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Vol 22 No 3 December 2023ISSN 2752-3918Offi cial Journal of the Institute of Animal Technology and European Federation of Animal TechnologistsIAT JournalAnimal Technology and Welfare●Congress 2023 posters ● Rat tickling ● 3Rs implementation by review and regulatory approval

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167IAT Education, graduation special Steven CubittPOSTERSAn assessment of rat hammocks as enrichment Megan Hickman, Lorraine Miller and Ruth MacDonaldMouse behaviour core at the Francis Crick Institute Lydia Colgate and Nadia UnalCooped up: refi nement of a Gallus gallus domesticus environment Billy Matthews, Rachel New, Yamini Mahesh Kanse, Jess Want, Sophie Bentley, Michael Collett, Victoria Chantler and Paul SmithThe Zebrafi sh health and welfare database, where we are now Nicola Goodwin, Rosemary Keeble and Michael PriceThe 3Hs – home, hammocks and happiness Sylvie Mehigan and John HobbsVol 22 No 3 December 2023Editorial Diane Hazlehurst, Chair of the Editorial BoardThe role of the review and regulatory approvals processes for animal research in supporting implementation of the 3Rs Frances RawleABSTRACT TRANSLATIONSTECH-2-TECH An exploratory interview study of researchers and technicians’ perception of rat tickling Sam Beechener, Sarah Brown, Vincent Bombail, Megan LaFollette, Ignacio Vinuela-Fernandez and Alistair LawrenceEditorialJas Barley, Chair of the Editorial BoardReport of the 2019 RSPCA/UFAW RodentWelfare Group meetingChloe Stevens, Emily Finnegan, Jasmine Clarkson,Charlotte Burns, Sonia Bains, Colin Gilbert,Caroline Chadwick, Samantha Izzard, Charlotte Inman,Penny Hawkins (Secretary) and Huw GolledgeReduction of the negative effects ofmethionine on bone parameters in broilers’embryos by intra-egg injection of Vitamin B12Mohammad Naser Nazem, Shima Tasharofi,Negin Amiri and Sepideh SabzekarThe care of the Childr en’s Python(Antaresia children)Alexander Hosking and Gary MartinicFeline-assisted therapy: a promising part of animal assisted therapy (AAT)Eliska Mičková and Krityna MachovaThe care of Central and Pygmy Bearded DragonsAlexander Hosking and Gary MartinicPAPER SUMMARY TRANSLATIONSFrench, German, Italian, SpanishLOOKING BACKPhysical hazards in the laboratory animal houseR.T. CharlesThe incidence of a pathogenic strain of pseudomonas in a rabbit colonyG.R. Alpen and K. MaerzTECH-2-TECHDevelopment of a sifting cage change method for rats to improve welfareSeonagh HendersonVol 1 9 No 2 A ugust 2020CONTENTSiAugust20:Animal Technology and Welfare 4/8/20 10:48 Page i201224233228175173221231211189

vOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richard Gardner MA PhD FRSBFIAT (Hon) FRSVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS,Stephen Barnett BA MSc FIAT (Hon) CBiol FRSBRAnTech, Miles Carroll PhD, Paul Flecknell MA Vet MBPhD DLAS DipLECVA MRCVS FIAT (Hon), PennyHawkins PhD BSc, Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.) DipECLAMFRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&SMSc MRCVS, Tim Morris BVetMed PhD DipACLAMDipECLAM CBiol FRSB CertLAS MRCVS, Clive PageOBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky RobinsonCBE BSc PhD, Paul Sanders MIAT RAnTech, DavidSpillane FIAT, Gail Thompson RLATG, RobertWeichbrod PhD RLATGLife MembersKen Applebee OBE FIAT CBiol FRSB RAnTech,Charlie Chambers MIAT RAnTech, Roger Francis MScFIAT RAnTech, Pete Gerson MSc FIAT RAnTech,Cathy Godfrey FIAT RAnTech, John Gregor y BSc (Hons)FIAT CBiol FRSB RAnTech, Patrick Hayes FIAT DipBARAnTech, Robert Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon)RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Sarah Lane MSc FIAT,Sue McHugh BSc FIAT, Norman Mortell BA (Hons)MIAT RAnTech, Wendy Steel BSc (Hons) FIATMembers of CouncilMatthew Bilton, Kally Booth, Steven Cubitt,Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Alan Graham, Linda Horan, Sam Jameson,Elaine Kirkum, Adele Kitching, Theresa Langford,Sylvie Mehigan, Steve Owen, Alan Palmer, AllanThornhill, John Waters, Lynda Westall, Carole Wilson,Adrian WoodhouseCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary:Simon Cumming BSc FIAT RAnTechHonorary Treasurer: Glyn Fisher FIAT RAnTechChair of Board of Educational Policy:Steven Cubitt MSc FIAT RAnTechChair Registration & Accreditation Board:Glyn Fisher FIAT RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATATW/Bulletin Editorial Board:IAT REPRESENTATIVESAugust20:Animal Technology and Welfare 4/2/21 13:19 Page vvOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richar d Gardner MA PhD FRSBFIAT (Hon) FRSVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS,Stephen Barnett BA MSc FIAT (Hon) CBiol FRSBRAnTech, Miles Carroll PhD, Paul Flecknell MA Vet MBPhD DLAS DipLECVA MRCVS FIAT (Hon), PennyHawkins PhD BSc, Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.) DipECLAMFRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&SMSc MRCVS, Tim Morris BVetMed PhD DipACLAMDipECLAM CBiol FRSB CertLAS MRCVS, Clive PageOBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky RobinsonCBE BSc PhD, Paul Sanders MIAT RAnTech, DavidSpillane FIAT, Gail Thompson RLATG, RobertWeichbrod PhD RLATGLife MembersKen Applebee OBE FIAT CBiol FRSB RAnTech,Charlie Chambers MIAT RAnTech, Roger Francis MScFIAT RAnTech, Pete Gerson MSc FIAT RAnTech,Cathy Godfrey FIAT RAnTech, John Gregor y BSc (Hons)FIAT CBiol FRSB RAnTech, Patrick Hayes FIAT DipBARAnTech, Robert Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon)RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Sarah Lane MSc FIAT,Sue McHugh BSc FIAT, Norman Mortell BA (Hons)MIAT RAnTech, Wendy Steel BSc (Hons) FIATMembers of CouncilMatthew Bilton, Kally Booth, Steven Cubitt,Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Alan Graham, Linda Horan, Sam Jameson,Elaine Kirkum, Adele Kitching, Theresa Langford,Sylvie Mehigan, Steve Owen, Alan Palmer, AllanThornhill, John Waters, Lynda Westall, Carole Wilson,Adrian WoodhouseCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary:Simon Cumming BSc FIAT RAnTechHonorary Treasurer: Glyn Fisher FIAT RAnTechChair of Board of Educational Policy:Steven Cubitt MSc FIAT RAnTechChair Registration & Accreditation Board:Glyn Fisher FIAT RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATATW/Bulletin Editorial Board:IAT REPRESENTATIVESAugust20:Animal Technology and Welfare 4/2/21 13:19 Page vRefining the welfare of immunocompromised mice receiving carbon tetrachloride (CCl4) to induce liver fibrosis Vicky Walczak-Gillies, Olivia Matthews, Alex Fyfe, Anna Hoy and Phillip Starkey LewisThe hard hitting tooth – does a schedule teeth check reduce the number of mice found dead with malocclusion? Jessica CiupaExploratory view into abdominal fat necrosis in mice Melissa HewsonThe benefit of study plans to facility management Floyd Laniyan, Claire Pearce, Jolene Hammonds, Mike Williams, Lynsey Cashnella, Bruno Correia da Silva, Jayne Morgan, Nadine Lebrasse, Enriqueta Navarro, Thomas Pitcher, Andrew Whatcott and Julie KeebleTo wheel or not to wheel Penny Ives and Jade CaygillContainment of Mycobacterium marium within a containment level 2 aquatics area Nicola Goodwin237POSTER PRESENTATIONSAssessing pain in models of Rheumatoid ArthritisSamuel Singleton, Meriam Nefla, Ngaire Dennison, Simon Arthur and Tim HalesRefinements to health monitoringHannah Jones and Rebecca KingBiosecurity risks and the pre-implantation embryo; lessons from the mouseJean Cozzi, Mendy Verrier and Jimmy MancipEnvironmental enrichment for a small colony of ratsNick Blackburn, Gemma Cronshaw and Mike MitchellOestr us checking – increasing productivity and embracing the 3RsSamantha Hoskins and Jack BrownUsing habituation to reduce str ess for rats being transported short distancesSarah TaylorShining a light on rearing pigmentless ZebrafishJacqueline Glover, Thom Berriman, Dimitra Mantzorou, William Havelange,Sam Berry and Bruno Correia da SilvaThe jacket with pulling power – a novel approach to early stage evaluationof magnetic nanoparticlesAlison Ritchie, James Dixon, Phil Clarke and Anna GrabowskaiiCONTENTSIndex to AdvertisersABPI ..................................................................x,xi LBS ..................................................................iiAS-ET ...............................................................OBC Somni Scientific ................................................ivDatesand Ltd......................................................IFC Special Diets Ser vices .....................................viiiInstitute of Animal Technology ...............................vii Tecniplast UK Ltd .............................................xiiIPS Product Supplies Ltd.....................................IBCAugust20:Animal Technology and Welfare 12/8/20 07:54 Page ii240242244248246ATW PROFILEATW aims to be the medium for Animal Technologists and all those concerned with the care and welfare of animals used for research purposes to communicate ‘good practice’.ATW especially aims to promote and develop the 3Rs particularly in respect of Refinement. More importantly, ATW promotes the generally accepted 4th R, Responsibility. The responsibility that all Animal Technologists have in ensuring dissemination of ‘good practice’ to every institution using animals in research.ATW enjoys a unique position as the scientific publication for the leading organisation for the welfare of animals used in research.ATW seeks to publish peer-reviewed articles, technical notes and reviews allied to animal science and technology, management and education. Particular encouragement is given to authors submitting papers leading to improvements in environmental enrichment and the care and welfare of genetically altered animals.A commemorative plaque, a cheque for £250 and free registration at the IAT Congress awaits the winner of the Marjorie (Sandiford) Whittingham Memorial Prize awarded annually to the best original article.Editorial– Peer-reviewed papers– Translations of paper summaries into 4 European languages– Tech-2-Tech informal short articles on l new or refined techniques l discussion forum reports l commercial submissions welcome (specific equipment advertorials will not be accepted)– Posters from international meetings– Book reviews– Letters to the editorMember StatesEFAT comprises of Member States of the European Union and Council of Europe https://www.efat.org/AFSTAL Association Francaise des Sciences et Techniques de L’Animal de Laboratoire DALAS Dutch Association for Laboratory Animal ScienceSECAL Sociedad Española para las Ciencias del Animal de LaboratorioATWAnimal Technology and Welfarewww.atwjournal.com

169vOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richar d Gardner MA PhD FRSBFIAT (Hon) FRSVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS,Stephen Barnett BA MSc FIAT (Hon) CBiol FRSBRAnTech, Miles Carroll PhD, Paul Flecknell MA Vet MBPhD DLAS DipLECVA MRCVS FIAT (Hon), PennyHawkins PhD BSc, Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.) DipECLAMFRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&SMSc MRCVS, Tim Morris BVetMed PhD DipACLAMDipECLAM CBiol FRSB CertLAS MRCVS, Clive PageOBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky RobinsonCBE BSc PhD, Paul Sanders MIAT RAnTech, DavidSpillane FIAT, Gail Thompson RLATG, RobertWeichbrod PhD RLATGLife MembersKen Applebee OBE FIAT CBiol FRSB RAnTech,Charlie Chambers MIAT RAnTech, Roger Francis MScFIAT RAnTech, Pete Gerson MSc FIAT RAnTech,Cathy Godfrey FIAT RAnTech, John Gregor y BSc (Hons)FIAT CBiol FRSB RAnTech, Patrick Hayes FIAT DipBARAnTech, Robert Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon)RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Sarah Lane MSc FIAT,Sue McHugh BSc FIAT, Norman Mortell BA (Hons)MIAT RAnTech, Wendy Steel BSc (Hons) FIATMembers of CouncilMatthew Bilton, Kally Booth, Steven Cubitt,Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Alan Graham, Linda Horan, Sam Jameson,Elaine Kirkum, Adele Kitching, Theresa Langford,Sylvie Mehigan, Steve Owen, Alan Palmer, AllanThornhill, John Waters, Lynda Westall, Carole Wilson,Adrian WoodhouseCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary:Simon Cumming BSc FIAT RAnTechHonorary Treasurer: Glyn Fisher FIAT RAnTechChair of Board of Educational Policy:Steven Cubitt MSc FIAT RAnTechChair Registration & Accreditation Board:Glyn Fisher FIAT RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATATW/Bulletin Editorial Board:IAT REPRESENTATIVESAugust20:Animal Technology and Welfare 4/2/21 13:19 Page vvOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richar d Gardner MA PhD FRSBFIAT (Hon) FRSVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS,Stephen Barnett BA MSc FIAT (Hon) CBiol FRSBRAnTech, Miles Carroll PhD, Paul Flecknell MA Vet MBPhD DLAS DipLECVA MRCVS FIAT (Hon), PennyHawkins PhD BSc, Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.) DipECLAMFRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&SMSc MRCVS, Tim Morris BVetMed PhD DipACLAMDipECLAM CBiol FRSB CertLAS MRCVS, Clive PageOBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky RobinsonCBE BSc PhD, Paul Sanders MIAT RAnTech, DavidSpillane FIAT, Gail Thompson RLATG, RobertWeichbrod PhD RLATGLife MembersKen Applebee OBE FIAT CBiol FRSB RAnTech,Charlie Chambers MIAT RAnTech, Roger Francis MScFIAT RAnTech, Pete Gerson MSc FIAT RAnTech,Cathy Godfrey FIAT RAnTech, John Gregor y BSc (Hons)FIAT CBiol FRSB RAnTech, Patrick Hayes FIAT DipBARAnTech, Robert Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon)RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Sarah Lane MSc FIAT,Sue McHugh BSc FIAT, Norman Mortell BA (Hons)MIAT RAnTech, Wendy Steel BSc (Hons) FIATMembers of CouncilMatthew Bilton, Kally Booth, Steven Cubitt,Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Alan Graham, Linda Horan, Sam Jameson,Elaine Kirkum, Adele Kitching, Theresa Langford,Sylvie Mehigan, Steve Owen, Alan Palmer, AllanThornhill, John Waters, Lynda Westall, Carole Wilson,Adrian WoodhouseCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary:Simon Cumming BSc FIAT RAnTechHonorary Treasurer: Glyn Fisher FIAT RAnTechChair of Board of Educational Policy:Steven Cubitt MSc FIAT RAnTechChair Registration & Accreditation Board:Glyn Fisher FIAT RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATATW/Bulletin Editorial Board:IAT REPRESENTATIVESAugust20:Animal Technology and Welfare 4/2/21 13:19 Page vCouncil OfficersChair: Linda Horan BSc (Hons) MIAT RAnTechVice Chair: Glyn Fisher FIAT RAnTechHonorary Secretary: Simon Cumming BSc FIAT RAnTechHonorary Treasurer: Sam Jameson MIAT RAnTechChair of Board of Educational Policy: Robin Labesse MIAT RAnTechChair Registration & Accreditation Board: Ken Applebee OBE FIAT CBiol FRSB RAnTech ATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATATW/Bulletin Editorial Board: Diane Hazlehurst MIAT RAnTech (Chair), Jas Barley, Patrick Hayes, Elaine Kirkum, Carole Wilson, Lynda WestallBranch Liaison Officer: Lynda Westall FIAT RAnTechEFAT Representatives:Glyn Fisher, Robin Labesse, Toby SandersWebsite Coordinator: Allan Thornhill FIAT RAnTechWebsite Support: Sam JamesonAnimal Welfare Group:Carmen Abela (Chair), Kally Booth, Diane Hazlehurst, Sam Jameson, Sylvie Mehigan, Steve Owen, John WatersBoard of Educational Policy:Robin Labesse (Chair), Adele Kitching (Secretary), Steven Cubitt (Deputy Chair), Diane Hazlehurst, Theresa Langford, Tina O’MahoneyCommunications Group:Adrian Woodhouse (Chair) Carmen Abela, Kally Booth, Hannah Easter, Sam Jameson, Wendy Jarrett, Elaine Kirkum, Theresa Langford, Sylvie Mehigan, Toby Sanders, Allan Thornhill, Lynda WestallVice-PresidentsSenga Allan MIAT RAnTech, David Anderson MRCVS, Stephen Barnett BA MSc FIAT (Hon) CBiol FRSB RAnTech, Miles Carroll PhD, Penny Hawkins PhD BSc, Wendy Jarret MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.) DipACLAM DipELAM CBiol FRSB CertLAS MRCVS, Clive Page OBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky Robinson CBE BSc PhD, David Spillane FIAT, Gail Thompson RLATG, Robert Weichbrod PhD RLATGLife MembersKen Applebee OBE FIAT CBiol FRSB RAnTech, Charlie 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 Hayes FIAT DipBA RAnTech, Robert Kemp FIAT (Hon), Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon) RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Stuart Mackrell FIAT RAnTech, Wendy Steel BSc (Hons) FIAT Members of CouncilCarmen Abela, Ken Applebee, Kally Booth, Steven Cubitt, Simon Cumming, Haley Daniels, Glyn Fisher, Alan Graham, Diane Hazlehurst, Linda Horan, Sam Jameson, Elaine Kirkum, Adele Kitching, Robin Labesse, Theresa Langford, Sylvie Mehigan, Tina O’Mahony, Claire Pearce, Toby Sanders, Allan Thornhill, John Waters, Lynda Westall, Carole Wilson, Adrian Woodhouse

170BRANCH SECRETARIES 2022Cambridge: Tony Davidge cambridgebranch@iat.org.ukEdinburgh: Kery-Anne Lavin-Thomson edinburghbranch@iat.org.ukHuntingdon, Suffolk & Norfolk: Jo Martin hssbranch@iat.org.ukIreland: Lisa Watson irelandbranch@iat.org.ukLondon: Rebecca Towns londonbranch@iat.org.ukMidlands: Ian Fielding midlandsbranch@iat.org.ukNorth East England: Zoe Smith and John Bland northeastbranch@iat.org.ukNorth West: Nicky Windows cheshirebranch@iat.org.ukOxford: Adam Truby oxfordbranch@iat.org.ukSurrey, Hampshire & Sussex: Francesca Whitmore shsbranch@iat.org.ukWest Middlesex: Josefine Woodley westmiddxbranch@iat.org.ukWales & West: Rhys Perry waleswestbranch@iat.org.ukWest of Scotland: Joanne King westscotlandbranch@iat.org.ukIAT OFFICERS M AY BECONTACTED VIA:IAT Administrator:admin@iat.org.ukOR VIA THE IAT WEBSITE AT :www.iat.org.ukOR THE REGISTERED OFFICE:5 South Parade, Summertown,Oxford OX2 7JLAdvertisement Managers:PRC Associates LtdEmail: mail@prcassoc.co.ukAlthough every effort is made to ensure that no inaccurate or misleading data, opinion or statement appear in thejournal, the Institute of Animal Technology wish to expound that the data and opinions appearing in the articles,poster presentations and advertisements in ATW are the responsibility of the contributor and advertiser concerned.Accordingly the IAT, Editor and their agents, accept no liability whatsoever for the consequences of any suchinaccurate or misleading data, opinion, statement or advertisement being published. Furthermore the opinionsexpressed in the journal do not necessarily reflect those of the Editor or the Institute of Animal Technology.© 2023 Institute of Animal TechnologyAll rights reserved. No part of this publication may be reproduced without permission from the publisher.CPD Officer: Alan Palmer MIAT RAnTechRegistration and Accreditation Board:Glyn Fisher (Chair), John Gregor y,Cathy Godfrey, Kathy Ryder (Home Office),Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee:Alan Graham (Chair), Haley Daniels, Adele Kitching,Allan Thornhill, John WatersDiversity Officer:Haley Daniels MBA MSc MIAT RAnTech CIPDUK Biosciences ASG Representative/Home Office:Alan Palmer MIAT RAnTechviAugust20:Animal Technology and Welfare 12/8/20 07:54 Page viRegistration and Accreditation Board:Ken Applebee (Chair), Glyn Fisher (Secretary), Charlie Chambers, John Gregory, Cathy Godfrey, Kathy Ryder, Stuart Stevenson Observer: Ngaire Dennison (LAVA)Congress Committee:Alan Graham (Chair), Haley Daniels, Adele Kitching,Claire Pearce, Allan ThornhillEquity, Diversity and Inclusion Offi cer:Haley Daniels MBA MSc MIAT RAnTech CIPDIndex to AdvertisersBRANCH SECRETARIES 2022Cambridge: Tony Davidge cambridgebranch@iat.org.ukEdinburgh: Kery-Anne Lavin-Thomson edinburghbranch@iat.org.ukHuntingdon, Suffolk & Norfolk: Jo Martin hssbranch@iat.org.ukIreland: Lisa Watson irelandbranch@iat.org.ukLondon: Rebecca Towns londonbranch@iat.org.ukMidlands: Ian Fielding midlandsbranch@iat.org.ukNorth East England: Zoe Smith and John Bland northeastbranch@iat.org.ukNorth West: Nicky Windows cheshirebranch@iat.org.ukOxford: Adam Truby oxfordbranch@iat.org.ukSurrey, Hampshire & Sussex: Francesca Whitmore shsbranch@iat.org.ukWest Middlesex: Josefine Woodley westmiddxbranch@iat.org.ukWales & West: Rhys Perry waleswestbranch@iat.org.ukWest of Scotland: Joanne King westscotlandbranch@iat.org.ukIAT OFFICERS M AY BECONTACTED VIA:IAT Administrator:admin@iat.org.ukOR VIA THE IAT WEBSITE AT :www.iat.org.ukOR THE REGISTERED OFFICE:5 South Parade, Summertown,Oxford OX2 7JLAdvertisement Managers:PRC Associates LtdEmail: mail@prcassoc.co.ukAlthough every effort is made to ensure that no inaccurate or misleading data, opinion or statement appear in thejournal, the Institute of Animal Technology wish to expound that the data and opinions appearing in the articles,poster presentations and advertisements in ATW are the responsibility of the contributor and advertiser concerned.Accordingly the IAT, Editor and their agents, accept no liability whatsoever for the consequences of any suchinaccurate or misleading data, opinion, statement or advertisement being published. Furthermore the opinionsexpressed in the journal do not necessarily reflect those of the Editor or the Institute of Animal Technology.© 2023 Institute of Animal TechnologyAll rights reserved. No part of this publication may be reproduced without permission from the publisher.CPD Officer: Alan Palmer MIAT RAnTechRegistration and Accreditation Board:Glyn Fisher (Chair), John Gregor y,Cathy Godfrey, Kathy Ryder (Home Office),Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee:Alan Graham (Chair), Haley Daniels, Adele Kitching,Allan Thornhill, John WatersDiversity Officer:Haley Daniels MBA MSc MIAT RAnTech CIPDUK Biosciences ASG Representative/Home Office:Alan Palmer MIAT RAnTechviAugust20:Animal Technology and Welfare 12/8/20 07:54 Page viAvid plc ........................................................172Institute of Animal Technology .........168, 210,0BCIPS Product Supplies Ltd ................................IBCLBS Serving Biotechnology Ltd .........................IFCTechnician Wellbeing Workshop ......................171Tecniplast UK Ltd ..........................................174BRANCH SECRETARIES 2023Cambridge: Tony Davidge cambridgebranch@iat.org.ukEdinburgh: Gordon Melville edinburghbranch@iat.org.ukHuntingdon, Suffolk & Norfolk: Thomas McCamley hssbranch@iat.org.ukIreland: Lisa Watson irelandbranch@iat.org.ukLondon: Louise Fisher londonbranch@iat.org.ukMidlands: Ian Fielding midlandsbranch@iat.org.ukNorth East England: Zoe Smith northeastbranch@iat.org.ukNorth West: Emma Owens cheshirebranch@iat.org.ukOxford: Adam Truby oxfordbranch@iat.org.ukSurrey, Hampshire & Sussex: Francesca Whitmore shsbranch@iat.org.ukWest Middlesex: Hannah Easter westmiddxbranch@iat.org.ukWest of Scotland: Joanne King westscotlandbranch@iat.org.uk

NOT SO ‘BLUE MONDAY’The date may be termed the ‘bleakest day of the year’but we guarantee to brighten it via virtual meeting platformon 15th January 2024 from 13.00 – 16.00 hrs for a TECHNICIAN WELLBEINGWORKSHOPhaving a Culture of Care shouldn’t stop with the animalsJoin us to discuss how we can improve wellbeing within our industryGuest Speakers:Penny Hawkins RSPCAHaley Daniels IATJonathon Wood DatesandAngela Kerton The Learning CurveMaizy Jenner University of ManchesterAnneke Keizer CopeplusDon’t miss out on this unique opportunity to learn aboutthe latest advancements in Technician WellbeingYou will also have the opportunity to ask questionsduring the Q&A sessionsFor more information or to book your place, please use this QR code or visithttps://www.eventbrite.co.uk/e/technician-wellbeing-workshop-tickets-746961220117?aff=oddtdtcreator

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173August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareEditorialJas BarleyChair of the Editorial BoardLooking back over issues of the Journal through its various identities, one thing is apparent and that is the contribution thatoverseas authors have made to the content. Topics have varied from dealing with exotic species, lack of sophisticated equipment,different attitudes to everyday problems, staff training and education and disease outbreaks. However, the resolute that has beenconstant throughout, despite the differences across the world, is the love and concern for the animals being cared for.Many include interesting photographs but I unfortunately am unable to use them as the quality of images is so poor whenrepr oduced, to the extent in some cases, they become worthless.Obviously, things have changed over seven decades and the technology described in contributions from overseas is less differentfrom what we use in the UK. This issue welcomes contributions from Australia, the Czech Republic and Iran as well, of coursefrom the UK. Since ATW became an Open Access publication and is being published electronically, it is enjoying a wider audienceand is attracting more contributions than usual. Not all are relevant to our profession, but knowledge is transferable so whatseems ‘off beat’ today may become useful in the future. However, as Editor I will always strive to maintain the quality of ourpublications and the usefulness to our readers.In this issue we include the RSPCA 2019 Rodent and Rabbit Welfare group meeting report. The 26th meeting that the RSPCA haveorganised focussed on ‘sentience, positive welfare and psychological well being’. The report contains contributions from 11presenters as well as notes on the interactive discussion session on sentience that closed the meeting.A paper from Iran, a first as far as I can see for the Journal, on reducing the negative effects of methionine on bone parametersin broilers’ embryos may seem of little relevance but it offers a better understanding of how methionine affects bone structurewhich is important to most species. Similarly, Feline Assisted Therapy as described by the team at the University of Life SciencesPrague does not appear to fall into the realms of Animal Technology but it gives us a better understanding of how animals can havea positive effect on some people, which in the current situation may be of significant benefit to a wider population. Our final paperfrom the team at Western Sydney University, details the care of the Children’ Python and two species of Bearded Dragons. Notperhaps the run of the mill laboratory animals but just as important to many Animal Technologists globally as mice and rats. If youkeep reptiles at home or know of someone who is contemplating one as a pet these papers make useful reference documents. Wealso offer two papers from previous issues of the Journal which were very different in appearance and content than today’s Journalof Animal Technology and Welfare and not only because of the change of title. Issues were printed in black and white and in the veryearly days were produced by hand. The paper from France on Physical Hazards in the laboratory animal house will bring back manymemories for some of the older technicians, myself included, but not necessarily good ones. The use of ether as an anaestheticwhich I know is still used in some countries where resources are limited, for human surgery, presented a very real danger to bothanimals and staff. Disease in laboratory animal units was often a recur ring problem, bacterial infections such as Pseudomonas asdescribed in the reprint of the article were still presenting Animal Technologists with problems as late as the end of the 1980s. Whenimporting animals and tissues from overseas it is important to realise that they may be carrying disease not seen in the UK forseveral decades. In r ecent times, Ectromelia was introduced into a unit in the USA via antibodies produced overseas. Precautionsmust be taken until such time as you are sure that the animals and tissues are clear of any underlying infections.We are also able to offer in this issue an interesting Tech-2-Tech article by Seonagh Henderson of the University of Glasgow, ona novel technique of cage cleaning which hasa positive effect on the welfare of laboratory rats. Finally, we included several postersprepared for AST2020 but sadly at the moment remain unpresented.Thanks again to all of our authors, past and present, both internationally and here in the UK. There would not have been 70 yearsof the Journal without you. Here is to the next seven decades and beyond.THE INSTITUTE OF ANIMAL TECHNOLOGYETHICAL STATEMENT“In the conduct of their Professional duties, Animal Technologists have a moral and legalobligation, at all times, to promote and safeguard the welfare of animals in their care,recognising that good laboratory animal welfare is an essential component of goodlaboratory animal technology and science.The Institute recognises and supports the application of the principles of the 3Rs(Replacement, Reduction, Refinement) in all areas of animal research.”ixAugust20:Animal Technology and Welfare 12/8/20 07:54 Page ixWe hope that you enjoy the content of this journal. We all continue to investigate ways to refine animal welfare throughout the animal science industry. We identify new ways of caring for the animals and try to enrich their lives as much as possible but also ensuring the care provided still permits them to remain suitable experimental models.In this edition we have an article on the role of the review and regulatory approvals processes for animal research in supporting the implementation of the 3Rs advances. The reviews mostly took place during the beginning of 2022 against a background of significant changes in ASRU processes and the government’s independent review of research bureaucracy. We have included an IAT graduation special article which was celebrated on the 4th July 2023 at the MRC Mary Lyon Advanced Training centre in Oxford for students who had successfully completed the IAT Diplomas Levels 4, 5 and 6 of Laboratory Animal Science and Technology. The Tech-2-Tech article provides details of both researcher and Animal Technician perceptions of rat tickling which includes the drivers and barriers to the uptake of doing this.There are posters from IAT Congress 2023 which took place during March that make very interesting reading material. The posters include different types of environmental enrichment for rats and the refinement of chicken environments. There are two posters which focus on Zebrafish, one is the Zebrafish health and welfare database and the other details how to contain Mycobacterium marinum within a containment level 2 aquatics facility. An animal facility introduced running wheels to overweight mice to establish that if by doing this would their obesity reduce and another poster discussing the introduction of scheduled teeth checking of mice. Are study plans a benefit to the management of animal facilities, this poster demonstrates the benefit of its introduction. There is another poster that provides an exploratory view into abdominal fat necrosis in mice and one which discusses how to refine the welfare of immunocompromised mice that are given carbon tetrachloride to induce liver fibrosis.Remember to use our journals as reference sources for both your independent learning and your IAT educational journey. We are here to provide you with the opportunity to publish your work as we are always looking for new material for ATW. The assistance that the ATW editorial board can give you will support you by turning your ideas and concepts into realities.EditorialDiane HazlehurstChair of the Editorial Board Email: info@atwjournal.com December 2023 Animal Technology and Welfare

900cm2standard rat cage floor area150cm2additional floor area+find out morewww.tecniplast.it/ukUNLOCK EXTRA CAGE SPACEExtra animal comfortIncreased animal densityLARGER FLOOR AREAReach up to 8 rows in lessINCREASED CAGE DENSITYthan 2 metres of heightthanks to HD racks

175August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareDecember 2023 Animal Technology and WelfareThe role of review and regulatory approvalsprocesses for animal research in supportingimplementation of the 3RsFRANCES RAWLEA report by Dr Frances Rawle, commissioned by the National Centre for the Replacement, Refi nement and Reduction of Animals in Research (NC3Rs), February 2023Correspondence: Frances.Rawle@nc3rs.org.uk IntroductionThe NC3Rs commissioned this work towards the end of 2021, based on a concern that advances in the 3Rs often take a long time to come into routine use. Most research in the academic sector using animals is reviewed three times, by the funder, by the local Animal Welfare and Ethical Review Body (AWERB) and by the Animals in Science Regulation Unit (ASRU) and my brief was to fi nd out the extent to which these reviews were supporting the adoption of 3Rs advances, or could be more effective at doing so. Most of the interviews took place during the fi rst half of 2022, against a background of signifi cant changes in ASRU processes and the government’s Independent Review of Research Bureaucracy and there was a concern that changes in review processes may inadvertently lead to the loss of opportunities to promote improvements in the 3Rs. It was a privilege and a pleasure to talk to so many people actively involved in the review processes for animal research and hear their honest views of what was working well (and less well) alongside their ideas for possible improvements. Many of the recommendations in the report were based on these ideas. I make no apology for straying beyond the review processes in my recommendations as I did not want constructive thoughts to be lost. The recommendations were designed to be practicable and not overly burdensome but I recognise that implementation against a background of signifi cant change will not be a trivial task and many stakeholders in addition to the NC3Rs itself will need to be involved to ensure the recommendations have an impact. The report was well received when it was initially published and I have given several presentations and held discussions with various stakeholder groups about how the recommendations might best be implemented. The NC3Rs itself is focussing its efforts initially on the recommendations directed at funders with a workshop planned for later this year. Frances Rawle, July 2023About Dr Frances RawleFrances has extensive experience of managing scientifi cpeer review and of research policy, research ethics and governance throughout a long career at the Medical Research Council (MRC). For many years Frances was responsible for policy onanimal research and the MRC’s funding of the NC3Rs andfrom 2018 until 2021 she represented the MRC on theNC3Rs Board. Frances has a particular interest in researchintegrity and reproducibility. Frances now works as an independent consultant and was on the Steering Group for the Animals in Science Regulation Unit (ASRU) Changeprogramme in 2021/22.ForwardIn the United Kingdom (UK) oversight of the use of animals for research purposes in the academic sector is undertaken by various bodies and at various stages.This includes review by public and charitable funding bodies, the national regulator and locally by ethics committees. The 3Rs are on the face of it at least an important consideration in the review by all these organisations. This means that inevitably there is the

176Animal Technology and Welfare August 2020potential for overlap and duplication of efforts. That said, there is often a long lag between the development of 3Rs approaches and their use in routine practice, even for simple advances that benefit animal welfare. The current oversight mechanisms should support the NC3Rs mission but it is not clear that this is happening to the extent it should be despite the academic community’s long-standing commitment to the 3Rs. To try to address this and to identify gaps and overlaps the NC3Rs commissioned Dr Frances Rawle to undertake a detailed and independent review, including engagement with key stakeholder groups.Dr Vicky Robinson, NC3Rs Chief ExecutiveBackgroundThe 3Rs principles – replacement, reduction and refinement – are the widely accepted ethical framework for the use of animals in research and compliance with these principles is a legal requirement in the UK under The Animals (Scientific Procedures) Act 1986 (ASPA).Obtaining a project licence (PPL) under ASPA requires review by an Animal Welfare and Ethical Review Body (AWERB) at a Home Office licenced establishment and by the Animals in Science Regulation Unit (ASRU) Inspectors to establish (amongst other things) whether the proposed research complies with the 3Rs principles. In addition, most academic research involving animals is subject to peer review by public sector or charitable funders. Most funders are committed to promoting the 3Rs and their peer review covers relevant areas but the extent to which implementation of the 3Rs is explicitly considered varies. The focus of funder peer review is mainly on the quality of the proposed research and the likelihood of achieving significant scientific advances.Aims of the projectThe project had three main objectives focussing on academic-led research involving animals in the UK:1. To map in detail what the various regulatory and review processes and bodies currently do to ensure compliance with 3Rs principles and to promote adoption of 3Rs advances.2. To identify any current variations in review processes, any gaps (or overlaps) in coverage and any lessons to be learned from examples of particularly effective practice.3. To explore opportunities for adjusting current processes and responsibilities to cover any gaps, remove unnecessary duplication and more effectively promote adoption of 3Rs advances.Project approachInformation was obtained from interviews (~40) with stakeholders1 involved in regulatory and review processes. These included chairs and members of AWERBs, Establishment Licence Holders (ELHs), Named Animal Care and Welfare Officers (NACWOs), Named Veterinary Surgeons (NVSs), Named Information Officers (NIOs), former and current Inspectors, Animals in Science Committee members, representatives of charitable and public sector funders, and senior scientists with experience as reviewers on funding panels and as holders of PPLs and personal licences (PIL). ASRU provided written responses to questions and three AWERB meetings were observed.Summary of findingsReplacementReplacement does not seem to be covered well by any of the review processes. AWERBs and ASRU inspectors rarely suggest use of replacements. They do not (and could not) have sufficiently detailed knowledge of the full breadth of the scientific areas they need to cover to know for every application whether appropriate and practicable replacement technologies are available. AWERBs may assume that by the time a licence application is submitted to them for review the researcher and the funder have considered the options for replacement and concluded that animal use is necessary.Funders’ peer review involves more specialist scientific expertise but their review tends not to focus explicitly on whether suitable replacements might be available but rather on whether the applicants’ chosen models will allow them to answer the scientific questions posed. Where the research is disease focussed, the key question for peer reviewers is the relevance of the animal model to the human disease and how likely the results are to translate rapidly into clinical benefits. All funders require applicants to justify the need to use animals and their choice of species but the extent to which this is challenged by reviewers varies between funders.ReductionBoth AWERBs and funders report paying closer attention to experimental design and statistics in their reviews over recent years. Although AWERBs report a shortage of people with the necessary expertise to review this area. The funders’ aim is to ensure the research they are funding is robust and reproducible which should lead to reductions in overall animal use, although paradoxically the review of experimental design often indicates that more animals are required for each experiment to achieve sufficient statistical power.The role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rs

177August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3RsIt is not possible to review the design of every experiment covered by a grant or a PPL application covering 3 to 5 years at the outset and both PPL and grant reviews focus on typical or early experiments. ASRU reviews the basic principles of experimental design but does not undertake a detailed assessment of the proposed statistical methods. As part of the new audit process ASRU inspectors will also evaluate the systems in place at licensed establishments to promote the use of appropriate experimental designs and statistical methods and the availability of local expertise. The NC3Rs experimental design assistant was designed to address the shortage of expert advice in this area but is not yet widely used in grant or PPL submissions. Some establishments review experimental design as part of individual study plans but the shortage of available expertise is likely to prevent this being done more widely.Efficient colony management and breeding are proven means of reducing animal use and it is important for reproducibility to avoid genetic drift. These aspects are rarely covered as part of project review by either AWERBs or funders, but oversight of colony management and breeding strategies for genetically altered animals at a facility wide level should be included in the AWERBs other functions.ASRU asks for information in PPL applications to assess efficiency of breeding and use of best practice in breeding is reviewed as part of the recently introduced audit process. AWERBs may also oversee a local system to make best use of tissues from culled animals in teaching and research as part of their wider role in promoting the 3Rs.Neither AWERBs nor funders reported much discussion in their reviews of the potential to use methodological advances such as in-cage monitoring, microsampling, or use of imaging techniques to enable more information to be obtained from fewer animals.RefinementRefinement is the area in which AWERBs are most confident to challenge when they review PPL applications and feel that their input adds most value. NACWOs and NVSs usually provide input on the refinement of protocols both in the preparation of licence applications and as a project progresses.Refinement in housing and husbandry, such as environmental enrichment is usually not covered as part of the PPL review but is overseen by the AWERB, the NACWO and the NVS on a facility wide basis.Funders’ grant reviews occasionally look at refinement (for example with protocols involving severe levels of animal suffering or for specially protected species when the NC3Rs normally provide an additional welfare review). A small proportion of PPL applications are referred to the Animals in Science Committee (ASC) and their reviews may cover refinement. Funders rarely consider housing and husbandry, except in cases where it is critical to the experiment (for example, in studies of the gut microbiome). They felt they could and should rely on AWERBs and ASRU to ensure appropriate refinements were in place.Barriers to uptake of 3Rs advancesReasons mentioned by stakeholders for slow uptake of 3Rs advances included:1. The time and cost involved in setting up new techniques in a laboratory and lack of access to expert help. The laboratories which have developed new techniques do not have the time and resources to help everyone who wants to try them. Researchers may be concerned that their lack of expertise may make a grant application involving a new method uncompetitive and that delays in producing data and publications while they get a new model established will negatively affect their career. 2. Lack of published data on how results using replacement technologies compare to established animal models and concerns about acceptance for publication or (in work to develop treatments) by the regulator, for example if there is an accepted gold standard animal model in the field. Many researchers think that they must use an animal model because a paper using a new in vitro model on its own will not be accepted by the scientific journals.3. Concerns that introducing refinements to experimental protocols will result in a lack of compatibility with earlier data.4. Poor access to information on 3Rs advances. Many stakeholders highlighted the need for better availability of credible sources of information on advances in all 3 ‘Rs’ for researchers, committee members, reviewers and named persons, including information on evaluation and validation of new methods and on approaches that had been tried and not proved useful. The need to better define and resource the role of the NIO was highlighted, to help researchers and AWERBs to access information.Recommendations1. Funders should make best use of their access to highly specialist scientific peer reviewers to ensure that possibilities for use of replacements or new approaches to obtain more information from fewer animals are identified and implemented where

178Animal Technology and Welfare August 2020appropriate. This could be facilitated by using more specific questions for reviewers on whether there are available alternatives and/or reduction strategies.2. Funders could introduce more targeted questions for applicants to elicit information on replacement and reduction, and guidance for applicants on expectations with the assumption that in most cases2 optimising refinement will be ensured by ASRU and AWERB oversight.3. Funders should be prepared to provide additional funding to allow grant holders to explore and validate the use of new alternatives alongside their established models and to facilitate dissemination of new methods3 by supporting laboratories which have developed them to provide access to the technology and train others to use it.4. It should be made clear in a PPL application what parts of the work have already been funded (including date of award and duration) and by whom so that AWERBs and ASRU are clear what has been externally peer reviewed and what has not. Funders should be willing to share information on whether their expert review has explicitly considered whether replacements are available.5. Establishments should ensure that their processes allow the use of animals to be challenged early in the research planning process. AWERBs should ask questions about whether/how an applicant has searched for information on possible replacements or reduction strategies. They should expect a clear explanation of what replacements have been considered and why they are not suitable and whether approaches to get more information from a group of animals have been considered. This could be facilitated by guidance to AWERBs on questions to ask and what should reasonably be expected of applicants.6. Best practice for induction for AWERB members should include training in the 3Rs and the principles of experimental design. The introduction of audit processes in ASRU’s new ways of working provides an opportunity to clarify expectations for training of AWERB members and to confirm via audit that these are being followed. In the longer term the requirement for continual professional development (CPD) for all AWERB members should be considered by the sector in line with the research ethics committees which cover projects involving human participants.7. AWERBs should be clear on the expectations for their role in promoting the 3Rs on a facility-wide basis outside the process of PPL review, including the importance of spending enough time and attention on this part of their role and what constitutes good practice. Areas to cover include refinement of housing and husbandry, efficient colony management and breeding, good experimental design, tissue sharing and sharing of 3Rs advances.8. The expectations of the NIO role should be set out clearly at each establishment in line with ASPA and IAT/LASA guidance.4 Establishments must ensure that NIOs have the expertise, time and appropriate resources and training to effectively support researchers, AWERB members and animal facility staff in accessing information on 3Rs advances. They should be well trained in approaches to search for information and have time to support researchers to fulfil their responsibility to look for alternative approaches. ASRU should cover the effectiveness of the NIO role in their audits.9. To facilitate access to information about 3Rs advances, the NC3Rs, scientific or learned societies and/or funders should convene expert groups to review information on 3Rs advances available in particular scientific areas or for commonly used animal models of disease, to produce authoritative, up to date and easily accessible information for researchers, peer reviewers and AWERBs. Funders should ensure that this information is considered in their funding decisions.10. All AWERBs and funder review panels should have access to expertise in statistics and experimental design. Inventive solutions may be necessary to make best use of available expertise for reviewing given the shortage. The NC3Rs experimental design assistant (EDA) should be more widely used in applications; this may require further development to make it more accessible. With the current focus on improving reproducibility across the life sciences, funders and universities should explore means to support development of more experts in statistics and experimental design, both to help and train researchers on the ground and to participate in expert review.11. ASRU and AWERBs should ensure that information on 3Rs advances obtained from retrospective reviews and retrospective assessments of PPLs is available to the research community whether via publication or some other means.512. To reduce unnecessary bureaucracy funders can rely on AWERBs and ASRU for checking implementation of refinement and on ASRU to monitor compliance with ASPA (for example, it is not necessary to include this in funder assurance checks or to ask for formal confirmation of licences before grant funds are released). However it remains important for funders to check that AWERBs have reviewed any animal research that falls outside of the ASPA such as work taking place overseas.The role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rs

179August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3RsBackground and context for the projectThe 3Rs principles – replacement, reduction and refinement are the widely accepted ethical framework for the use of animals in research and compliance, with these principles is a legal requirement in the UK under ASPA.Obtaining a project licence (PPL) under ASPA requires review by an Animal Welfare and Ethical Review Body (AWERB) at the establishment where the research will take place and by the Animals in Science Regulation Unit (ASRU) Inspectors to establish (among other things) whether the research complies with the 3Rs principles.NACWOs and NVSs have local responsibility for animal welfare and a key role in promoting the 3Rs (especially refinement) and NIOs have a responsibility to help researchers and animal facility staff access information about the 3Rs that might be relevant to their work.Assessment of 3Rs compliance used to be covered in ASRU inspections. ASRU is currently undergoing a major change programme with the aim of improving its efficiency and effectiveness as a regulator;Inspectors are no longer assigned to specific establishments and inspections have been replaced by a programme of facility, systems and thematic audits. This represents both an opportunity and a risk:– The opportunity for ASRU to set out clear expectations of establishments for what they should be doing to promote the 3Rs which they will audit against.– The risk that the focus is on box ticking rather than ensuring a culture of genuine commitment to advancing the 3Rs.Most academic research involving animals is externally funded from public sector or charitable bodies. The research covered by a PPL and by a research grant are rarely the same – one PPL often covers work funded from several different grants and the time periods covered by licences and grants are normally different. Funding is usually subject to peer review processes focussed on research quality and the likelihood of achieving significant scientific advances, although research funded from internally managed resources may not be subject to such detailed scrutiny.Although funders have no legal responsibility under ASPA to promote the 3Rs, all funders interviewed for this study have a commitment to do so. However the extent to which the 3Rs are explicitly considered in their peer review processes varies. There are concerns that the ASRU and AWERB review processes may involve assumptions that funders’ scientific peer review addresses aspects of the 3Rs such as experimental design or the potential for replacement.Conversely, funders may assume that implementation of the 3Rs is ensured by the AWERB and regulatory review processes. There is also concern that the predominance of well established senior researchers in peer review may lead to a bias towards use of well established and familiar animal methods. This project aimed to explore whether these concerns were justified.With the publication of the Independent Review of Research Bureaucracy6 funders will be reviewing their application processes to reduce unnecessary bureaucracy; it is important that any forthcoming changes to either funder reviews or ASRU regulatory processes do not result in gaps in coverage of the 3Rs.MethodologyInterviews (~ 40) with people7 involved in AWERB, funder and ASRU review processes were conducted via video conferencing and lasted about an hour. They followed a topic guide that included questions about the details of the review processes and specific questions about how each of the 3Rs was dealt with.Interviewees were also asked about barriers that they had encountered (actual or perceived) to implementation of 3Rs advances. Questions were modified to suit the experience an interviewee had of the various review processes. Any interesting observations relevant to the project were followed up by further ad hoc questions before returning to the prepared question set. Interviewees were selected to cover a variety of perspectives and experiences of the review of grant and PPL applications. Funder interviews included seven charities of varied sizes (annual research budgets ranging from £2.5m to >£1bn) and the two UK Research Councils that fund the most animal research, the Medical Research Council (MRC) and Biotechnology and Biological Sciences Research Council (BBSRC). They involved staff with experience of peer review processes, panel discussions and funder policies on animal research. Interviews with senior researchers provided an alternative perspective on expert peer review processes.All the AWERB members (lay and expert) and chairs interviewed had experience in academic establishments, several with more than one AWERB and a few also had experience of AWERBs in private sector establishments. Many people interviewed had experience of several aspects of these review processes – for example academics who undertook peer review for funders and had their own PPLs reviewed by AWERBs, or people with experience as ASRU inspectors, researchers and NVSs.The head of ASRU and the head of the new Animals in Science Policy and Coordination Unit contributed at the start and towards the end of the project, and ASRU provided responses to questions in writing. Three AWERB meetings at which PPLs’ applications and amendments were reviewed and reports relevant to the implementation of the 3Rs were considered were also observed.

180Animal Technology and Welfare August 2020Findings – who is doing what?Funder peer reviewContextAll funders point researchers to the document Responsibility in the use of animals in bioscience research8 and emphasise in their guidance the requirement for researchers to implement the 3Rs. All the charities interviewed are members of the Association of Medical Research Charities (AMRC) and subscribe to their position statement9 on animal research which includes the 3Rs.All funders include specific questions for grant proposals involving the use of animals which cover information relevant to the 3Rs and many mentioned trying to standardise these questions across funders facilitated by the NC3Rs. Most funders said they have guidance and/or specific questions for reviewers and panel members related to aspects of the 3Rs.All funders use a combination of written reviews and panel meetings for their expert peer review and decision making processes but exact details vary between funders and between different schemes (for example, written reviews may be sought before or after a short listing step, sometimes it is panel members that provide written reviews, fellowship awards often involve interviews while project grants usually do not). Most funders mentioned having increased their focus on experimental design and statistics in recent years requiring more information on this from applicants and more scrutiny by reviewers and/or panels. Several funders reported they had experienced increased demand for funding for animal research overseas recently either for academic collaborations or for preclinical testing by contract research organisations based overseas. They require researchers and their local AWERB to satisfy themselves that welfare standards are equivalent to the UK and most mentioned use of the NC3Rs checklist10 for that purpose. A few described requiring specific justification for doing work overseas rather than in the UK to ensure researchers are not just looking for a way of doing work more cheaply or to do experiments that would not be permitted under UK regulations.General pointsThe extent to which the 3Rs are covered in peer review is dependent on the quality of written reviews – although there are questions relevant to the 3Rs not all reviewers answer them. Time constraints are an issue in Panel/Board meetings; around 10 to 15 minutes per application is generally allowed for discussion, which does not allow time to cover all 3Rs issues in detail. Often the focus is more on scientific ideas and how they will advance the field.ReplacementMost funders reported that peer review focusses on whether the applicants are using an appropriate model to address the question they want to answer where the use of animal is proposed, rather than on the availability and suitability of replacement technologies per se. All funders rely on reviewers and panel members to identify potential replacements based on their knowledge of the field.None reported any systematic searches for possible replacements. Disease-focussed charities said that the key issue for their panels is the relevance of the animal model to human disease and how likely the results are to translate rapidly into benefits for patients.All funders ask applicants to justify the need to use animals and their choice of species. Answers varied as to how frequently the need to use animals is challenged in their review processes, from ‘often’ or ‘rigorously’ to ‘rarely’. Some funders report challenge from panels on whether all the animal experiments are necessary. Some disease focussed charities reported regular discussions of whether the research should be done in humans or using human tissue or induced pluripotent stem cell-based models rather than animals.Some funders highlighted that panels would look carefully at whether animals are needed for each part of a programme and may decide animal use is not appropriate for certain parts or may ask for the in vitro or in silico part of the project to be done first before agreeing to fund subsequent in vivo work. Most funders reported increasing numbers of applications wanting to use organoids and in silico modelling.ReductionMost funders reported an increased level of scrutiny of experimental design and statistics in their peer review. Some use statistics experts on panels or as reviewers to scrutinise this area specifically and others rely on panel members with experience of animal models to do this. Several funders point to the NC3Rs EDA in their guidance for applicants but the use of its outputs in funding applications is not yet common.Some funders commented that it was common for scrutiny of experimental design to raise concerns about under powered studies and identify the need for more animals per experiment to achieve robust results. Funders recognise that for 3 or 5 year programmes of work it is impossible to scrutinise the design of every experiment. They are looking for evidence from a design for an early or typical experiment that applicants know what they are doing and how they will use information from early experiments to inform the design of later ones.The role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rs

181August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3RsMost funders do not scrutinise breeding strategies or colony management in their peer review (some funders said they did so for applications involving the development of new transgenic lines). In general funders felt that this is best done locally overseen by the AWERB. Some funders said that applicants mention sharing of tissues from otherwise unused animals in transgenic breeding programmes as a reduction strategy, or specifically said that they have funded grants using such material. There was very little mention of other methodological advances that could reduce animal use, such as longitudinal imaging or in-cage monitoring.RefinementRefinement is rarely covered in funder peer review. The exception is for specially protected species (and in one case, pigs) where funders use the NC3Rs review service to obtain a welfare and 3Rs review. Some funders also use this service for overseas work or where there are specific concerns identified (such as very large numbers of animals or protocols likely to cause severe suffering). One scientist panel member said that these NC3Rs detailed reviews were very helpful in ensuring 3Rs advances were implemented and should be done for all applications involving animals. However staff from funders said that the workload involved in doing this would be impracticable due to the high volume of applications involving animals. All funders consider that responsibility for ensuring good practice in housing and husbandry and environmental enrichment should sit with AWERBs and ASRU and should not form part of funder review. The only instances where housing and husbandry might be discussed during grant review would be where this is critical to the experimental design, for example in studies of the gut microbiome.Project/protocol specific refinement is rarely, if ever discussed by funder panels. The exceptions mentioned were for work taking place overseas (and thus not covered by UK regulation), disease models rated ‘severe’ and areas of research where there are newer animal models being developed to replace ones with more severe harms.Several funders specifically said they relied on AWERB and ASRU review and local processes to ensure refinement of experimental protocols was optimised. Most thought that this is where responsibility for promoting refinement should lie.AWERB review and associated local processesContext and general pointsFlexibility in the implementation of the AWERB functions and the fact that responsibilities are not set out in detail in law means there is a lot of variation in practice, both in how PPLs are reviewed and in how other tasks of the AWERB are carried out.11 Apart from the smallest establishments, a commonly expressed concern was that the high workload of PPL review meant that AWERBs did not have sufficient time for other functions related to promoting the 3Rs. One establishment reported trialling separate meetings for PPL and other AWERB business, to ensure the latter got adequate time and attention.Several people commented that the length of PPL applications meant a very high reading workload for committee members and that there was little recognition or reward for being an AWERB member in academic establishments. The high workload is also a factor in the difficulty AWERBs experience in recruiting lay members who are independent of the establishment (many lay members are university staff from departments not involved in animal research). Establishments covered in interviews for this project ranged from ~15 to >150 PPLs in place at any one time, such that the annual workload of licence review varied widely. The extent to which AWERB work was delegated to subcommittees varied but this was not always related to the size of the establishment. There were some suggestions that scientists who are PPL holders sitting on the AWERB may be reluctant to challenge their colleagues robustly because they know their own licences will be coming round for review in due course.Several people commented that the NIO role is not well defined and often not well resourced. Many NIOs do the job part time alongside other busy roles and may lack sufficient training in how to search effectively for information to be able to support licence applicants in finding information on potential replacements, reduction strategies or refinements that may be appropriate in their research.AWERB review processes differ depending on a range of factors but a number of themes emerged during the interviews. The common elements of all AWERB PPL reviews are pre-review input, member comments and a process for applicants to respond to the comments. However there is a lot of variation in how these elements are implemented. All establishments offer some form of input from the NVS, NACWO and director of the animal facility prior to submission of a PPL application to the AWERB. Processes range from provision of written comments on a draft of the application, through informal meetings, to formal meetings with a subcommittee of the AWERB including an NVS and a NACWO.

182Animal Technology and Welfare August 2020In some establishments these preparatory meetings are a requirement for all licence applications including renewals, while in others the focus is more on new licences. In some places the pre-submission input is optional while in others there is a formal process for all applications. The timescale mentioned was usually to start preparation at least 6 months before submission to ASRU. One person said that the AWERB chair meets prospective new licence applicants before starting the application process and one mentioned that new applicants do a presentation on their work to the AWERBbefore preparing their application to enable the AWERB to identify any ethical concerns. A few people mentioned that pre-application meetings with the assigned inspector had been a useful part of the preparation process prior to the ASRU reforms.A few establishments reported having a formal AWERB subcommittee to review PPL applications, or putting together a subcommittee for each application, with expedited discussions in the AWERB meetings focussed on minutes or reports from that subcommittee. Some AWERBs use an online discussion forum or email to comment on applications before the AWERB meeting and applicants can review comments in preparation to respond at the meeting or even respond online. AWERBs vary in whether the applicant is required to attend the AWERB or the PPL subcommittee meeting in person and if they do attend, whether they give a formal presentation or simply answer questions. Sometimes only new applicants are required to attend in person.In some establishments there is a requirement to submit a study plan for each new study under a licence. This process is generally managed by the animal facility and not by the AWERB. The focus varies; sometimes there is a detailed scrutiny of the experimental design. In many cases the NVS reviews the protocols to ensure refinements are appropriate, including humane endpoints and facility staff usually check for compliance with licences and/or whether the facility has the resources and appropriately trained staff available to support the planned experiments.One person mentioned that the information required in this study plan is based on the ARRIVE guidelines to make sure all requirements for publication have been thought about before an experiment starts. Establishments that require individual study plans find them valuable and one person said it means they worry less about detailed scrutiny of experimental design at the licence application stage. However one person commented that their establishment had decided not to introduce them because they are a lot of work to prepare and review and there was no evidence that they lead to fewer instances of non compliance.ReplacementAWERBs rarely challenge animal use per se. Several people reported that by the time a PPL application comes to AWERB the need to use animals is a given though they may challenge the need for animal use for certain experiments within the licence. Some mentioned that it is particularly difficult to challenge animal use when it is an ongoing programme being renewed. Some people commented that the time at which animal use should be challenged is in the preliminary discussion for a new licence with the facility manager, the NACWO and the NVS, although they are unlikely to have sufficiently detailed knowledge of replacement methods to do this.Some AWERB members commented that due to the expense and difficulty of doing animal experiments they would expect the applicants themselves to have carefully looked for any possible replacements. Similarly, there was an expectation that funder peer review would have identified opportunities to replace animal use. Many people highlighted that detailed specialist knowledge of a scientific field is required to know whether suitable replacement technologies are available, validated and practicable to implement, and that AWERB members rarely have this level of knowledge, certainly not in all areas of research undertaken in an establishment. Similarly, NVSs said that they do not have sufficient expertise in replacement as their expertise is primarily with animal methods. Therefore when AWERBs review a PPL application they rarely suggest use of replacements and find it difficult to challenge applicants if they say possible replacements are not suitable.Many people also commented on the difficulty of accessing information about possible replacements and the role of NC3Rs (via the regional programme manager where there is one) in this and whether NIOs have sufficient skills to help applicants search for information.ReductionA shortage of biostatistical and experimental design expertise available to support licence applicants and to review PPL applications was mentioned by many people. Not all AWERBs include members with this expertise. AWERBs that do have access to statistics and experimental design expertise find it very useful for their PPL reviews. Some people mentioned the need for additional funding for statistical support for researchers.AWERB members noted that scrutiny of experimental design is easier now that there is a section for relevant information on the licence form. It is not however possible to include the detailed design of experiments to be done 5 years ahead, so the review must look for evidence that the design of early or typical experiments is robust and information on how the data from earlier experiments will inform the design of later ones.The role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rs

183August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3RsSome licence applicants use the EDA in preparing their applications but others find it too time consuming, difficult to use or not appropriate for their experiments. AWERB members pointed out that it is insufficient for applicants to say they have used the EDA, they need to provide an example of an experiment they have designed using it. People commented that individual study plans (where used) are the appropriate level for effective scrutiny of experimental design but this is only possible where sufficient expert resource is available.AWERBs rarely review colony management and breeding strategies as part of the PPL review but sometimes have an oversight mechanism for this as part of their wider role in promoting the 3Rs. Opinion was divided as to whether more attention to this would result in significant reductions in numbers of animals. Many thought high animal maintenance costs were a powerful driver towards efficient breeding and colony management and overbreeding was no longer a significant problem while others thought there were still gains to be made. One person commented that too much pressure to reduce numbers can be unhelpful because it encourages researchers to cut corners with quality controls in breeding to prevent genetic drift (for example not back-crossing lines) leading to problems with reproducibility.It was pointed out that some wastage of animals arises when lines needed to be kept on the shelf during the process of writing and review of scientific papers in case additional experiments are requested by reviewers.Methodological advances such as in cage monitoring, microsampling and use of imaging techniques were mentioned as ways to enable more information to be obtained from fewer animals. One person said AWERBs should look for possibilities to use these during PPL review. However these technologies are not always available in establishments and the equipment can be expensive.RefinementIn most places a lot of work is done on protocol specific refinement such as anaesthesia, analgesia and humane endpoints by NVSs and NACWOs before an application gets to the AWERB. Refinement is the area on which AWERBs are most confident to challenge when they review licence applications and feel that their input adds value. Refinement should be an ongoing process and AWERBs expect to see evidence of ongoing refinement at mid term and retrospective reviews. However, at least one researcher stated that when a laboratory has been using a model for some time and done a lot of work on refinement already, finding further refinements is difficult. Housing, husbandry and enrichment are mainly dealt with outside the PPL review process by facility staff and NACWOs, and in many establishments this is overseen by a 3Rs committee (in some cases this is a formal subcommittee of the AWERB). These committees may be involved in setting up studies to evaluate welfare refinements and developing establishment wide policies and standards for approval by the AWERB. Where there are establishment standards, the AWERB expects any proposed deviation from these to accommodate experimental requirements to be justified in a PPL application or amendment.Many people commented on the need for better sharing of knowledge and good practice in relation to both refinements specific to particular animal models and to housing, husbandry and other welfare improvements. Of note was a comment on the lack of shared information on approaches to refinement which had been tried and shown not to be effective.ASRU reviewContext and general pointsASRU is bringing in fundamental changes to its operating model underpinned by strategic shifts that are aligned with the Regulators’ Code. The new ways of working include a greater focus on the assessment of the suitability of all licence holders, including standards for licence holder training, and an increased focus on legal requirements in the assessment of PPLs. Inspectors are no longer assigned to establishments and PPL applications and amendments are reviewed on a first come, first served basis through a team of dedicated inspectors. As part of regulatory reform ASRU soon expects to develop and publish new quality standards for licence review.Licensing functions and compliance assurance are now separated with the latter including provision of facility, systems and thematic audits, enforcement activities investigating potential cases of non compliance and review of reports required as part of licence conditions, such as retrospective assessments. As part of the audits an inspector’s role is to assess the systems that an establishment has in place to implement the 3Rs and advise where they are not adequate.ReplacementASRU has always found replacement the most difficult of the 3Rs to deal with. ASRU inspectors are required to have a broad general knowledge of the life sciences and 3Rs issues but are not required to be technical experts in all 3Rs approaches. It would not be possible to maintain up to date knowledge about all available replacement technologies and their suitability across the full range of research areas for which an inspector reviews licence applications.Applicants are required to explain what steps they have taken to research alternatives and whether they have fully considered practicable alternative approaches.

184Animal Technology and Welfare August 2020They are also asked what in silico, in vitro or ex vivo techniques are used in the project overall and how they integrate with the proposed animal use.Inspectors assessing PPL applications will look for good answers to these questions but do not necessarily have the expertise to suggest replacements or challenge if the applicant says that replacements that have been considered are either not available or not suitable.ReductionInspectors are not required to be experts in statistics and experimental design. When reviewing a PPL application they review the basic principles of experimental design but do not carry out a detailed assessment of the proposed statistical methods. They evaluate as part of an audit, the systems in place at establishments to ensure that correct experimental design and statistical methods are used and whether local expertise is available.Licence applicants are required to provide information on various aspects of experimental design but it is not possible to predict 5 years ahead exactly what experiments will be done and thus to provide all the information to allow detailed scrutiny of the experimental design. Inspectors look for mentions of randomisation, masking/blinding, the use of appropriate controls and a credible explanation for the estimated numbers supported by power calculations if appropriate. They also check licence applications to ensure that any known duplication of procedures is justified.Problems with experimental design, such as lack of masking/blinding or inappropriate experimental units may have been noticed at site inspections but this is less likely with ASRU’s new ways of working.Licence applicants must explain how they will ensure any breeding of genetically altered (GA) lines is as efficient as possible and genetic integrity is maintained. Use of good practice in breeding is also reviewed during the audit of establishments. Unusually high numbers of animals culled without being used in experimental procedures may indicate potential problems with colony management.RefinementASRU finds refinement the easiest of the 3Rs to deal with. Licence applicants are asked to explain the choice of animals, models, and methods, why they are the most refined available, and how suffering will be minimised. Inspectors check that any form of animal suffering is justified in a licence application and relevant to the proposed programme of work. Specific aspects of refinement that are explored during licence review are aseptic surgery, the use of non-recovery anaesthesia and the use of anaesthetics and analgesia. However, it is the NVS’s responsibility to advise what anaesthetics or analgesic drugs are most appropriate.Inspectors look carefully for appropriate humane endpoints and challenge whether experiments could be stopped sooner. The inspector’s role is to assess (as part of audit) the systems that the establishment has in place to ensure the most refined techniques are used and advise where these systems are not adequate. Refinement of housing and husbandry is predominantly assessed during facilities audits.Does having peer-reviewed funding affect AWERB/ASRU decisions on project licences (or vice versa)?There were varied views on the extent to which the outcome of AWERB review is influenced by whether the research has already been peer reviewed and funded. Some people said that having peer reviewed funding in place made AWERBs more confident in the scientific benefits of the proposed work and thus influenced the weighing of harms to the animals used and likely benefits of the proposed research.Others mentioned that it was rare to see a licence application for which there was not at least some peer reviewed funding already in place and was difficult to identify any influence. People were conscious of the high cost of animal research and felt that if funders were willing to pay it showed that non animal replacements were not practicable and the most appropriate animal models were being used. However as discussed previously, funders may not be explicitly looking at replacement options in their reviews.Some people commented that where the research had already been peer reviewed and funded it was difficult for AWERBs to challenge aspects of the plans that they were unhappy with. A few people said that they were aware of specific occasions where pressure had been put on AWERBs to approve licence applications they were unhappy with because the work had already attracted big grants or that the AWERB felt they were under time pressure related to the availability of grant funding. Others said specifically that they had never seen this happen.ASRU requires PPL applicants to provide information on how they plan to fund their work. This is to provide assurance that research can be completed and that benefits will be realised from the use of the animals. Peer reviewed funding also gives some assurance of the quality of the research and that scientific advances will be made. Thus having peer reviewed funding influences the harm/benefit analysis inspectors are required to undertake before granting a PPL.The role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rs

185August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3RsIn contrast, none of the funders thought that having a PPL in place before the grant application was considered influenced the decision about whether using animals was appropriate. However if a researcher already has the animal model established in their laboratory it does give reviewers greater confidence that they can achieve their objectives for the grant.What are the barriers to the uptake of 3Rs advances and what might help to overcome them?In discussing the reasons for slow uptake of 3Rs’ advances and any experience interviewees had of people being reluctant to try new methods, some common themes emerged.The time and cost involved in setting up new techniques in a laboratory together with a lack of access to expert help is clearly an issue. People need access to equipment and expert help to enable them to try out new techniques to see if they are suitable for their own research. The laboratories which have developed new techniques do not have the time and resources to help everyone who wants to try them. Researchers may be concerned that if they apply for a grant which involves introducing new methodology their lack of expertise might make their grant application uncompetitive and delays in producing data and publications while they get a new technique established will negatively affect their track record and career prospects.Lack of published studies on how replacement technologies compare to established animal models can be a problem and academic researchers are concerned this will prevent their work being accepted for publication. Introducing new replacement methods may be particularly difficult when there is an accepted gold standard model in the field.For researchers developing new treatments there may also be concerns whether the regulator will accept the evidence to support moving into clinical studies. Many researchers think they must use an animal model because a paper using only in vitro model(s) will not be accepted by the major international journals as editors will ask them to demonstrate their results are valid in an in vivo model before a paper can be published.Where researchers are used to working with a particular model there may be concerns that introducing refinements to experimental protocols will introduce a source of variability or result in a lack of compatibility with earlier data that has already been published.Many people highlighted the need for better availability of information on advances in all 3 ‘Rs’ for researchers, committee members, reviewers and named persons, including information on evaluation and validation of new methods, and signposting of new methods for which the evidence base is strong. It was suggested that specialist scientific societies would be well placed to curate information on replacements and refinements to commonly used models in their field and to challenge the status quo.Sharing of information on approaches that had been tried unsuccessfully would also be extremely useful. The need to better define and resource the role of the NIO was highlighted, to help researchers and AWERBs to access information.ConclusionsReplacement is the area least well covered by existing review processes. The possibility for replacement is best considered at an early stage of the research planning process as AWERBs find it difficult to challenge once funding is in place. AWERBs and ASRU rarely have the detailed scientific expertise to determine whether replacements are available and suitable, the best strategy for improving this situation would be to ensure that the expert peer review organised by the funders explicitly covers this area.Improving the availability of information on replacements and how they compare to established animal methods, the ability of NIOs to help researchers fulfil their responsibilities to search for replacements and the access to expert help and funding to try out new methodologies should help speed uptake of replacement methods.Review processes scrutinise experimental design and statistical analysis to ensure the numbers of animals used is optimised to obtain robust and reproducible results and avoid the waste of animals that occurs when experiments are under- (or over-) powered.This is the area where there is the greatest potential for overlap between AWERB, ASRU and funder review and it is important to make the most efficient use of the limited specialist expertise available for reviewing.More attention should be paid to considering the suitability of new methodologies that allow more data to be obtained from fewer animals as a reduction strategy. Ensuring efficient colony management and GA animal breeding are important roles for the AWERB and best done at a facility-wide level rather than as part of PPL review by AWERBs and ASRU. This would remove the need for information on breeding strategies to be included in PPL and grant applications.Refinement is the area that is covered best by AWERB and ASRU PPL reviews and where NACWOs, NVSs and facility staff are most confident to provide challenge.

186Animal Technology and Welfare August 2020Funders rarely consider this area except in particularly ethically sensitive situations where they involve the NC3Rs in the review, or where housing, husbandry or animal stress levels may have a particular influence on experimental outcomes.There is insufficient evidence from this project as to what practices for AWERBs are ‘particularly effective’ (see project aim 2), but practices that some AWERBs or establishments might wish to consider trying are:– Presentations to the AWERB by people wanting to apply for a new licence before they start drafting their PPL application, to allow a chance for a proper ethical discussion.– Requiring all applicants to meet the NVS, NACWO, NIO and facility manager to gain input at the drafting stage for all licence applications and significant amendments.– Requiring applicants to attend the AWERB meeting when their PPL application or major amendment to an existing licence is considered.– Online posting of comments so applicants can think about their response before the AWERB meeting (but this should not replace discussion at the meeting).– Effective use of subcommittees to ensure the AWERB covers its full remit.– Review of individual study plans before each study starts, with study plan templates informed by the ARRIVE guidelines.– Standard housing and husbandry protocols for the establishment prepared by a 3Rs subcommittee, approved by the AWERB and reviewed annually.Recommendations1. Funders should make best use of their access to highly specialist scientific peer reviewers to ensure that possibilities for use of replacements of new approaches to obtain more information from fewer animals are identified and implemented where appropriate. This could be facilitated by using more specific questions for reviews on whether there are available alternatives and/or reduction strategies.2. Funders could introduce more targeted questions for applicants to elicit information on replacement and reduction, and guidance for applicants on expectations, with the assumption that in most cases12 optimising refinement will be ensured by ASRU and AWERB oversight.3. Funders should be prepared to provide additional funding to allow grant holders to explore and validate the use of new alternatives alongside their established models, and to facilitate dissemination of new methods13 by supporting laboratories which have developed them to provide access to the technology and train others to use it.4. It should be made clear in a PPL application what parts of the work have already been funded (including date of award and duration) and by whom, so that AWERBs and ASRU are clear what has been externally peer-reviewed and what has not. Funders should be willing to share information on whether their expert review has explicitly considered whether replacements are available.5. Establishments should ensure that their processes allow the use of animals to be challenged early in the research planning process. AWERBs should ask questions about whether/how an applicant has searched for information on possible replacements or reduction strategies. They should expect a clear explanation of what replacements have been considered and why they are not suitable, and whether approaches to get more information from a group of animals have been considered. This could be facilitated by guidance to AWERBs on questions to ask and what should reasonably be expected of applicants.6. Best practice for induction for AWERB members should include training in the 3Rs and the principles of experimental design. The introduction of audit processes in ASRU’s new ways of working provides an opportunity to clarify expectations for training of AWERB members and to confirm via audit that these are being followed. In the longer term the requirement for CPD for all AWERB members should be considered by the sector, in line with the Research Ethics Committees which cover projects involving human participants.7. AWERBs should be clear on the expectations for their role in promoting the 3Rs on a facility wide basis outside the process of PPL review, including the importance of spending enough time and attention on this part of their role and what constitutes good practice. Areas to cover include refinement of housing and husbandry, efficient colony management and breeding, good experimental design, tissue sharing and sharing of 3Rs advances.8. The expectations of the NIO role should be set out clearly at each establishment in line with ASPA and LASA/IAT guidance.14 Establishments must ensure that NIOs have the expertise, time and appropriate resources and training to effectively support researchers, AWERB members and animal facility The role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rs

187August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3Rsstaff in accessing information on 3Rs’ advances. They should be well trained in approaches to search for information and have time to support researchers to fulfil their responsibility to look for alternative approaches. ASRU should cover the effectiveness of the NIO role in their audits.9. To facilitate access to information about 3Rs’ advances, the NC3Rs, scientific or learned societies and/or funders should convene expert groups to review information on 3Rs’ advances available in particular scientific areas or for commonly used animal models of disease, to produce authoritative, up to date and easily accessible information for researchers, peer reviewers and AWERBs. Funders should ensure that this information is considered in their funding decisions.10. All AWERBs and funder review panels should have access to expertise in statistics and experimental design. Inventive solutions may be necessary to make best use of available expertise for reviewing given the shortage. The NC3Rs Experimental Design Assistant (EDA) should be more widely used in applications; this may require further development to make it more accessible. With the current focus on improving reproducibility across the life sciences, funders and universities should explore means to support development of more experts in statistics and experimental design, both to help and train researchers on the ground and to participate in expert review.11. ASRU and AWERBs should ensure that information on 3Rs advances obtained from retrospective reviews and retrospective assessments of PPLs is available to the research community, whether via publication or some other means.1512. To reduce unnecessary bureaucracy funders can rely on AWERBs and ASRU for checking implementation of refinement and on ASRU to monitor compliance with ASPA (for example, it is not necessary to include this in funder assurance checks or to ask for formal confirmation of licences before grant funds are released). However, it remains important for funders to check that AWERBs have reviewed any animal research that falls outside of the ASPA, such as work taking place overseas.Professor Cathy Abbott University of EdinburghDr Caroline Aylott Versus ArthritisDr Kate Chandler Animals in Science Regulation UnitDr David Coutts Multiple Sclerosis SocietyLauren Cresser Pirbright InstituteAndrew Cunningham University of SussexNgaire Dennison University of DundeeDr Colin Dempsey WellcomeProfessor David Dexter Parkinsons UKDr Megan Dowie MRCHelen Emery University of LeicesterDr Anne-Marie Farmer University of CambridgeGlyn Fisher King’s College LondonDr Richard Francis Stroke AssociationAnnex 1IntervieweesI am very grateful to the following people for contributing to this project by participating in interviews or providing written input.Thank you also to the AWERBs at the University of Bristol, King’s College London and the Royal Veterinary College for allowing me to observe meetings, and to Professor Cathy Abbott, Linda Horan, Professor Nick Plant and Dr Sally Robinson from the NC3Rs Board for advice on drafting the report.References1 Exceptions might be for specially protected species and/or severe protocols or work to be done in another country.2 There are opportunities for partnership funding with the NC3Rs.3 Guiding Principles for Named Persons, LASA.4 For example, a repository that is easily accessible and searchable. 5 Independent Review of Research Bureaucracy: final report.6 In a few cases more than one person participated in an interview, so the number of people is slightly higher than the number of interviews. One interviewee chose to remain anonymous.7 Annex 1 In a few cases more than one person participated in an interview, so the number of people is slightly higher than the number of interviews. One interviewee chose to remain anonymous.8 Responsibility in the use of animals in bioscience research/NC3Rs.9 Position statement on the use of animals in research/ AMRC.

188Animal Technology and Welfare August 202010 New checklists to support the assessment of welfare standards in overseas research/NC3Rs.11 Guiding principles on good practice for AWERBs/RSPCA and LASA.12 Exceptions might be for specially protected species and/or severe protocols or work to be done in another country.13 There are opportunities for partnership funding with the NC3Rs.14 Guiding Principles for Names Persons/LASA.15 For example a repository that is easily accessible and searchable.AMRC Association of Medical Research CharitiesASPAAnimals (Scientific Procedures) Act (1986)ASRU Animals in Science Regulation UnitAWERB Animal Welfare and Ethical Review BodyBBSRC Biotechnology and Biological Sciences Research CouncilCRUK Cancer Research UKEDA NC3Rs Experimental Design AssistantELH Establishment Licence HolderGA Genetically AlteredIAT Institute of Animal TechnologyLASA Laboratory Animal Science AssociationMRC Medical Research CouncilNACWO Named Animal Care and Welfare OfficerNC3Rs National Centre for the Replacement, Refinement and Reduction of Animals in ResearchNIO Named Information OfficerNVS Named Veterinary SurgeonPPL Project LicenceRSPCA Royal Society for the Prevention of Cruelty to Animals3Rs Replacement, Reduction and RefinementThe role of review and regulatory approvals processes for animal research in supporting implementation of the 3RsGlossary

189August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePAPER SUMMARY TRANSLATIONSCONTENU DE LA REVUERôle des processus d’examen et d’approbation réglementaire pour la recherche animale dans le soutien de la mise en œuvre des 3RRapport du Dr Frances Rawle, commandité par le Centre national pour le remplacement, le raffinement et la réduction des animaux en recherche (NC3Rs), février 2023Correspondance: Frances.Rawle@nc3rs.org.uk Résumé Ce travail a été commandé par le NC3Rs vers la fin 2021, les progrès en matière de 3R semblant souvent prendre beaucoup de temps avant d’être couramment mis en œuvre. La plupart des recherches menées dans le secteur universitaire en utilisant des animaux sont examinées trois fois, par le bailleur de fonds, par l’Organisme local de protection des animaux et d’examen éthique (AWERB) et par l’unité de réglementation des animaux en science (ASRU). Mon mandat consistait à de savoir dans quelle mesure ces examens appuyaient l’adoption des avancées réalisées en matière de 3R, ou s’ils pourraient être plus efficaces pour le faire. La majorité des entretiens ont eu lieu au cours du premier semestre 2022, dans un contexte de changements importants concernant les processus ASRU et dans le cadre de l’ examen indépendant de la bureaucratie de la recherche du gouvernement. On craignait que des changements dans les processus d’examen ne mènent par inadvertance à la perte d’occasions de promouvoir des améliorations dans les 3R. ★ ★ ★December 2023 Animal Technology and Welfare

190Animal Technology and Welfare August 2020Étude exploratoire menées par entretiens avec des chercheurs et des techniciens concernant leur perception du chatouillement du ratSAM BEECHENER, SARAH BROWN, VINCENT BOMBAIL, MEGAN LAFOLLETTE, IGNACIO VINUELA-FERNANDEZ ET ALISTAIR LAWRENCECorrespondance: Alistair.Lawrence@sruc.ac.uk Résumé Cet article met en évidence les principaux thèmes qui ont émergé d’une étude menée avec des techniciens et des chercheurs animaliers pour mieux comprendre les perceptions du chatouillement chez le rat, les facteurs potentiels et les obstacles à l’adoption du chatouillement chez le rat dans un environnement de laboratoire. Les personnes interrogées ont indiqué qu’elles avaient des attitudes positives envers les rats et l’idée de les chatouiller. Elles ont fourni des commentaires positifs sur le comportement social des rats, leur intelligence et leur capacité à interagir, par exemple en se faisant chatouiller par les techniciens animaliers et les chercheurs.Les participants ont indiqué que les obstacles à une plus large adoption du chatouillement chez les rats comprenaient notamment les contraintes de temps, un manque de formation sur les spécificités du chatouillement chez les rats et la façon d’interpréter les réponses des rats au chatouillement.En outre, il a été fait mention de préoccupations concernant le chatouillement car il affecterait l’intégrité expérimentale puisqu’il est nécessaire de maintenir un détachement professionnel des rats en tant qu’animaux de laboratoire. ★ ★ ★Évaluation des hamacs de rat comme enrichissement MEGAN HICKMAN, LORRAINE MILLER ET RUTH MACDONALD Correspondance: Megan.hickman@astrazeneca.com Résumé La recherche a montré que loger les rats dans des milieux plus enrichis et plus complexes sur le plan environnemental que dans des logements conventionnels revêtait de nombreux avantages pour le bien-être et la science. Un prototype de hamac a donc été conçu pour les cages de rat à deux étages afin d’augmenter la complexité environnementale en offrant des possibilités d’escalade et d’exploration et en fournissant une couverture supplémentaire. Le hamac a été testé au sein d’un groupe de rates pour évaluer la durabilité, l’impact sur le comportement et la préférence par rapport à d’autres éléments d’enrichissement de l’abri: • tunnels • maisons de coin pour ratsLe nombre d’interactions d’enrichissement observées a indiqué une forte préférence pour le niveau inférieur du hamac, suggérant que le hamac était un bon élément d’enrichissement.Paper Summary Translations

191August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsBase de données sur la santé et le bien-être des poissons-zèbres, où en sommes-nous ?NICOLA GOODWIN, ROSEMARY KEEBLE ET MICHAEL PRICECorrespondance: ntg22@cam.ac.uk Résumé La première base de données sur les termes de santé et de bien-être du poisson-zèbre a été créée courant 2019. En collaboration avec des spécialistes du poisson-zèbre et des vétérinaires, nous avons établi des scores de gravité, des indicateurs de bien-être, des définitions et des images pour former une ressource destinée à la communauté du poisson-zèbre. Depuis lors, la base de données sur le bien-être du poisson-zèbre a fourni une plateforme permettant une approche normalisée garantissant la cohérence du langage et de la notation de la gravité utilisés dans toutes les installations. Elle a permis de sensibiliser les gens aux problèmes de santé potentiels propres au contexte génétique ou aux impacts environnementaux sur la santé des poissons. En normalisant les termes et les scores de gravité utilisés, les exigences en matière d’élevage peuvent être partagées en tant que communauté afin d’assurer le bien-être des poissons, de créer une cohérence entre les installations et de se conformer à la loi de 1986 sur les animaux (procédures scientifiques) (The Animals (Scientific Procedures) Act 1986 ou ASPA). Nous discutons ici de la façon dont nous utilisons la base de données au sein des installations aquatiques de l’Université de Cambridge et des avantages que nous avons pu constater.

192Animal Technology and Welfare August 2020Paper Summary TranslationsINHALTVERZEICHNISDie Rolle der Überprüfungs- und Genehmigungsverfahren für die Tierforschung im Rahmen der Unterstützung der Umsetzung der 3REin Bericht von Dr. Frances Rawle im Auftrag des National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), Februar 2023Korrespondenz: Frances.Rawle@nc3rs.org.uk Abstract Diese Arbeit wurde Ende 2021 vom NC3Rs vor dem Hintergrund in Auftrag gegeben, dass es oft lange dauert, bis bei den 3R erzielte Fortschritte routinemäßig in der Praxis Anwendung finden. Die meisten Untersuchungen im akademischen Sektor, bei denen Tiere verwendet werden, werden dreimal überprüft – vom Geldgeber, vom örtlichen Animal Welfare and Ethical Review Body (AWERB) und von der Animals in Science Regulation Unit (ASRU). Meine Aufgabe bestand darin, zu ermitteln, inwieweit diese Überprüfungen die Einführung von 3R-Fortschritten in die Praxis unterstützen oder inwieweit sie effektiver sein könnten. Die meisten Interviews erfolgten im ersten Halbjahr 2022 vor dem Hintergrund wesentlicher Änderungen bei den ASRU-Verfahren und der staatlichen Independent Review of Research Bureaucracy, wobei es die Befürchtung gab, dass Änderungen von Überprüfungsverfahren ungewollt zum Wegfall von Möglichkeiten zur Förderung von Verbesserungen bei den 3R führen könnten. ★ ★ ★

193August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsEine explorative Interviewstudie zu Auffassungen von Forschern und Technikern über das Kitzeln von RattenSAM BEECHENER, SARAH BROWN, VINCENT BOMBAIL, MEGAN LAFOLLETTE, IGNACIO VINUELA-FERNANDEZ UND ALISTAIR LAWRENCEKorrespondenz: Alistair.Lawrence@sruc.ac.uk Abstract In diesem Artikel werden die wesentlichen Erkenntnisse einer Studie vorgestellt, die mit Tiertechnikern und Forschern durchgeführt wurde, um eine bessere Kenntnis der Auffassungen über das Kitzeln von Ratten sowie potenzieller Faktoren und Hindernisse für die Akzeptanz des Rattenkitzelns in einer Laborumgebung zu erlangen.Die Befragten gaben an, dass sie eine positive Einstellung gegenüber Ratten und der Idee des Rattenkitzelns haben. Sie äußerten sich positiv über das Sozialverhalten von Ratten, ihre Intelligenz und ihre Fähigkeit zur Interaktion, z. B. beim Kitzeln durch Tiertechniker und Forscher.Die Teilnehmer verwiesen darauf, dass Hindernisse für eine breitere Akzeptanz des Rattenkitzelns u. a. in Form von Zeitmangel, mangelnder Schulung in Bezug auf das genaue Vorgehen beim Rattenkitzeln und Unsicherheiten bei der Interpretation der Reaktionen von Ratten auf das Kitzeln bestehen.Außerdem wurden Bedenken geäußert, dass das Kitzeln die experimentelle Integrität beeinträchtigen könnte und dass es notwendig sei, eine professionelle Distanz zu Ratten als Versuchstiere zu wahren.★ ★ ★Eine Bewertung von Hängematten für Ratten als AnreicherungselementMEGAN HICKMAN, LORRAINE MILLER UND RUTH MACDONALDKorrespondenz: Megan.hickman@astrazeneca.comAbstract Forschungsergebnisse haben gezeigt, dass die Unterbringung von Ratten in einer angereicherten und komplexeren Umgebung im Vergleich zur herkömmlichen Haltung zahlreiche Vorteile für Tierschutz und Wissenschaft bietet. Mit dem Ziel, die Komplexität der Umgebung zu erhöhen, indem Möglichkeiten zum Klettern und Erkunden sowie zusätzliche Rückzugsorte geschaffen werden, wurde ein Hängematten-Prototyp für den Einbau in doppelstöckige Rattenkäfige entworfen. Die Hängematte wurde an einer Gruppe von weiblichen Ratten getestet, um ihre Strapazierfähigkeit, Auswirkungen auf das Verhalten und die Präferenz im Vergleich zu anderen Unterschlupfmöglichkeiten zu bewerten: • Tunnel • RatteneckenDie Anzahl der bei Anreicherungen beobachteten Interaktionen deutete auf eine starke Präferenz für die untere Ebene der Hängematte hin, was darauf schließen lässt, dass die Hängematte ein wirksames Anreicherungselement ist.

194Animal Technology and Welfare August 2020Paper Summary TranslationsDatenbank zu Gesundheit und Tierschutz von Zebrafischen – aktueller Stand NICOLA GOODWIN, ROSEMARY KEEBLE UND MICHAEL PRICEKorrespondenz: ntg22@cam.ac.uk Abstract Im Jahr 2019 wurde die erste Datenbank mit Begriffen zum Thema Gesundheit und Tierschutz von Zebrafischen erstellt. In Zusammenarbeit mit Zebrafischspezialisten und Tierärzten haben wir Schweregradbewertungen, Tierschutzindikatoren, Definitionen und Bildmaterial erarbeitet, um eine Informationsquelle für die Zebrafischgemeinschaft zu schaffen. Seitdem bietet die Datenbank zum Tierschutz von Zebrafischen eine Plattform für einen standardisierten Ansatz, der die Einheitlichkeit von Begriffen und der Schweregradbewertung in allen diesbezüglichen Einrichtungen sicherstellt.Damit wurde die Möglichkeit geschaffen, das Bewusstsein für potenzielle Gesundheitsprobleme zu schärfen, die im Zusammenhang mit dem genetischen Hintergrund oder Umwelteinflüssen auf die Gesundheit der Fische stehen. Durch die Standardisierung der verwendeten Begriffe und Schweregradbewertungen können die Haltungsanforderungen einheitlich umgesetzt werden, um das Wohlergehen der Fische zu gewährleisten, Einheitlichkeit zwischen den Einrichtungen zu schaffen und die Einhaltung des Animals (Scientific Procedures) Act 1986 (ASPA) zu gewährleisten. Im Folgenden erörtern wir, wie wir die Datenbank in den Einrichtungen der aquatischen Wissenschaften der Universität Cambridge einsetzen und welchen Nutzen wir dabei festgestellt haben.

195August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsINDICE DELLA REVISTAIl ruolo dei processi di revisione e approvazione regolamentare per la ricerca sugli animali come promotori dell’implementazione delle 3RRelazione della Dott.ssa Frances Rawle, commissionata dal National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), febbraio 2023Corrispondenza: Frances.Rawle@nc3rs.org.uk Abstract Questo lavoro è stato commissionato dal NC3Rs verso la fine del 2021 ispirato dalla consapevolezza che, spesso, gli avanzamenti nelle 3R richiedono molto tempo a trasformarsi in procedura ordinaria. Gran parte della ricerca condotta nel settore accademico con l’uso di animali è stata sottoposta a revisione tre volte: dall’ente finanziatore, dall’Animal Welfare and Ethical Review Body (AWERB) locale e dall’Animals in Science Regulation Unit (ASRU). La richiesta era quella di stabilire in che misura queste revisioni promuovevano l’adozione degli avanzamenti delle 3R o qualora necessitassero di maggiore efficacia. La maggioranza delle interviste aveva avuto luogo nella prima metà del 2022, sullo sfondo di notevoli cambiamenti nei processi dell’ASRU e della pubblicazione della relazione governativa Independent Review of Research Bureaucracy. Ci si preoccupava, inoltre, che i cambiamenti nei processi di revisione potessero involontariamente tradursi nella perdita di opportunità di promozione dei miglioramenti del principio delle 3R. ★ ★ ★

196Animal Technology and Welfare August 2020Paper Summary TranslationsUno studio basato su interviste esplorative della percezione di ricercatori e stabularisti in merito al solletico ai ratti SAM BEECHENER, SARAH BROWN, VINCENT BOMBAIL, MEGAN LAFOLLETTE, IGNACIO VINUELA-FERNANDEZ E ALISTAIR LAWRENCECorrispondenza: Alistair.Lawrence@sruc.ac.uk Abstract Questo articolo mette in evidenza i principali temi emersi da uno studio condotto in collaborazione con stabularisti e ricercatori per comprendere meglio le percezioni relative al solletico ai ratti, nonché potenziali fattori e ostacoli all’adozione del solletico in un ambiente di laboratorio. Gli intervistati hanno dimostrato un atteggiamento positivo nei confronti dei ratti e dell’idea del solletico, con commenti positivi sul comportamento sociale di questi mammiferi, sulla loro intelligenza e sulla loro capacità di interazione (ad es., tramite il solletico attuato da stabularisti e ricercatori).I partecipanti hanno indicato che gli ostacoli a un’adozione più ampia del solletico ai ratti includevano vincoli temporali, una mancanza di formazione nelle basi del solletico ai ratti e come interpretare le risposte dei ratti al solletico.Inoltre, sono state segnalate preoccupazioni sull’effetto del solletico sull’integrità sperimentale, evidenziando l’importanza di mantenere un certo distacco professionale dai ratti visti come animali da esperimento.★ ★ ★Una valutazione delle amache per ratti come strumento di arricchimentoMEGAN HICKMAN, LORRAINE MILLER E RUTH MACDONALD Corrispondenza: Megan.hickman@astrazeneca.comAbstract La ricerca ha mostrato numerosi benefici scientifici e di benessere associati alla stabulazione dei ratti in stanze più arricchite e complesse sotto il profilo ambientale rispetto agli stabulari tradizionali. Per accrescere la complessità ambientale attraverso l’opportunità di arrampicarsi ed esplorare e offrire un riparo aggiuntivo, è stato progettato il prototipo di un’amaca da usare nelle gabbie per ratti a due piani. L’amaca è stata messa alla prova in un gruppo di ratte femmine per valutare la durabilità, l’impatto sul comportamento e la preferenza rispetto ad altri strumenti di arricchimento dello stabulario: • tunnel • alloggi ad angolo per rattiIl numero di interazioni di arricchimento osservate ha evidenziato una forte predilezione per il piano inferiore dell’amaca, indicando che è uno strumento di arricchimento valido.

197August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsIl database sulla salute e il benessere del pesce zebra: stato attuale NICOLA GOODWIN, ROSEMARY KEEBLE E MICHAEL PRICECorrispondenza: ntg22@cam.ac.uk Abstract Nel 2019, è stato creato il primo database di termini sulla salute e il benessere del pesce zebra. In collaborazione con specialisti del pesce zebra e veterinari, abbiamo definito punteggi di gravità, indicatori di benessere, definizioni e immagini al fine di realizzare una risorsa a vantaggio della comunità di pesci zebra. D’allora, il database sul benessere del pesce zebra ha funto da piattaforma per un approccio standardizzato con l’obiettivo di garantire coerenza nel linguaggio e nei punteggi di gravità in tutte le strutture. Ha consentito di sensibilizzare su potenziali problemi di salute associati specificamente al background genetico del pesce o sull’impatto ambientale sulla sua salute. Grazie alla standardizzazione dei termini e dei punteggi di gravità utilizzati, i requisiti di zootecnia possono essere condivisi a livello di comunità per garantire il benessere del pesce, favorire la coerenza in tutte le strutture e promuovere la conformità alla legge inglese The Animals (Scientific Procedures) (ASPA) del 1996. In questa sede, viene discusso l’uso del database presso le strutture acquatiche dell’Università di Cambridge, unitamente ai benefici osservati.

198Animal Technology and Welfare August 2020INDICE DE LA REVISTALa función de los procesos de revisión y aprobación regulatoria en la investigación animal para respaldar la implementación de las 3RInforme de la Dra. Frances Rawle, encargado por el National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), febrero de 2023Correspondencia: Frances.Rawle@nc3rs.org.uk Resumen Este proyecto fue encargado por el NC3Rs a finales de 2021 debido a la preocupación de que los avances en las 3Rs a menudo tardan mucho tiempo en convertirse en uso rutinario. La mayor parte de la investigación en el sector académico en la que se utilizan animales es revisada tres veces: una por el financiador, otra por el Organismo de Revisión Ética y Bienestar Animal (AWERB) local y otra por la Unidad de Regulación de Animales en la Ciencia (ASRU). Mi misión consistía en averiguar en qué medida estas revisiones estaban fomentado la adopción de los avances de las 3R o podrían ser más eficaces a la hora de hacerlo. La mayoría de las entrevistas tuvieron lugar durante la primera mitad de 2022, en medio de cambios significativos en los procesos de ASRU y la Revisión Independiente de la Burocracia de la Investigación del gobierno. Asimismo, existía la preocupación de que los cambios en los procesos de revisión pudieran llevar inadvertidamente a la pérdida de oportunidades para promover mejoras en las 3R. ★ ★ ★Paper Summary Translations

199August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsUn estudio exploratorio de entrevistas sobre las percepciones de investigadores y técnicos acerca del acariciado de ratas SAM BEECHENER, SARAH BROWN, VINCENT BOMBAIL, MEGAN LAFOLLETTE, IGNACIO VINUELA-FERNANDEZ Y ALISTAIR LAWRENCECorrespondencia: Alistair.Lawrence@sruc.ac.uk Resumen Este documento destaca los principales temas que surgieron en el marco de un estudio realizado con técnicos de animales e investigadores para comprender mejor las percepciones sobre el acariciado de ratas (rat tickling), así como los posibles impulsores y obstáculos para la adopción de esta técnica en un entorno de laboratorio. Los entrevistados indicaron que tenían actitudes positivas hacia las ratas y la idea de hacerles caricias, con comentarios positivos sobre el comportamiento social, la inteligencia y la capacidad de interaccionar de las ratas, por ejemplo mediante las caricias de los técnicos en animales y los investigadores.Los participantes indicaron que los obstáculos para una mayor aceptación del acariciado de ratas estaban relacionados con las limitaciones de tiempo, la falta de formación en los aspectos específicos del acariciado de ratas y la forma de interpretar las reacciones de las ratas ante las caricias.Asimismo, se mencionó la preocupación de que las caricias afectaran a la integridad experimental y la necesidad de mantener un distanciamiento profesional de las ratas como animales de experimentación.★ ★ ★Una evaluación de las hamacas para ratas como enriquecimiento ambientalMEGAN HICKMAN, LORRAINE MILLER Y RUTH MACDONAL Correspondencia: Megan.hickman@astrazeneca.com Resumen La investigación ha demostrado que el alojamiento de las ratas en instalaciones más enriquecidas y complejas desde el punto de vista ambiental presenta numerosas ventajas científicas y de bienestar en comparación con un alojamiento convencional. Con el objetivo de aumentar la complejidad del entorno, incluyendo oportunidades para trepar y explorar y proporcionando cobertura adicional, se diseñó un prototipo de hamaca para implementarlo en las jaulas de dos pisos para ratas. Se hizo una evaluación de la hamaca en un grupo de ratas hembra para comprobar la durabilidad, el impacto en el comportamiento y la preferencia sobre otros elementos de enriquecimiento del refugio: • túneles • casas esquineras para ratasEl número de interacciones de enriquecimiento observadas indicó una considerable preferencia por la parte inferior de la hamaca, lo que sugiere que la hamaca es un elemento de enriquecimiento satisfactorio.

200Animal Technology and Welfare August 2020Paper Summary TranslationsLa base de datos sobre salud y bienestar del pez cebra: situación actual NICOLA GOODWIN, ROSEMARY KEEBLE Y MICHAEL PRICECorrespondencia: ntg22@cam.ac.uk Resumen Durante 2019 se creó la primera base de datos en materia de salud y bienestar del pez cebra. En colaboración con especialistas en pez cebra y veterinarios, establecimos puntuaciones de gravedad, indicadores de bienestar, definiciones e imágenes con el objetivo de crear un recurso para la comunidad de pez cebra. Desde entonces, la base de datos sobre bienestar del pez cebra ha proporcionado una plataforma para un enfoque normalizado que garantice la coherencia del lenguaje y la puntuación de gravedad utilizados en todas las instalaciones. Asimismo, la base de datos ha proporcionado la capacidad de concienciar sobre los posibles problemas de salud específicos de los antecedentes genéticos o las repercusiones ambientales en la salud de los peces. Al estandarizar los términos y las puntuaciones de gravedad utilizados, ahora la comunidad puede compartir los requisitos de cría para garantizar el bienestar de los peces, favorecer la coherencia entre las instalaciones y cumplir con la Ley de Procedimientos Científicos con Animales (Animals Scientific Procedures Act) de 1986 (ASPA). Aquí explicamos cómo utilizamos la base de datos en las instalaciones acuáticas de la Universidad de Cambridge y los beneficios obtenidos.

201August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareDecember 2023 Animal Technology and WelfareKey pointsThis paper highlights the main themes which emerged from a study carried out with Animal Technicians and researchers to better understand:• perceptions of rat tickling• potential drivers and barriers to the uptake of tickling in a laboratory environment The interviewees indicated they had positive attitudes towards rats and the idea of rat tickling with positive comments about rats’ social behaviour, their intelligence and their capacity to interact with Animal Technicians and researchers.The participants indicated that barriers to wider uptake of rat tickling including time constraints, a lack of training in the specifics of rat tickling and how to interpret rat responses to tickling.In addition, there was mention of concerns over tickling affecting experimental integrity and the need to maintain professional detachment from rats as experimental animals.49Haven’t the time to write a paper but want to have something published? Then read on!This section offers readers the opportunity to submit 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.What 3Rs idea have you developed?EMMA FILBYMira Building, University of Cambridge, University Biomedical Services,Charles Babbage Road, Cambridge CB3 0FSCorrespondence: emma.filby@admin.cam.ac.ukBased on an ar ticle written for the National Centre for the 3RsApril 2020 Animal Technology and We lfareTECH-2-TECHBackgroundEmma was invited to write an article as a 3Rschampion in NC3Rs ‘Tech 3Rs’ Issue 5, November2019.Here is her response describing how she has used anautomated system to reduce how frequently mousecage bedding is changed without compromisingcleanliness.IntroductionOur unit opened in 2017, during the procurement ofnew equipment we had the opportunity to purchase adigital ventilated rack system from Tecniplast UK. Thecages are referred to as the Digitally Ventilated Cage orDVC. This system uses the data collected by sensorsbelow the cage to flag when to clean out based on thechange in an electromagnetic signal. To have thisfunctionality we first needed to create an algorithmduring a learning phase.The learning phase: devising analgorithmWe held a meeting to agree what warranted a cage basechange based on pictures to avoid being subjective. Wereferred to the Home Office Codes of Practice for thehousing and care of animals bred, supplied or used forscientific purposes (HOCoP) for advice on husbandr ypractices to set our criteria, balancing hygiene and theimportance of olfactory cues to rodents and their needfor control over their environment.1We started the trial, noting when the cage reached thepoint it required a base change. We assessed airquality, what proportion of the cage base was wet andwhether the animals still had choice over theirenvironment and their ability to show spatial separationof different behaviours such as nesting and excretion,for example their nest was free of faeces. During the‘learning phas e’ we as ke d our Named VeterinarySurgeon (NVS) and Home Office inspector (HOI) tocheck that they agreed with our assessment.APRIL_1-628207435_4-628196990.e$S:Animal Technology and Welfare 24/9/20 06:51 Page 49TECH-2-TECHAn exploratory interview study of researchers’ and technicians’ perceptions of rat ticklingSAM BEECHENER1, SARAH BROWN2, VINCENT BOMBAIL3, MEGAN LAFOLLETTE4, IGNACIO VINUELA-FERNANDEZ5 and ALISTAIR LAWRENCE2,31 Centre for Epidemiology & Planetary Health (CEPH), Dept of Veterinary and Animal Science, An Lòchran, Inverness IV2 5NA 2 The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh EH25 9RG3 Animal Behaviour & Welfare, Department of Animal & Veterinary Sciences, Roslin Institute Building EH25 9RG4 The North American 3Rs Collaborative, Denver, CO, 80202, USA5 Bioresearch & Veterinary Services, The University of Edinburgh EH16 4SBCorrespondence: Alistair.Lawrence@sruc.ac.uk

202Animal Technology and Welfare August 2020Introduction Rat tickling was fi rst developed by neuroscientists to mimic rat play behaviour using the human hand to play with the rat1 (Figure 1).Letters indicate rat behaviours that share similar or the same physical characteristics: • pouncing• nape contact• pinning• chasing• boxing•fl ippingDuring social play behaviours can occur in any order and do not always occur in each play bout. During tickling the sequence is always B, F, C. Re-drawn from4 Original drawings by Tayla Hammond. These motions were similar to human tickling, hencethe term rat tickling. Rat tickling subsequently becameof interest in Animal Welfare as a practical approach toprovide social enrichment to and reduce handling stressfor laboratory rats.3,5 Rat tickling is also relevant withincreased interest in positive Animal Welfare indomesticated animals (https://www.positiveanimalwelfare.net/). Finally, rat tickling has even become known in thepopular press (e.g https://www.youtube.com/watch?v=d-84UJpYFRM); one reason for this wider interest may bethat rats emit ultrasonic vocalisations (USVs) which havebeen interpreted as rats laughing when being tickled6https://www.positiveanimalwelfare.net/research/rat-tickling/). (Figure 1) These drawings illustrate the behaviours seen in rats during: • social play (Pellis and Pellis, 2013) • during the standard version of tickling (Cloutier et al., 2018) Letters indicate rat behaviours that share similar or the same physical characteristics: • pouncing Figure 1. Figure 2 The work reported here was an addition to a National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) funded project on refining rat tickling (figures 3 and 4) (https://www.nc3rs.org.uk/our-portfolio/refinement-tickling-protocols-improve-positive-animal-welfare-laboratory-rats). Our aim was to extend the previous social science research on rat tickling by interviewing United Kingdom (UK) based Animal Technicians and researchers who Figure 2.These drawings illustrate the behaviours seen in rats during: • social play2•during the standard version of tickling3An exploratory interview study of researchers’ and technicians’ perceptions of rat tickling

203August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAn exploratory interview study of researchers’ and technicians’ perceptions of rat ticklingThere has been previous social science research on rat tickling led by Megan LaFollette (fi gure 2) https://www.linkedin.com/in/megan-lafollette/) including: a cross-sectional survey of laboratory animal personnel in the United States of America (USA) and Canada including their implementation of and perceptions towards rat tickling7 as well as a longitudinal study of the effects of training laboratory animal personnel in rat tickling (US based).8The work reported here was an addition to a National Centre for the Replacement, Refi nement and Reduction of Animals in Research (NC3Rs) funded project on refi ning rat tickling (fi gures 3 and 4) (https://www.nc3rs.org.uk/our-portfolio/refinement-tickling-protocols-improve-positive-animal-welfare-laboratory-rats). Our aim was toextend the previous social science research on rat tickling by interviewing United Kingdom (UK) based Animal Technicians and researchers who use rats in their studies. The questions also allowed expansion into moregeneral attitudes to the animals and human-animal interactions in the research environment.ApproachIn these exploratory investigations into perceptions of rat tickling, we conducted twelve interviews with a mixture of researchers and Animal Technicians (6 of each) working with rats in a laboratory environment. Interviews took place virtually and for consistency followed an outline topic guide (Appendix A). Fieldwork took place either virtually or face-to-face between October 2022 and January 2023. Participants were identifi ed through university veterinary services and invited via an introductory email to participate in the study. All the interviews were carried out by the same interviewer and varied between 20 minutes and 30 minutes duration. Interviews started with a recap of thestudy’s aims and sought informed consent (Appendix B)to proceed with the interview and to record the discussion. Recordings were subsequently transcribed and given the technical nature of some content, draft transcripts were reviewed for accuracy by the interviewer. A simpledeductive thematic analysis9,10 was followed whereby scripts were analysed according to the issues beinginvestigated, that is: • participants’ general experience of rats • their understanding of rat tickling • perceived benefi ts and disbenefi ts of rat tickling (for the animal, its handler, and wider research activities)• barriers to uptake of rat tickling A summary of emerging themes was shared with participants for their feedback and their responses were incorporated in a fi nal draft.FindingsWorki ng environmentIn a refl ection of their different roles and responsibilities, contact between researchers and the rats in their studiestended to be more intermittent whereas Animal Technicianswere in day to day contact with the rats in their care. Researchers were variously investigating rats as models for behavioural neuroscience or aging with variation in whether they were working with pups, juveniles or adult rats. In some cases, rats were housed individually. In this case efforts were made to ensure contact with rats use rats in their studies. The questions also allowed expansion into more general attitudes to the animals and human-animal interactions in the research environment.Figure 3Figure 3.Figure 4 Approach: In these exploratory investigations into perceptions of rat tickling, we conducted twelve interviews with a mixture of researchers and Animal Technicians (6 of each) working with rats in a laboratory environment. Interviews took place virtually and for consistency followed an outline topic guide (Appendix A). Fieldwork took place either virtually or face-to-face between October 2022 and January 2023. Participants were identified through university veterinary services and invited via an Figure 4.

204Animal Technology and Welfare August 2020in neighbouring cages to reduce isolation. Otherwise the rats tended to be kept in small groups. Among researchers the levels of interaction with the rats in their studies varied. For example one respondent described daily handling over the last 2 years whilst another researcher was interacting with the rats in their study twice per week for purposes of checking, weighing and handling. Interactions between the other researchers and the rats in their studies were more intermittent or cyclical with daily handling during their experiments but zero handling at other times as their focus turned to analysis and reporting.Animal Technicians were responsible for the day to day care and general husbandry of the rats including, routine feeding, watering, cleaning and marking as well as having some recording responsibilities. Their roles involved considerable handling and interaction with the rats, checking for wellbeing and signs of ill health and liaising as necessary with the veterinarian. There were also mentions of supervising the mating/breeding programmes, administering injections and maintaining cages of handling rats to help familiarise new Animal Technicians and researchers.ThemesSimilar and yet different perspectives from researchers and Animal TechniciansOverall there was a sense of responsibility to the rats with words reflecting respect, some affection and a sense of pride in the scientific work being conducted. Four of the researchers mentioned they had previously worked with mice and for two, working with rats was felt to open up new and exciting opportunities given the rich social repertoire of rats and the opportunity to establish relationships with them based on their friendly and inquisitive nature. One researcher mentioned they were often surprised by the intelligent behaviours demonstrated by rats that left them wondering “...wow, how is it possible that you learn how to do that?”. Another researcher spoke of forming a particular bond with some of their rats, noting that “...there are always three or four that are my favourites ...they are more friendly, enjoy human contact and seek out interactions”.Among Animal Technicians, some worked only with rats and some worked with rats and mice. Levels of experience varied from those that had started relatively recently to those with medium (4-5 years) and longer term (12-20 years) experience. One Animal Technician commented on the challenge of recruiting younger staff to work with rats observing that “...the workload, the size of the rats, the size of the cages puts them off …and a fear of being bitten given the size of the teeth”.There was a shared concern to enrich the laboratory environment. For example, one Animal Technician spoke of the play tunnels and chew sticks they saw as important for example to avoid development of “...stereotypical behaviours that can be detrimental”. Two Animal Technicians mentioned rats’ well-developed sense of smell, speaking of being ‘welcomed’ when they come into the room (by the same token, noting that rats may be apprehensive of newcomers). In common with participating researchers, Animal Technicians spoke of rats’ intelligence that makes working with them potentially highly rewarding. Knowledge and experience of rat ticklingAmong participants, existing levels of knowledge and personal experience of rat tickling were varied. Some respondents were less confident of their knowledge while others felt better informed. Both researchers and Animal Technicians acknowledged that even if colleagues in the same facility were applying tickling practices the nature of their work meant it would not always be visible to others “...it’s such a solitary life doing behavioural experiments, we’re just each in our own rooms. I don’t see my lab-mates do their experiments…” (researcher). Tickling was described by some as ‘an enrichment’ with interest expressed in the vocalisations that accompany tickling by way of developing a better understanding of its impact. Among Animal Technicians, one mentioned how tickling was embedded as a core function in a previous role and the challenge they faced in making time to incorporate tickling in their current post. Another three participants described implementing their own interpretation of tickling as a way of interacting with the rats, taking their cue from the rats as to how much or how little was appropriate. On being asked how they might describe rat tickling to a lay person, responses varied. Researchers tended to mention aspects of the interaction and enjoyment of the rat. One likened the approach to “...finding the rat’s sweet-spot in the same way that you might with a pet to encourage positive behaviours”. Another referred to the process of “...turning the rats over to tickle their tummies”. A third spoke of familiarising the rat with the hand through a process of touching and interaction with a view to“...forging a relationship”. A fourth researcher described tickling as: – An interaction that is pleasurable for the rat as it mimics natural play.– From a scientific point of view, an intervention that can be controlled and better controlled than other enrichments such as free play that is also of value in measuring neural responses.An exploratory interview study of researchers’ and technicians’ perceptions of rat tickling

205August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAn exploratory interview study of researchers’ and technicians’ perceptions of rat ticklingAmong technicians, one spoke of tickling as “…an enrichment that is beneficial for the rats and helps to give them a better life”. Another reflected on tickling as a kind of “...replication of social behaviour” by recreating the passive aggressive turning over of one rat by another. Other interpretations emphasised the interaction between the handler and their rats. One suggested that it was perhaps more about “...play-fighting rather than tickling, per se” and making time to handle the rats more than usual. Another Animal Technician described the motion of the hands and movement of the fingers but emphasised also the importance of ensuring that the “...rat understands it’s not threatening” (demonstrated by the rat coming back for more and not showing signs of avoidance e.g. stiffening-up or cowering). A third framed tickling as a behavioural approach to “finding out more about the animal” (and they likened it to interactions between mother and offspring) and by getting to know rats better and “...improving its well-being within the confines of the lab”. There was also mention of tickling as a ‘two way street’ speaking of the benefits for the rat and the handler akin to the pet owner relationship.Perceived benefits/disbenefits of tickling for the ratTickling was broadly perceived as a positive interaction for most but not all rats. There was an emphasis on taking time to introduce the practice ideally to young rats and being sensitive to the level of interaction being sought by the rat. One researcher recalled having read that “...five minutes of tickling is better than, say, half an hour of interacting with no clear purpose” and felt this was borne-out by their own experience of pups seeking to engage by “running to my hand ... it’s subjective but it’s telling me something”.In the context of the laboratory environment, researchers identified several potential benefits for rats that may be housed individually, in helping them become accustomed to the blue gloves that have to be worn, by reassuring them that fingers ‘...are not trying to hurt them’ and thereby helping to reduce levels of stress, keeping animals ‘happy and friendly’ and through this interaction, helping handlers be alert to any changing ‘mood or condition’. Together, this was seen as increasing the likelihood of obtaining good results from the laboratory work itself. Animal Technicians reflected on the differences between handling and tickling, reasoning that the latter may help to habituate rats more quickly and in a more enjoyable way, encouraging them to seek out interaction and reducing associated stress. Animal Technicians sought guidance on tickling and were concerned about getting it right and to comply with the requirements of different experiments so as not to risk compromising the results. Reflecting on their own workloads and day to day pressures on their time, they were also concerned about the consequences of habituating rats to tickling if resources were not there to support ongoing interaction over a rat’s lifetime. Perceived benefits/disbenefits for researchers and Animal TechniciansResearchers and Animal Technicians saw potential benefits from tickling. However there were concerns about experimental integrity and accommodating tickling in day to day routines given the competing demands on their time. Most agreed that more information about tickling would be of value:• In helping them to understand how it could impact on experiments.• In how it might be applied consistently to permit an informed choice to be made.Among researchers, some expressed concerns about the potential for tickling to interfere with or confound the integrity of experimental studies. Others felt that given the number of existing variables, tickling was unlikely to have a significant impact. Among those that were concerned, some wondered if it might best be introduced into breeding stock rather than the rats used in experiments. One spoke of the behaviour trials they were conducting and the importance of avoiding the introduction of anything that might influence the outcomes they were observing. They wondered if some form of self administered approach that avoids human interaction, e.g. back-scratchers, may have an application. As regards the need for consistency, there were queries over whether the varying amount or dose of tickling between rats might affect experimental outcomes. One researcher reflected on the Animal Technicians’ relationship with their rats and a tendency to have favourites, wondering how this might impact on the results of the wider study. Two of the researchers challenged the assumption that tickling was always positive and queried how to allow for rats that do not enjoy the experience. One of the participating researchers highlighted the significant levels of investment required to allow their experiments to go ahead and the associated risk that comes with compromising the integrity of this work. Among other researchers, tickling was associated with reducing stress. In one case (working with older rats) it was felt that “...allowing the rats to become inactive/obese represents a greater risk to the integrity of a study”.In the second case it was argued that as much as researchers try to control variables, the experience of each rat varies and in this context “...anything that helps the rats to feel comfortable and display natural behaviours has to be positive”.

206Animal Technology and Welfare August 2020In the third case, the researcher saw tickling, along with efforts to improve the housing and the habitat of rats as areas of continually evolving improvement. Crucially, believing that “I think it has to be better if the animals are less stressed”. One researcher spoke of the potential benefits of tickling for their experiments with rats that they recovered more quickly from surgery and were able to proceed sooner to the next stage. They were less apprehensive when confronted by new tasks and able to move more quickly into performing these tasks. Another commented on their perception that tickled rats were habituating more quickly to them, with less variability between animals, noting that “...on the first day there’s some that don’t seem that keen, on the second day none of them seem to mind any more judging by how floppy and relaxed they seem”. A third researcher expressed their wish for rats to have a good quality of life, with benefits for rats and Animal Technicians in having a good relationship and making the routine tasks, such as taking weights and monitoring blood pressure, less unpleasant for all concerned. Nevertheless, there was recognition that tickling is not risk free with the possibility of receiving a ‘really nasty bite’ from any interaction with a rat.From the Animal Technicians’ perspective, more ‘tame’, more ‘approachable’, more ‘amenable’ and more ‘interactive’ rats were perceived as a good thing for those handling them. Procedures involving tickled rats were described as going more smoothly with reduced risk of being scratched or bitten by rats that are more easily handled. Some concerns were expressed that rats will start seeking interaction, for example by pressing their noses up against the cage door, and although “...attention seeking is not necessarily a bad thing, it could become an issue”. Overall, applying tickling in practice, comes down to a ‘question of time’. One Animal Technician acknowledged the privilege of working in a specialist unit with fewer animals that allowed them sufficient time to “...say ‘hello’ to each of them, every day”. Others however, described competing demands on their time. One Technician noted “...there’s not time to do that [tickling] in some of the other units where I provide weekend cover but that’s just how it is”. Another Animal Technician spoke of the ad-hoc approach they had developed when time permits. They were spending 5 to 10 minutes tickling cages at weaning in the belief that the rats seemed to be more interactive thereafter. This sense of prioritising was echoed elsewhere with two Animal Technicians recognising that some strains seem to be more aggressive than others, prompting them to wonder if these rats should be the more immediate priority. Another mentioned cage-changing or single-housed animals as the prompts for tickling so that eventually you would work your way around the entire house. There was also consideration about who should do the tickling and this prompted one Animal Technician to suggest for the researchers to be more involved as “...even with some animals that are very relaxed with me, there can be something of a tense atmosphere between them and the researchers”. Animal Technicians were pragmatic about the need to balance interactions with individual animals with the day-to-day challenges of caring for the wider group. The majority of participating Animal Technicians were positively disposed, time permitting, to the idea of doing more tickling in the future and it was broadly seen as a positive thing. That said, they would welcome guidelines as to its application. One respondent summed it up by saying “I’m always up for change. That’s what science is all about, I suppose, isn’t it?”.If it improves Animal Welfare then it should be encouraged” with another observing “I think it’s a good thing and I think improving the interactions with the rats will help to encourage staff to work with rats e.g. the rats coming to you instead of running to the back of the cage and I would love to see that happen”.DiscussionKnowledge of ticklingKnowledge of tickling was in some cases related to use of tickling, but this was not always the case. Across all levels of knowledge about tickling there seemed to be some uncertainty about how to specifically carry out tickling and whether the hand movements that interviewees were currently using in practice could be termed tickling. There was recognition from some that tickling should not be forced on rats and that it may not be appropriate in all circumstances e.g. post-surgery; with some genetic strains. There were also variable levels of confidence about how to interpret rats’ response to tickling: rats engaging in the activity and actively seeking out the hand and coming back for more were mentioned as potential indicators of positive responses of rats to tickling. Another aspect that was mentioned on several occasions was the use of rats’ vocalisations to assess rat welfare. Both researchers and Animal Technicians expressed interest in these vocalisations which are mainly in the ultrasound and require specialised equipment to monitor them (https://www.positiveanimalwelfare.net/research/ rat-tickling/). An exploratory interview study of researchers’ and technicians’ perceptions of rat tickling

207August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAn exploratory interview study of researchers’ and technicians’ perceptions of rat ticklingTraining in tickling Linked to knowledge of tickling, these interviews also suggest the development of training to impart knowledge and confidence in how to tickle. Training materials on rat tickling do exist, for example on the NC3Rs and the 3RsC websites (https://nc3rs.org.uk/3rs-resources/rat-tickling; https://www.na3rsc.org/rat-tickling-faq/). There is also evidence that training can improve key outcomes for rat tickling such as increased self-reported knowledge and self efficacy.11 This raises the question of how best to incorporate training on rat tickling in the UK (this could be seen as a broader issue in how to create ‘behavioural change’ with respect to rodent welfare (e.g. stress free handling)). Possibilities include raising more awareness of rat tickling and other pro welfare approaches into Personal Licence (PIL) accreditation with more direction towards available training materials. Rat tickling (and other pro welfare approaches) could also be highlighted in Guidelines such as PREPARE.12 Barriers to uptake of ticklingThese interviews reinforce existing understanding of the barriers that prevent further uptake of rat tickling.7 The main constraint mentioned here was the time availability for tickling and the practical considerations of how to fit rat tickling into the busy day to day routine of both Animal Technicians and researchers. Several interviewees asked for guidance on how much and how often tickling needs to be administered to create a positive response in the rats. This links again to the need for more knowledge and training on rat tickling. Previously11 investigated the question of time and determined only 15 seconds of tickling for 3 days was enough to be effective. In addition there were also concerns about the effects of tickling on the repeatability of the research and that the act of interacting with rats through tickling may be at odds with a perceived need for professional detachment from experimental animals. This last point was referred to, for example, at the end of the experiment when the animals which had been interacted with would need to be culled. There was also reference to potential divergence of views on this point between researchers and their supervisors suggesting the need for more discussion and mentorship on the development of bonds between researchers and their rats. AcknowledgementsWe acknowledge the funding from NC3Rs grant NC/W001209/1 which supported this work. We would also like to acknowledge the time given freely by the Animal Technicians and researchers interviewed for the study and all the insights they provided. References1 Burgdorf, J., & Panksepp, J. (2001). Tickling induces reward in adolescent rats. Physiology & Behavior, 72(1–2), 167–173. https://doi.org/10.1016/s0031- 9384(00)00411-x2 Pellis, S.M., Pellis, V.C., Ham, J.R., & Stark, R.A. (2023). Play fighting and the development of the social brain: The rat’s tale. Neuroscience & Biobehavioral Reviews, 145, 105037. https://doi.org/10.1016/j.neubiorev.2023.1050373 Cloutier, S., LaFollette, M.R., Gaskill, B.N., Panksepp, J., & Newberry, R.C. (2018). Tickling, a Technique for Inducing Positive Affect When Handling Rats. Journal of Visualized Experiments, 135. https://doi.org/ 10.3791/571904 Bombail, V., Brown, S.M., Martin, J.E., Meddle, S.L., Mendl, M., Robinson, E.S.J., Hammond, T.J., Nielsen, B.L., LaFollette, M.R., Vinuela-Fernandez, I., Tivey, E.K.L., & Lawrence, A.B. (2022). Stage 1 Registered Report: Refinement of tickling protocols to improve positive animal welfare in laboratory rats. F1000 Research, 11, 1053. https://doi.org/10.12688/ f1000research.125649.25 LaFollette, M.R., O’Haire, M.E., Cloutier, S., Blankenberger, W.B., & Gaskill, B.N. (2017). Rat tickling: A systematic review of applications, outcomes, and moderators. PLOS ONE, 12(4), e0175320. https://doi.org/10.1371/journal.pone.01753206 Panksepp, J., & Burgdorf, J. (2000). 50-kHz chirping (laughter?) in response to conditioned and unconditioned tickle-induced reward in rats: effects of social housing and genetic variables. Behavioural Brain Research, 115(1), 25–38.7 LaFollette, M.R., Cloutier, S., Brady, C., Gaskill, B.N., & O’Haire, M.E. (2019). Laboratory animal welfare and human attitudes: A cross-sectional survey on heterospecific play or “rat tickling.” PLOS ONE, 14(8), e0220580. https://doi.org/10.1371/journal.pone. 02205808 LaFollette, M.R., Cloutier, S., Brady, C.M., O’Haire, M.E., & Gaskill, B.N. (2020). Changing Human Behavior to Improve Animal Welfare: A Longitudinal Investigation of Training Laboratory Animal Personnel about Heterospecific Play or “Rat Tickling.” Animals, 10(8), 1435. https://doi.org/10.3390/ani100814359 Vaismoradi, M., Turunen, H., & Bondas, T. (2013). Content analysis and thematic analysis: Implications for conducting a qualitative descriptive study. Nursing & Health Sciences, 15(3), 398–405. https://doi.org/ 10.1111/nhs.1204810 Braun, V., & Clarke, V. (2006). Using thematic analysis in psychology. Qualitative Research in Psychology, 3(2), 77–101. https://doi.org/10.1191/1478088 706qp063oa11 LaFollette, M.R., O’Haire, M.E., Cloutier, S., & Gaskill, B.N. (2018). Practical rat tickling: determining an efficient and effective dosage of heterospecific play. Applied Animal Behaviour Science. https://doi.org/ 10.1016/j.applanim.2018.08.00512 Smith, A.J., Clutton, R.E., Lilley, E., Hansen, K.E.A., & Brattelid, T. (2017). PREPARE: guidelines for planning animal research and testing. Laboratory Animals, 52(2), 135–141.https://doi.org/10.1177/002367721 7724823

208Animal Technology and Welfare August 2020Appendix A The question schedule used to organise the interviews (potential follow up questions indicated*): General questions– Are you a regular handler of rats?* How would you describe your role (e.g., technician, user of rats).– How familiar are you with rat tickling?– If you were asked to describe rat tickling to a friend how would you describe it? – Have you used rat tickling previously? * (Answer will determine how the next questions are framed – see under Yes or No below).* if Yes can you describe how you have previously tickled rats. * if Yes can you tell me for what reason you tickled rats.Positives– If Yes to previous question: Could you tell me about the positives you have experienced from rat tickling? * What do you think might be other positives e.g. to the rat or to the researcher?– If No: Could you tell me what positives you think could come from rat tickling? * What do you think might be other positives e.g. to the rat or to the researcher? Negatives– If Yes: Could you tell me about the negatives you have experienced from rat tickling? * What do you think might be other positives e.g. to the rat or to the researcher? – If No: Could you tell me what negatives you think could come from rat tickling? * What do you think might be other positives e.g. to the rat or to the researcher? Facilitating use of tickling– If Yes: Reflecting on your current use of rat tickling, what do you think would increase/ decrease your use of tickling? – If No: Reflecting on what you know about rat tickling what do you think would increase/ decrease your use of tickling?Balance of positives and negatives– Do you think you will be more likely or less likely to use rat tickling in the next 6 months?* Do you think others should be encouraged to use rat tickling? Appendix B Information and Consent Sheet:You are being asked to participate in a short interview and/or participant in a focus group discussion to examine your thoughts on Rat Tickling protocols to improve positive Animal Welfare in laboratory rats being carried out by SRUC and the University of Edinburgh and funded by NC3Rs.Background to projectThis is part of a larger experimental study funded by NC3Rs https://www.nc3rs.org.uk/our-portfolio/refinement-tickling-protocols-improve-positive-animal-welfare-laboratory-rats. The aim of the project is to improve the way that rats are tickled to enhance their welfare. As a part of this, we would like to understand more about how people who use rats in their research perceive rat tickling. We hope that this understanding could help in improving the uptake of rat tickling in practice. Aim of this studyTo explore and better understand how Animal Technicians and individuals who use rats in their research perceive rat tickling and what may constrain or encourage the uptake of rat tickling in laboratory contexts. ParticipationYour participation in this research is voluntary. You can refuse to take part and you can change your mind and withdraw from this study at any time without having to give an explanation. Ensuring your anonymity– All information about you will be treated in strict confidence. – No potentially identifiable information will be used. – When project progress is discussed with others and when findings are reported and published, only anonymised information and quotations will be used.The privacy policy that will cover this work can be found here: https://www.sruc.ac.uk/connect/about-sruc/policies-compliance/compliance/privacy-policy-gdpr-cookies/#:~:text=Information%20we%20collect%20and%20use&text=Special%20category%20data%20(or%20sensitive,in%20your%20relationship%20with%20SRUC What taking part will involveYou will be asked to take part in short one-to-one research interviews online (on MS Teams) with Prof. Alistair Lawrence. This is a longitudinal study so we would potentially like to talk to you at more than one point in time i.e. we would aim to interview you at the start, middle and end of this study. This is expected to take about 30 minutes but not last more than 45 An exploratory interview study of researchers’ and technicians’ perceptions of rat tickling

209August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAn exploratory interview study of researchers’ and technicians’ perceptions of rat ticklingminutes. In these interviews you will be asked several questions relating to what you think about the use of rat tickling and what you think may constrain or encourage it’s use. We may also conduct a group focus group session with other participants, which you may also be asked to attend and share your views and experiences of rat tickling. This is expected to take no more than one hour. Data collection, analysis and findingsThis project will use Teams as the secure environment to collect and store data. With your consent, an audio recording using Teams will be made of your answers which will be given an anonymous identifier. These recordings will be stored on Teams and from there downloaded to Otter.ai which will be used to transcribe the recordings into a text file. These text files will be corrected on Otter.ai before being uploaded back to Teams for secure storing prior to analysis at which point the files on Otter.ai will be deleted. The audio recording will be deleted at the end of the project. If you decide to withdraw any of your data (whether from individual interviews or focus groups) will be deleted immediately. The overall findings of this study will be reported to NC3Rs. Insights and perspectives you provide may be included in publications (e.g. academic papers, media articles, and policy reports). Contact If you have questions at any time about the study or the procedures, you may contact Alistair Lawrence at alistair.lawrence@sruc.ac.uk By signing the below you agree that• You have read the above information• You voluntarily agree to participate• You understand that participation will involve a recorded interview (of about an hour). • You agree that your responses will be used for the purposes of the research outlined above and reported in publications (e.g. media reports, policy briefs and academic papers), which may include your anonymised information and quotations (No information which could identify me you will be shared). • You are 18 years of age or olderI freely give my consent to participate in this research study and have been given a copy of this form for my own information.Signature: ___________________________Date: ___________________________

Copies can be ordered via the IAT website www.iat.org.uk/booksDo you need a reference book? If so, have you thought about ILASTW? This is an excellent entry level learning resource, ideal for those studying for first level qualifications. The book has been thoroughly revised into its third edition and is available to purchase as a download file and as a hard copy. This book is competitively priced – the hard copy at £20.00 (packaging and postage are included) and at £15.00 for the pdf download version.If you are seeking multiple copies of the book for your facility or department, a bulk purchase price is available. Details and an order form can be found on the IAT website – www.iat.org.uk/booksl Paperback: 232 pagesl Publisher: Institute of Animal Technology (November 2017)l Language: Englishl ISBN-13: 978-1-9999168-0-0l Cost: £20.00 (includes free p&p within the UK – overseas postage will be charged)Are you undertaking IAT Level 2 and 3 qualifications?

211August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareIAT graduation special: Congratulations to all the successful HE studentsThe 2023 Graduation CeremonyIAT Diploma Levels 4, 5 and 6 Laboratory Animal Science and Technology Awarded by the Institute of Animal TechnologySTEVEN CUBITTCorrespondence: info@iateducation.co.ukDecember 2023 Animal Technology and WelfareEducationSponsored by:Please email Robert Orvis: robert.orvis@astrazeneca.com to book your team and their meal option. Choices are: Beef or Chicken or Spicy-Bean burger meal.6 persons per team at £10.00 each as Agenda Life Sciences is generously sponsoring the remaining £8.95 per person.Note: Teams of fewer than 6 in a team are welcome, but the minimum cost is £60. Prizes for the highest scoring team and individual.Booking & Payment - in advance.Latest booking date: 10th May BACS payment to: HSBC - Cambridge I.A.T. branch 40-40-45 41322745 Reference: Your team nameWednesday 17th May 2023Tenpin Cambridge Leisure Park, Clifton Way, Cambridge, CB1 7DYArrive at 5.30 - Bowling starts at 6pmIAT Council would like to congratulate all the IAT Higher Education (HE) students for their outstanding achievement in passing their exams.

212Animal Technology and Welfare August 2020On Tuesday 4th July 2023 the graduation ceremony took place at the MRC Mary Lyon Advanced Training Centre for students who had completed the IAT Diploma Levels 4, 5 and 6 of Laboratory Animal Science and Technology. It was a wonderful day enjoyed by students, their families and friends who could share in their success. Most important of all, IAT Council, College of Laboratory Animal Science and Technology (CLAST) lecturers and trustees congratulate the Higher Education (HE) students for their hard work and dedication. It was great to see some newly elected Fellows of the IAT.For the day we are indebted to the IAT’s good friend Professor Sir Richard Gardner FRS for being the Guest of Honour, Mary Lyon Advance Training Centre for hosting and organising the graduation. A special thanks also to the MLC Advance staff who looked after everyone so well throughout the day. Ken Applebee for being Announcer and Linda Horan as Chair of the IAT for their roles in the ceremony. The following Students were awarded the IAT Diploma Level 4 Laboratory Animal Science and Technology:Bruna Delfini 2019Dimitra Mantzorou 2019Emma Mustafa 2019Kyle O’Sullivan 2019Aleksandra (Ola) Czerniak 2020Chelsea Cavanagh 2020Gemma Dykes 2020Jane Holby 2020Emily Thorpe 2020Athanasios Vouzas 2020Rachel Blackburn-Stout 2021Susan Fotheringham 2021Kelly Frater 2021Justyna Glegola 2021Jamie Kelly 2021Emma Scott 2021Jenny Bartley 2022Nicola Evans-Bailey 2022Kat Gowan 2022Amy Ferreira 2023Busisiwe Mogodi 2023Sian Murphy 2023IAT graduation special: Congratulations to all the successful HE students

213August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe following Students were awarded the IAT Diploma Levels 5 Laboratory Animal Science and Technology:Victoria Hearn 2019Victoria Lacey 2019Chelsea Cavanagh 2020Casey Dalling 2020Sarah Drummond 2020Louis Kitchenham 2020Bruna Delfini 2022Justyna Glegola 2022Jamie Kelly 2022Emma Mustafa 2022Kyle O’Sullivan 2022Emma Scott 2022The following Students were awarded the IAT Diploma Level 6 Laboratory Animal Science and Technology:Carmen Abela 2019Richard Berks 2019Karen Fry 2019Ian Bagley 2020Matthew Bilton 2020Emma Hamilton 2020Jolene Hammonds 2020Ania Sobczak 2020Emma Tozer 2020Francesca Flack 2021Lisa Watson 2021Cheryl Yalden 2021Kerry Lavin-Thomson 2022Michelle Scutter 2022Lisa Wright 2022Georgina Orosz 2023IAT graduation special: Congratulations to all the successful HE students

214Animal Technology and Welfare August 2020We are in exciting times with our completely revised Higher Education Continuing Professional Education system that begun delivery in September 2023. The new HE system has been developed in conjunction with CLAST and Mary Lyon Advance Training Centre, who have put incredible efforts in to developing the new syllabus based on their previous experience and student feedback which is world leading in our field. In addition, thanks go to: – The new teaching staff for developing the new lesson plans and online material.– The Board of Education Policy for overview and input into syllabus.– Jock Fraser for coordination of new units with Ofqual.– A special thanks to Brian Lowe as outgoing CLAST programme lead. Overview of the IAT HE programmeThe IAT acts as the professional body for persons engaged in laboratory animal science, supporting the interests of Animal Technologists in the United Kingdom (UK) and abroad. As a recognised Ofqual Awarding Organisation the IAT develops, manages and quality assures qualifications that are widely recognised within academia and the biotechnology industry in the UK and abroad.The IAT HE programme aims to provide an internationally recognised education programme that ensures Animal Technologists and other laboratory science professionals are equipped to meet their legal and ethical responsibilities in providing high standards of care and welfare for laboratory animals. It also provides the knowledge and skills for the future advancement of Animal Welfare, good science and to ensure Animal Technologists are suitable for personal and career advancement.How can you benefit from our courses?There are lots of benefits to be gained from studying with us at any level and the further you progress through our qualifications the more you stand to gain. Each of our units has been designed to develop your skills profile and will give you the chance to:– Learn about complex biological processes related to animals in research.– Develop and refine your critical thinking and writing abilities.– Maintain and improve Animal Welfare within your facility.– Develop your soft skills in management, negotiation and communication skills.– Perform experimental procedures safely and humanely.– Increase your job satisfaction and chance of career progression.– Provide you with valuable professional networking opportunities.– Learn to deliver confident and informative presentations and reports.– Graduate with an IAT HE certificate that can be used towards a full HE qualification in the future.How can your employer benefit from your studies?As an employer you want to know that you are making a worthy investment in your employees’ education and training. We strive to train laboratory professionals to the highest standards and work closely with many large employers of laboratory staff including the Universities of Edinburgh, Oxford, Cambridge, King’s College London, AstraZeneca and Cape Town. The benefits of higher education and training in animal science and technology are not just for your employees, your organisation stands to gain just as much, including:– Increased staff loyalty and sense of being valued by the employer.– Greater staff retention, especially whilst they are studying.– Be safe in the knowledge that your staff are experts in Animal Welfare, helping to maintain a Culture of Care.– Students will develop interpersonal skills to facilitate working effectively with a diverse range of stakeholders in a team environment.– Students learn cutting edge approaches from industry experts, equipping staff with a range of practical skills to bring to the workplace.– Our wide range of modules meet Continual Professional Development (CPD) needs and can be incorporated into staff appraisals.– Staff members who are confident public speakers with improved communication and presentation skills.– Efficient employees who can prioritise and manage their time effectively.– Employees with developed scientific writing skills for use in the company.Animal Technology requires integration with a broad range of disciplines. Senior Animal Technologists are expected to communicate effectively with a range of key personal including scientists, veterinarians, Named Persons under the Animal (Scientific Procedures) Act (ASPA), building engineers and other technical professions (for example histology and pathology). They require a broad knowledge of many disciplines and the ability to understand the key points of scientific practice, building design and management, good animal management and welfare, disease recognition and control and a thorough understanding of the key legislation controlling the use IAT graduation special: Congratulations to all the successful HE students

215August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfareof animals for research. The HE programme provides a wide range of skills, knowledge and behaviours which help the students integrate effectively with their professional colleagues.As science and Animal Welfare are rapidly evolving areas there is a responsibility on all Animal Technologists to continually update their skills and knowledge base. This is something all of these HE students have demonstrated very effectively over the course of their studies.Studying at any level is hard but when you must balance your studies with a full time job and commitments to family and friends it can become very difficult. Although the HE units are now delivered through a mix of formal taught sessions, tutorials, group work and practical activities, most of the work is done by the students independently, in their own time. This requires high levels of commitment and effort by the student and support from their employers for successful outcomes. It requires the students to develop excellent time management and prioritisation skills to juggle their professional and personal commitments successfully.Objectives of assessment processThe HE qualifications are designed to:– Recognise learning and prepare the learner for further learning and training and to develop knowledge and skills in Animal Technology.– Prepare learners for further learning and training by establishing a broad and relevant base of knowledge and skills in Animal Technology.– Develop knowledge and skills in laboratory animal science and technology specialist areas which enhance the learners’ abilities to take independence and autonomy in their work role.– Encourage updating of skills and knowledge and CPD.– Relates knowledge and skills to legal, technical, process or good practice requirements in the field of laboratory animal science and technology.– Develop knowledge and skills in order to gain recognition at a higher level or in a different role.– Develop knowledge and skills relevant to a particular specialisation within an occupation or set of occupations.The broad objectives of the assessment process are to:– Provide a sound theoretical knowledge and/or the practical abilities of the principles that underpin the practice of laboratory animal sciences and technology, and to improve the welfare of animals and the quality of the animal work necessary to fulfil the increasingly demanding role of an Animal Technologist under the legislation protecting animals used for experimental or other scientific purposes.– Assist learners effectively to demonstrate competence in the execution of practical skills.– Foster a spirit of independent learning, enquiry and continuing professional development (CPD).– Externally assess the learning that has taken place and its application to the practice of laboratory animal science and technology.– Provide the basis of career progression as a qualified Animal Technologist.– Provide a coherent and balanced education appropriate to the level of professional membership.– Provide a qualification that is recognised by employers and competent authorities.– Provide a portable qualification preparing learners for further professional development transferable between jobs and countries.IAT graduation special: Congratulations to all the successful HE students

216Animal Technology and Welfare August 2020We would like to thank and recognise all the CLAST teaching staff for all their support of students.Lecturers for the HE ProgrammeWe would like to thank the commercial sponsors of the graduation day.IAT, The Learning Curve, Tecniplast, CCTech, IPS, Agenda, a-tune, Aston Pharma, Charles River and Castium.David Anderson Paul FlecknellSimon Anderson Glyn FisherKen Applebee Judith FowlstonJas Barley Nicky GentPilar Bowne Susie KnightJames Bussell Matt LeachGary Childs Brian LoweSteven Cubitt Carol LuckAuriol Lamb Cubitt Michelle StewartHayley DanielsIt’s all about you so become a HE student and share the success!College of Laboratory Animal Science and TechnologyA non-profit course provider of Higher Education (HE) and CPD qualifications designed for those working with animals in research, including Animal Technologists, Registered Veterinary Professionals, and Senior Researchers.CLAST was established to provide the specialist training and education required at this level of study. CLAST is a non-profit course provider that has been set up to deliver HE qualifications in Animal Technology. The IAT continues to award the qualifications and CLAST now delivers and assesses all the Level 4, 5 and 6 units and qualifications.Brian Lowe Carol Luck Michelle Stewart We would like to thank the commercial sponsors of the Gradation Day. IAT, The Learning Curve, Tecniplast, CCTech, IPS, Agenda, A Tunes, Aston Pharma, Charles River and Castium. It’s all about you so become a HE student and share the success! IAT graduation special: Congratulations to all the successful HE students

217August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareCLAST is managed by a board of fi ve trustees who have the responsibility to ensure the provision and delivery of high quality education and training at a reasonable cost. The trustees are also responsible for ensuring the trust complies with its legal responsibilities as a charity and that fi nancial and other records are submitted to the Charity Commission as required.Board of TrusteesChair – Ken Applebee Secretary – Gary Childs Members – Ngaire Dennison, Paul Flecknell, David Spillane and Sara Wells.CLAST continues to draw upon the support of industry experts who teach and assess the qualifi cations.CLAST will also be looking to support the industry in developing and delivering new CPD short courses over the coming years.Unable to access higher education because of fi nancial reasons?The HE programme would have struggled to continue without attracting suffi cient funding and student numbers. AS-ET has played a big part in supporting the students who wanted to study but needed some fi nancial support.AS-ET is a charity, registered under the UK Charity Law with the objective to advance education and promote excellence in the care and welfare of animals used in science.AS-ET is a charity to advance education and promote excellence in the careand welfare of animals used in science. Sponsors of AS-ET are listed below and to find out more please visit thewebsite www.as-et.org.ukAS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15AS-ET is a charity set up to advance education and promote excellence inthe care and welfare of animals used in science.To see how you can apply for a bursary visit our website (www.as-et.org.uk)The companies listed below sponsor AS-ET. If you would like to join themfind out how to support us by looking at our website.BulletinMay 2014• 15Bulletin18 •July 2020Jul20:IATB NEW 10/7/20 10:02 Page 18The Trust achieves its objective by providing bursaries for education and training courses, supporting the development of new courses, promoting the production of teaching package. We also provide prizes to encourage people to undertake further education and training. Three AS-ET prizes were presented at this year’s graduation ceremony.The AS-ET Trustees would like to congratulate the graduates who have successfully undertaken a demanding course, extending their knowledge and making them better able to promote excellence in the care and welfare of animals used in science. We hope many other technologists will follow their example. If any potential student has diffi culty in raising the registration fee AS-ET will be happy to consider an application for a bursary.For further details visit www.as-et.org.ukOur Diplomas –Level 4Laboratory Animal Science and Technology DiplomaOur Level 4 Diploma is the fi rst step in gaining higher education qualifi cations in Laboratory Animal Science and Technology (LAS&T). Level 4 | CLASTCollege of LaboratoryAnimal Science & TechnologyA non-profit course provider of Higher Education (HE) and CPD qualifications designed for those working with animals in research, including Animal Technologists, Registered Veterinary Professionals, and Senior Researchers.The College of Laboratory Animal Science & Technology (CLAST) has been set up to provide the specialist training and education required at this level of study. CLAST is a non-profit course provider that has been set up to deliver Higher Education qualifications in Animal Technology. The IAT continues to award the qualifications, but CLAST now delivers and assesses all of the Level 4, 5 and 6 units and qualifications.CLAST is managed by a board of five trustees who have the responsibility to ensure the provision and delivery of high-quality education and training at a reasonable cost. The trustees are also responsible for ensuring the trust complies with its legal responsibilities as a charity and that financial and other records are submitted to the Charity Commission as required.Board of TrusteesChair- Ken Applebee Secretary - Gary ChildsMembers - Ngaire Dennison, Paul Flecknell, David Spillane, Sara Wells.CLAST continues to draw upon the support of industry experts who teach and assess the qualifications.CLAST will also be looking to support the industry in developing and delivering new CPD shortcourses over the coming years.Browse Our DiplomasLevel 4 Laboratory Animal Science and Technology Diploma Our Level 4 Diploma is the first step in gaining higher education qualifications in Laboratory Animal Science & Technology (LAS&T). View courses - https://clast.org.uk/diplomas/level/level-4/ Level 5 Laboratory Animal Science and Technology Diploma Our Level 5 Diploma continues on from Level 4 and provides a more comprehensive look at subjects View courses - https://clast.org.uk/diplomas/level/level-5/ Level 6 Laboratory Animal Science and Technology Diploma Our Level 6 Diploma continues on from Level 5 and upon successful completion you will gain a full IAT HE Diploma. IAT graduation special: Congratulations to all the successful HE students

218Animal Technology and Welfare August 2020Level 5Laboratory Animal Science and Technology DiplomaOur Level 5 Diploma continues from Level 4 and providesa more comprehensive look at subjects. Level 5 | CLASTIAT FellowshipFor Level 6 graduatesStudents that have completed a Level 6 Diploma are eligible for IAT Fellowship. Become a Fellow to receive exclusive benefi ts and become available for election to council.Level 4 Laboratory Animal Science and Technology Diploma Our Level 4 Diploma is the first step in gaining higher education qualifications in Laboratory Animal Science & Technology (LAS&T). View courses - https://clast.org.uk/diplomas/level/level-4/ Level 5 Laboratory Animal Science and Technology Diploma Our Level 5 Diploma continues on from Level 4 and provides a more comprehensive look at subjects View courses - https://clast.org.uk/diplomas/level/level-5/ Level 6 Laboratory Animal Science and Technology Diploma Our Level 6 Diploma continues on from Level 5 and upon successful completion you will gain a full IAT HE Diploma. Level 6Laboratory Animal Science and Technology DiplomaOur Level 6 Diploma continues from Level 5 and uponsuccessful completion you will gain a full IAT HE Diploma.Level 6 | CLASTView courses - https://clast.org.uk/diplomas/level/level-6/ IAT Fellowship For level 6 graduates Students that have completed a Level 6 Diploma are eligible for IAT Fellowship. Become a Fellow to receive exclusive benefits and become available for election to council. Register for a diploma Before enrolling in courses for your chosen diploma, please make sure you have registered onto your chosen level first. Registering is made very simple by our registration form. Register for a diploma - https://clast.org.uk/register/ About our Diplomas We offer level 4 – 6 IAT Diplomas in Laboratory Animal Science and Technology (LAS&T). Our Diploma courses follow on from IAT levels 1 – 3. We have split our qualification into three Diplomas (level 4-6) to allow students the freedom to progress to the level that suits them best. You will receive an IAT Diploma certificate upon completion of each year, culminating in a gaining a full IAT HE Diploma ( https://clast.org.uk/diplomas/) in LAS&T on completion of your third year. There is no obligation to complete all 3 years and you can take a break between levels, returning to study when the time is right for you. What will I learn? Each level comprises 4 different learning units. Each unit will focus on a subject relating to animal research and/or the roles and responsibilities within an animal research facility. Each subject will help you develop key skills that will improve your performance in the work place, provide you with a detailed understanding of the topic at hand and teach you how to apply those principles to improve animal welfare. How long will I be studying? In part, how long you study for is up to you. Each Diploma is taught over 1 year, the 4 units will be taught in separate teaching blocks that typically run over 4 consecutive days. This Register for a diplomaBefore enrolling in courses for your chosen diploma please make sure you have registered onto your chosen level fi rst. Registering is made very simple by our registration form. Register | College of Laboratory Animal science andtechnology (clast.org.uk)IAT graduation special: Congratulations to all the successful HE students

219August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAbout our DiplomasWe offer level 4 to 6 IAT Diplomas in Laboratory Animal Science and Technology (LAS&T). Our Diploma courses follow on from IAT levels 1 – 3. We have split our qualifi cation into three Diplomas (level 4-6) to allow students the freedom to progress to the level that suits them best. You will receive an IAT Diploma certifi cate upon completion of each year, culminating in a gaining a full IAT HE Diploma in LAS&T on completion of your third year. There is no obligation to complete all 3 years and you can take a break between levels, returning to study when the time is right for you.What will I learn?Each level comprises 4 different learning units. Each unit will focus on a subject relating to animal research and/or the roles and responsibilities within an animal research facility. Each subject will help you develop key skills that will improve your performance in the workplace, provide you with a detailed understanding of the topic at hand and teach you how to apply those principles to improve animal welfare.How long will I be studying?In part, how long you study for is up to you. Each Diploma is taught over 1 year, the 4 units will be taught in separate teaching blocks that typically run over 4 consecutive days. This way you do not have to make a weekly commitment to attending a teaching session and instead, can take a short break from work to complete the teaching block in just a few days. Units are usually spaced out over the year and because our Diplomas are split into individual levels, you will not be committed to 3 consecutive years of study and will achieve a Diploma certifi cate for each year completed.Even though the taught classes are part-time, our expert tutors will always be available to support you in your studies through email, video calls or telephone.How will a Diploma benefi t me?Studying LAS&T diplomas will equip you with the knowledge needed to excel in the world of laboratory animal care and welfare. Our Diplomas are perfect for those working in animal research who want to take their career to the next step, especially if you aspire to achieve a supervisory or management position within a research facility. Our Diplomas cover topics ranging from animal biology and experimental design to the design and management of an animal facility. Complete all three levels in LAS&T to gain a full IAT HE Diploma and become eligible for IAT Fellowship.IAT graduation special: Congratulations to all the successful HE students

For details of how you can support AS-ET please visit:www.animalsinscienceeducationtrust.org.ukAS-ET Corporate Sponsorswww.animalsinscienceeducationtrust.org.uk“I would like to thank AS-ET for the financial support and consideration given to me, which has allowed me to embark on and complete my MSc degree. The new knowledge gained has let me become a more rounded and complete technician, with confidence to help others with their educational requirements, and to better support the research projects”“With rising prices everywhere, our departmental budget wasn’t able to stretch to me studying a Level 4 module. I turned to AS-ET for help and they willingly agreed to part fund the course. For anyone debating to give a Level 4 unit a try I would say go for it! You will learn so much about the science behind the work going on in your facility as well as essay writing techniques and working to deadlines”

221August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareDecember 2023 Animal Technology and WelfareAn assessment of rat hammocks as enrichmentMEGAN HICKMAN, LORRAINE MILLER and RUTH MACDONALD Animal Sciences and Technologies, AstraZeneca UK Correspondence: Megan.hickman@astrazeneca.com POSTER PRESENTATIONSOriginally presented at:IAT Congress 2023151IntroductionA hallmark symptom of rheumatoid arthritis in humansis painful swollen joints. Pain can manifest before anyinflammation is noticeable1,2as well as persist longafter inflammation has resolved.3In rodent models of arthritis, ank le or footpad width isa commo nly u sed surrogate marker of pain (seeFigure 1).Measur ing footpad wi dth assumes that increasedswelling is proportional to enhanced pain. A mildarthritis phenotype in which there is minimal swellingmay therefore inaccurately reflect the extent of painand discomfort.POSTER PRESENTATIONSOriginally presented at:IAT Congress 2019Assessing pain in models ofRheumatoid ArthritisSAMUEL SINGLETON,1MERIAM NEFLA,1NGAIRE DENNISON,1SIMON ARTHUR2and TIM HALES1School of Life Sciences, Division of Cell Signalling and Immunology, University of Dundee,Dundee, DD1 5EH, UK2MRSU and Institute of Academic Anaesthesia, Division of Systems Medicine, NinewellsHospital, University of Dundee, DD1 9SY, UKCorrespondence: s.z.singleton@dundee.ac.ukAugust 2020 Animal Technology and WelfareFigure 1. Footpad width as a surrogate measur e of pain in arthritis models. Commonly used methods to assess painare footpad width (A), ankle width (B) or footpad ankle length (C).BCAAim: We aimed to determine how well pain correlated to footpad widths using the collagen antibody arthritismodel.August20:Animal Technology and Welfare 12/8/20 07:54 Page 151AbstractResearch has shown there are numerous welfare and scientific benefits to housing rats in more enriched and environmentally complex housing opposed to conventional housing. With the aim of increasing environmental complexity providing opportunities for climbing and exploring and providing additional cover, a hammock prototype was designed to implement into the double decker rat cages. The hammock was trialled in a group of female rats to assess durability, impact on behaviour and preference over other shelter enrichment items:• tunnels• rat corner homesThe number of observed enrichment interactions indicated a strong preference for the bottom tier of the hammock, suggesting the hammock is a successful enrichment item.IntroductionProviding laboratory rats with appropriate cage enrichment is important to accommodate core needs, provide mental stimulation and allow rats to express species-specific behaviours. The current enrichment provided at our facility in double decker rat cages follows conventional housing recommendations consisting of a play tunnel, two woodblocks and sizzle nest. However there is significant evidence to suggest that housing rats in more enriched caging has many benefits.Numerous studies have shown that rats maintained in enriched cages demonstrate a higher level of exploratory activity and lower anxiety compared with rats housed in conventional caging, resulting in animals that are calmer and easier to handle.1,4It has also been shown to reduce obesity which is a recognised problem in laboratory rats with some strains achieving a fat mass that accounts for 30 to 50% of bodyweight.

222Animal Technology and Welfare August 2020Spangenberg et al (2005) investigated the effects of housing rats in large pens furnished with enrichment (inverted wire grid, a towel hammock, an inverted opaqueplastic box and cage divider) compared with rats housed in standard cages containing only a black plastic tube and found that enriched pen housed rats weighed 17% less than the standard housed rats and displayed a wider behavioural repertoire.Providing adequate shelter is important to allow rodentsto retreat from perceived threats, bright lights and ambient temperatures thus reducing stress. Studies haveshown that rats have a strong preference for enclosed nest boxes over open end tunnels which provide better cover and an additional elevated resting surface (3).A prototype rat hammock (fi gure 1) was designed with the aim of providing additional shelter increasing cage complexity and providing opportunities for climbing and exploring. The hammocks were constructed from old lab coats, made of 65% polyester, 35% cotton following a honey comb design with two layers which provides an enclosed lower level and top level for climbing and resting on. Hammocks were connected to the cage roof using a beaded chain which loops through the fi lter holes in the cage lid and the metal eyelets of the four corners of the hammock.MethodThe hammocks were trialled in a group of 4 female ratsto assess durability, impact on behaviour and preference over other shelter enrichment items which were tunnels and rat corner homes.Rats were observed over a 4 day period following theschedule in Table 1, days 1 to 2 were used to acclimatise the rats to the new enrichment items and the preference test carried out on days 3 to 4. Figure 1 hammock prototype Method The hammocks were trailed in a group of 4 female rats to assess durability, impact on behaviour and preference over other shelter enrichment items which were tunnels and rat corner homes. Rats were observed over a 4 day period following the schedule in Table 1, days 1 to 2 were used to acclimatise the rats to the new enrichment items and the preference test carried out on days 3 to 4. At approximately 8:00 each morning enrichment items were added to the cage with rats given 1 hour to acclimatise to the enrichment prior Figure 1. Hammock prototype.At approximately 8:00 each morning enrichment itemswere added to the cage with rats given 1 hour to acclimatise to the enrichment prior to recording observations.Animals were observed at hourly intervals between the hours of 09:00 to 15:00.At each interval the location or enrichment item that the animal is in contact with was recorded (hammock, tunnel, corner home, cage fl oor or balcony).An ethogram created by NC3Rs was used to record the behaviour of each animal at each timepoint (table 1).Table 1.ResultsWhen housed in the enriched cage and given access toall 3 enrichment items, the number of observed enrichment interactions indicated a strong preference for the bottomtier of the hammock (Table 2).The number of observations of rats sleeping on the cage fl oor or balcony reduced from 11 on Day 1 to 0 on Day 4 when given access to additional shelter enrichment. Sleeping/resting out in the open is a very unnatural behaviour in rats suggesting that a play tunnel is not suitable shelter enrichment.DAYLEFT SIDE OF CAGE RIGHT SIDE OF CAGEDay 1 Play TunnelDay 2 Rat Corner Home, HammockDay 3 Rat Corner Home, HammockPlay TunnelDay 4 Play Tunnel Rat Corner Home, HammockThe number of observations of rats sleeping on the cage floor or balcony reduced from 11 on Day 1 to 0 on Day 4 when given access to additional shelter enrichment. Sleeping/resting out in the open is a very unnatural behaviour in rats suggesting that a play tunnel is not suitable shelter enrichment. Table 2. Location Preference Test.Poster Presentations

223August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsThe rats spent a large proportion of the observation period sleeping (Table 3) so an additional behaviour study carried out during the dark light cycle period would give a better understanding on whether the increased cage complexity provided by the hammock encourages more physical activity and natural behaviours (climbing, exploring) compared to conventional housing. At theend of the study the hammock was still in good condition other than some slight fraying around the corners. Exploring more durable fabric alternatives that can bewashed and reused could be a more sustainable way of implementing them.ConclusionThe data indicates the hammock is the most preferred place to rest/sleep out of the 3 enrichment items, suggesting that the inclusion of hammocks in rat enclosures is an effective way of providing shelter however an additional study carried out in the dark cycle would be needed to determine the effects on behaviour and activity levels. The rats spent a large proportion of the observation period sleeping (Table 3) so an additional behaviour study carried out during the dark light cycle period would give a better understanding on whether the increased cage complexity provided by the hammock encourages more physical activity and natural behaviours (climbing, exploring) compared to conventional housing. At the end of the study the hammock was still in good condition other than slight fraying around the corners. Exploring more durable fabric alternatives that can be washed and re used could be a more sustainable way of implementing them. Table 3. Number of observed behaviours.References1 Buchheister, S.; Bleich, A. Health Monitoring of Laboratory Rodent Colonies – Talking about (R)evolution. Animals 11.5 (2021), 1410.1 Exploratory activity of rats in three different environments. Genaro, G., Schmidek, W.R.; Ethology (2000). 2 Housing-related activity in rats: effects on bodyweight,urinary corticosterone levels, muscle properties andperformance. Spangenberg, E.M.F., et al. Laboratory Animals. (2005). 3 The cage preferences of laboratory rats. Patterson, K., et al. Laboratory Animals. (2001). 4 Long-term effects of the periadolescent environment on exploratory activity and aggressive behaviour inmice: social versus physical enrichment. Pietropaolo, S., et al. Physiol. & Behav. (2004). 5 https://nc3rs.org.uk/sites/default/fi les/documents/Evaluating Environ mental Enrichment/General%20ethograms.pdf

224Animal Technology and Welfare August 2020AbstractThe Mouse Behaviour Core is a self serve facility locatedin one of our main experimental units. It provides adedicated area to perform behavioural testing of genetically altered and wildtype mice to determine the effect that gene alteration has on behaviour, memory, learning and motor function. The requirement for expanded Containment Level 3 facilities led to the relocation of our previous behaviour suite and provided the opportunity to create a new area specially designed for various behaviour tests. It has been open for just under a year and has allowed researchers to carry out a variety of behaviour related research, under Project Licence authority. In this poster we are going to cover how the Biological Research Facility and associated Animal Technicians support the researchers to carry out their work, whilst ensuring Animal Welfare is kept to a high standard.Why do we study behaviour of animals?Animal behaviour has been studied in laboratories sincethe early 20th century but has been increasingly used inthe last 2 decades. There are more than 100 behaviouraltests available in publication and many are still in development.We use mice in our behavioural research as they have a lot of similarities to humans: anatomical, physiological, and genetic. By understanding animal behaviour, we cancreate a better understanding of genes, as well as regions of the brain and its circuits. It can also be used to see howdrugs and/or treatments can affect mouse behaviour.These results can then be analysed/interpreted and translated into human understanding (Figure 1).Mouse behaviour core at the Francis Crick InstituteLYDIA COLGATE and NADIA UNALThe Francis Crick InstituteCorrespondence: lydia.colgate@crick.ac.uk and Nadia.Unal@crick.ac.uk What happens in the mouse behaviour core at The Crick?The Biological Research Facility (BRF) behaviour core (fi gure 2) is set up for various behaviour related tests which can be broken down into four main categories:• learning and memory• anxiety• social• motor functionFigure 1. Graph showing the increase of publications on mouse behavioural studies.Figure 2. Bird’s eye view of the behaviour core.Animal Technology and Welfare December 2023

225August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsUnder each category there are a number of tests that canbe performed using various mazes and set ups supplied by the BRF.Each individual area (Figure 3) is booked on the dedicated online booking system (PPMS) by the researchers to avoid disappointment and to help with planning experiments.AnxietyThe light/dark test apparatus (Figure 6) is made up of a dark chamber and a brightly lit chamber. Mice are allowed to move freely between the two chambers. This measures anxiety like behaviours using their aversion to open spaces as when anxious mice are more likely to prefer dark enclosed areas.Behavioural tests used in the behaviour coreLearning and memoryThe Barnes Maze which is a circular table (Figure 4) withcircular holes around the outside. The objective is forthe mouse to reach an escape box under one of the holes. The animal uses visual cues and prompts allowingassessment of memory and spatial learning.Figure 2 Birdseye view of the behaviour core Figure 3 Behaviour core setup Each individual area (figure 3) is booked on the dedicated on line booking system (PPMS) by the researchers to avoid disappointment and to help with planning experiments. Behavioural tests used in the behaviour core Learning and memory The Barnes Maze which is a circular table (figure 4) with circular holes around the outside. The objective is for the mouse to reach an escape box under one of the holes. The animal uses visual cues and prompts allowing assessment of memory and spatial learning. Figure 3.Behaviour core setup. Figure 4 Barnes Maze The Y Maze (figure 5) is a test used to assess working or short term memory. A mouse is placed in the centre and observed. Mice with intact working memory have been shown to more likely explore areas that it has not visited before. Figure 5 Y Maze Anxiety The light/dark test apparatus (figure 6) is made up of a dark chamber and a brightly lit chamber. Mice are allowed to move freely between the two chambers. This measures anxiety like behaviours using their Figure 4. Barnes Maze.The Y Maze (Figure 5) is a test used to assess working or short term memory. A mouse is placed in the centre and observed. Mice with intact working memory have been shown to more likely explore areas that it has not visited before.Figure 5. Y Maze.aversion to open spaces as when anxious mice are more likely to prefer dark enclosed areas. Figure 6 The light/dark test The open field test (figure 7) involves releasing a mouse into a new area called an area. Movement and reactions are recorded which provides information on anxiety related behaviours when in an unknown environment. Figure 7 The open field test Figure 6. The light/dark test.aversion to open spaces as when anxious mice are more likely to prefer dark enclosed areas. Figure 6 The light/dark test The open field test (figure 7) involves releasing a mouse into a new area called an area. Movement and reactions are recorded which provides information on anxiety related behaviours when in an unknown environment. Figure 7 The open field test Figure 7. The open fi eld test.The open fi eld test (Figure 7) involves releasing a mouseinto a new area called an ‘area’. Movement and reactions are recorded which provides information on anxiety relatedbehaviours when in an unknown environment.

226Animal Technology and Welfare August 2020Poster PresentationsSocialThe three chamber test (Figure 8) evaluates cognition and has shown that normally mice prefer to spend more time with another mouse. The equipment consists of threeseparate rooms and provides an insight into their thinkingprocesses and desires to interact with each other.The tube dominance test (Figure 9) is where mice are trained to walk forward in the tube. Two mice are then put into the tube at opposite ends and the animals walktowards each other, where they can interact in the middle. This test is typically used to test the hierarchy of a groupof mice living in the same cage.Figure 8. Three chamber test. Figure 9 Tube dominance test Motor function The Rotarod test (figure 10) is designed to monitor motor coordination, balance and fatigue. The apparatus consists of a circular rod that turns at a constant or increasing speed. The mice must continue to hold on and climb around the rod at it spins. This tests the effects of genetic alteration, drugs, brain damage or diseases. Figure 10 Rotarod test Figure 9. Tube dominance test.Motor functionThe Rotarod test (Figure 10) is designed to monitor motorcoordination, balance and fatigue. The apparatus consistsof a circular rod that turns at a constant or increasing speed. The mice must continue to hold on and climb around the rod at it spins. This tests the effects of geneticalteration, drugs, brain damage or diseases.Figure 10. Rotarod test.The Catwalk test (Figure 11) is when mice are put ontothe catwalk where they walk along the illuminated glassplatform. A camera measures and records the animal’s movement and their gait can be assessed and evaluated.Figure 11. Catwalk test.

227August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster Presentations Future plans for the behaviour core Ethovision video systemAll the mazes are set up under cameraswhich areconnected to the Ethovision videosystem (Figure 12). Ethovision XT is a video tracking software that tracks and analyses the behaviour, movement and activity of mice. The data recorded can then be stored and analysed/interpreted by the researcher later.Future plans for the behaviour coreWith the increasing usage of the area, plans have been put in motion for the BRF and the research offi cers (RO) working within, to be trained in performing behavioural tests for the researchers. This will allow the researchers to make more effi cient use of their time and will also allow some variety and progression in the RO role.To maintain the productivity of the area, the BRF will be sending out a 1 year survey to all researchers to ensure the area is fi t for purpose, the correct equipment is available and if further support can be offered.This will enable the BRF to have greater oversight of any welfare issues that may arise from the tests and maintain the best possible husbandry and welfare practices.AcknowledgementsCaroline Zverev, Clare Brazil-Adams, Helen Baily and Sam Cooper. Figure 12 Ethovision video systems The process of using the behaviour core Figure 12. Ethovision video systems.The process of using the behaviour core

228Animal Technology and Welfare August 2020IntroductionProviding an enriched species specifi c environment coupled with the principles of the 3Rs (Refi nement, Reduction and Replacement) is at the heart of the poultryteam at The Pirbright Institute. Periodically reviewed custom made cage designs (Raised Floor Pens) have allowed the team to combine bird welfare with staff requirements thus formulating an improved livable environment for the birds and a workable setting for Animal Technicians.Other aspects recognised as key to providing a healthy environment include: • temperature and humidity• lighting• space allowance• food and non food enrichment • human interactionWe show the refi ned conditions installed at The PirbrightInstitute based on Animal Technician experience and information from the Home Offi ce Code of Practice for the housing and care of animals bred, supplied or used for scientifi c purposes.Cooped up: refi nement of a Gallus gallus domesticus environment BILLY MATTHEWS, RACHEL NEW, YAMINI MAHESH KANSE, JESS WANT, SOPHIE BENTLEY, MICHAEL COLLETT, VICTORIA CHANTLER and PAUL SMITHThe Pirbright InstituteCorrespondence: enquiries@pirbright.ac.uk Animal Technology and Welfare December 2023Housing 2016 to 2018Raised Floor Pen version 1: Original design bespoke to The Pirbright Institute. Floor area: 11,745cm2. Optimal number of birds housed: up to 43. Bird numbers vary based on weight up to 600g.Housing 2016 to 2018Raised Floor Pen version 1: Original design bespoke to The Pirbright Institute. Floor area: 11,745cm2. Optimal number of birds housed: Up to 43. Bird numbers vary based on weight up to 600g. Housing 2018 to 2020 Raised Floor Pen version 2: Second design, version 1 refined to improve bird welfare and usability. Floor area: 10,125cm². Optimal number of birds housed: Up to 37. Bird numbers vary based on weight up to 600g. Housing 2018 to 2020Raised Floor Pen version 2: Second design, version 1 refi ned to improve bird welfare and usability. Floor area: 10,125cm². Optimal number of birds housed: up to 37. Bird numbers vary based on weight up to 600g. Housing 2018 to 2020 Raised Floor Pen version 2: Second design, version 1 refined to improve bird welfare and usability. Floor area: 10,125cm². Optimal number of birds housed: Up to 37. Bird numbers vary based on weight up to 600g.

229August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster Presentations2020 to presentRaised Floor Pen version 3: Third design, version 2 refi ned to further improve bird welfare and usability. Floor area: 10,854cm². Optimal number of birds housed: up to 40. Bird numbers vary based on weight up to 600g. 2020 to present Raised Floor Pen version 3: Third design, version 2 refined to further improve bird welfare and usability. Floor area: 10,854cm². Optimal number of birds housed: Up to 40. Bird numbers vary based on weight up to 600g. 2020 to present Raised Floor Pen version 3: Third design, version 2 refined to further improve bird welfare and usability. Floor area: 10,854cm². Optimal number of birds housed: Up to 40. Bird numbers vary based on weight up to 600g. EnrichmentA variety of food and non food enrichment is provided and rotated daily to promote the following:• natural foraging behaviours• positive reinforcement • boredom prevention Enrichment A variety of food and non food enrichment is provided and rotated daily to promote the following: • natural foraging behaviours • positive reinforcement • boredom prevention Enrichment A variety of food and non food enrichment is provided and rotated daily to promote the following: • natural foraging behaviours • positive reinforcement • boredom prevention Lighting Well managed lighting promotes healthy immune system, growth, mibility and natural behaviours LightingWell managed lighting promotes healthy immune system,growth, mobility and natural behaviours Space allowance

230Animal Technology and Welfare August 2020Space allowance Temperature and relative humitdy (rH%) Temperature and relative humitdy (rH%) References Home Office. (2014, December 17). Code of practice for the housing and care of animals bred, supplied or used for scientific purposes. https://www.gov.uk/government/publications/code-of-practice-for-the-housing-and-care-of-animals-bred-supplied-or-used-for-scientific-purposes References1 Home Offi ce. (2014, December 17). Code of practice for the housing and care of animals bred, supplied or used for scientifi c purposes. https://www.gov.uk/government/publications/code-of-practice-for-the-housing-and-care-of-animals-bred-supplied-or-used-for-scientifi c-purposesPoster Presentations

231August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareDecember 2023 Animal Technology and WelfareIntroductionIn 2019 the first Zebrafish health and welfare terms database was created. Working in collaboration with Zebrafish specialists and veterinarians we defined severity scores, welfare indicators, definitions and images to form a resource for the Zebrafish community. Since then the Zebrafish welfare database has provided a platform for a standardised approach ensuring the consistency of language and severity scoring used across facilities. It has provided the capability to raise awareness of potential health concerns specific to genetic background or environmental impacts on the health of the fish. By standardising the terms and severity scores used, husbandry requirements can be shared as a community to ensure the welfare of the fish, create consistency across facilities and compliance with The Animals (Scientific Procedures) Act 1986 (ASPA). Here we discuss how we use the database within the University of Cambridge aquatics facilities and the benefits we have seen.The Zebrafish health and welfare database, where we are nowNICOLA GOODWIN, ROSEMARY KEEBLE and MICHAEL PRICEUniversity of Cambridge, UBS DepartmentCorrespondence: ntg22@cam.ac.ukFigure 1. ParameterWelfare IndicatorsSynonyms Definitions Severity Score PictureSkin Swelling under skinMasses, raised areas, lumpsAbnormal appearance of masses or all descriptions (hard, soft, different shapes etc.)Mild: 1 mass under the skin which is not affecting natural behaviour. Moderate: Mulitple masses under the skin which is not affecting natural behaviour. Severe: Multiple masses under the skin and natural behaviour affected.General OedemaAn excessive accumulation of fluid in tissue spaces or a body cavity.Mild: Oedema – low level and highly localised fluid accumulation in tissue. Moderate: Oedema – increased fluid accumulation in one or many tissues. Severe: Oedema – high fluid accumulation in one or multiple tissues.General Distended Enlarged Abdomen appears curved outward or swollen.Mild: Abdomen marginally distended. Moderate: Abdomen distended and reddening. Severe: Abdomen distended, reddened, unable to maintain skilful and effective interaction of movements and maintenance of equilibrium.Accessing the database at the University of Cambridge1. The database is easily loaded from the ZFIN website onto a computer within the fish rooms using WiFi connection. Within the University of Cambridge we can readily see the database using desktop tablets.2. Where WiFi connection or workspace is limited, we have PDF copies which have been printed from the ZFIN website and held within the rooms.3. Where the ARMIS database is being used, these terms are integrated into this database, allowing for ease of drop down options for selection of welfare terms, severity and scoring. This data is automatically loaded into the Home Office (HO) returns component of the database, for accuracy and efficiency of the Home Office Return of Procedures.Zebrafish Health and Welfare DatabaseFigure 1 from the ZFIN website.Pictures providefurther clarity and can be used as a training tool fornew care staff Figure 1 The benefits of using the Zebrafish welfare terms and how the database can be used is seen in table 1. Figure 1 The benefits of using the Zebrafish welfare terms and how the database can be used is seen in table 1. Figure 1 The benefits of using the Zebrafish welfare terms and how the database can be used is seen in table 1.

232Animal Technology and Welfare August 2020The benefi ts of using the Zebrafi sh welfare terms and how the database can be used is seen in table 1.Where to fi nd the databaseBenefi ts of using the Zebrafi sh welfare termsHow we have used the database at the University of Cambridge Recognition of patterns of disease across tanks and colonies.Awareness of disease at the earliest time point.Awareness of potential crosscontamination across sites/ spread of pathogens.The University of Cambridge have 3 fi sh facilities, communication and consistency of the welfare language used is vital to ensure we see signs of disease patterns across facilities.Any animals found sick are recorded onto our database which allows for immediate fl agging to the NACWO and NVS for welfare support.We hold monthly meetings in which we discuss the number of fi sh which have been culled or found dead, any unusual health issues we have found, any patterns and what has/will be put in place to alleviate any further issues (if required) to improve animal welfare.Being aware of health concerns in one facility ensures we stop the movement of tanks between facilities, until we understand the cause, prevalence and potential contamination issues. Phenotype identifi cationWhen we fi nd any clinical signs, we record the welfare concern based on the welfare database details. This has ensured users are aware of any phenotypes quickly, expected or not within our colonies, amendment to husbandry care for optimal welfare, culling at earlier time points, awareness of health concerns developing at specifi c time points and for further details to be provided to anyone receiving these animals in future to assist with husbandry care.Adherence to humane end points/ compliance with ASPA Animal welfareProject Licences include humane end points for an experiment. We use the welfare terms severity score and images to assist with notifying the NACWO, personal licence holder and project licence holder of welfare concerns and severity reached. This has assisted with decision making regarding the continuation of the experiment and whether the animal needs to be euthanized for welfare and licence reasons.Severity scoringImagesThe severity scoring and images within the welfare terms has assisted us with training new members of staff, especially with severity scoring of animals with multiple clinical signs. It has ensured all of team members across all sites are using the same severity scores for consistency of scoring and identifi cation of concerns.Table 1.AcknowledgementsWe would like to thank Zoltan Varga of the Zebrafi sh International Resource Centre (ZIRC) and Jonathan Knight of ZFIN for their continued support with the creation and development of the database. The Wellcome Trust Sanger Institute, James Bussell and Natasha Karp for their support in the development of the database. We would like to thank the ARMIS database developers for including the welfare terms in their software. We would like to thank the Zebrafi sh Journal for publishing our manuscript discussing the welfare terms:Standardized Welfare Terms for the Zebrafi sh Community Standardized Welfare Terms for the Zebrafi shCommunity - PubMed (nih.gov)Where to find the database Acknowledgements We would like to thank Zoltan Varga of the Zebrafish International Resource Centre (ZIRC) and Jonathan Knight of ZFIN for their continued support with the creation and development of the database. The Welcome Trust Sanger Institute, James Bussell and Natasha Karp for their support in the development of the database. We would like to thank the Armis database developers for including the welfare terms in their software. Poster Presentations

233August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareDecember 2023 Animal Technology and WelfareAbout the facility• Academic Institution• 600 cages of 3,000 rodents• Zebrafi sh facility• Staff: director/designated veterinarian, biomedical advisor, chief technical offi cer, 4 senior technicians and 2 laboratory attendants• 25-30 research groups with multiple researchers pergroup at any one timeImplementation frameworkThe 3Hs – home, hammocks and happinessSYLVIE MEHIGAN and JOHN HOBBSUniversity College DublinCorrespondence: sylvia.mehigan@ucd.ie and john.hobbs@ucd.ie Misson statementIt is our mission to continue to improve the environment and enrich all animals in our care. We continue to fi nd better ways to interact with animals daily. To this end we started a programme to improve the experience and environment of our animals and give them a better life and to aid research and all those who work with animals in our facility.Enrichment programmeIn University College Dublin’s (UCD) biomedical facility we always strive to improve our animal’s welfare and Figure 1.environment. We endeavour to enrich the lives of our research animals, promote a Culture of Care and implement continuous improvements insupport of the 3Rs. Standard environmental enrichment for our rodents has always included some or all the following; • perspex red house• opaque tubes• running wheels• nesting material • chew sticksDuring 2017 tunnel handling was implemented for all mice and we commenced our current environmental enrichment programme in 2019. Since then we have added hammocks, vet beds, climbing ropes, play balls, sand pits, pools and additional nesting material to our standard environmental enrichment. In addition to this ourhabituation programme has been expanded to include rat tickling.Engaged and active Animal Welfare BodyLeadership and top down Culture of Care promotion

234Animal Technology and Welfare August 2020PlaypensPlaypens are easy to set up and are inexpensive. The majority of our rats are given 30 minutes 3 times per week inthe playpens. Inside is an assortment of toys which include ropes, balls, bath, sand pit, seesaws and hammocks.HammocksHammocks heavenProvides a comfortable, warm shelter for animals. Evenwhen sick the animals will often retreat to their hammocks.Poster PresentationsFigure 2. Mouse.Figure 3. Mouse hammock.Figure 4. Peak aboo.Figure 5. Male rat playpen.Figure 6. Female rat playpen.

235August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareTail cleanliness difference when a bath is provided. Rats enjoy their bath (Figure 9) and clean themselves from head to tail. A clear difference can be seen between rats that have access to the bath (Figure 7) and those that do not (Figure 8).HabituationThese rats are keen to say hello when people come into the animal room. They greet us every morning.Poster PresentationsFigure 7. Star clean tail.Figure 8. Dirty tail.Figure 9. Rats enjoy their bath.Figure 10. Rats greet us every morning.Figure 11. Rats are keen to say hello.

236Animal Technology and Welfare August 2020Poster PresentationsRat ticklingWe aline this with other activities such as cage cleaning and when the rats are being placed into their playpen once or twice per week. This does add an additional 2 to 3 minutes of time to cage changing time per cage. The rats are less stressed and more relaxed with being handled.The rats have been trained to associate a positive experience with being on a vet bed e.g. rat tickling. During procedure habituation training the same vet bed is used.Benefits– Further promoted our Culture of Care for animals, staff and researchers.– A more relaxed environment in which to work and conduct research.– Increased job satisfaction from knowing the animals have a better quality of life.– Better life and less stress for the animals in our care.– Benefits to research by reducing stress for both the researcher and the animal.– Less confounding variables and improved reproducibility.Figure 12. Rat tickling.Future directionsWe are always looking out for new refinements for us to trial for both rodents and fish. Currently under investigation are:• continue to monitor and update current practice• micro pipetting for mice and rats• playpen benefits and possibilities for mice• encouraging further engagement from researchers in our enrichment programmeAcknowledgementsBiomedical facility staff and our animals.Further thanks to Emer Conroy and Mark d’Alton.

237August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareDecember 2023 Animal Technology and WelfareIntroductionCarbon Tetrachloride (CCl4) is a highly toxic substance. When administered consecutively at a low dose CCl4 can model liver fibrosis in rodents and be used in the assessment of new anti-fibrotic therapies. RTx use immunocompromised mice which have a lower tolerance to CCl4 than wild type mice.We will discuss how we refined the care of the immunocompromised mice following each dose of CCl4.The mouse strains that were used are • NSG – NOD.Cg-PrkdcSCIDIl2rgtm1Wjl/SzJ• NOD-SCID – NOD.CB17-Prkdcscid/NcrCrlWhich were sourced from Charles River (UK).The experiments were conducted in compliance with The Animals (Scientific Procedures) Act 1986 (ASPA). Mice were housed under standard conditions of 12:12 hour light-dark cycle, temperature 21 +/-2°C, with free access to chow (Irradiated RM3 and sterilised water) in sterilised individually ventilated cages (IVCs).MethodMice were administered CCl4 using the intraperitoneal (IP) injection route twice weekly. All of the mice had their bodyweight (BW) monitored and were checked for any signs of ill health before each injection. Refining the welfare of immunocompromised mice receiving carbon tetrachloride (CCl4) to induce liver fibrosisVICKY WALCZAK-GILLIES1,2, OLIVIA MATTHEWS2, ALEX FYFE2, ANNA HOY2 and PHILLIP STARKEY LEWIS21 Bioresearch & Veterinary Services, The University of Edinburgh 2 In Vivo Pharmacology, Resolution Therapeutics, Centre for Regenerative Medicine, Edinburgh Correspondence: vicky.walczak-gillies@ed.ac.uk and vicky.walczak-gillies@resolution-tx.comWe reference the dosing criteria (Table 1) to determine when each mouse will receive a full dose, a half dose or if they will miss a dose. CCl4 is diluted 1:4 with olive oil under sterile conditions. All humane endpoints are as described in the specific Project Licence that the work is being carried out under.Table 1. Original dosing criteria.CONDITIONS TO TRIGGER A HALF-DOSE1Last three weeks of the protocol2Recent anesthetic exposure3Losing 0.5g BW in two successive weighings4Losing 10% BW in a two-week period5If all mice in a cage lose 0.5g BW simultaneouslyCONDITIONS TO MISS A DOSE1An animal loses > 20% BW, allow 48 hours recovery and proceed

238Animal Technology and Welfare August 2020In addition, all mice under new dosing criteria:– Must be > 23g bodyweight at the start of experiment.– Have access to a gel pot post injection to increase hydration.– Placed into a heating cabinet (26°C) for up to 5hrs post-injection.– Are checked regularly throughout the day.– Scored by body condition1 and the NC3Rs’ grimace scale.2 Figure 1 New dosing criteria action tree In addition, all mice under new dosing criteria: • Must be > 23g bodyweight at the start of experiment. • Have access to a gel pot post-injection to increase hydration. • Placed into a heating cabinet (26 °C) for up to 5 hrs post-injection. • Are checked regularly throughout the day. Figure 1. New dosing criteria action tree.ResultsOur method provides evidence that we can alter CCl4 dosing and implement recovery techniques to enhance Animal Welfare, whilst maintaining sufficient levels of fibrosis in the liver.Consequently, this has improved both Animal Welfare and scientific outputs, which in turn reduces the need to repeat studies and the overall number of animals required.Poster Presentations

239August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsConclusionWe have successfully refined the care and welfare of the animals without compromising the science. This method can easily be adapted for any study.The smallest actions can have the biggest impact.References1 Burkholder, Tanya., et al. Health Evaluation of Experimental Laboratory Mice. Current protocols in mouse biology vol. 2 (2012): 145-165. doi:10.1002/ 9780470942390.mo110217.2 Langford, Dale J., et al. Coding of facial expressions of pain in the laboratory mouse. Nature methods vol. 7,6 (2010): 447-9. doi:10.1038/nmeth.1455.Table 2. Difference between the original and new dosing regimen.• Scored by body condition1 and the NC3R's grimace scale2. Results Table 2 Difference between the original and new dosing regimen 012345010203040Original dosingand monitoring regimenCCl4 pre-dose checkBody weight (g)012345010203040New dosing andmonitoring regimenCCl4 pre-dose checka bFigure 2. Representative image of PSR stained liver from a NOD-SCID mouse following a 12-week CCl4 regimen.• Scored by body condition1 and the NC3R's grimace scale2. Results Table 2 Difference between the original and new dosing regimen 012345010203040Original dosingand monitoring regimenCCl4 pre-dose checkBody weight (g)012345010203040New dosing andmonitoring regimenCCl4 pre-dose checka b

240Animal Technology and Welfare August 2020IntroductionMalocclusion is a common dental disorder for laboratory mice. Characterised with improper alignment of teeth, if the incisors of the mouse have become overgrown or are missing it can significantly impact Animal Welfare. (NC3Rs, 2021).The hard hitting tooth – does a scheduled teeth check reduce the number of mice found dead with malocclusion?JESSICA CIUPACancer Research UK, Cambridge InstituteCorrespondence: jessica.ciupa@cruk.cam.ac.uk Malocclusion is evident around the first 2-5 weeks of age for mice (NC3Rs, 2021). Therefore, it is hypothesised that a regular welfare check, the POST WEAN CHECK (PWC) at 5 weeks, would:– Significantly increase the health concerns (HC) found as more mice would be diagnosed and caught before being found dead.– Therefore a significant decrease in mice found dead with malocclusion is expected after the PWCs implementation.Figure 1. Example of malocclusion mice (NC3Rs, 2001).Introduction Malocclusion is a common dental disorder for laboratory mice. Characterized with improper alignment of teeth, if the incisors of the mouse have become overgrown or are missing it can significantly impact Animal Welfare. (nc3rs., 2021). MethodsA retrospective data analysis of the HCs from September 2020 to March 2021 were compared to September 2021 to March 2022. The implementation of the PWC was the end of March 2021.Using the AniBio database, key word searches were carried out on the HC, producing reports which had mentioned; malocclusion, malformed, misaligned, overgrown, top, bottom and teeth. To make sure no HC was missed a ➡➡How to counter?If malocclusion is still prevalent after solid diet, appropriate environmental enrichment and users not using the mice for breeding – regular welfare checks should be carried out (NC3Rs, 2021)Why is it common?Mice have monophyodont dentition, defined as possessing one set of teeth in their lifetimes (Diagnosis, 2007). Their rootless teeth (incisors & molars) can grow indefinitely. If their jaws are not aligned, or the appropriate environmental enrichment is not provided/used – it may occur! (NC3Rs, 2021)Animal Technology and Welfare December 2023

241August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster Presentationssimilar search for the common misspelling of the words was additionally carried out. For the MCMS database health concerns under the bracket of ‘misaligned teeth’ and ‘weight loss’ were used. Lines that had stated to be prone to malocclusion were omitted. The data is analysed using a two sample Poisson rate test and a Chi-squared test.whether an implementation of PWCs can improve a technician’s ability to identify malocclusion. Therefore providing an explanation for a lack of significant decrease in found dead reported after the implementation – as the result is unexpected as a significant increase in diagnosis was seen.Additionally, further analysis into relative proportions, of number of births to cases of malocclusion, can suggest a possible reasoning for a lack of significance found for the second hypothesis.AcknowledgementsBRU Core, and in particular, Sally Ashworth, Nicky Jacobs, Brendan Doe and Tony Davidge.References1 Diagnosis. (2007January). Severe prognathic malocclusion. Laboratory Animals. 36, 22–23.2 Mouseland. (2017, April 6). Dentist. Retrieved from Mouse House: http://mouseshouses.blog spot.com/ 2017/04/dentist.html.3 NC3Rs. (2021, September). Malocclusion in mice. Retrieved from NC3Rs: https://www.nc3rs.org.uk/ 3rs-resources/malocclusion- miceResultsThere was a significant increase in HCs of malocclusion found after the PWC implementation. As (Comparative error = 15.47) < (Difference = 66)Therefore, the two values are significantly different.No significant decrease of found dead mice reported after the implementation of PWC. A chi-squared goodness of fit test was performed, and the proportions did not differ between the life status of the mice found with malocclusion, X2(1, 139) = 0.537*, p= .386DiscussionThe results show that more cases of malocclusion were identified after the implementation of the PWC. There were 36 malocclusion cases before the PWC and 103 after the implementation. This suggests that more mice were diagnosed and thus given a humane endpoint/teeth clipping before severe suffering seen in maloccluded mice found dead. However, a chi-squared test analysis to determine whether a proportional difference in found dead reported after the implementation of the PWC found no significance. The assumptions were violated due to the low number of found dead reported prior and post the PWC.To investigate the second hypothesis, further research can compare two facilities: (1) with a regular PWC and (2) without a regular PWC. A timed experiment with images of maloccluded dead mice, should be used to determine Figure 2. A bar chart to show the impact of PWC on reported cases of malocclusion. Results There was a significant increase in HCs of malocclusion found after the PWC implementation. As (Comparative error = 15.47) < (Difference = 66) Therefore, the two values are significantly different. No significant decrease of found dead mice reported after the implementation of PWC. A chi-square goodness of fit test was

242Animal Technology and Welfare August 2020BackgroundAbdominal fat necrosis is a condition found in a variety of species including mice. It is a problem of mature animals, more commonly occurs in cattle and is characterised by the formation of necrotic fat masses in the abdominal cavity. Fat necrosis has historically been termed lipomatosis. However this term is no longer appropriate because the masses are neither neoplastic nor hyperplastic.Animal and Plant Health Agency (APHA) undertakes research into Transmissible Spongiform Encephalopathies (TSEs) part of which uses a mouse bioassay of various transgenic mouse lines. TSEs are a group of slowly progressive fatal neurodegenerative disorders which include Bovine Spongiform Encephalopathy (BSE) in cattle, Scrapie in sheep and goats and Chronic Wasting Disease (CWD) in deer, moose and reindeer. BSE is zoonotic and was linked to the emergence of variant Creutzfeldt-Jakob Disease in humans.In APHA’s mouse colonies, the mice affected are female tgSHPXI mice, a transgenic line bred, maintained and intracerebrally inoculated for prion disease studies. On post mortem the Fat Necrosis has been shown to affect fatty tissue surrounding the uterine horns.ObservationsAll mice are observed daily (Figure 1 healthy mouse) per standard procedure and mice inoculated with a prion disease are closely monitored for clinical signs indicating the onset of the disease to ensure the humane endpoint in the project licence is observedAll signs of ill health in inoculated mice are recorded on the animals’ cards and a named veterinary surgeon is requested to look at those animals and advise on next action if required.Exploratory view into abdominal fat necrosis in miceMELISSA HEWSONPathology and Animal Sciences Department, APHA, WeybridgeCorrespondence: Melissa.Hewson@apha.gov.uk Clinical signs of abdominal fat necrosis observed are: weight loss and/or sunken sides. If suspected these mice will be weighed (to provide an objective baseline) and offered wet diet on the floor of the cage.The mice suspected of Abdominal Fat Necrosis are closely monitored for the condition deteriorating. Clinical endpoints for these mice have been a sudden loss in weight and/or disinterest in food and piloerection (Figure 2 and 3).Figure 1. Necrosis has been shown affects fatty tissue surrounding the uterine horns Observations All mice are observed daily (figure 1 healthy mouse) per standard procedure and mice inoculated with a prion disease are closely monitored for clinical signs indicating the onset of the disease to ensure the humane endpoint in the project licence is observed Figure 1 Images 4 and f show fat necrosis in tgSHPXI female mice Results Figure 2.Figure 2 and Figure 3. TgSHPXI females with clinical signs of fat necrosis.Figure 3.Animal Technology and Welfare December 2023

243August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsResultsBetween 01/2019 and 365/2022, out of 498 tgShpXI mice there were 7.83% suspect abdominal fat necrosis suspects of which 4.23% so far have been confirmed by histology. The remainder are awaiting screening. TgSHPXI’s which have been suspected of abdominal fat necrosis have typically shown symptoms at around 8 months old.When samples of abdominal fat necrosis were sent to histology it was initially thought that these masses were tumours; only after histological assessment they were characterised as fat necrosis.Through performing a post-mortem examination, we have been able to see what areas are affected by Abdominal Fat Necrosis. In figures 4 and 5 you can see intraperitoneal, demarcated, white masses. Sample collection of affected tissues and organs is ongoing alongside observations for clinical signs.DiscussionThere are 3 known causes of abdominal fat necrosis found in other species, one of which is the ongoing consumption of feeds containing high concentration of fatty acids.1 In 2021 sunflower seeds for environmental enrichment were discontinued for all mice.Original signs of abdominal fat necrosis were seen in the previous tgSHPXI line. This mouse line has been refreshed and named TgSHPXIG. Unfortunately fat necrosis has also been identified in this rederived line.It is still undetermined as to how the tgSHPXI mouse line is affected by abdominal fat necrosis. Investigations are still ongoing to help determine the causes of the identified fat necrosis.AcknowledgementsAnna Roberts, Daniel Strawbridge, Timm Konold, John Spiropoulos, Janet Hills, Nicola Fletcher and Jo Moore. Funded by APHA, DEFRA (TS5001, SE1961/62).References1 The MSD Veterinary by Peter D. Constable. Last review/ revision Apr 2021 | Modified Oct 2022. https://www.msdvetmanual.com/digestive-system/abdominal-fat-necrosis/abdominal-fat-necrosis-in-animals Images 4 and f show fat necrosis in tgSHPXI female mice Results Image 6 shows both necrotic fat tissue and normal adipose tissue Discussion There are 3 known causes of abdominal fat necrosis found in other species one of which is the ongoing consumption of feeds containing high concentration of fatty acids1; in 2021 sunflower seeds for environmental enrichment were discontinued for all mice. Original signs of abdominal fat necrosis were seen in the previous tgSHPXI line. This mouse line has been refreshed and named TgSHPXIG. Unfortunately fat necrosis has also been identified in this rederived line. Figure 6. Shows both necrotic fat tissue and normal adipose tissue.Figure 4.Figure 5.Figure 4 and Figure 5. Shows fat necrosis in tgSHPXI female mice.

244Animal Technology and Welfare August 2020Why?Study plans were part of a contingency plan during the COVID-19 pandemic. They provided a way to ensure that all Named Persons had knowledge of the work being carried out. This was especially important with fewer people on the campus.The introduction of this contingency plan meant that researchers across all sites at King’s College London (KCL) were required to complete and submit a study plan before carrying out work across the animal facilities.This requirement was introduced quickly to permit research to resume and allow them to access the animal facilities within the limits of the COVID-19 restrictions.An additional welcomed outcome of the study plan process was the ability to identify experiments that The benefit of study plans to facility managementFLOYD LANIYAN, CLAIRE PEARCE, JOLENE HAMMONDS, MIKE WILLIAMS, LYNSEY CASHNELLA, BRUNO CORREIA DA SILVA, JAYNE MORGAN, NADINE LEBRASSE, ENRIQUETA NAVARRO, THOMAS PITCHER, ANDREW WHATCOTT and JULIE KEEBLEKing’s College LondonCorrespondence: Julie.keeble@kcl.ac.uk and Floyd.laniyan@kcl.ac.uk would have been non-compliant with the Home Office regulations and the process also highlighted protocols which required refinement. This resulted in positive outcomes that included reduced adverse effects to laboratory animals.AimsThe aims of a study plan are to:– Promote collaboration between project licence holder (PPL), personal licence holder (PIL) and Named Persons regarding the regulated procedures animals will undergo during an experiment.– To enable an experiment to be assessed to ensure Home Office compliance (e.g. PIL and PPL authorities and training records) preventing unnecessary non-compliance and potential Animal Welfare issues.Study plansWhen study plans were first introduced they were submitted as a word document (Figure 1).However to integrate study plans and animal management better, the process is now managed by the web based laboratory animal management system we use.Using the software reduced the administrative burden involved with completing study plans as they can be duplicated/copied easily and with alerts automatically being sent to the people responsible for approval. This has created a centralised transparent and easy to audit record for interested parties.outcomes that included reduced adverse effects to laboratory animals. Aims The aims of a study plan are to: • Promote collaboration between project licence holder (PPL), personal licence holder (PIL) and Named Persons regarding the regulated procedures animals will undergo during an experiment. Animal Technology and Welfare December 2023

245August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsStudy plan steps1. Study plans are prepared by the researchers and submitted on the animal management system.2. PPL holder receives notifi cation to check and approve the study plan and ensure the study is compliant withthe PPL authorities.3. The Named Animal Care and Welfare Offi cer (NACWO) receives notifi cation the PPL holder has approved it and will check and authorise it as the facility sign off. The NACWO will ensure the study plan is compliant with the project licence and that endpoints and actionsto be taken are clearly stated and Animal Welfare costs have been minimised.As the study plan process evolved it has become much more than a tool to approve and manage researchers plans of work. The information from study plans help to review the training and competency records of researchers. This also helps with health and safety by checking occupational health status and reporting of any substances which would be hazardous to health.Figure 1. Snapshot of study plan.1. Details of Personal Licensee responsible for animals on this studyFull name including titlePositione-mail addressContact phone no.Name & address of research group and/or departmentPersonal Licence no.Faculty (*select as appropriate)Life Sciences & Medicine Dental Institute IoPPN, Psychology & NeuroscienceBSU in which the animals will be held (*select as appropriate)Franklin-Wilkins St Thomas’ Guy’s – Hodgkin Guy’s – New Hunt’s HouseInstitute of Psychiatry Maurice Wohl InstituteHodgkin Fish Facility 2. Who can be contacted regarding your animals if you are unavailable?Full name including titlePositione-mail addressContact phone no.Personal Licence no.3. Project Licence detailsWhich Project Licence will your study be conducted under?Project Licence holderWhich Protocol no. will your study be conducted under?Please note that if your study involves more than one protocol on the Project Licence then a separate form should be completed for each protocol.Which aim/objective of the Project Licence does your study align with?Biological ServicesRegulated procedures study plan formThis form should be used to apply for:- Obtaining NACWO approval for a new study to involving regulated procedures be carried out in a BSUConclusionThe introduction of the study plans has been a great positive for research at KCL by ensuring compliance andhelping to promote the principles of the 3Rs.They have helped prevent non-compliance and given the animal facility staff easy access to the study plans and a better understanding of the experiments being performed on the animals.3RsReplacement: avoiding or replacing the use of animals in areas where they would otherwise have been used.Refi nement: minimising the number of animals used which is consistent with scientifi c aims.Refi nement: minimising the pain, suffering, distress or lasting harm that research animals may experience.There have been many different refi nements made to the study plans due to the approval process. Examples are:– Refi nement of sepsis scoring system and endpoints for a severe protocol.– Experiment involving myocardial infarction that was going to be performed as a recovery model and as a severe protocol. This led to it being carried out as a non recovery procedure instead.– The change of the duration of the time course for mice on a severe protocol, reducing the time the micewould have potentially been suffering due to the administration of the drug.–Non regulated instead of regulated experiments beingcarried out.that endpoints and actions to be taken are clearly stated and Animal Welfare costs have been minimised. As the study plan process evolved it has become much more than a tool to approve and manage researchers plans of work. The information from study plans help to review the training and competency records of researchers. This also helps with health and safety by checking occupational health status and reporting of any substances which would be hazardous to health. Conclusion

246Animal Technology and Welfare August 2020IntroductionOur study into the implementation of running wheels in overweight prone strains of mice and the potential integration of this practice into our mouse protocols.Aims and objectives– To monitor the health and welfare of our overweight laboratory mice.–Induce and enhance animal enrichment whilst reducingobesity.– Document the weight, food and water consumption of all mice involved (both in the cages with wheels and in the control cages without the wheels).– Implement the 3Rs by determining that the use of wheels is benefi cial to the mice.HypothesisH1 cages with the implementation of running wheels willhave signifi cant weight difference in comparison to the control cages.To wheel or not to wheelPENNY IVES and JADE CAYGILLUniversity of ManchesterCorrespondence: penny.ives@manchester.ac.uk Null cages with the implementation of running wheels will not have a signifi cant weight difference in comparison tothe control cages (Figure 1).MethodologyThe cages were given aspen, sizzle nest, tunnel, transbreeddiet and wheel in 4 out of 8 cages.Mice were taken at weaning age of 21 days old and using arandomised approach to wheel and no wheel, determinedwhich cages were given a wheel or not to avoid bias.Once half of the cages were allocated a wheel, the rest were placed as control.Every week each mouse was weighed and the following components were weighed:• water• food left• food replacedThe condition of each nest was recorded to help give an idea on stress levels within each cage.Each base was cleaned weekly. This study trialled for 16 weeks with a total of 30 mice.Figure 1. Figure 1MethodologyThe cages were given aspen, sizzle nest, tunnel, transbreed diet and wheel in 4 out of 8 cages.Mice were taken at weaning age of 21 days old and using a randomiser the phrase, wheel and no wheel determined which cages were given a wheel or not to avoid bias.Once half of the cages were allocated a wheel the rest were placed as control.Every week each mouse was weighed and the following components were weighed:Figure 1MethodologyThe cages were given aspen, sizzle nest, tunnel, transbreed diet and wheel in 4 out of 8 cages.Mice were taken at weaning age of 21 days old and using a randomiser the phrase, wheel and no wheel determined which cages were given a wheel or not to avoid bias.Once half of the cages were allocated a wheel the rest were placed as control.Every week each mouse was weighed and the following components were weighed:Figure 1MethodologyThe cages were given aspen, sizzle nest, tunnel, transbreed diet and wheel in 4 out of 8 cages.Mice were taken at weaning age of 21 days old and using a randomiser the phrase, wheel and no wheel determined which cages were given a wheel or not to avoid bias.Once half of the cages were allocated a wheel the rest were placed as control.Every week each mouse was weighed and the following components were weighed:Animal Technology and Welfare December 2023

247August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsResultsFrom the data, we found that on average the cages with wheels had a weight gain of only 1.5g (Figure 2). The cages with wheels drank on average an extra 10.6g of water per week and ate 3g more of diet.Although the mice in the cages with wheels gained weight,the nests were found to be neater and more consistent in the cage. This could be an indicator of less stress due to increased enrichment in the cage.Throughout the study we only had 2 cages that developed stereotypical behaviour. One cage did not have a wheel and one cage did. The cage without a wheel started showing this behaviour from week 14 and the cage with a wheel from week 15.ConclusionFrom this study although it did not meet the hypothesis set out at the start, the fi nal results showed that providing a wheel in the cages decreased stereotypical behaviour and increased metabolic needs e.g. food and water intake.Figure 2. Figure 2 Although the mice in the cages with wheels gained weight, the nests were found to be neater and more consistent in the cage. This could be an indicator of less stress due to increased enrichment in the cage. Throughout the study we only had 2 cages that developed stereotypical behaviour. Once cage did not have a wheel and one cage did. The cage without a wheel started showing this behaviour from week 14 and the cage with a wheel from week 15. Conclusion

248Animal Technology and Welfare August 2020IntroductionWith the use of fish in research continuing to rise, the requirements to adapt facilities for the housing of multiple different species has grown.In many instances the introduction of new aquatics species requires room alterations, building management system amendments, compliance with regulations differing to that of the mammalian species and structural modifications.The University of Cambridge created 2 Containment Level 2 (CL2) aquatics facilities (Figures 1 and 2), housing multiple species of fish infected with Mycobacterium marinum. Here we define the PPE used to prevent the spread of Mycobacterium marinum in a multi roomed facility, the CL2 system design solutions, training, fish feeding and screening of fish within the facility.Containment of Mycobacterium marium within a containment level 2 aquatics areaNICOLA GOODWINUniversity of CambridgeCorrespondence: ntg22@cam.ac.uk Personal Protective EquipmentPotential routes of contamination are ingestion, inhalation and inoculation with particular risk of infection through contact of skin, eyes, and mucous membranes. To ensure the safety of the staff within the CL2 area and prevent contamination to our other fish rooms, the following PPE has been implemented in line with Health and Safety Executive (HSE) recommendations, this is donned within a clean zone of CL2 and removed before leaving CL2:– Conducting procedures not requiring the removal of a tank from the rack – CL2 specific shoes, 2 pairs of gloves, disposable lab coat, face shield.– Conducting procedures requiring removal of a tank from the rack – CL2 specific shoes, 2 pairs of gloves, Tyvek with boots due to the increased risk of splashing contaminated water, face shield.Figure 1. CL2 Room 1. Figure 1 CL2 Room 1 Figure 2. CL2 Room 2. Figure 1 CL2 Room 1 Animal Technology and Welfare December 2023

249August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsTraining facility technicians and usersWithin our facilities, staff movement is permitted between rooms and CL2 without cross contamination. This has been achieved by:– All staff must be trained and assessed with in depth knowledge of the CL2 control measures, health and safety risks and legislation.– Where possible perform tasks at the end of the day.– Standard operating procedures and risk assessments must be read and understood by each member of staff before working in the area.– Code of Practice provided detailing; work conducted in the area, hazards, disinfection processes, control of contamination, process to handle spills, waste disposal, approved disinfectants and key contacts.Disinfection and cleaning procedures– Water disinfection: collection within storage tanks and provided chemical treatment before entering drains.– Tank cleaning: cleaning of non infected tanks first. All tanks cleaned using tissue and Reverse Osmosis water (RO) before being autoclaved at 134°C using a validated, efficacy tested autoclave.– Non autoclavable tank components: baffles are cleaned with 70% ethanol for 2 minutes before rinsing with RO water.– Nets: almost all are autoclavable, those which are not are cleaned with 70% ethanol. Training facility technicians and users Within our facilities, staff movement is permitted between rooms and CL2 without cross contamination. This has been achieved by: • All staff must be trained and assessed with in depth knowledge of the CL2 control measures, health and safety risks and legislation. • Where possible perform tasks at the end of the day.

250Animal Technology and Welfare August 2020Fish feedingRisk: Cross contamination of bacteria from infected tank to clean tank.Solution:• separate feeding equipment (one for non infected and one for infected)• feed uninfected fish first, then infected• unused food should be discarded after each group is fed• cleaning of feeding equipment after each group is fed• colour coded feeding equipment and tanks for ease of differentiation between control and infected animals and equipmentScreening of fish and swabs for Mycobacterium marinumScreening for pathogens potentially harmful to fish health is routinely performed in an aquatic facility. Within a CL2 facility, the screening performed is not to confirm what pathogens you have, but rather to confirm the processes you have in place are reliably preventing cross contamination.Procedure at University of Cambridge• screen control fish every 3 to 6 months• screening of fish by PCR and histology• screening of swabs of tank biofilm pre and post filtration• swabs of room surfaces to confirm room hygiene standards are maintainedFurther readingZebrafish Journal – Design and Husbandry Considerations for a Containment Level 2 Aquatic Facility – Nicola Goodwin and Lynda Westall.Poster Presentations

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