionalcage bases already in place. Shelving was added to theroom to replace the cage racks – this shelving is costeffective, adjustable and can be dismantled when notin use to maximise space in the room.The new design of the cages resulted in a total heightof 35cm, which should allow an adult rat to standcomfortably. These taller cages also allow space fordeeper litter to encourage digging and a shelter toprotect from light. Enrichment designed to encourageplay behaviour can be suspended from the bars toincrease floor space and can allow individual rats tospend time away from one another. These changes,promoting natural rat behaviours, are likely to result indata obtained from the rat, being of improved quality.Furthermore, from a practical perspective, adjustingthe cage height rather than designing a larger cagebase means that cages are not significantly heavierthan before, making it easier for staff to move andclean them.The changes made to the current caging are likely to behighly beneficial for rats, as well as practical and cost-Figure 1. Images of rat pups scattered throughout thenest made from paper wool (left) and pups clusteredtogether in nest made of hay (right).Credit: Demi Minhinnett.Figure 2. Conventional rat cage (left) and rat cage withmodified hopper (right). The modified hopper allowsspace for more enrichment and for rats to rear to fullheight.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 86

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting87effective for staff. Tecniplast have therefore produceda prototype based on the changes made and furtheradjustments will be made to the design before they aredeveloped. This demonstrates that relatively simplechanges can be made which are likely to greatlyimprove the quality of life of laboratory animals.Severity classification of repeatedanaesthesiaKatharina Hohlbaum1,2, Bettina Bert2,3Silke Dietze2, Heidrun Fink2, Christa Thöne-Reineke11Institute of Animal Welfare, Animal Behavior andLaboratory Animal Science, Department of VeterinaryMedicine, Freie Universität Berlin, Germany2Institute of Pharmacology and Toxicology, Departmentof Veterinary Medicine, Freie Universität Berlin,Germany3German Federal Institute for Risk Assessment (BfR),German Centre for the Protection of LaboratoryAnimals (Bf3R), GermanyWithin the concept of the 3Rs of Russell and Burch18one strategy to reduce the number of laboratoryanimals is the repeated use of a cohort of animalsover the course of an experiment. For example, inimaging studies, animals are anaesthetised each timeimaging is carried out to avoid ar tefacts caused byunpredictable movements. However, little is knownabout the effects of repeated anaesthesia, which mayhave a greater impact on the wellbeing of the animalsthan a single anaesthetic episode.19The 3Rsadvantages of repeated animal usage are thereforeonly relevant if the animals involved do not experiencemore suffering, pain or distress than animals used forsingle procedures. Reflecting the lack of knowledge inthis area, Directive 2010/63/EU states that theseverity of general anaesthesia is mild but does notdifferentiate between single and repeatedanaesthesia. We therefore aimed to investigate thewelfare impacts of single and repeated anaesthesiaon mice.In order to examine the effects of repeatedanaesthesia, we explored the effects of two commonanaesthetic methods on adult C57BL/6JRj mice ofboth sexes: inhalation of isoflurane (induction: 4.0%;maintenance: 1.75–2.50%) in 100% O2for 45 minutesand injection with a combination of ketamine (80mg/kg) and xylazine (16 mg/kg) (KX). For each method,mice were randomly allocated to either control, singleanaesthesia, or repeated anaesthesia groups(anaesthesia every 3-4 days, a total of six times).Welfare was assessed after the last anaestheticepisode according to our protocol ‘systemicassessment of wellbeing in mice for procedures usinggeneral anaesthesia’, https://paperpile.com/c/EbWJuJ/racD20which includes the Mouse GrimaceScale (MGS), burrowing and nest building, the free-exploratory test for anxiety-related behaviour, homecage activity, food intake, and bodyweight, as well asthe analysis of faecal corticosterone metabolites (FCM)for acute stress (24 h post-anaesthesia). In addition,hair corticosterone concentrations were measured.We found that neither single nor repeated use ofisoflurane influenced nest building, home cage activity,bodyweight, FCM or hair cor ticosteroneconcentrations.21Isoflurane increased MGS scores infemale mice 30 minutes after anaesthesia comparedwith controls but scores did not differ between singleand repeated anaesthesia. Repeated anaesthesiareduced burrowing behaviour in both males andfemales and increased time before displayingexploratory behaviour in female mice, indicating greaterlevels of anxiety than those exposed to singleanaesthesia or controls.21Anaesthesia with KX did not affect nest building, homecage activity, or hair corticosterone concentrations.22Both single and repeated KX anaesthesia increasedMGS scores 150 minutes after anaesthesia comparedwith controls. Repeated KX anaesthesia increased thetime before displaying exploratory behaviour in femalemice one day after anaesthesia, although singleanaesthesia did not. However, after eight days, femalemice exposed to single or repeated anaesthesiashowed greater time before exploring than controls.Changes in food intake and FCM excretion indicated anincreased stress response in male mice after single KXanaesthesia, although there was no effect of repeatedanaesthesia.22Besides the degree of pain, suffering, distress orlasting harm, an understanding of the duration of thenegative effects is essential for severity classificationof any procedure. Although we saw behaviourssuggesting increased anxiety following repeatedisoflurane anaesthesia, these would be associatedwith mild, rather than moderate, levels of severity andthe wellbeing of the mice was affected for only a shortterm – mainly in the immediate post-anaestheticperiod.21In our view, therefore, the severity of repeatedisoflurane anaesthesia in C57BL/6JRj mice can beclassified as mild. This also applies for other protocolsusing a comparable anaesthesia regime. However,severity may deviate if a different anaesthesia regime,mice of a different age, other mouse strains, or othermouse disease models are used. Within the mildseverity category, repeated isoflurane anaesthesiawould clearly be of higher severity than a singleisoflurane anaesthesia.20For the final severityclassification of repeated KX anaesthesia, furtherinvestigations are needed in order to specificallydetermine the effects on anxiety and the duration ofthe mild distress indicated by changes in food intakeand FCM excretion.22LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 87

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting88Welfare implications of differentidentification methods for miceDominic Wells, Royal Veterinary CollegeA variety of methods for marking mice, both permanentand temporary, are used in UK laboratories, but little isknown about the animal welfare impacts of thesemethods. A survey of animal units showed that earpunching and notching are the most common mouseidentification methods, followed by marking with ink onthe tail.23We therefore chose these methods for furtherinvestigation into their welfare impacts on marked mice.We initially explored the effects of ear punching andnotching on male and female C57BL/6 and Balb/cmice. Mice were either i) ear punched, ii) ear notched,iii) restrained or iv) not handled, and behaviouralmeasures of welfare and faecal samples forcorticosterone measurement were collected. Miceshowed an immediate head star tle response topunching and notching compared with restrained miceas well as more grooming and freezing behaviour intheir home cages. However, no significant differencesin faecal corticosterone levels were found. Ear notchedmice also ate less novel food the following day,indicating higher levels of anxiety, than unrestrainedcontrol mice.Next, we examined whether marking mice with a markerpen or using local anaesthetic alongside ear punchingwould improve welfare indicators in marked mice. MaleBalb/c mice were housed in pairs, with one mousefrom each pair undergoing either ear punching, earpunching with the application of EMLA cream(lidocaine/prilocaine), marking with a permanentmarker pen or no marking. The second mouse in eachpair was unmarked. As marking mice with a pen wouldneed to be repeated regularly, marker pen was appliedweekly for the duration of the experiment, whereas earpunching was only done in week 1. The welfare of theanimals during marking was assessed by counting thenumber of animals which defecated during the markingprocess. In the first week, similar numbers of micedefecated during marking, regardless of the methodused. However, defaecation during marking with amarker pen significantly decreased by week 3,suggesting that mice had habituated to the method.Furthermore, mice that were ear-punched, whether ornot EMLA cream was applied, were more likely toreceive grooming from the unmarked mouse they werehoused with, whilst those that received an ear punchand EMLA cream application were more likely to groomtheir ears and less likely to eat novel food.In conclusion, we found that ear punching and notchingappear to cause short-term pain and anxiety to micebut that application of a local anaesthetic cream didnot help to alleviate these responses and causedgreater behavioural disturbance. Our results suggestthat regular use of a permanent marker pen is areasonable option which mice appear to habituate to.This may therefore be a good refinement option forthose needing to mark individual mice.How modern imaging techniquescontribute to the 3RsThomas Snoeks, The Francis Crick InstituteOver the last decade, imaging has made its way intomost academic animal facilities. At The Francis CrickInstitute, for example, we use bioluminescence andfluorescence, ultrasound, microCT, 9.4T MRI, PET/MRI,SPECT/CT and intra-vital microscopy. The use ofimaging and the wide range of techniques on offer, canmake a valuable contribution to the 3Rs in a number ofways, by helping to reduce the number of animals used,refining techniques and helping in the earlieranticipation of disease.One of the major benefits of using imaging techniquesis that they allow longitudinal measurements. Thismeans a single cohort of animals can be used overtime, reducing the total number of animals used.Longitudinal measurements can also improveexperimental design, as they yield paired data,meaning that studies have higher statistical power thandesigns which yield unpaired data. Imaging also allowsmore flexible time points to be used in experiments.However, there are potential welfare impacts toconsider, as repeated imaging sessions will involverepeated anaesthesia (see above), which requires acareful harm-benefit assessment.Although reducing animal use is a decided benefit ofusing imaging techniques, the impact of imaging onanimal research stretches beyond this straightforwardreduction in numbers. Imaging can also offerrefinement opportunities – for instance, by reducingintra-obser ver variation. Various image-guidedapproaches can also be used, such as image-guidedinjection or image-guided irradiation. For example,ultrasound imaging can be used to guide injections intothe pancreas, hepatic portal vein, or other organs ortissues without the need for additional surgery. Image-guided injection has been used at the FCI to producemice with lentiviral-mediated transgenic skin, leavingthe rest of the mouse unmodified and thereforeavoiding certain pathologies that may be associatedwith that transgenic model. Image-guided irradiationcan also make it easier to shield tissues that are notof interest and better target tissues that are of interest.Other areas in which imaging techniques can offerrefinement opportunities include the anticipation ofdisease. Early disease detection allows for refinedLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 88

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting89experimental protocols where animals are enrolled intoexperiments before the onset of over t clinicalsymptoms, which can reduce the degree of sufferingthe animal experiences. Researchers are also able tobetter identify the animals they want to include in theirexperiments – for example, imaging can be used todetect tumours and assess whether they are the rightsize to be included in a study. Imaging can also be usedin the phenotyping of new models – for example,imaging allows the contraction of the heart or bloodflow through the aorta to be compared in differentsubjects.In summary, the use of the wide variety of imagingtechniques available offers opportunities for bothreduction in the number of animals used, generatingpaired data and lower intra-observer reliability, andrefinement in procedures, through early diseasedetection, image-guided methods and phenotyping.Whilst the welfare impacts of repeated anaesthesiamust be contrasted with the alternative of using agreater number of animals, researchers can makechanges to their experiments to effectively contributeto the 3Rs by exploring and applying the imagingtechniques available at their own institutions.Home Office update: food andwaterJohn Marshall, Animals in Science RegulationUnit, Home OfficeThe freedom from hunger and thirst is the first of theFive Freedoms to which all animals are entitled (Box 1).Therefore, the provision of food and water toexperimental animals is a fundamental part of ensuringanimal welfare. Establishment licence (PEL) standardcondition 4(3) states that protected animals must beprovided with food and water unless authorised by theSecretary of State (i.e. as an experimental procedure),and PIL holders are entrusted with the primaryresponsibility for animals on whom they haveperformed regulated procedures. Failure to providefood and water causes unnecessary suffering andpotentially death of animals, while experimental dataand therefore the benefits of animal use are lost.However, failure to provide food and water does occurand is a major concern for ASRU.Failure to provide food and water as part of the normalcare and husbandry of animals represents a significantcause of non-compliance, accounting for approximately20% of non-compliance cases annually.25For many ofthese cases, the causes fit into certain themes, suchas changes in housing, lack of communication,occurrences over weekends and failure to identify theproblem over multiple checks.Changes in housing, whether these are followingtransportation or delivery of animals, due to the use ofweaning or splitting cages, or after procedures, are acommon cause of failure to provide food and water.This is particularly found to be the case where both aremissing. A lack of communication between facility staffand researchers and a lack of understanding ofresponsibility are linked to this – for example, after aprocedure where food or water has been withheld theremay be confusion as to which team member isresponsible for returning food or water to the cage.Another common feature of cases of non-compliancedue to lack of food and water is that issues tend tooccur over the weekend, perhaps due to changes instaffing or the checking schedule. Lack of food andwater is often identified on a Monday, meaning that theinitial incident leading to food and water not beingprovided has usually occurred the previous week. Inthese cases, where food and water have been absentfor several days, the most severe consequences foranimal welfare tend to occur. These cases are alsoparticularly concerning, as this usually means thatseveral different people have checked the animals andfailed to note the lack of food and water.There are a number of relatively easy interventions thatcan be done to reduce the risk of non-compliance. Forexample, within the animal unit, the set-up of animalhousing should make it easy to observe animals, andovercrowding should be avoided. It can also be helpfulto identify units that may be ‘at-risk’ – for example,isolators, cabinets, recovery areas or any roomsoutside the main facility – so that these can beappropriately addressed. Staff should also considerwhat the most effective system of checks would be fortheir unit. Technology inter ventions that haveautomated checking of food and water may also helphere but should be an addition to, not an alternative to,human checks. During the meeting, we askedparticipants to share their own good practice tips tohelp ensure animals do not go without food or water.These are listed below:1. Have multiple checks throughout the day – usuallyone in the morning and one in the afternoon.Box 1. The Five Freedoms.24G Freedom from hunger and thirstG Freedom from discomfortG Freedom from pain, injury or diseaseG Freedom to express normal behaviourG Freedom from fear and distressThey were originally set out for farmed animals bythe Farm Animal Welfare Council but are oftenapplied in other contexts.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 89

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting90Ideally, each check should be done by a differentperson.2. When checking for water, make sure that waterbottles are touched and not just observed.3. Make sure labelling is informative, especially ifanimals have special requirements. For example,weaning mice are clearly labelled at anestablishment where these are uncommon.4. Ensure the NACWO is made aware of any absenceof food or water, even if this is short term.5. Carry out random audits of rooms to check generalperformance across a unit, as well as to flag anyspecific racks or rooms that might be higher risk.6. Ensure that the roles and responsibilities of AnimalTechnologists are clearly defined – for example, byassigning individual rooms or areas to differentpeople, or by ensuring clear instructions arepresent as to who should return food and water(and when) if these have been restricted as part ofa procedure.7. Consider using a timer when animals are on diet orwater restriction, so that when the timer goes off itserves as a reminder to return the food or waterbottle.8. Although automation may help avoid situationswhere there is a lack of food or water, or otherundesirable situations, be aware that it is not aper fect solution, and automation should becombined with checking from staff as well.Editors Note. The IAT Animal Welfare Group haspublished an advice notice on the Feeding and Wateringof Laboratory Animals. and is available to downloadfrom the IAT website www.iat.org.uk/news. Thisdocument aims to provide advice with a focus on therole of Animal Technologists, on the steps to be takento ensure compliance with the terms of ASPA standardcondition 4.Meeting action pointsThe following is a list of action points, based on all thepresentations and discussions, which may be of use toyou in your facility:– If your facility is being refurbished, or a new facilityis being built, ask the NACWO and/or NVS to ensurethat all Named Persons and Animal Technologistshave appropriate input into the design, so that newthinking about refinement can be fully incorporated.– Encourage researchers to visit the facility regularly(if they do not already), to encourage greaterawareness of the animals’ welfare needs.– Be aware that (like many other species) rabbits canhave different personalities, which may haveimplications for welfare assessment, evaluatingrefinement, and day-to-day care.– Ask for a review of the amount and type of nestingmaterial provided for rodents at your establishment,especially rats. If nests have not been of goodquality to date, consider trying different materials oroffering a combination of materials.– If rats are housed in cages that do not allow themto stand up at your facility, ask for this to bediscussed at the AWERB (using the paper byMakowska & Weary, reference [16] below). Could aplan be drawn up to change to taller caging?– If you are caring for animals undergoing repeatedgeneral anaesthesia using gaseous agents, discussthe potential for increased anxiety with theresearcher and include indicators of anxiety, suchas reduced exploratory behaviours, in welfareassessment protocols.– Ask for a discussion and review of the methodsused to mark mice at your facility, using the sectionon ‘Welfare implications of different identificationmethods for mice’, above.– If imaging is used within scientific protocols at yourestablishment, ask the researcher(s) to give apresentation to animal unit staff, so that you canlearn more about different imaging techniques andhave a discussion on further opportunities toimplement the 3Rs.– Initiate a review of protocols in place to ensureanimals do not go without food or water at yourestablishment – and watch out for forthcomingguidance from the Institute of Animal Technology onthis topic.AcknowledgementsThank you to all the speakers and delegates for theirtalks and discussions.References1Campbell, K.H., McWhir, J., Ritchie, W.A. and Wilmut, I.(1996). Sheep cloned by nuclear transfer from a culturedcell line. Nature, 380 (6569), 64-66.2Wilmut, I., Schnieke, A.E., McWhir, J., Kind, A.J. andCampbell, K.H. (1997). Viable offspring derived from fetaland adult mammalian cells. Nature, 385 (6619), 810-813.3Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W. andPrasher, D.C. (1994). Green fluorescent protein as amarker for gene expression. Science, 263 (5148), 802-805.4Morrissey, B., Blyth, K., Carter, P., Chelala, C., Jones, L.,Holen, I. and Speirs, V. (2017). The Sharing ExperimentalAnimal Resources, Coordinating Holdings (SEARCH)Framework: Encouraging Reduction, Replacement, andRefinement in Animal Research. PLoS Biology, 15(1),e2000719.5UK Home Office. (2018). Statistics of scientificprocedures on living animals, Great Britain 2017.Download from: https://www.gov.uk/government/statistics/statistics-of-scientific-procedures-on-living-animals-great-britain-20176Cressey, D. (2015). Poorly designed animal experimentsin the spotlight. Nature News.7Leach, M.C., Bowell, V.A., Allan, T.F. and Morton, D.B.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 90

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Report of the 2018 RSPCA/UFAW Rodent and Rabbit Welfare Group meeting91(2002). Aversion to gaseous euthanasia agents in ratsand mice. Comparative Medicine, 52(3), 249-257.8Animals (Scientific Procedures) Act. (1986). Available at:https://www.legislation.gov.uk/ukpga/1986/14/contentsAccessed November 22, 20189Briffa, M. and Weiss, A. (2010). Animal personality.Current Biology, 20 (21), R912-4.10Highfill, L., Hanbury, D., Kristiansen, R., Kuczaj, S. andWatson, S. (2010). Rating vs. coding in animalpersonality research. Zoo Biology, 29(4), 509-516.11Gosling, S.D. (2001). From mice to men: what can welearn about personality from animal research?Psychological Bulletin, 127(1), 45-86.12Van Loo, P.L.P. and Baumans, V. (2004). The importanceof learning young: the use of nesting material inlaboratory rats. Laboratory Animals, 38(1), 17-24.13Deacon, R.M.J. (2006). Assessing nest building in mice.Nature Protocols, 1(3), 1117-1119.14RSPCA. (2011). Rats: Good practice for housing andcare. Download from: https://science.rspca.org.uk/sciencegroup/researchanimals/repor tsandresources/housingandcare.15NC3Rs. (2013). Housing and husbandry of rodents.NC3Rs. Available at: https://www.nc3rs.org.uk/3rs-resources/housing-and-husbandr y/rodents. AccessedNovember 20, 2018.16Makowska, I.J. and Weary, D.M. (2016). The impor tanceof burrowing, climbing and standing upright for laborator yrats. Royal Society Open Science, 3(6), 160136.17Burn, C.C. (2008). What is it like to be a rat? Rat sensor yperception and its implications for experimental designand rat welfare. Applied Animal Behaviour Science,112(1), 1-32.18Russell, W.M.S and Burch, R.L. The principles of humaneexperimental technique. Pub. Universities Federation forAnimal Welfare, 1992. Special edition ISBN 09007677829780900767784.19Hawkins, P., Prescott, M.J., Carbone, L., Dennison, N.,Johnson, C., Makowska, I.J., Marquardt, N., Readman,G., Weary, D.M. and Golledge, H.D.R. (2016). A GoodDeath? Report of the Second Newcastle Meeting onLaboratory Animal Euthanasia. Animals, 6(9).20Hohlbaum, K., Bert, B., Dietze, S., Palme, R., Fink, H.and Thöne-Reineke, C. (2018). Systematic assessmentof well-being in mice for procedures using generalanesthesia. Journal of Visualized Experiments, (133).21Hohlbaum, K., Bert, B., Dietze, S., Palme, R., Fink, H.and Thöne-Reineke, C. (2017). Severity classification ofrepeated isoflurane anesthesia in C57BL/6JRj mice –Assessing the degree of distress. PLOS One, 12(6),e0179588.22Hohlbaum, K., Bert, B., Dietze, S., Palme, R., Fink, H.and Thöne-Reineke, C. (2018). Impact of repeatedanesthesia with ketamine and xylazine on the well-beingof C57BL/6JRj mice. PLOS One, 13(9), e0203559.23Mazlan, N.H., Salesansky, N.L., Burn, C.C. and Wells,D.J. (2014). Mouse identification methods and potentialwelfare issues: A survey of current practice in the UK.Animal Technology and Welfare, 13(1).24Home Office. (2018, March 12). Animals in ScienceRegulation Unit Annual Report 2016. GOV.UK. GOV.UK.Available at: https://www.gov.uk/government/publications/animals-in-science-regulation-unit-annual-report-2016. Accessed November 22, 2018.25Farm Animal Welfare Council. (2009). Farm animalwelfare in Great Britain: past, present and future.Download from: https://assets.publishing.ser vice.gov.uk/government/uploads/system/uploads/attachment_data/file/319292/Farm_Animal_Welfare_in_Great_Britain_-_Past__Present_and_Future.pdf.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 91

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93A comparison of enrichment items for thepromotion of natural gnawing behaviour inlaboratory miceIRENE LOPEZ JUARISTIUniversity College London, Biological Services, Cruciform Building, Gower Street, LondonWC1E 6BTCorrespondence: irene.lopez@ucl.ac.ukBased on a report to the UCL Collective Laboratory Animal Welfare Society (CLAWS)AbstractAppropriate housing and husbandr y, includingenvironmental enrichment, must take the naturalhabitat, biology and behaviour of each species intoconsideration.1The Guide for the Care and Use ofLaboratory Animals (1996)2states that the goal shouldalways be to maximise species-specific behaviours andminimise stress-induced behaviours.This report is a Collective Laboratory Animal WelfareSociety (CLAWS) initiative to test different types ofchew blocks to promote gnawing behaviour and seewhich one is the most enriching for mice. UniversityCollege London (UCL) is constantly considering newinitiatives that challenge current standards of goodanimal welfare. This trial was completed by twocollaborating UCL Biological Ser vices facilities:Institute of Ophthalmology (IoO) and the Cruciform.Key words: Mice, natural behaviour, species-specificbehaviour, environmental enrichment, chew blocks.Introduction and aims of the reportHousing and husbandry have a major impact onlaboratory animal health and welfare. Determining theamount or form of environmental enrichment needed toproduce a positive impact without causing harm is acomplex task.3It is also important to recognise andpromote positive/good welfare; for example, whenevaluating the effects of refinement techniques suchas environmental enrichment.4Malocclusion is a common problem seen in laboratorymice. It occurs when the incisors overgrow because thejaws are misaligned5, and the mandibular and maxillaryteeth do not properly occlude (Figure 1). Basicrequirements for good rodent housing and husbandryare to provide material to gnaw (e.g. wooden blocks).August 2019 Animal Technology and WelfareFigure 1. Mice have one upper pair and one lower pairof incisors at the front of their mouths which they useto grip and bite their food or other objects.Malocclusion occurs when the incisors overgrowbecause the mandibular and maxillary teeth are notnormally aligned.5This prevents the teeth from overgrowing, encouragesexploration behaviour, reduces abnormal behaviour(such as aggression and apathy) and maintains generalwellbeing.6The purpose of this trial was to carr y out a comparisonof enrichment items for the promotion of naturalgnawing behaviour in laboratory mice. The aim of thestudy was to analyse different chew items available forlaboratory mice to decide which one was most suitablefor fulfilling the needs of this behaviour.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 93

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A comparison of enrichment items for the promotion of natural gnawing behaviour in laboratory mice94Material and methodsThe IoO and the Cruciform Biological Ser vices facilities,tested 5 wooden chew blocks over 28 days (Figure 2):– Aspen balls (1)– Aspen chew sticks (2)– Flat chew stick (3)– Aspen bricks (4)– Lolly sticks (5)The following table contains the specifications of eachproduct used in the study:Each trial group consisted of four cages of the samestrain of mouse (twenty cages in total) with a similarnumber of mice housed. The gender used was basedon availability of existing stock. Each cage wasprovided with an item in the morning, which wasweighed, replaced if completely gnawed and thisinformation was recorded on the data sheet.The trial was constructed in two parts.Every day, for a period of 14 days, the item wasweighed and recorded on the data sheet (if an item hadbeen completely gnawed, it was replaced).Original cages were kept, and groups randomised. Oldwooden chews from the first part of the trial wereremoved and the new item was weighed and recordedfor 14 days.ResultsA simple way to quantify gnawing is by weighing thechewing item. We hypothesised that, the higher theweight reduction of the product, the more it was gnawedon (graph A). Using weight loss as a reference, the orderof preference was from favourite to least favourite:1st Aspen chew stick2nd Flat chew stick3rd Aspen brick4th Lolly stick5th Aspen balls (*)* In this study the Aspen ball group had to be removedfrom the analysis due to lack of evidence of usage asa chew item.These results do not contain information for the Aspenball, leaving four items to compare: Aspen chew sticks,Aspen bricks, flat chew stick and lolly sticks.In the Aspen bricks and lolly sticks groups, one itemlasted for the whole 14 days of the trial whereas, theAspen chew sticks needed to be replaced in 70% of thecages (sometimes more than once) and the flat chewin just one cage (graph B).Table 1. Products used during trial.Item Dimensions Material Supplier––––––––––––––––––––––––––––––––––––––––––––––––––––––––1 Aspen 30mm (diameter) aspen DatesandBall x 100mm (perimeter) wood––––––––––––––––––––––––––––––––––––––––––––––––––––––––2 Aspen 112mm (length) x 10mm aspen LabodiaChew (width) x2mm (height) woodStick––––––––––––––––––––––––––––––––––––––––––––––––––––––––3 Flat 150mm (length) birch AstonChew x18mm (width) wood PharmaStick x 1.6mm (height)––––––––––––––––––––––––––––––––––––––––––––––––––––––––4 Aspen 50mm (length) x10mm aspen DatesandBrick (width) x10mm (height) wood––––––––––––––––––––––––––––––––––––––––––––––––––––––––5 Lolly 112mm (length) x10mm unknown AmazonStick (width) x2mm (height) woodFigure 2. Types of wooden chew items used.Graph A. Four wooden chew blocks: aspen chew sticks,aspen bricks, flat chew stick, and lolly sticks. 14 daysweight loss measurements per item.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 94

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A comparison of enrichment items for the promotion of natural gnawing behaviour in laboratory mice95Each chew block had different properties such as size ormaterial made of, making it difficult to make a directcomparison of use. As our purpose was to compare theusage, an average between the four cages was measuredand recorded each day (one test group), and calculated.Graph B. Amount of times aspen chew sticks and flatchew sticks were added due to the complete use of theprevious item.Graphs 1-3. Graph 1: Four wooden chew blocks: aspenchew sticks, aspen bricks, flat chew stick, and lollysticks, weight loss measurements per item for 14 days.Graph 2: Weight loss of items during the first week.Graph 3: Weight loss of items during the second week.Graph 1 shows chew block item usage/weight loss over14 days. Graphs 2 and 3 demonstrate and comparethe difference in usage during the first and secondweek of the trial.Graph 3 suggests that during the second week theusage rate (or weight loss) in all items slowed down.A single-way analysis of variance (ANOVA) showed thatthere is a statistically significant difference betweenthe four groups tested (p-value (=0.004108) < 0.05).However, the ANOVA does not tell you where thedifference lies. A Tukey HSD (“honestly significantdifference” or “honest significant difference”) test wasthen conducted to ascertain where the difference lies.When comparing each pair of chew blocks, the TukeyHSD showed that there were honest significantdifferences between:– Aspen chew stick vs aspen brick (graph 4a).– Aspen chew stick vs lolly stick (graphs 4b).Figure 3. Visual difference in wooden chew itemsbefore and after a week trial.Graphs 4a and 4b.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 95

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A comparison of enrichment items for the promotion of natural gnawing behaviour in laboratory mice96On the other hand, no honest significant differenceswere found between:– Aspen chew stick vs flat chew stick (graph 5a)– Flat chew stick vs aspen brick (graph 5b)– Flat chew stick vs lolly stick (graph 5c)– Aspen brick vs lolly stick (graph 5d)DiscussionThe Guide for the Care and Use of Laborator yAnimals (1996)2states that the goal should always beto maximise species-specific behaviours and minimisestress-induced behaviours. According to NC3Rs,4if asick animal needs to be euthanatised, the researchproject will need to use more animals to obtain thesame research information. Therefore, it will fail on twoof the basic 3Rs’ principles for animal welfare,refinement and reduction.The trial results posed a few questions:1. Should we rotate chew items during weekly cleaningprocess to keep the animal’s interest in gnawing?2. What about the costs for the unit?3. What about the practicality in terms of how often thetechnologist needs to replace chew items?The statistical analysis indicates that there weresignificant differences between the four groups and theTukey test confirmed that the Aspen chew stick wasthe most chewed. However, as the usage of Aspenchew sticks was very high, 70% of the cages had a newblock added (sometimes more than once) as it hadbeen completely gnawed. This could lead to theconclusion that Aspen chew sticks are the best option.On the other hand, in terms of cost, the animal facilitywill need to hold more of this product in stock,considering the amount needed in just one cage duringtwo weeks of observations. In means of practicality, thetechnologist must monitor the amount of stick left inthe cage and replace it when it is completely gnawed,therefore disturbing the animals more frequently.The cost factor and practicality are the same for thethree groups left to consider. Statistically speaking, thesecond-best option was the flat chew stick, there isconstant usage along the two weeks (graphs 5b and5c). Finally, both the Aspen brick and the Lolly stickwere highly used until day 6 of observations. However,from day 7 there was a reduced use potentially due toa loss of interest (graph 5d).It is important to consider another factor: the supplier.The health of the animal must always be considered.Although during the trial no animals showed signs ofillness, the lolly sticks (supplier: Amazon) had not beentested on animals and the type of wood is unknown.Therefore, considering both the Aspen bricks and theLolly sticks usage was similar, and there were nosignificant differences between groups, it is safer todiscard the Lolly sticks as a possible chew item.Conclusions and recommendationsThe aim of the study was to compare different chewitems available for laboratory mice to decide which oneis the most suitable as part of cage enrichment forfulfilling animals’ natural needs such as gnawing(promoting teeth wearing).Graphs 5a-d.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 96

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A comparison of enrichment items for the promotion of natural gnawing behaviour in laboratory mice97According to the results we should rotate chew itemsduring weekly cleaning process to keep animal’sinterest on gnawing to prevent habituation toenrichment and any health impact. Novelty is animportant factor to consider in this matter. When a newitem is provided as part of the enrichment, weencourage the animal’s natural behaviour of exploringthe new territory. This behaviour promotes gnawingactivity as part of the discovery of their territory.Once the novelty factor disappears, habituation to theenrichment occurs with all items.Rotating between two different items may help to keepan active interest. The difference in shape andmaterials are decisive factors. The flat chew stick andaspen brick are different in both shape and wood. Thefirst one is bigger and flat birchwood, while the brick isthicker, smaller and made of Aspen wood.Constant gnawing activity has a high positive healthimpact on laboratory animals. Signs of poor healthwould interfere with the studies they are involved in.Considering the cost and disturbance factor therecommended strategy for best animal welfare will be:1. Rotate the chew blocks between the flat chew stickand the aspen brick.2. Change the enrichment when cleaning cage to avoidunnecessar y disturbance, unless the item iscompletely gnawed before cleaning.AcknowledgementsI am particularly grateful for the assistance given byZoe Windsor for her constructive recommendations andher willingness to give her time so generously. Specialthanks to Kirsty Butler for her professional guidanceand enthusiastic encouragement during the planning ofthis study. I would also like to express my deepgratitude to Danny Contillo from The Institute ofOphthalmology (IoO) and Lizzie Steptoe from TheCruciform Building for their help in collecting the data.I wish to acknowledge the help provided by Cortex Labfor kindly providing the mice for the study. Finally, I wishto extend my thanks to all the staff at Cruciform BSUfor their help and efforts in running this trial.References1Royal Society for the Prevention of Cruelty to Animals(RSPCA). Housing, husbandry and care. https://science.rspca.org.uk/sciencegroup/researchanimals/ethicalreview/functionstasks/housingandcare2Guide for the Care and Use of Laboratory Animals (1996).8th edition. Chapter: 2 Animal Environment, Housing, andManagement. National Academies Press. ISBN 13: 978-0-309-15400-0 (Book) ISBN -10: 0-309-15400-6 (Book)ISBN 13: 978-0-309-15401-7 (PDF) ISBN 10: 0-309-15401-4 (PDF)3Toth, L.A., Kregel, K., Leon, L. and Musch, T.I. (2011).Environmental Enrichment of Laboratory Rodents: TheAnswer Depends on the Question. Comparative Medicine61(4): 314-3214National Centre for the Replacement, Refinement andReduction of Animals in Research (NC3Rs).https://www.nc3rs.org.uk/the-3rs5Garcia-Arocena, D. (2016). How to spot and managemalocclusion in research mice. The Jackson Laboratories.6Baumans, V. and Van Loo, P.L. (2013). How to improvehousing conditions of laborator y animals: the possibilitiesof environmental refinement. The Veterinary Journal 195:24-32.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 97

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99Legal and ethical aspects of using animalsin research: A NACWO’s guideMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road W12 0NNCorrespondence: m.bilton@imperial.ac.ukBased on an IAT Higher Education Level 5, Law and Animal Welfare module assessment submissionAbstractUnder the provisions of the Animal (ScientificProcedures) Act 1986 (ASPA)5at least one NamedAnimal Care and Welfare Officer (NACWO) must beincluded in the membership of each establishment’sAnimal Welfare and Ethical Review Body (AWERB). Inorder to fulfil their responsibilities NACWOs have a goodunderstanding of both the law protecting animals usedin scientific procedures and the ethics of such use.The author discusses the history of the use of animalsin science, the implementation of ASPA and guidesNACWOs through the stages of ethical assessment ofresearch proposals.Key words: Law, Ethics, Morals, Animals, ScientificProceduresIntroduction and history of animalsused in researchAnimal experimentation, commonly known asvivisection, has a long history and dates back to theancient Greeks.1The philosophers and physicians ofthose times wanted to increase their knowledge aboutthe way in which organisms such as humans andanimals functioned.2Over the next 2000 years the useof animals in research increased. As the use ofanimals increased, so did opposition to vivisection.3During the nineteenth centur y there was a gradualbuild-up of demands for legislation to protect animalsused in research. The Society for the Prevention ofCruelty to Animals (later the Royal Society for thePrevention of Cruelty to Animals – RSPCA) was foundedin 1824 by the 1880s the anti-vivisection society hadbeen established. Due to their pressure and a petitionsigned by Thomas Huxley, Edward Jenner and Owen,the presidents of the Royal Colleges of Physicians andSurgeons and scientists including Dar win, called forthe control of animal experimentation.4Laboratory animal lawIn the United Kingdom the main piece of legislationrelated to animal research is the Animals (ScientificProcedures) Act 1986 (ASPA).5ASPA has come a longway from the older legislation regarding animals suchas the Mar tin Act 1822, the Cruelty to Animals Act1876 and the Protection of Animal Act 1911.4ASPA is an enabling act which means it is an Act ofParliament that gives specified powers to individuals ororganisations. ASPA is an enabling Act because it givespowers to the Secretary of State to make amendmentsto it with having to pass a new Act through Parliament.4ASPA permits, through the issuing of licences, peoplemeeting the relevant criteria, to carry out research thatwould normally be punishable by other animalprotection acts.Kolar (2006) says that animal experimentation isunique as, in animal experiments, pain, suffering anddistress are deliberately inflicted on animals, whereasin other fields this would be regarded an illegalmistreatment.6ASPA has a three-tier licencing system in place, theestablishment (a section 2c licence or Establishmentlicence), the person (a personal individual licence) andthe project (a project individual licence). ASPA section2C (5) specifies ‘a person to be responsible foroverseeing the welfare and care of the animals kept atthe place specified in the licence’. Up until 1997 thisperson was known as the person responsible for theday to day care of the animals since 1997 as theNamed Animal Care and Welfare Officer (NACWO).Under ASPA the NACWO has several responsibilities. Incarrying out these responsibilities the NACWO mustdecide what is morally and ethically the right course ofaction regarding the impact that decision will have onanimal welfare.Morals and ethicsDolan (1999) describes ethics as ‘The philosophicalAugust 2019 Animal Technology and WelfareLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 99

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Legal and ethical aspects of using animals in research: A NACWO’s guide100study of the moral value of human conduct and therules and principles that ought to govern it’.7Dolan(1999)7goes on to say that ethics is an exploration oftheories about right and wrong, while morality tends tobe prescriptive about what is right and wrong.Wolfensohn and Lloyd (2013) state that the word‘ethics’ can be used in many contexts and that it is anexamination of the acceptability of the motives thatdrive the behaviour of people.3Garrett (2012)describes an ethical issue as one that challengespeople to apply to concepts of right, wrong, good andbad to a situation.8All these statements are relevant tothe NACWO when carrying out their roles. The NACWOcould at times have to justify whether a decision is rightor wrong not only in legal terms but ethically. Thedistinction between what is legal and what is ethical iscrucial as described by Dolan (2007) writing in the IATManual of Animal Technology, the distinction betweenethics and law is that ethics is ‘exhortation withoutenforcement’ while law is ‘an order, command ordirective by authority backed by a sanction, penalty orpunishment’.9Kolar (2006) states that legislation onanimal experimentation mostly stipulates that anethical decision is made whether an animal experimentis to be authorised and lays down the basic parametersfor that decision.6The NACWO is not alone in makingdecisions regarding ethics and law. In the UK the holderof a section 2C licence is required to establish andmaintain an Animal Welfare and Ethical Review Body(AWERB). The AWERB must consist of at least oneNACWO.Animal Welfare and Ethical ReviewBodyThe AWERB must carr y out the tasks mentioned inArticle 27.1 of Directive 2010/63/EU.10This includesreviewing project licences and whether that therequested research is both ethical and legal. Everyproject approved will impact on the day-to-day duties ofthe NACWO – as the NACWO would need to be aware ofthe humane endpoints, adverse effects and controlmeasures specified and assess animal welfare.Project licence reviewA project licence will propose a programme of work tobe applied to animals for the following purposes;– Basic research.– Translational or applied research.– The development, manufacture or testing of thequality, effectiveness and safety of drugs,foodstuffs and other substances.– Protection of the natural environmental in theinterests of the health and welfare of human beingsor animals.– Research aimed at preservation of the species.– Higher education, or training for the acquisition,maintenance or improvement of vocational skills.– Forensic inquiries.(Article 5 Directive 2010/63/EU)10ProposalAn example could be using 50 Guinea pigs to establishwhether antiviral compounds can influence theoutcome of herpes virus (HSV) infection the nature andlevel of immune responses of the animals to a varietyof viral vaccine preparations.The Guinea pigs will be challenged with a live virusintravaginally. Blood samples will be collected atintervals and vaginal / swabbing carried out to checkantibody responses. It would be anticipated that mostGuinea pigs will fully recover from HSV challengeinfection. The Severity is considered to be Moderate;this would reflect the vaginal lesions and urinaryretention which can occur after challenge with the virus.In most Guinea pigs some urinary retention will occurduring the first two weeks or until successful treatment.This will be relieved by emptying the bladder withabdominal massage once or twice daily as necessary.Some Guinea pigs may develop localised immuneresponses which will be difficult to treat.The Nuffield Council on Bioethics (2005)2states that itis important to consider the ethical issues raised byanimal research and take into account:– The impact on the lives and welfare of animals thatdifferent uses have.– The broader consequences if there were a ban onusing animals in specific circumstances.– A comparison of the benefits arising from thedifferent uses of animals: and– The numbers of animals involved.The Nuffield Council on Bioethics (2005)2also statesthat there are five features which may be consideredapplicable to all animals these are;– Sentience– Higher cognitive capacities– The capacity to flourish– Sociability and– The possession of a lifeAll of these will need to be considered when reviewinglicences and the impact the research will have on thewelfare of the animals. The NACWO will need toconsider, question and offer advice regarding:SpeciesHome Office Annual Statistics of Scientific Procedureson Living Animals Great Britain 2017 state that a totalof 22,5602 Guinea pigs were used in scientificLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 100

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Legal and ethical aspects of using animals in research: A NACWO’s guide101procedures.11When compared with the number of miceused which was 1,094,867, Guinea pigs make up asmall percentage of animals used. The choice ofspecies is an interesting consideration for a NACWO asthere is thought that some species of animals can beused because they are less sentient than others.7Public opinion polls and legislation reflect the fact thatwe are more concerned about the use and welfare ofsome species than others.1Since 1999, the pollingcompany Ipsos MORI has been asking the public in theUK about their views on animal research. The findingssuggest that more than 80% of respondents acceptedthe need for animal research provided certainconditions were met such as there is no unnecessarysuffering, the research is for serious medical or life-saving purposes and that there is no alternative(Wolfensohn and Lloyd 2013).3Sentience is associatedwith Jeremy Bentham (1748-1832). Benthamunderstood sentience as the capacity to feel pleasureand pain.12The ascription of such states is not alwaysstraightforward, it is uncontested that to cause pain ismorally problematic and needs to be taken into accountin moral reasoning.2Peter Singer’s view is that allanimals are equal7as does Tom Regan (1983)13writingin the case for Animal rights.The 3RsIt is an obligation to constantly seek more acceptableexpressions of the 3Rs in practice it is not only ethicalbut is also grounded in ASPA. The concept of the 3RsReplacement, Reduction and Refinement wasdeveloped by Russell and Birch and the termsrepresent widely accepted principles of humaneexperimental technique.14Regarding reduction, wouldthe use of 50 Guinea pigs provide sufficient scientificdata compared to using a larger number of mice?Research questions should always be clearly defined. Itwould also be important to consider whether changesof the various elements of a project might preventwastage of animals.1This view is backed up by theNuffield Council on Bioethics (2005)2who state thatanimals will suffer needlessly if they are used inresearch where scientific methodology is poor. TheCouncil go on to state that the application of the 3Rsshould begin with a careful assessment of the initialexperimental design and a number of questions beaddressed such as, is the animal model relevant to thescientific question being asked or health problem understudy? Is there a genuine scientific basis for using aparticular animal model? Could the scientific objectiveof the work be modified to avoid the use of an animalmodel?Pain & Welfare assessmentsDolan (1999) states that pain is one of the most vividforms of awareness.7He goes on to say that pain is atthe heart of any ethical discussion of the use ofanimals is research.7Under ASPA pain is described inthat carr ying out regulated procedures has thepotential to cause ‘pain, suffering, distress or lastingharm’. Rollin (1989)15states that it is claimed thatworr ying about animal pain is misplacedanthropomorphism and that in circumstances in whichhumans would be screaming and writhing, manyanimals show very few signs of extreme pain. However,work carried out by Langford et al. (2010)16havedemonstrated that pain could be shown in mice via themouse grimace scale but this is not going to be helpfulin assessing pain regarding the Guinea pig in theproposal. It is clear from the proposal that theseGuinea pigs will experience pain and that the NACWOwill need to consider/question how this pain will becontrolled. The Nuffield Council on Bioethics (2005)2do state that humans will have to apply concepts suchas pain, suffering and distress, which are usedsuccessfully in human-human interactions, whendealing with assessments of animals. The idea that theGuinea pigs will need abdominal massage to empty thebladder would be a cause for concern as this would bea painful experience if it was done to a human. TheNuffield Council on Bioethics (2005)2do consider theconcept of critical anthropomorphism as a usefulstarting point as the experience of humans to alleviateanimal suffering by combining the perception of theanimal’s situation.TransportWill these Guinea pigs require transport? If so, whatwill be the requirements? When an animal istranspor ted there is a complete dependence ofanimals on their handlers.7Wolfensohn and Lloyd(2013)3state that stress caused by transport may lastup to 5 days and affects physiological processes suchas immunity and reproduction. While Hubrecht (2014)1states that transport stressors may include disruptionof social groups, unfamiliar housing and stimuli,changes in husbandr y and handlers, changes intemperature and humidity, exposure to noise, vibrationand movement, all of which may cause physiologicalchanges and adverse mental states such as fear andfrustration. White et al. (2010) states that transientperiods of stress will occur during transportation.17Allof which the NACWO will need to consider.In asking questions regarding the sections above theNACWO will be safeguarding the welfare of thoseanimals to be used in research. As Garrett (2012)8states that there is a challenge with people who carryout animal research as they must provide a compellingcase for the unique benefits offered by animal researchand they must provide a rationale for why thesebenefits justify treating animal subjects in ways thatwould be unacceptable for human subjects. The AWERBis one of the ways in which this is done. Using animalsin research is not considered lightly. The NACWO is onlyone of a number of members of the AWERB, before acase is presented at AWERB a cost-benefit analysis iscarried out usually by the Home Office and is aLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 101

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Legal and ethical aspects of using animals in research: A NACWO’s guide102balancing act as described by Dolan (1999).7This is adifficult thing to do as predicting beneficial outcomes ofexperiments before they have been conducted isdifficult as results may be unexpected.3Wolfensohnand Lloyd (2013) also go on to state that measuringharms is a judgement of the quality of life ofexperimental animals. That would be considered themain question regarding the whole process of theAWERB, it is the quality of life the animal will haveduring the proposed research. Nuffield Council onBioethics (2015) consider that animals in research dohave a possession of a life.2Experience and quality oflife in a research setting does bring about its issues,Frey (2002) that defending animal experimentation isby means of the augment form benefit. Are the totalbenefits of the experiment to humans greater than thetotal costs to the animals?18The debate on whetheranimal experimentation is right or wrong will rage on insuch books as Tom Regan’s ‘The case for AnimalRights’ (1983),13Animal Rights and Human obligationsedited by Tom Regan and Peter Singer (1989)19whichgive both for and against, however the case for oragainst the use of animals in research is not one forNACWOs or the AWERB to consider. That is forGovernments to decide but for as long as animals areused in research it is impor tant that people who carefor animals are involved in the ethical review process tooptimise the welfare of the animals in a researchsetting. That is why NACWOs and AWERBs are a vitalresource in a scientific establishment.References1Hubrecht, R.C. (2014). The Welfare of Animals used inresearch: Practice and ethics. West Sussex. UFAW.2Nuffield Council on Bioethics (2005). London. NuffieldCouncil on Bioethics.1Wolfensohn, S. and Lioyd, M. (2013). Handbook ofLaborator y Animal Management and Welfare FourthEdition. West Sussex. Wiley-Blackwell.3Dolan, K. (2000). Laborator y Animal Law. OxfordBlackwell Science Ltd.4Home Office. Animals (Scientific Procedures) Act 1986Amendment Regulations 2012.5Kolar, R. (2006). Animal Experimentation. ScienceEngineering Ethics. 12, 111-122.6Dolan, K. (1999). Ethics, Animals and Science. Oxford.Blackwell Science Ltd.7Garrett, J.R. (2012). The Ethics of Animal ResearchExploring the controversy. Massachusetts USA.Massachusetts Institute of Technology.8Dolan, K. (2007). The Ethical Implications of the use ofAnimals in Scientific Procedures in: Barnett S W edManual of Animal Technology. Oxford. Blackwellpublishing.9Directive 2010/63/EU of The European Parliament andof the Council of 22 September 2010 on the protection ofanimals used for scientific purposes.10Home Office. Annual Statistics of Scientific Procedureson Living Animals Great Britain 2014.11Bentham, Jeremy (1879). [This edition first published1823]. An Introduction to the Principles of Morals andLegislation (A New Edition, corrected by the Author).Clarendon Press.12Regan, T. (1983). The Case for Animal Rights. California.University of California Press.13Russell, W.M.R. and Burch, R.L. (1959). The Principles ofHumane Experimental Technique. Wheathampstead:Universities Federation for Animal Welfare.14Rollin, B. (1989). Animal Pain in: Regan, T., Singer, P. ed:Animal Rights and Human Obligations Second Edition.New Jersey. Prentice-Hall.15Langford, D.J. (2010). Using the Mouse Grimace Scale torevaluate the efficacy of postoperative analgesics inlaboratory mice. Nature Methods. 7, 447-449.16White, W.J. et al. (2010). Transportation of laboratoryanimals in: Hubrecht, R., Kirkwood, J. ed: The UFAWHandbook on the Care and Management of Laboratoryand Other Research Animals. West Sussex. Wiley-Blackwell.17Frey, R.G. (2002). Justifying Animal Experimentation.Society. (6) 39 pp37-47.18Animal Rights and Human Obligations (2nd edition)Editors Tom Regan and Peter Singer. Publishers. Prentice-Hall. ISBN-13: 978-0130368645 ISBN-10: 013068644LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 102

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103PAPER SUMMARYTRANSLATIONSINHALTVERZEICHNISINHALTVERZEICHNISBericht zur Tagung der RSPCA/UFAW-Tierschutzgruppe für Nagetiere und Kaninchen 2018CHLOE STEVENS,1PENNY HAWKINS (Schriftführerin),1ROBIN LOVELL-BADGE,2ROBERT HUBRECHT,3HUW GOLLEDGE,3ANNA SLAVIERO,4CLARE ELLIS,5DEMI MINHINNETT,6REBECCA TERRY,7KATHARINA HOHLNAUM,8,9DOMINIC WELLS,10THOMAS SNOEKS,2JOHN MARSHALL111Research Animals Department, Science Group, RSPCA, Wilberforce Way, Southwater,West Sussex RH13 9RS, UK2The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK3UFAW, The Old School, Brewhouse Hill, Wheathampstead, Hertfordshire AL4 8AN, UK4University of Surrey, Guildford, Surrey GU2 7AL, UK5University of Northampton, University Drive, Northampton, Northamptonshire NN1 5PH, UK6Durham University, Stockton Rd, Durham, Northumberland DH1 3LE, UK7University College London, Cruciform Building, Gower Street, London WC1E 6BT, UK8Institut für Tierschutz, Tierverhalten und Versuchstierkunde am Fachbereich Veterinärmedizin,Freie Universität Berlin, Deutschland9Institut für Pharmakologie und Toxikologie am Fachbereich Veterinärmedizin, Freie UniversitätBerlin, Deutschland10Department of Comparative Biomedical Sciences, Royal Veterinary College, Royal CollegeStreet, London, NW1 0TU, UK11Home Office Animals in Science Regulation Unit, 14th Floor, Lunar House, 40 WellesleyRoad, Croydon CR9 2BY, UKKorrespondenz: penny.hawkins@rspca.org.ukAbstractDie RSPCA/UFAW-Tierschutzgruppe für Nagetiere (und inzwischen auch Kaninchen) veranstaltet seit 25 Jahren jedenHerbst ein eintägiges Treffen, bei dem die Mitglieder über Themen der aktuellen Tierschutzforschung diskutieren undErfahrungen und Meinungen über nagerrelevante Tierschutzfragen sowie über die Umsetzung der 3R-PrinzipienVermeidung, Verringerung und Verbesserung des Einsatzes von Nagern und Kaninchen austauschen können. EinHauptziel der Gruppe ist es, zum Nachdenken über die Gesamtheit der Lebenserfahrungen von Nagern undKaninchen als Laborversuchstiere anzuregen und sicherzustellen, dass jede potenzielle Beeinträchtigung ihresWohlergehens geprüft und auf ein Minimum gesenkt wird.Im Rahmen der diesjährigen Präsentationen erfolgte ein Rückblick auf die Entwicklung der Tiertechnologie in denletzten 25 Jahren und auf die Auswirkungen dieser Entwicklungen auf den Tierschutz von Labornagetieren und –kaninchen. In einem weiteren Vortrag wurden mögliche wissenschaftliche und praktische Entwicklungen desAugust 2019 Animal Technology and WelfareLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 103

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Paper Summary Translations105Ein Vergleich von Anreicherungselementen zurFörderung des natürlichen Nageverhaltens vonLabormäusenIRENE LOPEZ JUARISTIUniversity College, London, Biological Services, Cruciform – North Wing, Cruciform Building,Gower Street, London WC1E 6BT, UKKorrespondenz: irene.lopez@ucl.ac.ukBasierend auf einem Bericht an die UCL Collective Laboratory Animal Welfare Society (CLAWS)AbstractEine angemessene Unterbringung und Haltung, einschließlich Umweltanreicherung, muss den natürlichenLebensraum, die Biologie und das Verhalten der einzelnen Arten berücksichtigen.1Laut Guide for the Care and Useof Laboratory Animals (1996)2muss das Ziel immer darin bestehen, artenspezifische Verhaltensweisen maximal zufördern und stressbedingte Verhaltensweisen auf ein Minimum zu senken.Dieser Bericht beschäftigt sich mit einer Initiative der Collective Laboratory Animal Welfare Society (CLAWS) zumTesten verschiedener Arten von Kaublöcken, um das Nageverhalten von Mäusen zu fördern und zu ermitteln, welcherfür sie am bereicherndsten ist. Das University College of London (UCL) entwickelt ständig neue Ansätze undEmpfehlungen, die die aktuellen Standards optimalen Tierschutzes hinter fragen. Diese Studie wurde inZusammenarbeit zwischen zwei UCL-Abteilungen durchgeführt: dem Institute of Ophthalmology (IOO) und denCruciform Units.Schlagwörter: Mäuse, natürliches Verhalten, artenspezifisches Verhalten, Umweltanreicherung, KaublöckeLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 105

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August 2019 Animal Technology and Welfare107CONTENU DE LA REVUECONTENU DE LA REVUERapport issu de la réunion du RSPCA/UFAW Rodentand Rabbit Welfare Group en date de 2018CHLOE STEVENS,1PENNY HAWKINS (Secretary),1ROBIN LOVELL-BADGE,2ROBERT HUBRECHT,3HUW GOLLEDGE,3ANNA SLAVIERO,4CLARE ELLIS,5DEMI MINHINNETT,6REBECCA TERRY,7KATHARINA HOHLNAUM,8,9DOMINIC WELLS,10THOMAS SNOEKS,2JOHN MARSHALL111Research Animals Department, Science Group, RSPCA, Wilberforce Way, Southwater,West Sussex RH13 9RS2The Francis Crick Institute, 1 Midland Road, London NW1 1AT3UFAW, The Old School, Brewhouse Hill, Wheathampstead, Hertfordshire AL4 8AN4University of Surrey, Guildford, Surrey GU2 7AL5University of Northampton, University Drive, Northampton, Northamptonshire NN1 5PH6Durham University, Stockton Rd, Durham, Northumberland DH1 3LE7University College London, Cruciform Building, Gower Street, London WC1E 6BT8Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Service demédecine vétérinaire, Freie Universität Berlin, Allemagne9Institut de Pharmacologie et de toxicologie, service de médecine vétérinaire, Freie UniversitätBerlin, Allemagne10Service des sciences biomédicales comparatives, Royal Veterinary College, Royal CollegeStreet, London, NW1 0TU11Home Office Animals in Science Regulation Unit, 14th Floor, Lunar House, 40 WellesleyRoad, Croydon CR9 2BYAdresse e-mail: penny.hawkins@rspca.org.ukLe groupe de bien-être du RSPCA/UFAW en charge des rongeurs (et désormais des lapins) se réunit durant unejournée à chaque automne depuis 25 ans afin de permettre à ses membres de débattre de l’état de la recherche enmatière de animal, d’échanger leurs points de vue sur ce sujet et de par tager leur expérience personnelle concernantles « 3 R » que sont le « replacement, reduction and refinement » (« remplacement, réduction et amélioration ») pourl’utilisation des rongeurs et des lapins. L’un des principaux objectifs du groupe est de favoriser la réflexion surl’ensemble de l’expérience de vie des rongeurs et des lapins en laboratoire afin de garantir que tout aspect négatifpour leur bien-être soit détecté et réduit.Les présentations comprenaient un regard rétrospectif sur la façon dont la technologie animale s’est développée aucours des 25 dernières années et sur les conséquences produites par ces développements sur le bien-être desrongeurs et des lapins de laboratoire. Parmi les contributions se trouvait également un exposé sur la façon dont lascience et les pratiques en matière de bien-être animal pourraient évoluer au cours des 25 prochaines années. Uneprésentation por tait sur les manières de favoriser la construction de leur nid par les rats de laboratoire via lafourniture de matériaux de construction appropriés, une autre fournissait des conseils sur la conception de nouvellesinstallations pour le bien-être des lapins et une dernière consistait en un débat sur la façon dont les techniquesd’imagerie peuvent être utilisées pour améliorer les procédures expérimentales et réduire le nombre d’animauxutilisés dans les études. La journée s’est achevée sur une présentation du Home Office Animals in ScienceRegulation Unit et une séance de débat interactif portant sur le thème « s’assurer que les rongeurs et les lapins delaboratoire ne soient jamais privés de nourriture ou d’eau ». Ce rapport résume la réunion et se conclut sur une listede mesures que les lecteurs peuvent envisager d’appliquer dans leurs établissements.Mots clés: Logement, Personnalités de lapins, Construction du nid, Gravité, IdentificationLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 107

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Paper Summary Translations108Comparaison des éléments d’enrichissementfavorisant un rongement de type naturel chez lessouris de laboratoiresIRENE LOPEZ JUARISTIUniversity College, London, Biological Services, Cruciform – North Wing, Cruciform Building,Gower Street, London WC1E 6BT, UKAdresse e-mail: irene.lopez@ucl.ac.ukSur la base d’un rapport soumis à l’UCL Collective Laboratory Animal Welfare Society (CLAWS)RésuméAfin que le logement et l’élevage soient adaptés – notamment en matière d’enrichissement de l’environnement –ceux-ci doivent prendre en compte le milieu naturel, la biologie et le comportement de chacune des espèces.1LeGuide for the Care and Use of Laboratory Animals (Guide pour la prise en charge et l’utilisation des animaux delaboratoire, 1996)2stipule que l’objectif doit toujours consister à favoriser les comportements naturels et à réduireautant que possible les comportements induits par le stress.Ce rapport est le fruit d’une initiative du Collective Laboratory Animal Welfare Society (CLAWS) visant à tester lesdifférents types de blocs à mâcher favorisant le rongement et à déterminer lequel d’entre eux est le plus enrichissantpour la souris. L’University College of London (UCL) fournit régulièrement de nouvelles approches et de nouvellesrecommandations qui remettent en cause les normes établies concernant le bien-être animal. Ces essais ont étémenés en collaboration par deux équipes de l’UCL : L’Institute of Ophthalmology (IOO) et les unités Cruciform.Mots clés: Souris, comportement naturel, comportement naturel des espèces, enrichissement environnemental,blocs à mâcher.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 108

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Paper Summary Translations109Aspects éthiques et juridiques de l’utilisation desanimaux dans le cadre de la recherche: un guideNACWOMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road, London W12 0NN, UKAdresse e-mail: m.bilton@imperial.ac.ukSur la base de la soumission d’une évaluation du module Droit et bien-être animal de niveau 5 de l’enseignementsupérieur de l’IATRésuméEn vertu des dispositions de l’Animal (Scientific Procedures) Act 1986 (ASPA)5, au minimum un membre du NamedAnimal Care and Welfare (NACWO) doit être intégré dans chaque établissement de l’Animal Welfare and EthicalReview Body (AWERB). En vue d’assurer cette responsabilité, les représentants de NACWO doivent posséder à la foisune connaissance approfondie des lois en matière de protection des animaux utilisés dans le cadre de recherchesscientifiques et de l’éthique qui se rattache à cet usage.L’auteur examine l’histoire de l’utilisation des animaux en science, la mise en œuvre de l’ASPA et offre un guide auxmembres de NACWO concernant les étapes de l’évaluation éthique des propositions de recherche.Mots clés: Législation, éthique, morale, animaux, protocoles scientifiquesLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 109

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110INDICE DE LA REVISTAINDICE DE LA REVISTAInforme sobre la reunión del Grupo para elbienestar de roedores y conejos RSPCA/UFAW 2018CHLOE STEVENS,1PENNY HAWKINS (Secretaria),1ROBIN LOVELL-BADGE,2ROBERT HUBRECHT,3HUW GOLLEDGE,3ANNA SLAVIERO,4CLARE ELLIS,5DEMI MINHINNETT,6REBECCA TERRY,7KATHARINA HOHLNAUM,8,9DOMINIC WELLS,10THOMAS SNOEKS,2JOHN MARSHALL111Departamento de Investigación Animal, Science Group, RSPCA, Wilberforce Way, Southwater,West Sussex RH13 9RS, UK2The Francis Crick Institute, 1 Midland Road, Londres NW1 1AT, UK3UFAW, The Old School, Brewhouse Hill, Wheathampstead, Hertfordshire AL4 8AN, UK4University of Surrey, Guildford, Surrey GU2 7AL, UK5University of Northampton, University Drive, Northampton, Northamptonshire NN1 5PH6Durham University, Stockton Rd, Durham, Northumberland DH1 3LE, UK7University College London, Cruciform Building, Gower Street, Londres WC1E 6BT, UK8Instituto de Bienestar Animal, Comportamiento Animal y Ciencia de Animales de Laboratorio,Departamento de Medicina Veterinaria, Freie Universität Berlín, Alemania9Instituto de Farmacología y Toxicología, Departamento de Medicina Veterinaria, FreieUniversität Berlin, Alemania10Departamento de Ciencias Biomédicas Comparativas, Royal Veterinary College, Royal CollegeStreet, Londres NW1 0TU, UK11Home Office Animals in Science Regulation Unit, 14th Floor, Lunar House, 40 WellesleyRoad, Croydon CR9 2BY, UKCorrespondencia: penny.hawkins@rspca.org.ukResumenEl Grupo de trabajo para el bienestar de roedores (y ahora también conejos) RSPCA/UFAW ha celebrado una reuniónde un día cada otoño durante los últimos 25 años para que sus miembros puedan mantener un debate acerca dela investigación actual sobre el bienestar, intercambiar opiniones sobre temas relacionados con el bienestar deroedores y compartir su experiencia respecto a la implementación de las 3 R (reemplazo, reducción y refinamiento)en relación con el uso de roedores y conejos. Uno de los objetivos primordiales del Grupo es fomentar que laspersonas piensen sobre la experiencia vital de los roedores y conejos de laboratorio de forma que se garantice quecualquier repercusión negativa en su bienestar sea revisada y reducida.Las presentaciones incluían una visión retrospectiva sobre cómo la tecnología con animales ha ido evolucionandoen los últimos 25 años y cómo estos desarrollos han influido en el bienestar de los conejos y roedores delaboratorio. Las contribuciones también incluían un debate sobre cómo la ciencia del bienestar animal y las prácticaspueden cambiar en los próximos 25 años. Otras presentaciones trataron sobre formas de fomentar la creación denidos entre las ratas de laboratorio dándoles el material adecuado, consejos sobre el diseño de nuevas instalacionespara conejos a fin de promover su bienestar y un debate sobre cómo las técnicas de toma de imágenes puedenutilizarse para refinar procedimientos experimentales y reducir el número de animales utilizados en los distintosestudios. El día terminó con una presentación de la Unidad de regulación del uso de animales en la ciencia (Animalsin Science Regulation Unit) del Ministerio del Interior del Reino Unido y un debate interactivo, ambos sobre medidaspara garantizar que los conejos y roedores nunca se queden sin agua ni comida. Este informe resume la reunión yfinaliza con una lista de puntos de acción que los lectores pueden presentar en sus propios centros.Palabras clave: alojamiento, personalidades de conejos, creación de nidos, gravedad, identificaciónAnimal Technology and Welfare August 2019LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 110

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Paper Summary Translations111Comparación de medidas de enriquecimiento para lapromoción del comportamiento natural durante laacción de roer de los ratones de laboratorioIRENE LOPEZ JUARISTIUniversity College London, Biological Services, Cruciform – North Wing, Cruciform Building,Gower Street, Londres WC1E 6BT, UKCorrespondencia: irene.lopez@ucl.ac.ukBasado en un informe para la Asociación colectiva del bienestar de los animales de laboratorio (CollectiveLaboratory Animal Welfare Society o CLAWS) de la UCLResumenPara la cría y el enjaulamiento adecuados, incluido el enriquecimiento ambiental, se deben tener en cuenta el hábitatnatural, la biología y el comportamiento de cada especie.1En la Guide for the Care and Use of Laboratory Animals(1996)2se indica que el objetivo siempre debe ser maximizar los comportamientos específicos de cada especie yminimizar los comportamientos inducidos por el estrés.Este informe proviene de una iniciativa de la Collective Laboratory Animal Welfare Society (CLAWS) con la que sepretende testar distintos tipos de bloques para roer con el fin de fomentar el comportamiento durante la acción deroer y ver cuál es el más enriquecedor para los ratones. La University College London (UCL) contribuyeconstantemente con nuevos métodos y recomendaciones que cuestionan las normas actuales para que el bienestaranimal sea el mejor posible. Este ensayo se hizo en colaboración con dos unidades de la UCL: El Institute ofOphthalmology (IOO) y las unidades Cruciform.Palabras clave: ratones, comportamiento natural, comportamiento específico de cada especie, enriquecimientoambiental, bloques para roer.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 111

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Paper Summary Translations112Aspectos legales y éticos sobre la utilización deanimales para la investigación: guía sobre losNACWOMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road, London W12 0NN, UKCorrespondencia: m.bilton@imperial.ac.ukCon base en la presentación de una evaluación de un módulo de IAT Higher Education Nivel 5, Ley y bienestaranimalResumenCon arreglo a las disposiciones de la Ley británica sobre Animales (procedimientos científicos) 1986 (ASPA)5almenos un responsable del cuidado y el bienestar de animales (Named Animal Care and Welfare Officer o NACWO)deberá formar parte de la afiliación del Organismo del bienestar de los animales y la revisión ética (Animal Welfareand Ethical Review Body o AWERB) de cada establecimiento. Para poder cumplir con sus responsabilidades, losNACWO deben asimilar en profundidad la ley de protección animal aplicable a procedimientos científicos y la éticaque esta comprende.El autor hablar sobre la historia del uso de animales en la ciencia, la implementación de ASPA y las guías de NACWOa través de las fases de evaluación ética de propuestas de investigación.Palabras clave: ley, ética, moral, animales, procedimientos científicos.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 112

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113April 2019 Animal Technology and WelfareINDICE DELLA REVISTAINDICE DELLA REVISTAResoconto dell’incontro del RSPCA/UFAW Rodentand Rabbit Welfare Group del 2018CHLOE STEVENS,1PENNY HAWKINS (Segretaria),1ROBIN LOVELL-BADGE,2ROBERTHUBRECHT,3HUW GOLLEDGE,3ANNA SLAVIERO,4CLARE ELLIS,5DEMI MINHINNETT,6REBECCA TERRY,7KATHARINA HOHLNAUM,8,9DOMINIC WELLS,10THOMAS SNOEKS,2JOHN MARSHALL111Research Animals Department, Science Group, RSPCA, Wilberforce Way, Southwater,West Sussex RH13 9RS, UK2The Francis Crick Institute, 1 Midland Road, Londra NW1 1AT, UK3UFAW, The Old School, Brewhouse Hill, Wheathampstead, Hertfordshire, AL4 8AN, UK4University of Surrey, Guildford, Surrey GU2 7AL, Regno Unito5University of Northampton, University Drive, Northampton, Northamptonshire NN1 5PH, UK6Durham University, Stockton Rd, Durham, Northumberland DH1 3LE, Regno Unito7University College London, Cruciform Building, Gower Street, Londra WC1E 6BT, UK8Istituto di Benessere Animale, Comportamento Animale e Scienze per gli Animali daLaboratorio, Facoltà di Medicina Veterinaria, Università libera di Berlino, Germania9Istituto di Farmacologia e Tossicologia, Facoltà di Medicina Veterinaria, Università libera diBerlino, Germania10Department of Comparative Biomedical Sciences, Royal Veterinary College, Royal CollegeStreet, Londra, NW1 0TU, UK11Animals in Science Regulation Unit del Ministero degli Interni, 14th Floor, Lunar House,40 Wellesley Road, Croydon CR9 2BY, UKCorrispondenza: penny.hawkins@rspca.org.ukOgni autunno, da ormai 25 anni, il RSPCA/UFAW Rodent and Rabbit Welfare Group (Gruppo sul benessere dei roditori(e ora anche) dei conigli) organizza un incontro di un giorno per consentire ai suoi membri di discutere degli studiattuali di ricerca sul benessere, di scambiarsi opinioni sulle questioni legate al benessere e di condividere esperienzedi applicazione del principio delle 3 R, ovvero sostituzione (replacement), riduzione (reduction) e perfezionamento(refinement), in relazione all’uso di roditori e conigli. Uno degli obiettivi primari del Gruppo è quello di invitare aprendere in considerazione l’intero percorso di vita dei roditori e dei conigli da laboratorio, accertandosi che vengavalutato e minimizzato ogni possibile impatto negativo sul loro benessere.Le presentazioni hanno incluso una valutazione retrospettiva dell’evoluzione della stabulazione negli ultimi 25 anni,esaminando l’impatto di questi sviluppi sul benessere di roditori e conigli da laboratorio. Si è tenuta, inoltre, unadiscussione sul possibile cambiamento delle scienze e delle prassi di benessere animale nell’arco del prossimo quartodi secolo. Altre presentazioni si sono incentrate sui metodi da utilizzare per incoraggiare i ratti da laboratorio a costruireil proprio nido mettendo a loro disposizione i materiali adatti e su nuove idee per la progettazione di strutture per coniglial fine di promuovere al meglio il loro benessere, oltre a esplorare l’uso delle tecniche di imaging per perfezionare leprocedure sperimentali e ridurre il numero di animali usati negli studi. La giornata si è conclusa con una presentazionedella Animals in Science Regulation Unit del Ministero degli Interni e con una sessione interattiva, entrambe mirate afar sì che i roditori e i conigli da laboratorio non siano mai privi di cibo o acqua. Il presente resoconto riassume l’incontroe termina con un elenco di punti di intervento che i lettori possono portare all’attenzione delle loro strutture.Parole chiave: alloggio, personalità dei conigli, costruzione dei nidi, gravità, identificazioneLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 113

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Confronto tra oggetti di arricchimento per lapromozione dell’abitudine naturale di rosicchiarenei topi da laboratorioIRENE LOPEZ JUARISTIUniversity College, London, Biological Services, Cruciform – North Wing, Cruciform Building,Gower Street, Londra WC1E 6BT, UKCorrispondenza: irene.lopez@ucl.ac.ukBasato su un resoconto della Collective Laboratory Animal Welfare Society (CLAWS) dell’UCLAbstractUna stabulazione e un allevamento appropriati, tra cui l’arricchimento ambientale, devono prendere in considerazionel’habitat naturale, la biologia e il comportamento di ciascuna specie.1La Guide for the Care and Use of LaboratoryAnimals (1996)2afferma che si dovrebbe mirare sempre a massimizzare i comportamenti specie-specifici, riducendoinvece al minimo quelli indotti dallo stress.Il presente resoconto si basa su un’iniziativa della Collective Laboratory Animal Welfare Society (CLAWS), il cuiobiettivo era provare vari tipi di blocchetti da masticare per promuovere l’abitudine di rosicchiare e scoprire quelloche comportava maggiore arricchimento per i topi. La University College of London (UCL) introduce costantementenuovi approcci e raccomandazioni che sfidano gli standard attuali di garanzia del miglior benessere animale. Questostudio è il risultato della collaborazione tra due unità dell’UCL: l’Institute of Ophthalmology (IOO) e l’unità Cruciform.Parole chiave: topi, abitudine naturale, comportamenti specie-specifici, arricchimento ambientale, blocchetti damasticare.Paper Summary Translations114LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 114

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115Aspetti etici e legali dell’uso di animali per scopi diricerca: guida per NACWOMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road W12 ONN, UKCorrispondenza: m.bilton@imperial.ac.ukBasata sulla presentazione di una valutazione relativa al modulo ‘La legge e il benessere animale’ del Livello 5 diformazione superiore IATAbstractAi sensi delle disposizioni della legge inglese Animal (Scientific Procedures) Act del 1986 (ASPA)5, ciascuna strutturadeve includere almeno un Named Animal Care and Welfare Officer (NACWO – Responsabile della tutela del benessereanimale) durante la formazione di un Animal Welfare and Ethical Review Body (AWERB). Per adempiere alle lorofunzioni, i NACWO devono avere una buona conoscenza delle leggi a tutela degli animali usati in procedurescientifiche e dei principi etici alla base di tale uso.L’autore discute la storia dell’uso di animali nella scienza, l’attuazione delle legge ASPA e guida i NACWO lungo lefasi di valutazione etica delle proposte di ricerca.Parole chiave: legge, etica, principi morali, animali, procedure scientifichePaper Summary TranslationsLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 115

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117The world of biomedical research changed forever in1959. It was in that year that Drs. Russell and Burchpublished their much acclaimed book The Principles ofHumane Experimental Technique.1While their publication was a highly regarded treatiseon a variety of topics relevant to what they described asthe distinguishing factors between ‘humanity’ and‘direct’ and ‘indirect inhumanity’ within the context ofconducting animal research, their work in thatpublication is most remembered for introducing theconcept of the ‘3Rs’. While the precise definitions ofthe 3Rs have morphed considerably since theirinception, today, somewhat loosely interpreted,perhaps the most widely accepted definitions are asfollows:2– Replacement: methods which eliminate the need touse animals at all for the scientific purpose(s) athand.– Reduction: methods requiring fewer animals toobtain the objectives of the study or that allow moreinformation to be gleaned from the same number ofanimals.– Refinement: methods which reduce, minimise oralleviate animal suffering or distress.Those three principles have proven so sound, robustand universally palatable to scientists and non-scientists alike that they have become a foundationallodestar for nearly ever y major animal welfarelegislation adopted worldwide. A plethora of books,checklists, guidelines for establishing StandardOperating Procedures and instructions for writinganimal care and use protocols have leaned heavilyupon their precepts. Animal research oversightcommittees the world over spend countless hourscontemplating if proposed research projects adhere tothe standards promulgated as a result of those threeideas put forth by two scientists some six decadesago. This veterinarian, for one, believes whole-heartedly that the life of research animals and thequality of biomedical research has improveddramatically as a result.However, I believe that even Drs. Russell and Burchwould have understood that over time, the universe ofanimal based research would change dramatically, andwith that change, the need for refreshing ourunderstandings of the 3Rs would become self-evident.In particular, I would like to revisit the first of the 3Rs,‘replacement’, in the light of today’s complex andglobal research arena. Recall that the generallyaccepted definition in use today for ‘replacement’ ismethods which permit a given purpose to be achievedwithout conducting procedures on animals. Thisinterpretation (or re-interpretation) is heavily weightedtowards scientists writing protocols and conductingresearch, and animal welfare oversight bodies chargedwith assuring wellbeing. However, that definition, andthe context of its original intent, can only be trulyappreciated when considered in light of Russell andBurch’s observation and vision that ‘The greatestscientific achievements have always been the mosthumane and the most aesthetically attractive,conveying that sense of beauty and elegance which isthe essence of science at its most successful.’1Onecan only marvel at the “sense of beauty and elegance”that Russell and Burch would observe in today’s worldof science. Animal models are now selectively createdusing gene editing technologies, entire colonies ofanimals being maintained free of adventitiouspathogens using simple biochemistr y-basedmethodologies, valuable research animal lines beingpreser ved, derived, and rederived in laboratoriesacross the world using cryopreservation innovations,targeted genetic testing and strain background testingto assure the most optimised use of research animalmodels possible. And through all of that, their threefoundational principles designed to abolish inhumanityOPINION ARTICLEThe 3Rs and optimisation in a decentralisedresearch world: new perspectives on an establishedparadigmDARRELL HOSKINSTransnetyx, 8110 Cordova Road, Suite 119, Cordova TN 38016, USACorrespondence: sbugeon@transnetyx.comAugust 2019 Animal Technology and WelfareLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 117

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Opinion Article118in biomedical research has shaped the nature of theworld every step of the way.Conduct of global animal research today mandates alevel of model-sharing and outsourcing of experimentaltechniques and technologies that Drs. Russell andBurch could not have possibly foreseen. Theoutsourcing phenomenon has revolutionisedbiomedical research in countless ways but has alsopresented the research community with a need torevisit the impact of research service providers andoutsourcing companies on the welfare of animals asmeasured using the foundational 3Rs principles.This exercise in juxtaposing the 3Rs against modernbiomedical outsourcing processes could, and perhapsshould, be conducted for a wide variety of divergenttechnologies but for the purposes of this discourse, letus limit our considerations to the narrower fields ofoutsourced automated genotyping and relatedtechnologies.One common inefficiency in mouse research the world-over is insufficient housing space, made morecomplicated because litters of mouse pups reside incages longer than necessary – until research laboratorypersonnel can find time to set up for, conduct manualgenotyping, quality check results, and create cullinglists. Only then can animals with the desired genotypebe separated from those who lack the required geneticcharacteristics, finally allowing unusable animals to beculled. Russell and Burch would not have characterisedthis process, which once was the standard in theindustry, as giving one a ‘sense of beauty andelegance’. Consider, on the other hand, outsourcing ofgenotyping as a new norm. Tissue samples can becollected prior to weaning, can be shipped to arobotically automated genotyping laboratory within 24hours of collection, and can be processed, with theresultant data uploaded to the scientists’ animal colonydatabases within 72 hours, anywhere in the world. Pupscan be identified, culled, and new research housingspace created before the animals are old enough to beweaned from their mothers. Automated genotyping ismanyfold more time and cost efficient and significantlyless prone to human error than is possible to achieve byany manual genotyping process in laboratories aroundthe world – ‘beauty and elegance’ in action! Byidentifying the animals that need to be culled early, andproactively preventing unnecessar y down-streamactivities, outsourcing of genotyping is serving the 3Rsprinciple of ‘reduction’.Yet another example of outsourcing which is quicklybecoming a standard necessity for adhering to the 3Rsprinciples and for preventing issues with non-reproducibility of results is genetic background testing.It is now clear that a gene acts within the context of theentire genome of the animal in which it is situated.3Ifthe gene is studied without confirming and controllingfor the genetic background of the model being studied,it is very likely that any research data obtained will beerrant and will not be reproducible.4-7A simple test ona tissue sample sent to an automated outsourcinggenotyping lab can confirm both the gene(s) of interestand the genetic background upon which they exist. Thespeed, cost efficiency and confidence in the modelbeing used enhances the quality of the data producedand prevents the necessity of having to attempt torepeat studies on other strains of mice. It occurs to thisauthor that Drs. Russell and Burch would find suchoutsourced resources both desirable and necessary asa means of promoting animal welfare – ‘reduction’ and‘refinement’. I anticipate that in time many animalwelfare oversight bodies and publications will beginarriving at the same conclusions.Today we are faced with an even more elusive issuethat continues to challenge the 3Rs approach to animalwelfare – the data non-reproducibility crisis that muchof the research world now believes is associated withdivergent microbiome and microbiota across studiesand settings. Just as rodent pathogens were once anelusive source of variability in research results, it nowseems clear that as the sophistication of our researchendeavours increases, the microbiome represents thenext elusive, only modestly characterised, source ofscientific variability.8-14The microbiome is clearlyassociated with changes in phenotype, yet methods toefficiently characterise and manage it are just nowgaining significant traction. Clearly, issues related tothe microbiome and microbiota are playing havoc withthe 3Rs. Drs. Russell and Burch would surely beunsettled! But once again, highly automated, roboticlaboratories, with systems optimised to both identifyand characterise the microbiome of animal models arepoised to salvage the applicability of the 3Rs. A freshfaecal sample shipped to a specialised microbiomeoutsourcing laboratories can be used to generate highlyreliable data sets, optimised for interpretation byscientists and veterinarians in a matter of days toweeks. In time, we will learn to efficiently use thatmicrobiome data to fur ther minimise avoidablescientific variability and the associated need tounsuccessfully repeat studies. As a result, animals andanimal space will be used more humanely andefficiently, and we will continue to benefit from thetime-tested 3Rs paradigm for animal research.It seems clear that outsourcing of key scientificprocesses has, is, and will continue to provide sleek,rapid, reliable, reproducible, humane and scientificallyindispensable resources with which Drs. Russell andBurch would indeed approve.‘The greatest scientific achievements have alwaysbeen the most humane and the most aestheticallyattractive, conveying that sense of beauty and elegancewhich is the essence of science at its mostsuccessful.’ – Drs Russell and Burch.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 118

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Opinion Article119References1Russell, W.M.R. and Burch, R.L. (1959). The principles ofhumane experimental technique. Wheathampstead:Universities Federation for Animal Welfare.2Tannenbaum, J. and Bennett, B.T. (2015). Russell andBurch’s 3Rs Then and Now: The Need for Clarity inDefinition and Purpose. JAALAS, 54(2): 120-1323Geurts, N., Martens, E., Verhenne, S., Lays, N., Thijs, G.,Magez, S. et al. (2011).Insufficiently defined genetic background confoundsphenotypes in transgenic studies as exemplified byMalaria infection in Tlr9 Knockout Mice. PLOS One. Vol.6(11):e27131.4Fahey, J.R., Katoh, H., Malcolm, R. and Perez, A.V.(2013). The case for genetic monitoring of mice and ratsused in biomedical research. Mamm Genome. Vol. 24(3-4):89-94.5Fontaine, D.A. and Davis, D.B. (2016). Attention tobackground strain is essential for metabolic research:C57BL/6 and the International Knockout MouseConsortium. Diabetes. Vol. 65(1):25-33.6Gulati, A.S., Shanahan, M.T., Arthur, J.C., Grossniklaus,E., von Furstenberg, R.J., Kreuk, L. et al. (2012). Mousebackground strain profoundly influences Paneth cellfunction and intestinal microbial composition. PLOS One.Vol. 7(2):e32403.7Mahajan, V.S., Demissie, E., Mattoo, H., Viswanadham,V., Varki, A., Morris, R. et al. (2016). Striking immunephenotypes in gene-targeted mice are driven by a copy-number variant originating from a commercially availableC57BL/6 Strain. Cell Rep. Vol. 15(9):1901-9.8Brinkman, B.M., Becker, A., Ayiseh, R.B., Hildebrand, F.,Raes, J., Huys, G. and Vandenabeele, P. (2013). Gutmicrobiota affects sensitivity to acute DSS-induced colitisindependently of host genotype. Inflamm Bowel Dis. Vol.19:2560-2567.9Hansen, A.K., Hansen, C.H.F., Krych, L. and Nielsen, D.S.(2014). Impact of the gut microbiota on rodent models ofhuman disease. World J Gastroenterol. Vol. 20:17727-17736.10Hart, M.L., Ericsson, A.C. and Franklin, C.L. (2017).Differing complex microbiota alter disease severity of theIL-10-/-mouse model of inflammatory bowel disease.Front. Microbiol. Vol. 8:792.11Nielsen, D.S., Krych, L., Buschard, K., Hansen, C.H.F.and Hansen, A.K. (2014). Beyond genetics. Influence ofdietary factors and gut microbiota on type 1 diabetes.FEBS Lett. Vol. 588:4234-4243.12Ooi, J.H., Waddell, A., Lin, Y., Albert, I., Rust, L.T.,Holden, V. and Cantorna, M.T. (2014). Dominant effectsof the diet on the microbiome and the local and systemicimmune response in mice. PLOS One. Vol. 9:e86366.13Rune, I., Rolin, B., Larsen, C., Nielsen, D.S., Kanter, J.E.,Bornfeldt, K.E., Lykkesfeldt. J., Buschard, K., Kirk, R.K.,Christoffersen, B., Fels, J.J., Josefsen, K., Kihl, P. andHansen, A.K. (2016). Modulating the gut microbiotaimproves glucose tolerance, lipoprotein profile andatherosclerotic plaque development in ApoE-deficientmice. PLOS One. Vol. 11:e0146439.14Sasada, T., Hinoi, T., Saito, Y., Adachi, T., Takakura, Y.,Kawaguchi, Y., Sotomaru, Y., Sentani, K., Oue, N., Yasui,W. and Ohdan, H. (2015). Chlorinated water modulatesthe development of colorectal tumors with chromosomalinstability and gut microbiota in apc-deficient mice. PLOSOne. Vol. 10:e0132435.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 119

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121Haven’t the time to write a paper but want to get something published? Then read on!This section offers readers the opportunity to submit informal 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.Pro’s and pro’s of selective cleaningMARK WHITEUniversity College London, Biological Services, Kathleen Lonsdale Building, 5 Gower Place,London WC1E 6BSCorresponding author: m.white@ucl.ac.ukBased on a presentation at the 8th IAT West Middlesex Branch Technician Symposium 2019August 2019 Animal Technology and WelfareTECH-2-TECHIntroductionThe Kathleen Lonsdale Building (KLB), is a highspecification, high health status animal facility thatopened in 2011. It is a full IVC facility with an integralquarantine section. It houses transgenic (TG) mice,inbred, outbred and immuno-suppressed mice andoccasionally rats and hamsters.The reasons we decided to investigate includedimproving animal welfare with less disturbance to thecage through reduced cleaning and with a 60% increasein bedding, the ability for the mice to show some naturalbehaviours such as tunnelling, foraging and digging. Wealso wanted to work ‘leanly’, save the College money inproduct and labour time and create time for ourtechnicians to do other things and provide much neededplanned preventative maintenance on IVC racks andcages. Time did not allow this with the current systemof cleaning. Of course, we were aware that reducedcleaning may increase ammonia levels in the cage sowe assessed the environment of the IVC during thestudy with ammonia detectors to ensure cleaning tookplace at >25ppm but never exceeded >50ppm whichcould damage the nasopharynx regions of our mice.AimsIncreased animal wellbeing.– Mice do not enjoy a pristine environment.– Cleaning out male cages often causes aggression.– Disturbing new litters can increase pre weaningmortality.Creating time for new challenges.– New more detailed cost-recovery software.– Routine PPM of Total Cage Change.– IVC Rack PPM (spigots, plenums, etc.).– More forums, meetings and symposia.– Increasing workload.– Create time = less chance of mistakes.– Creating a ‘Leaner’ facility.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 121

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122Tech-2-TechCleaning regime prior to October2015– First week – FULL CLEAN WEEK (100%).– Second Week – PART-CLEAN WEEK (approximately40%).– Averaging 70% weekly.Pre-studyTrialled 4 different bedding products from 3 differentcompanies. Product of choice IPS Lignocel Select.**This was the best product and after a visit to thebedding plant in Germany I was assured of good quality(QA).** Important! Please note the findings of this studyare on Lignocel Select only. You will need to performyour own study if using a different bedding.Cleaning regime after October 2015– Monday – Thursday or Tuesday – Friday.– 3 day gap: 4 day gap. Technicians ask the question“Will the Cage go until then?”.Increase in amount of beddingWe wanted to encourage natural behaviours in our miceand decided to explore using more bedding toencourage this. We hypothesised that an increase inbedding would also improve absorbency of urine/Figure 1. Mouse and litter.Figures 2-3. Standard IVC cage used and bedding.Figure 4. Latrinearea has reachedtop of the bedding.Remember! Area of2 x 10p piecesconsistentlyshowed ammonialevels just as it ischanging frommedium (lightgreen) to high (darkgreen).faeces/ammonia. With bedding levels at 2.5cm thiswas now possible.OLD : 152.5g with a depth of 1.5cm.NEW : 244g WITH A DEPTH OF 2.5cm.= an increase of 60%.Initial concernsWould the cage flood? – No increase in cage floods.If they do occur, more of the water is absorbed ensuringanimals are not on soaked bedding.Study – November 2015 to January2016– Room 302 – 17% cleaned, Breeding/Stock,Technician MT.– Room 303 – 18% cleaned, Breeding/Stock,Technician MT.– Room 308 – 16% cleaned, Breeding/Stock/Experimental, Technician MT.– Room 309 – 29% cleaned, Breeding/Stock/Experimental, Technician PW.– Room 310 – 31% cleaned, Breeding/Stock,Technician MW.– Room 316 – 26% cleaned, Breeding/Stock/Experimental, Technician TJ.– Room 317 – 12% cleaned, Breeding/Stock/Experimental, Technician TC.Result assessmentBig percentage differences 12% to 31%. Is thisbecause of different stocking densities or the type ofwork?No! A subjective decision as everyone’s assessmentof a dirty cage was different. It was decided with theproof of the ammonia detectors that a cage is cleanedwhen:– Latrine areas that have reached the top up to thearea of two 10p pieces (still <10% of overall floordimensions).LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 122

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125Tech-2-TechDepth of bedding and sex of animals appear to affectammonia levels.We found with a bedding depth of 1.5cm ammonialevels can reach 25ppm in as little as 5 days. Thesame cage averaged 12 days on 2.5cm of bedding.Females = dispensing 152.5g with a depth of 1.5cm ina cage base.– Cage of 5 female adult mice averaged 5daysuntil ammonia level reached 25ppm.– The same cage averaged 12 days on 2.5cm.Males = dispensing 152.5g with a depth of 1.5cm in acage base.– Cage of 4 male adult mice averaged Only 6daysuntil ammonia level reached 25ppm.– The same cage averaged 10 days on 2.5cm.Dispensing 152.5g with a depth of 1.5cm in a cagebase.– Cage of 2 male adult mice averaged 20 daysuntil ammonia level reached 25ppm.– The same cage averaged 30 days on 2.5cm.RingtailConcerns regarding Ringtail? Only 1 cage of ringtail in20 months. We do not feel this is related to our study.Figure 19.Bedding andammoniasensorshowingeffect onammonialevels.Figure 20.Ringtail inyoung mouse(librarypicture).Other welfare benefits– Natural behaviour increased. Tunnelling andforaging.– Less cage changes = less stress = happy mice.– Reduced need to disturb females with new littersdue to soiled cages.Financial benefitAlthough the move to less frequent cleaning was notmotivated by financial gains, there were significantfinancial benefits.Bedding saving over 1 year of 22016 2017–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Jan 36 Jan 54–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Feb 54 Feb 36–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Mar 54 Mar 36–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Apr 36 Apr 36–––––––––––––––––––––––––––––––––––––––––––––––––––––––––May 36 May 18–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Jun 54 Jun 36–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Jul 36 Jul 18–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Aug 54 Aug 18–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Sep 54 Sep 54–––––––––––––––––––––––––––––––––––––––––––––––––––––––––TOTAL 414 TOTAL 306Further savingsReduced contract labour by 3 days a week leading to asaving of approximately £30,000 a year.ConclusionsNew regime has allowed– cleaning out reduced by approximately 66%– increased technician time– reduced bedding use by 26%– reduced capital equipment use by 40%– reduced ammonia levelsTime for new challenges– New more detailed software: LAMIS – LaboratoryAnimal Management Information System more datainput required than previous less detailed system.– Routine Planned Preventative Maintenance (PPM).– IVC PPM – spigots, plenums, full-cage, etc.– Time to attend more forums, meetings andsymposiums.– Ability to cope with units increasing workload.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 125

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Tech-2-Tech126– Create time = less chance of mistakes.– Created a more ‘Lean’ facility.– Technicians making a more intelligent and closerassessment of the cage environment results in lesstime spent in actual cleaning (lean!).– Technicians more aler t. No routine cleaning.Lessens boredom on full-clean weeks. Createsmore time (lean!).– Better for the mice: Less home cage changes.– Reduction in cross contamination – Increase in Bio-security!– Complete PPM on racks/cages. All rackcomponents including cages completely changedevery 3-4 months– 26% Less bedding used: 40% less chemicals used(lean!)– More storage space created (lean!) – Decreasedlabour time (lean!)– Reduction in use of equipment i.e. autoclaves, cage-wash (lean!)– Reduction in cleaning of up to 66% based on a fullclean/part clean.How was this achieved?This was just 5% of total cages changed duringChristmas 2018/New Year 2019.Just two small cleaning days where staff cleaned andwatered. Other days, checking and weaning. Similarly,for the first week of the New Year.Obviously during this time most IVC’s were opened forfeeding!AcknowledgementsKLB team – Pete, Toni, Mel, Tony for enthusiasticallyassisting with this study.Paul, Facility Manager for his support throughout thisstudy.IPS – For the quality of the bedding product and QA andplant visit in Germany.KLB PIL Holders for their support.Vet-Tech Solutions for importing the ammonia sensors.And most importantly the mice for not eating theammonia sensors and allowing us to play with theirenvironments.Figure 21. Did your cage wash area look as good or asempty as this after the Christmas break…? Thearrowed stacks are the only dirty cages changed! Just5%!!!Date Cleaned–––––––––––––––––––––––––––––––––––––––––––––––––––––––––W/C 10th Dec 2018 All 1 & 2’s (49%)–––––––––––––––––––––––––––––––––––––––––––––––––––––––––W/C 17th Dec 2018 All 3, 4, 5 & Br. (51%)–––––––––––––––––––––––––––––––––––––––––––––––––––––––––22nd Dec 2018 Selective cleans onlyto 1st Jan 2019 (5%)(11 days)Date Cleaned–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Back to work 02-01 to 6.5%04-01-19 (short 3 day week)–––––––––––––––––––––––––––––––––––––––––––––––––––––––––W/E 11-01-2019 13%(1st full week back afterChristmas)–––––––––––––––––––––––––––––––––––––––––––––––––––––––––W/E 18-01-2019 20%(2nd full week back afterChristmas)LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 126

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127Team awesome: Why we can be proudJOANNA KINGUniversity of Dundee, Medical School Resource Unit, Ninewells Hospital, Dundee DD1 9SYCorrespondence: j.w.king@dundee.ac.ukBased on a platform presentation at IAT Congress 2019IntroductionThe presentation I gave at IAT Congress 2019 wasbased on the changes and refinements the Universityof Dundee, Medical School Resource Unit (MSRU) hasmade over the past 18 months.New homes from oldThe first change we made was to the housing of ourGuinea pigs. The old cages (Figure 1) were small opentopped cages. Five animals were housed in thesecages, there was not much floor space for them to runaround and no room to add extra enrichment.One of our vacant rooms was used to create a floorpen for the animals using redundant materials lyingaround the unit, old sizzle nest boxes, tubes andboxes (Figure 2). The fronts of the old cages were usedto create a secure structure for the food hoppers andwater bottles. We housed 10 Guinea pigs in this roomand saw a difference in their behaviour almostimmediately: they were more inquisitive and wouldapproach technologists willingly apparently withoutfear and were easier to handle and appeared generallyhappier, we know this due to their persistent“popcorning” (“popcorning” – sudden and erraticjumping behaviour, resembles popcorn when cookingand is associated with excitement and happiness inthe Guinea pig), and happy ‘piggies’ equals happytechnologists.August 2019 Animal Technology and WelfareFigure 1. Shows old guinea pig cages.Figure 2. Shows new floor pen.Figure 3. Shows gridded floor time-mating cage.Refinement of breeding coloniesThe refinement and reduction of the breeding rats wassomething that we are particularly proud of, thischange made a huge impact on the lives of ouranimals.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 127

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Tech-2-Tech128The old system consisted of two separate colonies, onefor each researcher who used the rats. The femalesand males lived in separate cages, only comingtogether to mate. Figure 3 shows the timed mating gridboxes the animals were placed in. They could be inhere up to 7 days or until a plug had been found at thebottom of the cage.There were many problems with the old system:– Animals were singly housed.– Researchers refused to share one colony meaningthere was a huge surplus of pups.– Welfare issues with regards to the grid bottomedboxes.– Approximately 15 females breeding in each colony,with 3 males on rotation, 3 matings establishedeach week.– Animals were difficult to handle and aggressive.– Small litters and short breeding life.New breeding stock, both males and females, werepurchased and housed as monogamous pairs when itwas suitable. We did not charge the researchers forthis as we knew we had to first convince them it wasthe way forward and best course of action. Taking intoconsideration the date they were mated and also whenwe could see they were visually pregnant; we were ableto determine when the female would litter down withone to two days margin of error. This was satisfactoryfor the needs of the users. The litter sizes hadincreased and we found the animals in the pairs wereeasier to handle and friendlier.Although we had convinced the users to change themethod of breeding and to share one colony, weencountered a number of problems following thechange. The standard open top cages we use to houseour animals was not suitable for monogamous pairsand their large litters. At times there could be up to 21animals in the box meaning the cages needed to becleaned at least twice a week and there was a problemwith overcrowding and the females over-groomingthemselves, some had completely removed all hair ontheir upper body including their forelimbs.With the help of our ver y handy washroom supervisorJim, we were able to combine two standard cages usinga Standard red tube and drilling holes into the cages sothe tube can connect them (Figure 4).The animals now had a lot more space to move aroundthe cage and to care for their pups. The femalesstopped over-grooming, however despite being pairedat different time, all the females began to give birthwithin a day or two of one another, which was no use tous.It was then decided to remove the male before thefemale littered down in order to avoid post-partumoestrus with males being reintroduced later, to onceagain stagger parturition.We quickly learned that this was not ideal as the maleshowed aggression towards his pups and the femalemeaning that we could not place him back in his cageuntil all the pups had been weaned. This would haveresulted in our needing more breeders to compensatefor the gaps. To avoid this, we inserted a clear disc withholes drilled in the middle of the red tube connectingthe two boxes (Figure 5) which allowed us to separatethe male and female when needed but allow them tosmell and see each other whilst apart which in turnstopped the aggression.Eventually we arrived at a system which worked for usand the researchers. The animals were no longeraggressive or alone, they now have larger cages andare not being confined to the small timed-mating cages.The pairs are producing larger litters consistently andthe animals have better welfare, which of courseequals better science.Improved environmentalenrichmentAt the MSRU we are progressing and experimentingFigure 4. Shows boxes connected by tube.Figure 5. Shows Perspex insert for separation ofcages.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 128

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Tech-2-Tech129with enrichment. The code of practice provides nostandardisation, it strongly recommends enrichmentsuch as tunnels, boxes and climbing apparatus butdoes not enforce it. Which is why there are such largediscrepancies between units.The MSRU’s standard cage receives a house or tunnel,foraging diet, chew-sticks, sizzle nest and paper wool.The extra enrichment we provide includes tunnels orhouses on the roof, paper towels through the bars,aspen balls and cubes, swing or cable ties andnestlets. We add this mainly to reduce barbering andfighting between animals but we do provide it to otheranimals where and when we can.The cage in (Figure 6) shows a cage where the animalshave been heavily barbered: we placed almost every bitof extra enrichment we could in it. In this case theapproach gave good results, within a few weeks all themice had grown back their hair. However, not all cageswere as successful, in some cases the enrichmentmade no difference at all.Figure 6. An ‘enriched’ cage.Rotation of the enrichment seems to help, addingdifferent items each week to the cages at cage change.Once an item has been in the cage for a while itbecomes part of the standard cage, that is acceptableif the problem is fixed in a short amount of time. In thecase of fighting, this should stop in a very shortamount of time otherwise the animals must beseparated. Barbering it may take a little longer to seeany improvement so rotation of items in the cage isideal.Further investigation is needed to determine theeffectiveness of enrichment on barbering and fighting.Carrying out preference testing beforehand would be agood idea. As there is no point in adding enrichment tocages if the mice do not like it.Doing our bestOur goal at the MSRU is the same as all AnimalTechnologists, to make the lives of the animals underour care as painless and as comfortable as possiblewithin the scope of the scientific requirements. Animalsthat are kept for ageing purposes or in our case surplusLister hooded rats which we obtained and decided tokeep for training others in handling were given extraliving space. Using the same techniques as thebreeding rat cages we joined three cages with the redtubes. (Figure 7)Figure 7. Shows 3 combined cages.We came to realise through the efforts of other units inDundee that not all mice like the same enrichment.C57Blk/6 and Balb/c especially have very differentpreferences when it comes to swings and chew-sticksas well as the placement of their enrichment in thecages.We have also trialled new treats for the rats as we feelthe standard sunflower seeds provide little challengefor them, however, peanuts still in their shells (knownas monkey nuts) were a big hit (Figure 8) with them. Wehave started ordering them from our suppliers andadding them to the cages at cage change on a weeklybasis.Before we cannibalised the cages to give the rats morespace, we would place the rats in playpens for a fewhours a day, to give a little more exercise and room toplay. Now, the increased floor space in most of the rats’cages supersedes the playpen and we can add extraLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 129

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Changing our waysThe way we handle the animals is now changing and atthe MSRU all staff and users of the unit havesuccessfully moved from the traditional method of tailhandling to tubing and cupping. As with any change itwas met with some resistance but throughperseverance we prevailed. The initial change was timeconsuming for the technologists as neither they nor theanimals were used to the method of handling. Aftersome practice there was very little time differencewhen changing boxes when compared to the oldmethod. Once the animals were used to the newmethod, they too, were a lot more relaxed whenhandling, so much so that there was no need torestrain the tail when cupping (Figure 10). There is a lotmore interaction between the technologist and theanimals are now used to this method which is muchmore rewarding.Tech-2-Tech130enrichment, we mostly use them when we need toamalgamate groups of animals as playpens provide aneutral territory, a method which has proved extremelysuccessful. Again, the playpens were created by Jim,using materials we had lying around the unit (Figure 9)and some old toad tanks.Figure 9. Playpen made from old toad tank.Figure 10. Shows mouse calm and unrestrained.Figure 8. Rats enjoying ‘monkey’ nuts.As mentioned previously the rats receive two differenttypes of nesting material, paper wool and sizzle nest.Having learned from our Named Veterinar y Surgeon(NVS) that a recent paper highlighted that nestingmaterials used by the rats in the wild use coarsermaterials, such as sticks and foliage, we decided tointroduce some hay at cage change to see if they woulduse it in their nests. Most of the animals ate it, the onlyones to use the material in their nest were thebreeders. Therefore, we continued to only give the hayto the breeders at cage change.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 130

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Tech-2-Tech131Rehoming animalsAfter a behavioural study ended, we had 34 adolescentLister Hooded rats which were to be euthanised whichseemed inappropriate, we therefore made posters inthe unit asking if anyone would like to take them homeas pets. We received a lot of interest and eventually wewere able to rehome 30 of the animals, we kept theremaining 4 females for handling. The process took afew weeks, with Home Office approval and our NVShelping the recipients who did not have muchknowledge or experience with the rats. No one likeswasted animals, especially on this scale and we arevery proud of the fact that we rehomed 30 of the ratsand hear regularly from their owners that they are doingwell.Onwards and upwardsWe are a proactive unit, taking part in Tech Month,organised by the Institute of Animal Technology, earlierthis year where we had Tech Tea with all our users andran a raffle making over £200. We used this money forvarious things during Tech Month such as prizes for thetreasure hunt and animal trivia bingo for the users andtechnologists. We also had a little gathering in theseminar room where we had a presentation from aTecniplast representative who also sponsored a techdinner at a restaurant the same day.This year, we are also planning to carry out 2 studies:the first will be preference testing the enrichment andthe second will be, the effect enrichment has onaggression and barbering.We at the MSRU are most proud of our ability to changeand adapt. Change will never be easy and will never goto plan but the important thing is to not revert to oldhabits or to just be satisfied with the way things arebecause they have always been like that and it works.Just because it has worked for a long time does notmean there is not a better, more refined way to dothings. Whether it is changing handling methods,housing conditions or enrichment, change will alwaysbe difficult but with any problems which arise, solutionsmust be found. You must remember you are making thechange for a reason.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 131

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133NACWO my first year – dream job ornightmare?RHYS PERRYDepartment of JBIOS, Cardiff University, College of Biomedical and Life Sciences,College House, King George V Drive East, Heath Park, Cardiff CF14 4EPCorrespondence: r.perry4@exeter.ac.ukBased on a First Time Presenters Platform presentation at IAT Congress 2019IntroductionHello, I am Rhys Perry and I have been an AnimalTechnologist for the past 14 years, I am also thecurrent Named Animal Care and Welfare Officer(NACWO) for the Psychology Animal Facility at CardiffUniversity.I have been lucky enough to have been given theoppor tunity to work within several different facilitiesat Cardiff University over the years with differenthealth statuses and various caging systems includingconventional, isolators, IVCs and filter tops. I havealso worked with a mix of species during my time,genetically modified and wild type mice as well asrats, rabbits, Guinea pigs, tree shrews and cats.Reflecting on my career I believe it has been to myadvantage that whenever an opportunity has arisen towork with a different species, facilities or caging, Ihave accepted and looked to learn as much aspossible.As my career has developed I have continued myeducation by completing the IAT levels 2 and 3 andever y few years, when the opportunity has arisen Ihave attended the NACWO training course. These havehelped develop my knowledge of the job in hand andthe NACWO training course is a par ticularly brilliantcourse and I would highly recommend anyone, ifpossible, to take the oppor tunity to attend this.After a couple of years in the industry I felt I really hadsomething to offer and made a point of informing myline manager that I would like to be a NACWO. At thetime I was rather disappointed that the response tothis was that I need to get a few more years’experience under my belt before this would beconsidered. Looking back this could not have beentruer and having gone through my first year as aNACWO I would say that experience is absolutelyever ything and the more time you take to learnbeforehand, will benefit once you take that next step.I have always been the kind of person that has found itimportant to be happy and positive in work andespecially within this industry and the type of work thatwe perform. This naturally has led to some good,possibly daft times, for example when I once built atower from rat chew sticks from the floor to the ceiling(more than 6ft tall – big accomplishment)!Learning to handle peopleThis is not key to being approachable but for me I feelit has helped and that I have never lost my sense ofhumour and there must always be time to laugh andsmile, in what can at times be a tense and stressfulAugust 2019 Animal Technology and WelfareFigure 1. Learning key skills!LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 133

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Tech-2-Tech134environment. As a NACWO working with all ourlicensees with their own personality traits, the more Ibelieve it is critical that people feel they can approachus with any/all concerns when it comes to animalwelfare and have confidence in us, that we can helpensure that the right thing is done.One of my early challenges was when a colleaguedecided to not completely inform me of situation thatthey were aware of and a licensee opted to not take mysuggestion as seriously as they possibly should have. Iimagine I am not alone with situations like these but Ifound that it is important to take problems in yourstride, put together the correct response anddiscussed how I needed to be fully conversant with allour project licences and how we must work togetherwith licensees to ensure everything in the licence isdone correctly. Also that it is really important to fullyconvey all information regarding an animal’s conditionand that by not doing so they may potentially becausing an additional issue. This is a relationship thatmust be strong with trust and confidence working bothways and I found taking time to fully explain situationsmade a difference. I took almost an identical coursewith the licensee however, being a new NACWO I madethe decision to be firmer with my advice and thatalthough recommendations must be discussed to getto the best and correct outcome for both the animaland research, they must not be ignored and I made itclear I would not tolerate this in the future.Over the course of my first year it was always myintention to build strong positive relationships withproject and personal licence holders. It is my belief thatthis leads to building confidence in both parties andopens up very good lines of communication andultimately leads to improvement in welfare.Something that was mentioned to me on a fewoccasions is that I ‘could not be friends with everyone’,I understand this point of view but do not truly acceptit. While I would not say that each individual I work withis a friend, I would like to think there is a mutualrespect between all of these people. If we look at whatis required of the NACWO it may include having difficultconversations with the licensees and requiresindividuals informing the NACWO of all welfare relatedissues. This would be, I imagine, significantly moredifficult and less productive where there are poorrelationships and in my opinion would only improve asrelationships between these people become morepositive.Dealing with senior researchersAs a recent NACWO I found the thought of approachingexperienced project licence holders quite daunting,especially in the case of a ver y experienced PPL holderwho was also the head of research in one of the areasI took responsibility.I followed my decided path of being engaging andtaking the time to learn the individual’s research sothat I had some understanding of their requirements.This has led to a good relationship and the individualwas even happy to participate in a video I waspreparing to present at the IAT congress, outlining whatthey felt were important in the qualities of a NACWO,which turned out to be confidence and communication.They felt that having confidence that advice orrecommendations offered by a NACWO who hadknowledge of the work was important. All forms ofcommunication must be used in order to build thatpositive relationship, not just the occasional email. Idid initially feel it would be of benefit to arrangemeetings with all the PPL holders that would be workingin the area I would be responsible for in order tointroduce myself and my aims while in the position. Iquickly discovered that this can be difficult and what Ifound of more benefit was to wait until an issue orimprovement could be discussed regarding their workand that the individual was then more likely to take thetime to sit and discuss their work. The tunnel handlingimprovement project helped here as this affected thevast majority of licence holders.Introducing tunnel handlingDespite having been a senior technologist for 10 yearsprior to becoming a NACWO, there was always anelement of nervousness when dealing with PPL holdersand looking to push improvements I felt are important.I recall a ver y early conversation with a ver yexperienced PIL holder regarding tunnel handling inparticular where they had commented that they hadseen all the ‘fads’ come and go over the last 25 years.It can easily sound dismissive but this individual, I felt,certainly was not tr ying to be dismissive but they didhave a point that over the years some improvementscome in and become the norm however there arealways those that never last the distance.Personally these are the conversations I enjoy mostand I was quite keen to laugh this off and simply saidthat I could promise that this would not be a ‘fad’ andthat this was a necessary change that would be takingplace but that we would work hard together to ensure itfitted with individuals’ requirements. This seemed tohave the desired affect and so I realised that this reallyneeded to be the approach in the future, that workingtogether to find solutions is best. That simply insistingthat something is changing and the attitude of ‘toughluck’ is always going to put someone’s back up andmake for a difficult working relationship. In situationslike this, I was always worried at first I would saysomething stupid or ask a daft question and thatpeople perhaps may not then take me seriously. I thinkwhat I found is actually entirely the opposite. Peopleactually seemed to be a bit more comfortable speakingwith me and knowing that there was no judgement oneither side for saying something ‘a bit daft’. Stupid is aLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 134

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Tech-2-Tech135strong word and sometimes it is important thatconversations are of a lighter tone and for everyone tofeel comfortable being able to say what they think.As we are not scientists I imagine there will always besomeone who can sit down, start discussing their workand in doing so, completely baffle us and finish byasking if there are any welfare improvements we canrecommend. This happened to me relatively early in myrole as a NACWO. Someone was ver y enthusiastic tocome and speak with me and discuss the work theywould be carrying out and after 30 minutes I was notsure I had understood very much and I must admitwhen he finished speaking I think I actually laughed andsaid I was sorry but that I was quite new and that I hadnot understood much of what was said. I went awayand spoke with my colleagues and our NamedVeterinar y Surgeon (NVS) and we continued discussingthe work the researcher would be carrying out in morelay terms and now the whole team understands thework and this has benefitted everyone. However, itdemonstrated to me that it is really worth taking thetime to learn and understand and this is part of therequirements of the NACWO under The Animals(Scientific Procedures) Act 1986 (ASPA).ReflectionIf I could convey an important point I took from my firstyear, is that mistakes happen and I struggle to imaginea facility where they do not occur!We work hard as a team in Cardiff and we havepractices in place that minimise issues as much aspossible and catch mistakes as early as possible. Theimportant thing with these is to learn from them: to sitdown and assess what happened, why and what can wedo together to make sure it is not repeated andeveryone is happy moving forward. We are constantlylearning, constantly developing and so we need to takethe time for this to ensure that we learn from anymistakes.But it is not all about PPL holders and PIL holders,there are many named persons where it is crucial tobuild a good working relationship with. I think I havebeen lucky in that I walked into an establishment wherethere were regular meetings arranged and that issuesthat anyone might be having are regularly raised anddiscussed. This approach means any issues are lookedat by several people, who can all contribute to possibleapproaches and solutions. You are not always going toagree with each other but it is important that we alwaysget across what we feel is important, that we havemade sure we cover thoughts and processes so thateveryone can have a clear understanding of where weare coming from. No one person is always right and itis important to never get disheartened just becausesometimes other people disagree, we are all here towork together.This is also true when sitting on the Animal Welfare andEthics Review Board (AWERB). This is the opportunityto look over and discuss the work that people wish todo or have been doing at your establishment. We reallymust assess projects and ensure that the benefitsoutweigh the harms to animals and if you do not feelthis is true then it needs to be said. Again there arealways going to be different opinions and it may be thatthere is something that you missed or perhaps did notunderstand but all your concerns should always bediscussed. This is also an opportunity to look at theprotocols where you may notice improvements can bemade and it is such a great feeling when you cancontribute in this way. The very first time I noticedsomething that I felt could be improved I was terrifiedabout discussing it. However, I managed it and theperson took this on board and made the adjustmentwhich was wonderful! The whole time I felt like a rabbitin headlights. For some of us that will always be theway but as time goes on you become more experiencedand it gets better and you feel more comfortablediscussing your thoughts.ConclusionI will end this article by saying that there are so manywonderful people in this industr y and many have foundsometimes big, and sometimes little, ways of improvingnot only animal welfare but also technologist welfare. Ihave only in the last three years tapped into thisresource and it has brought so many improvements tothe way in which I work, I could not stress enough thatit is worth getting yourself out there and meeting the somany amazing people in this industr y! It is not aboutbeing the smartest person in the room but aboutworking together to improve what we can, when we can.This is a dream job for sure and like any job there aredifficult times that can knock you down but keep going,celebrate the positives and share your experiences!Figure 2. Still having fun.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 135

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137AbstractThe poultr y team at The Pirbright Institute reviewedexisting husbandr y practices and trialled severalrefinement practices. The objective was to provide thebirds used in research with an environment whichenables them to express their natural behaviours andhabituates them towards human contact, includingprocedures, with the potential to reduce cumulativesuffering. This is especially important for animals usedin research as stress is known to alter physiologicaland immunological responses, which could affect theresults from infectious disease research. This postersummarises refinement approaches, includingenrichment products used in the commercial poultryindustry and pet trade. Various items of enrichmenthave been introduced through obser ving naturalbehaviour and interest. This poster presentsrefinement approaches which have been subjectivelyassessed as being of benefit to the birds. Future workwill consist of implementing a more objectiveassessment of benefit of these refinementapproaches, such as quantitative behaviouralassessment.IntroductionIt is known that a barren environment can have anegative emotional impact on poultry welfare.1Inaddition, environmental enrichment can improveplumage and health conditions of laying hen pullets.2POSTER PRESENTATIONSOriginally presented at:IAT Congress 2019Refinement: promoting Gallus Galluswelfare in an experimental poultry unitKATIE HARRIS, RACHEL NEW, PAUL SMITH, BILLY MATTHEWSand JESS WANTThe Pirbright Institute, Ash Road, Pirbright, Woking GU24 0NFCorrespondence: katie.harris@pirbright.ac.ukWinner of the LBS Biotech Best Poster AwardAugust 2019 Animal Technology and WelfareFigures 1-3. Examples ofhuman interaction withchickens.Moreover, it has been demonstrated that broilers in amore varied environment have showed to be lessfear ful by reduced tonic mobility and had lowerbasophil levels when exposed to acute stressors.3Human interactionIn the hope to reduce stress, staff interact with thepoultry multiple times a day. This is achieved by:LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 137

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Poster Presentations138– Hand feeding enrichment to the birds.– In addition to routine husbandry practices, a radio isplayed and more time is spent in each room tohabituate birds to human voices.– Birds are handled daily as part of a new approachof collecting daily weights.Live foodGiving live food allows the birds to carry out naturalforaging behaviours, such as scratching and pecking.In past studies, live food rewards induced more of aninterest in quail after a negative stimuli had beenapplied.4Live food is now given after procedures to offerbirds distraction. Additionally, dried ‘live’ food can begiven autoclaved to promote good laboratory practice.Figure 4. Chicken enjoying live food enrichment.Figure 5. Silent Brown Cricket used as live food.(Gryllus assimilis).Figure 6. The Pirbright raised chicken pen.Figure 7. Chick wearing leg band.Raised floor pensThe raised floor pens at The Pirbright Institute (Figure6) allow a fully bedded environment to be provided.They also have an inbuilt ability to separate birds fromeach other when undertaking a variety of activitiesincluding regulated procedures and health checking.This allows birds to be given enrichment after regulatedprocedures as a form of positive reinforcement, as wellas, avoiding re-handling straight after regulatedprocedures.Leg bandingThe use of leg banding is being trialled instead of wingbanding for identifying the animals.Wing banding is an invasive process, which requires thepiercing of the skin of the wing with a permanent tag.– Initial results show that leg bands remain in placebut may need to be replaced as the bird growslarger – Alternative materials are being investigated.– Future work: to assess changes in weight gain ofwing banded vs leg banded birds.PerchesPerches have been provided for the birds in allenvironments in which they are housed at The PirbrightInstitute.Obser vations of bird perching behaviour havesuggested that there is a preference for a rectangularperch opposed to a circular one. This has also beenLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 138

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Poster Presentations139demonstrated in peer reviewed research whenexamining different perch shapes.5– A perch has been designed and improved which willfurther refine the bird’s environment.Sand bathingSand baths are provided as a standard enrichmentitem.A greater interaction with the sand has been observedwhen it has been dampened. Additions of smallamounts of food enrichment also appear to increasethe interaction with the sand/bathing behaviour.The futureFuture work will look to measure the impact of theseenrichment approaches using an objective, quantitativeset of parameters including measuring tonic immobilitywith or without different enrichment items, humancontact and enrichment items latency periods usingethograms and measuring stress hormones.References1Bassler, A.W.,C. Arnould, A. Butterworth, L. Colin, I.C.De Jong, V. Ferrante et al. (2013). “Potential risk factorsassociated with contact dermatitis, lameness, negativeemotional state, and fear of humans in broiler chickenflocks.” Poultry Science 92(11): 2811-2826.2Liebers, C.J., Schwarzer, A., Erhard, M., Schmidt, P. andLouton, H. (2019). “The influence of environmentalenrichment and stocking density on the plumage andhealth conditions of laying hen pullets.” Poultry Science.2019 Jun 1; 98(6):2474-24883Altan, Ozge, Seremet, Cigdem and Bayraktar, Özer.(2013). The effects of early environmental enrichment onperformance, fear and physiological responses to acutestress of broiler. Archiv fur Geflugelkunde. 1. 23-28.4Favreau-Peigné, A., Calandreau, L., Constantin, P.,Bertin, A., Arnould, C., Laurence, A. et al. (2016).Unpredictable and repeated negative stimuli increasedemotional reactivity in male quail. Applied AnimalBehaviour Science, 183, pp.86-94.5Duncan, E.T., Appleby, M.C. and Hughes, B.O. (1992).“Effect of Perches in Laying Cages on Welfare andProduction of Hens.” British Poultry Science 33(1): 25-35.Figure 8. Chicken using perch.Figure 9. Perch design.Figure 10. Immature chickens using sand bath.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 139

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140IntroductionAs Animal Technologists we are constantly looking atrefining and enriching the environmental conditions ofall our animals to give them both physical exercise andmental stimulation regardless of their size. However,animals can still experience stress when being movedfrom their home cage.The tunnelling and cupping methods have been widelyadvertised as positive alternatives for mouse handlingbut can be met with hesitation when used within highercontainment levels.The following information describes how weimplemented the tunnelling method within positivepressure isolators and what happened next.Animal handling in containmentCHRISTINE ZVEREV, SHAUN BAKER and DHRUTI PATELNational Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms,Potters Bar, Hertfordshire EN6 3QGCorrespondence: christine.zverrev@nibsc.orgAnimal Technology and Welfare August 2019Figure 1. Restraining mice using the tail handlingmethod.Materials and methodsUsing a clear Polycarbonate tunnel, we first introducedthe tunnel handling method as a small-scale trial withinan open shelf stock room. Over a period of months, theanimals were acclimatised to the new routine and theirbehaviour was compared between the new method oftunnelling mice and the existing method of handling bytheir tail.In the trial, we looked at time taken, aggression (ifany), behaviour and if there was any obviouspreference.– The time was unsurprisingly longer but not by toomuch. It was approximated to take 15 secondslonger per cage while the user is acclimatising tothe tunnelling method and then the time graduallydecreases as the handler becomes moreaccustomed to the technique.– Our transgenic mice (TG) are generally unaggressiveand this remained the case throughout the trial. Thetunnels were added to the cage as additional cagefurniture and although the males were choosing touse the tunnel as their toilet area, no additionalaggression or fighting over the tunnel was observed.– As a comparison for behaviour. It was noticed thatthe tailed animals were faster at seeking cover thanthose that were tunnelled, suggesting that thetunnelled mice were more likely to explore the newsurroundings. This, for most of the animals, mirrorsthe findings from studies held at LiverpoolUniversity.– Also, the tunnelled mice were not observed todisplay any ‘startle responses’ like that of the MoroReflex.– This appears to suggest that the likelihood was thatthese mice were less stressed and possiblypreferred the tunnel or that they were indifferent ofthe handling technique.Cupping these mice seemed to be equally as effectiveas tunnelling and this method appeared to be easierand less stressful for young mice around weaning ageFigure 2. Restraining mice using the tunnel method.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 140

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Poster Presentations141which are more likely to bolt through the tunnel than sitin it.However, in some cases, cupping can prove to be ahigher bite risk from more aggressive strains, forexample the B6 strain.Progression into containment froma technologist’s perspectiveShortly after the stock rooms had started their tunneltrial, I was working with the positive pressure isolatorshousing immunocompromised mouse models.While watching them scatter in all different directions,I put myself into the mouse’s ‘shoes’ and quicklydecided that I would not like being grabbed by amassive hand either!Figure 3. A possible ‘mouse eye’ view of handling!Consequently, the decision to add the tunnel seemedobvious even if it did appear to take a little longer.Surprisingly, I found that on many occasions the tunnelwas far quicker and easier to handle the mice than byhand. Most mice appear happy to wander in withoutmuch or, any encouragement at all.The mouse can be gently ushered into the tunnel byholding the tunnel against the side of the cage and theanimal can be easily encouraged to enter. In manyoccasions the mouse will walk freely into the tube.Once inside the tunnel, the mouse is free tocomfortably move the length of the tube allowing thetechnologist to easily view the mouse through the sidesand at all angles for obvious signs of illness or injurythat may be initially hidden through naturalpredator/prey responses from tail handling.When needed, the mouse can simply be slid out ontothe hand to then be scuffed or for a more thoroughhealth check before being placed into the new cage.Figure 4. Mouse entering tunnel.Figure 5. Mouse being slid onto technologist’s hand forcloser inspection.Figure 6. Mouse removed from tunnel.This method can easily be used pre-procedure andhelps to reduce stress while keeping the integrity of anycontainment level.Within the isolators, I found the tunnelling methodbenefits both the technologist and the animal by beingquicker, easier, presents a truer representation of theanimal’s health and reduces stress for the mice.ConclusionsIt has become clear that in our adult mice the tunnel orcup method is greatly dependent on the nature of theanimal or strain; some strains are commonly moreinquisitive and lively which make them more likely toventure into the tunnel with minimal or noencouragement. In contrast, slightly more relaxedLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 141

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Poster Presentations142strains are less forward in entering the tunnel, in thisscenario the cup method is better suited. It should bethe temperament of the individual animal thatdetermines the way in which we handle the animalregardless of containment level.When working with immunocompromised and/or highercontainment mice, we needed to pay attention todisinfecting the tunnels both before use and after thetest ends. These tunnels were treated with a stabilisedhydrogen peroxide chemical before use and did nothave any damage or clouding at all. However, withregular washing the clarity could potentially deteriorateand the tunnel may need replacing.When disinfecting from a higher containment level afterthe test concludes, autoclaving is usually performedfollowed by the cage wash process and again, nochange to the integrity of the tunnel was observedduring our decontamination cycle at 121°C.When in use as permanent cage furniture, tunnels mayget chewed at the edges but this does not inhibit theiruse in any way.AcknowledgementsThe research that inspired this trial and implementationof refined handling methods was carried out byProfessor Jane Hurst and Dr Kelly Gouveia of theUniversity of Liverpool. The trial was aided by thepublication of their work over several years and theirresults and conclusions being published on the NC3Rs’website along with all the informational tutorial videosthat enabled this trial.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 142

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143IntroductionIn-vivo imaging is a revolutionary technology offering apowerful tool for constructing studies involving mice. Inthe Biological Ser vices Department (BSD) at NIBSC,the IVIS®Spectrum In Vivo System model is used toperform our studies. This instrument is supplied byPerkinElmer and provides our scientists with valuabledata.Imaging has a wide range of capabilities, at NIBSC wehave used IVIS for:G tracking viral vectors and antibodiesG tumour growth monitoringThe Spectrum model in the BSD division has thecapability to work with bioluminescence.In order to create bioluminescence for the type ofstudies we perform, suitably genetically modified cellsof interest are required, particularly oxidized enzymesfor example luciferase and a reporter substrate e.g.luciferin (this can be found in organisms such as thefirefly). When these are combined, they emitbioluminescence which can be detected by IVIS.Using the in vivo imaging systemFirstly, a well-plate should be used with differentstrengths of cells together with the added luciferase.This allows the user to determine the optimalparameters preventing any potential waste of mice.When you are ready, you apply the luciferin into thedesired well and then place into the imager. Luciferinoriginally comes in powder form and made up to thesupplier’s instructions. Being light sensitive, it shouldalways be stored in the dark and when transported, itneeds to be wrapped up with aluminium foil to preventexposure to light.The standard screenshot the software provides can beseen (Figure 1). This is provided in two layers, theoriginal photo at the back and on the top layer, thefeedback on the location where it was received. TheIn-vivo imaging at NIBSC and how we useit to promote the 3RsLUKE GURNEYNational Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms,Potters Bar, Hertfordshire EN6 3QGCorrespondence: luke.gurney@nibsc.orgAugust 2019 Animal Technology and WelfareFigure 1. Screen shot from software.strength is taken from strongest to weakest and thenchanged into the colour that reflects this. The exampleabove shows that red is the strong signal and purple isthe weaker. From an image like this, we get a good ideaof the substances abilities before commencing in vivowork.The miceWhen selecting mice, it is important to take their coatinto consideration. Imaging mice with black fur requiresa little more preparation as the fur can block the signal.The signal can be improved by shaving the area whichyou are expecting to see signal. The best choice wouldbe either nude or white strains as the instrument canimage these without any problem.Working with IVIS has provided us with a greaterknowledge as to the identifying the mouse.– Nude mice – ear tags can be used together withpermanent marker pen on the tail.– White furred mice – staining pens (Red and Blue)are used employing our own marking system. Eartags have been used however, there were a fewproblems af fecting the ears which requiredveterinary attention.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 143

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3Rs (Replacement, Reduction,Refinement)Recovery consists of placing the mice in their homecage on a heated mat with a wet mash to provide foodand water and a towel over the cage for darkness. Miceare monitored regularly and when they appear fullyrecovered, they are returned to the home cage.The aims of the project were to replace the animalswhere possible, reduce the numbers when needed andrefine the impact on the animals.– These non-invasive technologies allow us to usesmaller group sizes of animals, as there is no needto cull at set points to observe hidden tumourgrowth.– Imaging has allowed us to refine endpoints. Thiswas evident in a lung tumour model whichdeveloped in an unexpected location. Thanks to IVISwe could intervene before it became a welfareissue.– During imaging, mice are under anaesthesiapreventing them from moving, minimising time andimproving the data.– Prior to anaesthesia we give them Viscotears,reducing eye irritation.– One needle per mouse is used for the i.p. injectionof luciferin so there is no risk of discomfort fromusing a blunt needle. The subcutaneous (s.c.) routeis being considered for future work. This is a lessinvasive technique and therefore provides furtherrefinement.– There is potential to connect heart and respiratorymonitors inside the instrument, enabling mice to bemonitored during the procedure.– The studies have allowed us to gain fur therknowledge on effectively identifying the mice,minimising stress and reducing negative effects onthe mice.ConclusionsUsing imaging technologies has created a great tool forboth the scientist and the Animal Technologist. Thetechnology:– Provides the scientist with valuable data which ismore sensitive and robust.– Animal Technologists are enabled to monitordisease development non-invasively. The dataallows for effective health and welfare assessment,pinpointing the location and the strength of signal.AcknowledgementsAdvanced Therapies – Dr K. Zaki, S. Satkunanathan,TDI – Dr R. FrancisBSD divisionNC3Rs – Dr A. HolmesPoster Presentations144Black furred mice – we used an in-house ear notchsystem and tail marking with permanent marker. Eartags were tried with these mice; however, they tend topull them off, occasionally resulting in injury.Figure 2. Examples of mice used.The procedureAll work involving mice is per formed under theprevailing principals and authority of the Animals(Scientific Procedures) Act 1986.Luciferin dose is weight dependant therefore the firststep is to weigh the mice so that the dose of luciferincan be calculated. The dose of luciferin is administeredvia the intraperitoneal route (i.p.). Enough syringes toprovide one per mouse are required, as this will enableyou to fill the syringe with the correct dose and canthen be discarded (single use of needle). We found thatinsulin syringes were the most accurate as theincrements are in single units.The animals are required to be anesthetised as theinstrument is very sensitive and the body movementscan affect the quality of the images. There are twoseparate anaesthetic points, one anaesthetic inductionchamber and then 5 nose cones inside the machineitself.When the mice are inside the instrument, they areplaced into the head cones and laid upon a heatedstage. The luciferin/luciferase reaction takes 10-15minutes to it reach its optimum range. When thishappens, you can close the door making the box lighttight. The software is then used to take the desiredimage, following which, the mice will be ready forrecovery.Figure 3. Images of tumours.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 144

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145Abstract1. Animal unit users can be exposed to high levels oflaboratory animal allergens (LAAs).2. Allergens such as airborne mouse and rat urinaryproteins (MUP/RUP) can lead to sensitisation, longterm health issues and an inability to work withcertain species.3. Photocatalytic Oxidation (PCO) systems have beendeveloped to oxidise LAAs rendering them safe toinhale without becoming sensitised.4. As PCO systems work on the room levels they maybe a valuable alternative to PPE in high risk areas.Aims and objectiveAim – determine whether PCO systems can be used toreduce airborne animal allergen contaminants in a highrisk area.Objective – run a three-week trial of a photocatalyticoxidation system measuring the amount of airborneMUP and RUP.Methods– The PCO system (Figure 1) was turned off duringweeks one and three and switched on during weektwo.– Static LAA sampling was conducted by running aknown quantity of air through filters capturing theairborne MUP/RUP.– These samples were taken from the dirty side ofthe University of Manchester’s animal unit cagewash between 9:00 and 15:00 on weekdays forthree consecutive weeks.FindingsAn investigation of how a photocatalyticoxidation system can be used to reduceairborne allergensJOSEPH PENNOCK and HAYLEY BUCKLEBiological Services Facility, University of Manchester, Stopford Building, Oxford Road,Manchester M13 9PTCorrespondence: atweditor@iat.org.ukAugust 2019 Animal Technology and WelfareAverage urinary protein (ngUP/m3)MouseRatWeek 1 (Off)10.961.88Week 2 (On)1.20.24Week 3 (Off)4.541.36Table 1. Average nanograms of MUP/RUP in a m3of airduring each week of the trial.Figure 1. Contaminated air is drawn in and passedover a matrix of TiO2(Titanium dioxide) and ZrO2(Zirconium dioxide) whilst being irradiated by UltraViolet C lamps. The bound organic material is oxidisedresulting in CO2/ H2O as the only by-product.Graph 1. Urinary protein levels present each day.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 145

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147AbstractFor the past few years, as reported in the yearly HomeOf fice summar y for the UK, a consistent non-compliance is the failure to provide food and water.This poster will look at the importance of a healthyanimal for research and the implications otherwise toscience, legislative and ethical concerns. I will suggestways to address this issue by analysing commonpractices in animal units and concerns due to thedesign of caging. Finally, I will propose possibleimprovements to prevent future non-compliance andallow easier detection of food or water, includinglooking at innovative new equipment.Importance of healthy animalsWhy do we need healthy animals?Failure to provide food and water:implications and preventionMURAD MIAHThe Francis Crick Institute, 1 Brill Place, London NW1 1BFCorrespondence: murad.miah@crick.ac.ukAugust 2019 Animal Technology and Welfare– In order to achieve reliable scientific findings.– Compromised health and functionality may lead toinconclusive and unreliable results and affectreproducibility.´In the absence of food and water, mice release high levels of corticosterone (a hormone triggered in the presence of stress). Mice fasted for more than 16 hours had higher concentrations of corticosterone than a 15 minute exposure to a rat D SUHGDWRU LQ D FDJHµ -HQVHQ HW DO Both the Animals (Scientific Procedures) Act 1986Amendment Regulations 2012 and accompanying Codeof Practice for the Housing and Care of Animals Bred,Supplied or Used for Scientific Purposes requireanimals to be provided with food and water.Figure 1. Categories of non-compliance, by type, 2015-2017.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 147

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Poster Presentations148IAT advice note: the provision ofwater and food for laboratoryanimalsIAT Guidance on the provision of Water and Food forLaboratory Animals. Available from the Institute ofAnimal Technology at www.iat.org.uk (under media/documents).Root cause = the effectiveness of routine checks ofthe animals; both checking for the lack of provision andthe declining condition of animalsMajor risk = establishment of new cagesRecommends1. SOPs covering the provision of food and water andchecking for provision (daily health check).2. Staff induction process – impor tance andresponsibility of providing food and water.3. Highlight any newly set up cages with colouredcards to ensure they get an extra check.4. Consider using different people for morning andafternoon checks wherever possible.5. Spot check audits by a senior technologist.6. Physical touch check of bottles or functioning ofautomatic watering systems.Current practices – what is doneand the flawsCage design – good and bad pointsCage design has the potential to drastically reduce non-compliance by making the food and water in every cagemore visible. Designs from Tecniplast’s individuallyventilated cages (IVC) will be used to illustrate thepotential benefits and implications that may come withit.Cage design with rear diet hopperFigure 2. Cage with rear diet hopper. (Credit Tecniplast)– Water bottle slot at the front allowing for easy waterlevel detection.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 148

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Poster Presentations149– Hopper is at the back of the cage – it may bepossible to overlook the diet level or miscalculatethe amount present.– This design allows for better observation of theanimals.Cage design with front diet hopperFigure 3. Cage with front diet hopper. (Credit Tecniplast)– Both the water bottle and hopper are at the front ofthe cage – allowing for easier detection of food andwater levels.– Observation of the animals is more difficult.Proposed improvements – methodsof work and technologyConclusionHealthy animals are a fundamental requirement forsuccessful experiments. For the full functionality of ananimal it must have access to adequate and suitablefood and water.1. It is an ethical and legal requirement that everyanimal in any establishment have access to food andwater unless there is a programme of work statingotherwise. Failing to do so may result in a review andpotential revoking of a licence by the Home Office.2. Standard practices, including SOPs, should be inplace to stop this non-compliance. If they areadhered to, the risk of non-compliance isminimised. It should be noted that most non-compliance cases are accidental, due to humanerror rather than gross negligence.3. Systems, including the automated watering system,reduce the risk of non-compliance.4. Cage design can have a large impact on the ease ofchecking diet and water levels. This is generally atrade off as cages with diet in a more visible areagenerally restrict the view of the animals. This couldlead to a greater non-compliance – not identifyingpoor health or illness in an animal.5. Technology can be used to support animalhusbandr y – for example Tecniplast’s DigitalVentilated Cage (DVC). A simple yet effective way isto make different components of the cagetransparent to allow easy visibility of water and foodlevels – for example Innovive’s plastic food holder.My suggestions for future technology that may make apositive contribution:– Dials similar to car fuel tanks, on bottles indicatingthe level of water present, especially for thosebottles that are not fully transparent.– Cages to have a mechanism that weighs the dietcontent and has a similar kind of dial to alert thetechnician instantly of low levels of food.Finally, animals in research are ever dependant onthe technologists and researchers for all theirrequirements. Failing to do so only leads to pooranimal welfare and a possible compromise of anyscientific research carried out, potentiallyinvalidating the entire study.Figure 4. Plastic hopper. (Credit Innovive)Proposed improvements Details–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Methods of Improved through training ofwork technologists and researchers,including refresher training onbasic animal husbandry.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Animal checks Issues may be missed due todone by different complacency by the technologist,technologists especially during weekend checks.A fresh pair of eyes may spotthese issues.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Highlight new New cages from weaning,cages experiments or deliveries should behighlighted to place moreimportance on checking diet/waterlevels.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Improved rack Allows cages to slide out fordesign improved viewing whilst minimisinguser’s arm strain.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Improved cage Innovative development in rack anddesign cage design now incorporate infra-red sensors to detect food levels.Electronic notifications are sent onanything running low. Could lead toover-reliance on technology.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Improved hopper Innovive manufacture plasticdesign hoppers. Metal hoppers may givethe technologist a false perceptionof diet levels.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 149

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150AbstractFood and water intake of laboratory animals is oftenmeasured in various research fields such as behaviour,diabetes and obesity. The housing environment canaffect these parameters.1In this study, we aimed toinvestigate the effect of relative humidity (RH) on waterintake of C57BL/6J mice housed at RH controlledsteadily at cage level (55±3%), when compared to micehoused in cages with RH controlled at room level in aless stable fashion.MethodWe controlled the RH of one rack using the ScanClime(Scanbur, Denmark) at three different levels (45%, 55%and 65%); each level was tested for one month andcompared to a rack with RH controlled at room level.Over the course of the experiment, daily RH readings ofthe holding room, air handling units (AHUs) and insidethe cages were recorded. Two groups of C57BL/6J witha mix of female and male were compared (N=35) and2-4 animals were housed in each cage. All micemaintained under the ScanClime were firstacclimatised to the unit for two months. Water intakemeasurements were taken weekly and measured as anaverage per mouse in each cage.All animals were housed in IVC caging on aspenbedding with nesting material, cardboard tunnel andchew sticks that had been autoclaved. They were fedstandard rodent chow ad libitum and housed on a 12hday/night cycle. Both groups of animals were housed inthe same room but were on different racks withdifferent AHUs.Due to biosecurity restrictions the cages were cleanedon a Monday between 13.00-15.00 and to avoid doublehandling, the animals and water bottles were weighedduring this husbandry routine. This also reduced theaccidental loss of water through excess moving ofcages. To work out the water intake the bottles wereweighed and the following Monday they were weighedagain; the difference was then divided by the number ofanimals in the cage to find the average intake peranimal.The effect of relative humidity on waterintake in C57BL/6J miceREBECCA TOWNS1and KAREN EKKELUND PETERSEN21Biological Services, University College London, Gower Street, London2SCANBUR A/S, Silovej 16-18, 2690 Karlslunde, DenmarkCorrespondence:Animal Technology and Welfare August 2019Figure 1. Mice drinking from water bottle.ResultsIn Graphs 1-6 the results of the three tests of waterintake and RH can be seen. In Graph 1 it is clear thatthere is a significant difference in water consumptionbetween the two groups of mice with the group housedat a steady humidity drinking significantly less. FromGraph 2 it is evident that the room humidity fluctuatedduring the course of the test, sometimes dropping aslow as 32%, whereas the ScanClime maintained asteady humidity throughout.In the group that were housed at 55% humidity (Graphs3 and 4) it appears that the graphs of room-controlledand ScanClime-controlled RH are similar, as the roomhumidity did not fluctuate as often as in the other twomonths. In Graph 3 there is a significant difference inLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:19 Page 150

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Poster Presentations152DiscussionThe animals in this study were of different ages andsexes, although comparative sexes and ages were usedin each group where feasible. It is possible that thisvariety in ages could have caused some variability in theamount of water consumed, due to different bodyweights; however, rather than specifically ordering inanimals to complete this study, we used C57BL/6Janimals that were already available in the unit, whichfurther helped satisfy the reduction principle of the 3Rs.In most cases the same technician completed theweighing, but this was not always possible due tobiosecurity restrictions and technician availability. Thiscould possibly have influenced some results due to adifference in handling techniques. In addition, it ispossible that different results may have been observedif different strains were used or if the animals werebeing maintained on an alternative diet e.g. highprotein or high fat diet.1Although effect on water consumption was the mainpoint of this experiment, humidity can also have otherimpacts in mice. It has been shown in studies thatexposure to low humidity has an effect on tearsecretion, goblet cell density, and occurrence of dryeye-related ocular sur face signs.2Such impacts may be of particular relevance in long-term experiments, particularly those in the dementiaresearch field. Low humidity also has significanteffects on dehydration, which in turn can cause largephysiological changes in mice.33Rs going forward…Water restriction studies involve reducing wateravailability to either a specific daily ration, or toprovision for only a specified period of time each day.Most studies use the water restriction to induce amotivational stimulus to per form a behavioural task.With the results collected in this study, it ishypothesised that mice being used for behaviourresearch that are on water deprivation or restrictioncould benefit from being housed at a steady humidity.If an appropriate RH is chosen, it may be possible torestrict mice for shorter periods of time, or to allowaccess to larger volumes of water, without any negativeimpact on task performance.Future work will hopefully look at collaborating withvarious behavioural research groups, to investigatetask performance of animals housed at different RHand on various types of water restriction; thiscollaboration should facilitate development ofrefinements to this procedure.In addition, the increasing use of aged mice inexperiments in the dementia field means it may provebeneficial to explore the apparent increase in eyeissues that may be caused from long term housing invariable RH.4ConclusionThe results shown here suggest that RH cansignificantly affect the water intake of C57BL/6J mice.When tightly controlling RH, especially within the higherend of the regulatory requirements (65%), the micedrink significantly less than when the animals arehoused under room-controlled RH.To ensure accurate data results and reproducibility ofstudies, it would potentially be of great value to steadilycontrol the RH in the microenvironment, therebyminimising this otherwise uncontrolled variable instudies where water intake is measured, or where thirstis used as a motivator for behavioural studies.AcknowledgementsScanbur (Denmark) for supplying the ScanClime AHU,Technical staff at the UCL Central BSU for assistancewith data collection.References1Nicolaus, M.L. et al. (2016). Effect of Ventilated Cagingon Water Intake and Loss in 4 Strains of Laboratory Mice.American Association for Laborator y Animal Science.2Pelegrino, F.S.A. et al. (2011). Low humidityenvironmental challenge causes barrier disruption andcornification of the mouse corneal epithelium.Experimental Eye Research p150-1563Bekkovold, C.M. et al. (2013). Dehydration Parametersand Standards for Laboratory Mice. American Associationfor Laboratory Animal Science.4Suhalim, J.L. et al. (2014). Effect of Desiccating Stresson Mouse Meibomian Gland Function. The Ocular Surface12 (1)LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:20 Page 152

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153IntroductionThe Laboratory Animal Technology department atEnvigo were engaged to perform a toxicity study injuvenile Göttingen Minipigs. The test compound was ahuman milk formula additive. Therefore, to mimic theconditions of clinical administration, the dose wasgiven in commercial porcine milk replacer. The pigletswere removed from the dam on the third day after birth.Piglets were fed every three hours, and dose wasincluded in the feed at every second feeding occasion.The study was divided into a dose range findingpreliminary study and a main toxicology study.AccommodationA farrowing pen was designed to provide space, safetyand security for the sow and piglets, plus a creep areafor additional warmth and seclusion for the piglets. Thepen was retrofitted around existing minipig enclosures.Key components included anti-crush bars around theperimeter, Perspex on bars to prevent piglets escapingand fitting a hatch to an adjacent pen to create a creeparea.FarrowingIn preparation for farrowing and three hourly feeding, 24-hour on-site care was arranged for the duration of study.At least three technologists were always present, plus aveterinary surgeon on call to provide advice and attendif required. All sows farrowed with no, or minimal,intervention. All piglets were given a post-natal ironsupplement, as part of standard porcine husbandry.Preliminary studyDosing via bottleDosing on the preliminary study was performed viabottle. The dose was mixed into a small portion of thefood ration of milk replacer and fed to the piglet. If thiswas not readily accepted by bottle, the dose could begiven by syringe, or oral gavage. Once the full dose wasconsumed, more milk was then offered to appetite.After a few days, most piglets accepted bottle feeding.However, in the first few days of dosing, most pigletslost weight and subsequent growth was not as fast asexpected. Piglets often seemed erratic with feeding –drinking lots at one feed, then very little at next feed.Several piglets showed signs of possible milkaspiration. Therefore, a new method of dosing had tobe considered for the main study.Bottle or bowl? Feeding and dosing injuvenile Göttingen minipigsALICE McNAMARA and LUCY ALLENEnvigo, Woolley Road, Alconbury, Huntingdon, Cambridgeshire PE28 4HSCorrespondence: lucy.allen@envigo.comAugust 2019 Animal Technology and WelfareFigure 1. Farrowing pen and pregnant sow.Figure 2. Bottle feeding piglets.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:20 Page 153

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Poster Presentations154SparesDue to high piglet survival, the final litter of piglets inthe preliminary phase were not required on study.These animals were reared as stock animals and thenallocated to other studies, in line with the principles ofthe 3Rs. These Piglets were also used to assess otherpossible methods of feeding (and therefore dosing).Our animals took readily to pan feeding. However, theyoften swapped bowls and climbed into the milk. It wasdifficult to assess how much each piglet was drinkingand to prevent the piglets applying the milk topically!Figure 3. Sharing a meal.Graph 1. Growth of bottle versus pan fed piglets.Figure 4. Piglet feeding stall.Different methods of separating the piglets weretrialled. It was discovered that piglets found itdistressing to be completely separated from eachother, even for the short feeding period. The piglets fedbest when they were still in contact with their littermates.Main studyDosing via panFor the main study, a special designed stall wascreated to allow the piglets to be separated for panfeeding, while still allowing visual and tactile contactwith their cohort. A small bowl that attached to thestall, minimised the risk of the dose and milk beingspilled or applied topically.The stalls were kept in the piglet accommodation at alltimes to allow them to habituate to the equipment. ThePerspex back to the stall could be removed betweenfeeds to allow the piglets access to the enclosure.Piglets took a similar amount of time to adjust to panfeeding compared to bottle feeding. As before, thedose was mixed into a small portion of the milkreplacer. This was then offered in the bowl in the stall.If this was not fully consumed, the dose could be givenby syringe, or oral gavage. Once the full dose wasgiven, the remainder of the milk ration was given. In thepan fed cohort, the piglets did not lose weight afterbeing removed from the dam. The piglets continued togain weight more than the bottle-fed piglets. No casesof milk aspiration were seen. Feeding appeared moreconsistent in the pan-fed animals.ConclusionsThe above graph shows the growth of the bottle-fedpiglets compared to the pan-fed piglets. The growth isshown as the mean percentage change in body weight,compared to the initial weight (taken within 24 hours ofbirth). The low dose group in both the primary and mainstudy have been used for comparison (dose levels wereidentical between both groups). As can be seen, thebottle-fed animals initially lost weight compared to theirbirth weight. However, the pan-fed animals never fellbelow their birth weight. The rate of growth in thebottle-fed animals was slower that the pan-fed animals.Pan feeding was also significantly less demanding ofstaff time once the piglets had adapted to using thepan system. Therefore, based on this data, pan feedingis recommended as a method of oral dosing juvenileminipigs, and may be superior to bottle feeding.LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:20 Page 154

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155BackgroundThe production of transgenic animals is a team effortwithin The University of Edinburgh. Bioresearch andVeterinar y Services Technologists support the CentralTransgenic Core (CTC) in transgenic production/rederivation and cr yopreser vation ser vices fornumerous research groups.Female mice of breeding age are set up with males withthe expectation that 20-30% of them will plug. Femaletransgenic mice are also set up in this way so thatembryos can be collected at specific stages forscientific purposes (Figure 2).SummaryUsing oestrus selection, the females are mated at theirmost fertile stage. The oestrus cycle has 4 stageswhich lasts 4-5 days. By detecting oestrus efficientlywe can reduce the number of mice required. Therebyimproving the pregnancy rate after embryo transfer asall females used for embryo transfer will definitely bepseudo-pregnant. Therefore, the females are morereceptive, consequently refining animal welfare.ObjectiveTo reduce the number of mice used and increase thenumber of embryos available for transgenic productionand study requirements.Refined methodBy oestrus selecting females before mating we canreduce the number of females used and improve thenumber of plugged females for both use as pseudo-Refining oestrus selection in the animalunit to optimise transgenic productionCHRISTOPHER WILSON, MIKE WALLS, DAVIE BLACKand JULIE THOMSONUniversity of Edinburgh, Bioresearch and Veterinary Services (BVS), Hugh Robson Building,George Square, Edinburgh EH8 9XDCorrespondence: chris.wilson@ed.ac.ukAugust 2019 Animal Technology and WelfareFigure 1. Stages of oestrus.Figure 2. Numbers of female mice not oestrus selected.Figure 3. Number of females oestrus selected.Traditional methodLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:20 Page 155

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157Subjects considered for publication may include originalarticles, technical notes and reviews pertaining to allaspects of animal science and technology, managementand education. The Editorial Board wishes to offerparticular encouragement to papers leading toimprovements in environmental enrichment, the generalcare and welfare of the animals used, in particular thosespecies and strains exhibiting harmful genetic defects,and papers describing refinements in techniques, areduction in the number of animals that need to be usedor alternatives to animal use.Papers describing experimental procedures will only beaccepted for publication if authors clearly state that theprocedures conform to the prevailing principles andCodes of Practice of the Animals (Scientific Procedures)Act, 1986. Papers submitted from outside the U.K.,should state what legislation and/or ethical approvalthe work has been carried out under. In addition, authorswho describe surgical techniques with recovery shouldinclude details of post-operative care and any analgesictherapy provided. All submissions should follow theARRIVE (Animal Research: Repor ting of In VivoExperiments) guidelines (Kilkenny C, Browne WJ, CuthillIC, Emerson M, Altman DG (2010) Improving BioscienceResearch Reporting: The ARRIVE Guidelines forReporting Animal Research. PLOS Biol 8(6): e1000412.doi:10.1371/journal.pbio.1000412)The Editorial Board reser ves the right to seekindependent advice on any aspect of the content of anarticle but the final decision on acceptance or rejectionremains with the Board.SubmissionMaterial submitted for publication will be consideredprovided that it is contributed exclusively to AnimalTechnology and becomes the property of the Institute ofAnimal Technology.Articles may be submitted either electronically or by hardcopy as follows:ElectronicArticles should be submitted in Word format with doublespacing to the lines and all pages serially numbered.Any photographs or graphs must be submitted asseparate files and conform to the format in point 4)below.The relevant ar ticle must clearly indicate wherephotographs and/or graphs are to be inserted.Address for submission: atw@iat.org.ukHard copyThe original manuscript plus two copies should be sentto the address below together with a copy on disk (CD orDVD).All sheets should be typewritten on one side in doublespacing and serially numbered.Any photographs or graphs should be supplied asoriginals and conform to the format in 4) below.Address for submission: Journal Editorial BoardChairman, 5 South Parade, Summertown, Oxford OX27JL.No responsibility will be accepted for loss or damage tosuch articles.Electronic files of submissions are required together withseparate files of photographs and any graphics thatappear in the manuscript. Electronic submissions shouldbe sent via email via atw.iat.org.ukalternatively,manuscript plus two copies may be sent as hard copy tothe address below. All sheets should be typewritten onone side in double spacing with 4 cm margins andserially numbered. Additionally, a copy on disk should beprovided or sent by email via atw@iat.org.ukArticles for submission should be sent to: JournalEditorial Board Chairman, 5 South Parade, Summertown,Oxford, OX2 7JL.No responsibility will be accepted for loss or damage tosuch articles.Format1). The first sheet of the article should contain thefollowing:i. the full title of the paperii. the initials and last name of the author(s)iii. the full address of the depar tment(s) andinstitution(s) where the work was carried out.iv. the address for correspondence if different to above.2). For the remainder of the paper, the text should beclear and concise and, where appropriate, sub-dividedunder the following headings:i. Summaryii. Introductioniii. Methodsiv. Resultsv. Discussionvi. Acknowledgementsvii. References3). Measurements should be given in metric units – seeThe use of S.I. Units (1969) British Standards Institutionpublication and spelling should follow that of the OxfordInstructions to AuthorsAnimal Technology and Welfare August 2019LATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:20 Page 157

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158English Dictionary. Abbreviations must be defined in fullat their first appearance in the text. The 24 hour clockshould be used for times. Words to appear in italic typeshould be underlined. Designation of inbred strainsshould be in accordance with the International Index ofLaboratory Animals, 6th edition, compiled, edited andpublished by M.W. Festing, 1993.4). Photographs should have clear and well contrastedtone values and be in colour. All illustrations, charts (e.g.histograms and graphs) and photographs should besubmitted separately and bear on the reverse side theauthor’s name, a number corresponding to the order inwhich it appears in the text e.g., Figure 1, and an arrowpointing to the top.Illustrations, charts and photographs supplied on diskshould be in JPEG, TIFF or EPS formats and have aresolution of no less than 300dpi.The captions for illustrations, charts and photographsshould be typed in double spacing in numerical order ona separate sheet of paper.5). References: Only essential references should beincluded. Authors are responsible for verifying themagainst the original source material. ATW uses theVancouver referencing system: references should beidentified in the text by superscript Arabic numberse.g.12after any punctuation and numbered and listed atthe end of the paper in the order of when they are firstcited in the text. Automatic numbering should be avoided.References should include the names and initials of upto six authors. If there are more than six authors, only thefirst three should be named, followed by et al.Publications for which no author is apparent may beattributed to the organisation from which they originate.Simply omit the name of the author for anonymousjournal articles – avoid using ‘Anonymous’.References should be set out as follows:Journals:- Surname and initials of author(s) (date), title ofarticle. Name of journal in full, volume number, first andlast page numbers.e.g. Saigeman, S. (1998). Environmental enhancementof cats – what? why? how? Animal Technology, Vol 49,No.3, 145-154.Books:- Surname and initials of author(s) (date), title ofbook. Name of publisher, Town of publisher.e.g. Flecknell, P.A. (1987). Laboratory AnimalAnaesthesia. Academic Press, London.Chapter from a multi-author book:- Surname and initialsof chapter author(s) (date), title of chapter. In: title ofbook (surname and initials of book editors). Name ofpublisher, Town of publisher, first and last page numbersof chapter.e.g. Gregory, J.A. (1985). Principles of AnimalHusbandry. In: Laboratory Animals – An Introduction forExperimenters. Second Edition. (Tuffrey, A.A.). John Wiley& Sons Ltd., Chichester, 87-105.Papers accepted for publication but not yet publishedshould be included in the list of references followed by‘(in press)’. Papers in preparation, personalcommunications and unpublished observations shouldbe referred to as such in the text only.ContentPapers describing procedures involving the use ofanimals should always include full details of the animalsand husbandry conditions used. These would be asfollows:AnimalsSpeciesBreed or strainSexAge and weight at start of procedureGenetic status: inbred; outbred; hybrid; mutantSourceMicrobiological status: conventional; specifiedpathogen free (define which pathogens animals arefree from); gnotobiotic (define which micro-organismsare present)Quarantine or acclimatisation periodHusbandry during procedureType of housing: material; size; cage type if relevantNumber of animals per cage or unitBedding: type; quality; any pretreatmentType of system: conventional; barrier; ventilated rack;isolatorEnvironmental temperature (°C ± range)Relative Humidity (% ± range)Lighting: natural; artificial (state hours of light anddark)Ventilation: number of air changes per hourPeriod of acclimatisation before start of procedureFeed: type; composition; any pretreatment; amount;frequencyWater: type; quality; any pretreatment; amount;frequencyScientific procedureNumber of animals and any pretreatmentTime of day of procedure(s)Quantity and frequency of any samplesStatisticsTests used should be namedReprintsFree reprints are no longer provided but the ATW EditorialBoard are happy to provide PDF files of articles afterpublication. Use of these files is subject to Copyrightrestrictions.Instructions to AuthorsLATEST_6-648255753.e$S:Animal Technology and Welfare 6/8/19 07:20 Page 158

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INDEX TO ADVERTISERSINDEX TO ADVERTISERS August 2019Allentown Inc ........................................................................................................OBCAST2020 ...........................................................................................................xii, xiiiAvid (Labtrac) ............................................................................................................viBell Isolation Systems Ltd..........................................................................................ivDatesand Ltd ..........................................................................................................IFCInstitute of Animal Technology .......................................................................xv, xvi, xviiIPS Product Supplies Ltd.........................................................................................IBCLBS...........................................................................................................................vPFI Systems Ltd .......................................................................................................viiSomni Scientific.......................................................................................................viiiSurrey Diagnostics Ltd...............................................................................................xiTecniplast UK Ltd................................................................................................x, xviiiTransnetyx Inc ...........................................................................................................iiiVet-Tech Solutions Ltd ..............................................................................................xiv