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Vol 22 No 1 April 2023ISSN 2752-3918Offi cial Journal of the Institute of Animal Technology and European Federation of Animal TechnologistsIAT JournalAnimal Technology and Welfare●Andrew Blake Award winning entry ● Xenopus, Xenopus, Xenopus! ● Final Congress 22 posters

1Vol 22 No 1 April 2023Editorial Jas Barley, Chair of the Editorial BoardNorth African animal legislation: a review A.N. Other ABSTRACT TRANSLATIONSAndrew Blake Tribute AwardValidating the use of box training as a refi nement to rabbit handling Alicia Kinally, Lucy Onions and Sarah Glenn TECH-2-TECH Improving the welfare of mice used in the study of ageing mice: Collated Workshop Feedback – March 2021 and March 2022 Linda Horan and Michael Wilkinson Sensory enrichment for the common marmoset (Callithrix jacchus) Millie FullerHow to breed and raise African Clawed-Toed frogs (Xenopus laevis) Therese Jones-Green African Clawed frog (Xenopus laevis) diseases and clinical signs in photographs Therese Jones-Green POSTERSValidating in-cage mouse enrichment Chelsea Burbidge, Zoe Beresford and Valentina Serrano-GallegDigital photography and the care and welfare of African Clawed frogs (Xenopus Laevis) Therese Jones-GreenMinimising aggression in CD-1 and CD-1 background male mice with different enrichment types Amy Veness, Chis Coyle, Sian Murphy, Jamie Redden, Tina O’Mahony and Eleni M AmanitiA refi nement in hamster breeding: creating a sustainable colony for vaccine research K. Anderson, M. Davenport, J. Finney and S. MacCuishEditorialJas 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 i4135477463539744682277

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 Tel: +44 (0)1293 827940Email: sales@lbs-biotech.comContact LBS - your trusted supplier, servingthe needs of the Biotechnology Industry www.lbs-biotech.com
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3vOFFICERSPresidentDr 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: Glyn Fisher FIAT RAnTechChair of Board of Educational Policy: Steven Cubitt MSc FIAT 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: Jas Barley (Chair), Nicky Gent, Patrick Hayes, Diane Hazlehurst, Elaine Kirkum, Carole Wilson, Lynda WestallBranch Liaison Officer:Kally Booth MIAT RAnTechEFAT Representatives:Glyn Fisher, Robin Labesse MIAT RAnTech, Toby SandersWebsite Coordinator:Allan Thornhill FIAT RAnTechAnimal Welfare Group:John Waters (Chair), Carmen Abela, Kally Booth, Nicky Gent, Sam Jameson, Sylvie Mehigan, Steve OwenBoard of Educational Policy:Steven Cubitt (Chair), Adele Kitching (Secretary), Diane Hazlehurst, Robin Labesse, Tina O’Mahoney Communications Group:Adrian Woodhouse (Chair), Carmen Abela, Kally Booth, Hannah Easter, Sam Jameson, Wendy Jarrett, Elaine Kirkum, Teresa 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 Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.) DipECLAM FRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&S MSc MRCVS, Tim Morris BVetMed PhD DipACLAM DipECLAM CBiol FRSB CertLAS MRCVS, Clive Page OBE PhD BSc, Jan-Bas Prins PhD MSc, Vicky Robinson CBE BSc PhD, Paul Sanders MIAT RAnTech, David Spillane FIAT, Gail Thompson RLATG, Robert Weichbrod PhD RLATGLife 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) RAnTech, Phil Ruddock MIAT RAnTech, Ted Wills FIAT (Hon) RAnTechHonorary MembersMark Gardiner MIAT RAnTech, Sarah Lane MSc FIAT,Stuart Mackrell FIAT RAnTech, Sue McHugh BSc FIAT, Wendy Steel BSc (Hons) FIAT Members of CouncilCarmen Abela, Ken Applebee, Kally Booth, Steven Cubitt, Simon Cumming, Haley Daniels, Glyn Fisher, Nicky Gent, Alan Graham, Diane Hazlehurst, Linda Horan, Sam Jameson, Elaine Kirkum, Adele Kitching, Robin Labesse, Theresa Langford, Sylvie Mehigan, Tina O’Mahony, Toby Sanders, Allan Thornhill, John Waters, Lynda Westall, Carole Wilson, Adrian Woodhouse

4BRANCH 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, Charlie Chambers, John Gregory, Cathy Godfrey, Kathy Ryder, Wendy Steel, Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee:Alan Graham (Chair), Haley Daniels, Adele Kitching,Allan Thornhill, John WatersEquity, Diversity and Inclusion Officer: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 viBRANCH SECRETARIES 2023Cambridge: 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: Louise Fisher 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.ukWest of Scotland: Joanne King westscotlandbranch@iat.org.ukDatesand Ltd................................................ IFCAvid plc........................................................... 6College of Laboratory Animal Science and Technology (CLAST)..................................... 26Institute of Animal Technology................ 5, 62, 0BCIPS Product Supplies Ltd............................... IBCLBS Serving Biotechnology Ltd......................... 2Tecniplast UK Ltd............................................. 8

If the answer to the above two questions is yes then listen up:The Institute of Animal Technology (IAT) is actively seeking new Council members, so whether you are interested in welfare initiatives, communications, education and training or just want to have a say, then why not get more involved in your professional body and join Council?Our Mission: Advancing and promoting excellence in the care and welfare of animals in researchAre you currently a full Member or Fellow of the Institute with two years’ consecutive membership, who wants to make a difference? Do you want to be involved in developing the future of the Institute of Animal Technology?Visit our website www.iat.org.uk for further informationInstitute of Animal TechnologyYOUR COUNCIL NEEDS YOU!Why bother?• the opportunity to shape the future• great networking and personal development opportunities• choose the area that interests you on Council and join that group• expenses are paid to attend Council meetings• discounted Congress attendance• the Council Election form is simple to completeStill unsure?Contact us and we can talk you through the process, provide more information about Council activities and groups or you can visit the IAT website members’ section.You would need to be proposed and seconded by either two Members or Fellows of the Institute or nominated by a Branch. The IAT can assist with this too, if you have any difficulties.Interested?Nomination forms should be completed online from the IAT website using this link http://iatforms.org.uk/view.php?id=16312When the form is submitted it is sent to the IAT Administrator (admin@iat.org.uk). The closing date for nominations is Friday 3rd November 2023. Simon CummingHonorary Secretary

7August 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 differ ences 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 r epor t. 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 recurring problem, bacterial infections such as Pseudomonas asdescribed in the reprint of the article were still presenting Animal Technologists with problems as late as the end of the 1980s. Whenimporting animals and tissues from overseas it is important to realise that they may be carrying disease not seen in the UK forseveral decades. In recent times, Ectromelia was introduced into a unit in the USA via antibodies produced overseas. Precautionsmust be taken until such time as you are sure that the animals and tissues are clear of any underlying infections.We are also able to offer in this issue an interesting Tech-2-Tech article by Seonagh Henderson of the University of Glasgow, ona novel technique of cage cleaning which hasa positive effect on the welfare of 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 ixFirstly allow me to apologise for the late publication of this issue. The cause for this has been due to many reasons, namely the 2500 members plus of the Institute of Animal Technology who have failed to put the important developments in Animal Technology they have devised into words. I know that you are busy, when have Animal Technologists ever been able to sit around twiddling their thumbs but, for the past 70 plus years we have generally had a steady stream of material for the Journal from technical staff. Obviously the COVID-19 pandemic changed work patterns a great deal but the importance of the way Animal Technologists develop new techniques to improve Animal Welfare has not changed and you need to blow your own trumpets. So please support ATW and write up your work, the tradition of the Editorial Board of helping authors has not changed and we will do our best to enable you to achieve the recognition for your work that you deserve. This will be my last year as Editor of Animal Technology and Welfare and Diane Hazlehurst will be gradually taking over during the year. I have loved every minute (well most of them at least) of putting each issue together over the past 13 years but its time to go. Every well managed organisation has a succession plan and my decision to finally retire from IAT Council is part of that, I have new aspects of my life that I want to develop and to enjoy some free time. Hopefully you will support Diane and she will be able to continue the traditions of the IAT Journal but at the same time develop it to grasp the challenges of the future.The review of North African animal legislation looks back to the sadly, now defunct, Post-Graduate Diploma course in Laboratory Animal Technology, which led to several of the attendees gaining a MSc degree in Laboratory Animal Technology. The article is the result of a project that all the students were given as part of the law module, to review the legislation concerning animals in different areas of the world, It also, I think, demonstrates that this profession not only concerns itself in what happens in the UK and Europe but also much further afield. Its old work but is still relevant even though some of legislation has subsequently been repealed or developed. I am delighted to be able to include a Tech-2-Tech article from Alicia Kinally, Lucy Onions and Sarah Glenn from the University of Leicester based on their poster which was awarded the 2023 Andrew Blake Tribute Award. It explains an idea to improve the handling of rabbits in order to reduce stress for them, it has potential for all Animal Technologists to rethink rabbit handling and thus considerably improve the lives of rabbits. The issue also contains several other Tech-2-Tech articles based on Congress 22 posters, if you care for Xenopus frogs in your facility then the two Tech-2-Techs by Theresa Green should be essential reading for you. Similarly, the article from Linda Horan and Michael Wilkinson article on the welfare of ageing mice is also important reading for anyone working with this class of animal.The last of the Congress 22 posters are included covering topics which comprise work on minimising aggression in mice, validating mouse in cage enrichment and refining hamster breeding. EditorialJas BarleyChair of the Editorial BoardApril 2023 Animal Technology and Welfare

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9August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2023 Animal Technology and WelfareIntroductionThe following review was a student assignment as part of the Institute of Animal Technology Post Graduate Diploma (PGDip) course in Laboratory Animal Technology and Science, established in 1992 in conjunction with theUniversity of Middlesex. Students successfully gaining their PGDip were able to continue their studies to achievea Master of Science Degree, by dissertation, in LaboratoryAnimal Technology.As part of the law module, students were given the task of reviewing legislation relating to animals in different parts of the World. The original remit of this project was to survey legislation pertaining to animals in the African states north of the Sahara Desert. Unfortunately no one authority appears to agree on the current confi nes of the Sahara, therefore an arbitrary decision was made to survey those African states where the majority of the land mass lies north of the equator. In countries where the landmass lies equally either side of the equator, the position of the capital city has been used as the deciding factor, hence the Gabon has been included in this review while Kenya and the Congo has been classifi ed as South Africa. Using this rationale 30 countries were identifi ed as falling into the classifi cation ‘North Africa’. 19 UK embassies or consulates of the countries concerned were approached and a further 10 states were addressed via the United Nations. Only in the case of Cape Verde was contact not made, due to the author being unable to ascertain whether the islands were an independent state or North African animal legislation: a reviewA.N. OTHER Institute of Animal TechnologyCorrespondence: atweditor@iat.org.comdependency of another national power, (it has since beenascertained that whilst once a former colony of Portugal,it is now an independent state). Most of the approaches made to the appropriate UK embassy were unproductive and much of the information used in this review was gained from library sources. References were updated where possible in 2023. The legislation reviewed falls into fi ve major groups: – Animal in Research/Science. – Conservation Legislation. – Laws relating to Animal Protection.– Legislation affecting Animals in Agriculture.– Animal Health Legislation.3 the countries concerned were approached and a further 10 states were addressed via the United Nations. Only in the case of Cape Verde was contact not made, due to the author being unable to ascertain whether the islands were an independent state or dependency of another national power, (It has been it has since been ascertained that once a former colony of Portugal, it is now an independent state). Most of the approaches made to the appropriate U.K. embassy were unproductive and much of the information used in this review was gained from library sources.. References were updated where possible in 2023. Figure 1. Countries of Africa The legislation reviewed falls into five major groups: - Animal in Research/Science., - Conservation Legislation. - Laws relating to Animal Protection. Figure 1.Countries of Africa. 3 the countries concerned were approached and a further 10 states were addressed via the United Nations. Only in the case of Cape Verde was contact not made, due to the author being unable to ascertain whether the islands were an independent state or dependency of another national power, (It has been it has since been ascertained that once a former colony of Portugal, it is now an independent state). Most of the approaches made to the appropriate U.K. embassy were unproductive and much of the information used in this review was gained from library sources.. References were updated where possible in 2023. Figure 1. Countries of Africa The legislation reviewed falls into five major groups: - Animal in Research/Science., - Conservation Legislation. - Laws relating to Animal Protection. Untitled-3 1 02/02/2023 13:00:11

10Animal Technology and Welfare August 2020Legislation relating to animals use for scientific purposesOf the countries surveyed only Uganda had specific legislation relating to the use of animals in experiments. The Animals (Prevention of Cruelty) Act 1957,1 combines aspects of the English Protection of Animals Act 1911,2 and the now defunct Cruelty to Animals Act 1876.³ In addition to defining cruelty, etc., in a similar way to the 1911 Act,² (this aspect will be reviewed at a later point) the Act prohibits the use of live animals in experiments calculated to cause pain, the exception to this being the use in experiments performed with a view to the advancement, by new discovery, physiological knowledge or of knowledge that will be used in saving or prolonging life or alleviating suffering. These uses of animals are only permitted after the issuance of a licence to those concerned. The performance of experiments on animals without the use of anaesthetics is strictly controlled and the use of animals in experiments as part of a lecture is prohibited. However if in the opinion of the lecturer, such experiments are necessary to enable the students to gain physiological or life-saving knowledge then this condition is waived. Using animals in experiments to acquire manual skills is also prohibited. Provision is made for the destruction of animals if pain inflicted during the experiment is likely to continue following recovery from anaesthesia. Revocation of the licences in the instance of contravention of the Act is provided for.Other legislation affecting the scientific use of animals relates to conservation. where the capture, etc., of animals is permitted if it is for scientific purposes. Algeria,4-6 prohibits the taking of protected species except for scientific purposes and then only after the award of a permit. A Statutory Instrument (S.I),7 issued by Benin as part of the Protection of Nature and Hunting Act 1980,8 regulates the issuance of Scientific Purpose permits. Passed in connection with the Wildlife Conservation and Hunting Act 1919,9 Burkina Faso establishes conditions for the issuance of Scientific Purpose Permits in a S.I.10 Further legislation enacted in 1977,11 permits the capturing in exceptional circumstances even fully protected species. An Act and S.I.12,13 passed in 1985 re-opening hunting also established additional conditions for the issuance of scientific permits.The United Republic of Cameroon14,15 permits the taking a fully protected and partially protected species for scientific purposes. Similar legislation16 enacted by the Central African Republic establishes conditions for the issuance and validity capturing permits scientific or commercial purposes.The position regarding Ethiopia is somewhat confusing, the ambassador to the United Kingdom Ethiopian and the Eritrean United Nations representative both informed the author that there is no legislation pertaining to animals Ethiopian or Eritrean (personal communication 1992). However library sources reveal a statutory instrument issued in 197217 in accordance with the provisions of the Forest and Wildlife Conservation and Development Proclamation 198018 designating certain species as specially protected except for important scientific research. Provision for Ethiopian authorities to undertake scientific research with regards to forests and wildlife resources is also made under this Act.Ghanaian legislation19 empowers the Minister to authorise the taking of protected species for scientific purposes. Similar legislation enacted by Guinea-Bissau regulates the taking of wild animals for scientific purposes.20The transport of trophies of totally protected animals in the Ivory Coast is prohibited under a Statutory Instrument21 issued under the Wildlife and Hunting Act 196522 except by holders of scientific permits. A further S.I.23 regulating the issuance and validity of game licences in the Ivory Coast issued in 1966, includes regulations for the issuance, validity and use of scientific licences, etc.Much of the legislation of Nigeria was enacted during colonial rule and was incorporated into Federal Law on independence. This includes legislation concerning the areas of Anambra, Imo Rivers and Cross Rivers,24 which permits the hunting of protected animals for important scientific purposes under a permit issued by the Minister of State. This waiver was extended in 1985 to include additional species and other areas.25-28 Similar legislation exists for Senegal.29Conservation legislationThe Convention on International Trade in Endangered Species of Flora and Fauna, Washington 1973, often referred to as CITES or the Washington Agreement, was ratified by several of the countries reviewed here. The terms of the convention are put into effect by each country’s national legislation with the intention of regulating the trade in endangered species.In addition to legislation regulating the hunting of protected species,4 including the establishment of game reserves and the spraying of pesticides which may be harmful to game species, Algeria passed in 1983 legislation,5,6 to protect the environment. This deals with all aspects of environmental conservation including the control of air, freshwater and marine pollution, waste, radioactive substances, chemical substances, noise and the preparation of impact assessment studies. With regards to species and ecosystems it provides for the establishment of river reserves and the protection of animals and plant species.North African animal legislation: a review

11August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareNorth African animal legislation: a reviewBenin’s conservation legislation,8 protects all free-living mammals (except bats, rats and mice), all birds and crocodiles, monitor lizards, pythons, turtles and fishes. The Act provides for the designation of species according to the degree of protection afforded them. Types of hunting weapons and methods of hunting are regulated. Provision for the establishment of game reserves is also made and for the killing of protected animals in self-defence or in the defence of crops. The Act also provides for enforcement measures and penalties. A Statutory Instrument issued under this legislation provides for a register for Ivory workers,7 a further S.I. passed in ratification of CITES provides that no hunter may kill more than one elephant and one hippopotamus in any one year.30Also a signatory to CITES, Burkina Faso passed in its main conservation legislation in 1968,9,10 defining wildlife as including all mammals, birds, crocodiles, chelonians, monitor lizards and pythons and vests ownership of defined wildlife with the State. The Act prohibits certain hunting methods, protects females, juveniles, eggs and nests of species and, establishes protected areas and several categories of hunting licences.Provision for the enforcement measures and penalties is also made. A further S.I provides complete protection for the Golden Cat.11 Hunting was once again authorised by an S.I. which regulates hunting for the 1985-1986 season,12 lists fully and partially protected species and restricts the hunting of said species to the areas of Gourma and Tapoa. This S.I. sets bag limits and royalties in respect of the taking of such species. A fishing ordinance,13 regulates fishing gear and prohibits the use of certain nets, explosives, toxic substances, electric devices or firearms and prohibits, except under licence, the release of chemical substances into aquatic environments with the proviso that licences shall not be granted if such release is harmful to aquatic animals or plants. The introduction of exotic fish species is also banned, except under permit. Provision is made for enforcement measures and penalties.While not appearing to be a signatory to CITES, the United Republic of Cameroon has comprehensive legislation,14 regulating hunting, possession of and trade in wild animals and trophies. In addition the Act provides protection for certain species, restricts the importation of exotic animals and plants (dead or alive) and the export of protected animals. Animals protected include all terrestrial and aquatic animals. Fishing is regulated as are methods of fishing and fishing gear. Provision is made for enforcement measures and penalties. A decree passed in 1983,32 regulates traditional hunting and certain methods of hunting are prohibited. The trade in live animals is also regulated by this decree and enforcement measures and penalties are provided for. A Statutory Instrument,15 lists protected and partially protected species and permits the taking of protected species for game management and the protection of human life. The taking of other species is regulated through bag limits and seasons. The young of all species are protected, as are the eggs of fully protected species of birds. Originally passed in 1955 when Cape Verde was a province of Portugal, the conservation legislation of this group of islands covers nature conservation in general, soils, fauna, hunting and flora. The S.I was adopted into the law of the now independent state.33 The legislation establishes rules for the preservation of soils, flora and fauna, it regulates hunting and hunting methods and establishes the requirement for hunting permits including conditions for their issuance and validity. Trade in trophies is regulated as is fishing in inland waters. Enforcement measures are also established.The Central African Republic establishes that wild animals form an integral part of the national heritage of the State in an ordinance concerning protection of wildlife and regulating hunting passed in 1984.34 The legislation defines various categories of protected areas and lays down rules for their management. Species are classified as protected, partially protected and game species. The taking of fully protected species is completely banned, partially protected species may only be taken under permit. Customary hunting is regulated together with hunting methods and the Act establishes conditions for the issuance of licences. Game species may be taken by customary hunters or under a small game licence but the principle of taking only male animals is established. The Minister is empowered to establish open and close seasons and to regulate the trade in trophies. A further Act issued in connection with CITES,35 regulates the capture of live wild animals- fully protected species as listed in the previous Act may only be taken with the express permission of the Head of State and trade in such species is not permitted. Other species may be captured only by the Wildlife Department or under its control, a permit being required if capture is by an organisation other than the Wildlife Department. The definition of live wild animals includes the eggs of birds and reptiles. Additionally an Act passed in 1995 prohibits the hunting relevance throughout the territory of the Republic.36Egyptian conservation legislation,37-40 covers particular areas of the State and prohibits the taking of animals and birds in certain parts of these areas. In Sinai the fishing for all species of coloured fish is prohibited in Ras Mohammed area as is the removal of special specimens from the coral reefs.41Legislation providing for conservation in Equatorial Guinea,42 was passed by the Spanish Administration in 1953 but it is unclear if legislation is still in force. And S.I. It established protected areas and provided for the necessity of hunting licences. Species were

12Animal Technology and Welfare August 2020classified according to their degree of protection with the females of all species being singled out for special protection. Traditional hunters were not required to hold hunting permits providing that traditional methods and weapons were used. Similar confusion exists over conservation legislation in Ethiopia and Eritrea (see scientific legislation). A Statutory Instrument (S.I.),17 passed in 1972 classified as game, all wild animals including birds and reptiles which are not vermin. Full protection was given to females and their young. Specially protected species were listed and these could not be taken except for important scientific research. It has not been possible to ascertain if this legislation is still valid in any part of Ethiopia or Eritrea.In 1981, Gabon passed an S.I.43 suspending all big game and tourist hunting licences and banned the capture of certain species including the Mountain Gorilla, Chimpanzees, Elephants and Hippopotamus. A subsequent S.I.44 reopening hunting for small game and renewed the provisions concerning the species listed in the previous S.I.43 Protected areas were established in The Waters And Forest Act 1985,45 which additionally regulated activities in such areas, hunting methods and possession and, domestic or international trade in wildlife and trophies. Enforcement measures and penalties are specified.All vertebrates, except fish, are protected in the Gambia under the Wildlife Conservation Act 1977.46 It provides for the development of wildlife management plans together with the establishment of protected areas. Certain hunting methods are banned as is the trade in wild animals, meat and trophies with the exception of legally hunted Bush pig. An enabling act, it empowers the Minister to authorise trade in other species and to make regulations to further the implementation of the Act. In 1977 an S.I. prohibits the sale of wild animals and of meat and trophies except in respect to the sale of game meat by Indigenes (Indigenies are Gambians citizens, members of tribes indigenous to Gambia and the inhabitants of villages in the provinces). Vermin species are listed in a further S.I.48The Wild Animals Preservation Act 1961 of Ghana lists species in five schedules.19 It empowers the president to amend these schedules. Hunting using motor vehicles or aircraft is prohibited as is the surrounding of animals by fire. The Act empowers game officers to arrest without warrant. It prohibits the import into Ghana, except with certification of lawful export, of trophies exported from territories to which the Convention on the Protection of Fauna and Flora of 1933 applies. The 1971 Wildlife Conservation,49 regulations completely replace the list of protected species which appeared in the previous Act. It prohibits the hunting, capture or destruction or possession of protected species. Also prohibits the hunting, capture or destruction of young or animals accompanied by their young of partially protected species. It prohibits the export, alive or dead, of animals listed as protected, partially protected or game species.Hunting Regulations (Statutory Instrument) 1980 issued by Guinea Bissau prohibited capturing, collection, destruction or sale of birds, bird nests and eggs of protected species listed in the regulations.20 The State Commission for National Resources is empowered to prohibit the taking of any species throughout the national territory or any part of it. Certain hunting methods are banned under the S.I. Females of game animals when accompanied by their young are protected and hunters are required to generally avoid shooting female game animals. Bats, rats and mice are excluded from the provisions of the Ivory Coast conservation legislation.22 All other mammals, birds, crocodiles, monitor lizards and pythons are protected. The legislation deems species listed as being fully or partially protected or as game species. Licences are required for the taking of, in limited numbers, of partially protected species and game species.The only piece of legislation relating to animals that the author of this review has been able to find from Mauritania is the Hunting and Wildlife Protection Act 1975.50 Backed by enforcement measures and penalties, this Act provides for the requirement of a licence for hunting of wild animals. It empowers the appropriate government departments to make regulations regarding the issuance of licences, to declare open and close seasons and to designate totally or partially protected species. Commercial capturing, etc., is regulated. Certain hunting methods are prohibited under the Act as is the trade in game and game meat. Young animals and females accompanied by their young are afforded full protection, similarly wild birds and the eggs of wild birds (particularly the ostrich) which are not listed as pest species are also protected. However the taking of a protected wild animal in order to protect human life is authorised and the Act empowers the Minister to authorise the taking of a protected animal when they are causing damage.Several Acts and subsequent amendments and Statutory Instruments regulate hunting in Morocco. The main legislation, Hunting Act 1923,51 amended 17 times between 1932 and 1969, establishing conditions under which hunting rights may be exercised by private persons. Conditions for the issuance and validity of hunting licences are established by the Act as are enforcement measures and penalties. Protected and harmful species are listed and the Act empowers the competent authority to make regulations for hunting seasons, bag limits, the trade in game meat and the regulation of the hunting methods. An S.I.,52 enacted in 1962, authorises the destruction of pest species by landowners at any time, North African animal legislation: a review

13August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfareexcept by fire or bacterialogical methods. It also regulates the distruction of wild boar when they are causing damage to crops. Other aspects of the S.I. concern regulation of bag limits for species, the trade in game meat, etc. A second S.I. regulates the 1985 to 1986 hunting season.53 It lays down additional rules for the hunting of wild boar, prohibits the trade in certain types of game meat and provides that the sale of other types of game meat must be authorised by the Director of Waters, Forestry and Soil Conservation. Protected species on this and prohibits the taking and trade in such species.The conservation legislation of Niger,54-57 provides for the regulation of hunting and prohibition of the sale of game meat. Further regulations are to control such aspects of conservation as the possession, sale, importation and exportation of wild animals and trophies or products Females of game species are fully protected.In addition to implementing CITES, the Endangered Species (Control of International Trade and Traffic) Decree 1985,58 of Nigeria also implements the African convention on the Conservation of Nature and Natural Resources and the Agreement on the Joint Regulation of Fauna and Flora on the Lake Chad Basin. It prohibits the hunting, capture or trade in animal species threatened with extinction. The Act provides enforcement measures and penalties and empowers the Minister to make regulations to effect the provisions of the Act. As mentioned previously much of the legislation relates to specific areas of Nigeria,24-28,58 but generally applies to all animals whether vertebrate or invertebrate, normally found in the wild state or to any part, nest, eggs, eggshells and young thereof. Prohibited animals are listed in three schedules – prohibited, specially protected and protected animals. The hunting of females is given special consideration. All categories of animals may be killed if considered to be a threat to life or property. The Act also provides for enforcement and penalties. The oldest legislation relating to conservation in Nigeria dates back to 1916.59 Amended in 1976 the list of fully protected species is directly derived from the appendix to the London Convention of 1933 and contains several species not native to Nigeria. It requires game licences for hunting and regulates the trade in trophies. It specifically prohibits the sale of powdered rhino horn but makes provision for the granting of special licences to kill rhino or elephants. Enactment of this legislation was divided into several areas of Nigeria.59Conservation in Senegal is provided for by the Hunting of Wildlife Protection Code (Statutory Part) 1967.60 The Act provides for the regulation of hunting through licences with the exception of hunting on enclosed property by the owner of that property. Wild animals may be killed in defence of life or property. Infractions of the regulations are to be enforced and penalties are provided for. Species and numbers of animals which may be taken by hunters under a hunting licence are specified in an S.I., enacted in 1967 with further amendments made in 1969, 1973 and 1980.29 In order to hunt lion and hippopotamus a permit from the President is required. Human activities in protected areas are prohibited or regulated. Certain hunting methods and weapons are prohibited as is the trade and export of game meat. Legislation regarding the conservation of wild animals appears to be limited in Sierra Leone, rules passed in 1950,61 provide that no animal may be exported from Sierra Leone except through Freetown and regulate the conditions of transport and confinement of wild animals pending export. A further Act, The Wildlife Conservation Act 1972,62 provides for the establishment of protected areas and also the issuance of hunting licences and permits. It empowers the Minister to make regulations to give effect to the provisions of the Act and provides for enforcement measures and penalties. No further regulations seemed to have been enacted.Dated 1969, the Somalian Law on Fauna (Hunting) and Forest,63 provides for the establishment of game reserves, lists various categories of species protected by the Act and provides for requirement of hunting licences. The killing of protected animals is authorised if in defence of life or property and the hunting of young and females, with the exception of vermin species, is prohibited. Trade in trophies is closely regulated. The Minister is empowered to make regulations to implement the Act and to enforce it.The original Act to conserve wildlife in the Sudan was passed in 1935 and was still valid as of 1983.64 It lists protected species with specially protected species being listed in Schedule one to the Act; these species may only be taken by special permission of the Minister. Other schedules list different degrees of protection and include species which may not be hunted with firearms except under licence, the Minister is empowered to amend schedules as required. The hunting of young and females is given special consideration. The establishment of National Parks and other protected areas is provided for. Export of animals and trophies is regulated by permit, unusually permits are not required for the export of articles made from parts of elephant or hippopotamus but the sale of protected animals is prohibited. Elephant tusk and rhino horn must be marked before export. The Minister is empowered to make regulations for the carrying out of the Act.Generally applying to all animals (exceptions being bats, rats and mice), all birds, crocodiles, turtles, monitor lizards and pythons, the main Act relating to conservation in Togo,65 lists species as protected, predatory and game. Ownership of wild animals is vested with the State. Hunting licences are required except by customary hunters or in the case of defensive life or property. Customary hunting is regulated and the use of certain North African animal legislation: a review

14Animal Technology and Welfare August 2020weapons is prohibited. The Act provides for the establishment of protected areas and for enforcement measures and penalties. Implementation regulations,66,67 deal with trade in export and import of trophies in addition to other aspects of hunting such as totally prohibiting night hunting.Tunisia deals with conservation under the Forest Act 1966.68 Whilst mainly dealing with the conservation of forest and the exploitation of timber, the Act also provides for the requirement of hunting licences and for the declaration of hunting seasons. Certain hunting methods are prohibited, as is the trade of game animals and game meat during the close season. Trade in eggs, nests and broods of wild animals is prohibited. The Act lists certain protected mammals and provides for the establishment, by the Minister, of a list of protected birds.The Fish and Crocodiles Act 1950 regulates fishing and the taking of crocodiles in Uganda.69 The use of poison, explosives or electrical devices is prohibited. Taking, and trade in crocodiles or crocodile skin is regulated. The Minister is empowered to control fishing methods by making regulations. The introduction of exotic fish species is prohibited except under permit. Enforcement measures and penalties are provided for and conditions are established for the issuances of licences. Protection of other species is provided for in the Game (Preservation and Control) Act 1959.70 Protected species are listed as game species and the Act provides for the requirement of different categories of hunting licences. The legislation empowers the Minister to declare hunting seasons and to protect or restrict the hunting of certain species. Import and export of animals and trophies are regulated. An amendment to the Act adopted in 1970,75 provides that all domestically produced ivory belongs to the State and that all ivory in private hands was to be surrendered. Enforcement measures and penalties all provided for. Animal Welfare legislationThe majority of North African legislation protecting animals is related to conservation. The only Act relating specifically to protecting animals from cruelty that the author of this review could find listed, is the Animals (Prevention of Cruelty) Act 1957 of Uganda.1 As discussed previously the Act combines provision against general cruelty to animals and to restrict the use of live animals in experiments. The Act makes various definitions, in addition to terms relating to enforcement of the Act such as ‘authorised officers’, it also defines the offence of cruelty. As in the UK 1911 Act,3 it is considered an offence for any person to cruelly beat, kick, ill-treat, over-ride, over-drive, overload, torture or infuriate any animals or to cause, procure of if the owner permits any animal to be so used. The killing of an animal in an unnecessarily cruel manner is also considered to be in offence as is the performance of any operation on an animal performed without due care or humanity. The administration of drugs without reasonable cause or excuse is considered to be an act of cruelty and therefore an offence. An owner is considered under the Act to have permitted cruelty if he fails to exercise reasonable care and supervision in respect to the protection of the animal from cruel acts. The slaughter of animals or the preparation of animals for slaughter, if for human consumption, is not covered by this legislation providing that unnecessary suffering is not inflicted. Where an owner is convicted of cruelty, the Court may order the destruction of the animal if it is considered cruel to keep the animal alive. There is no appeal against a destruction order. If an owner is convicted of cruelty under the Act the Court is empowered to deprive the owner of the animal and may dispose of the animal as it thinks fit.The sale, use, etc., of poisons (including poisoned grain) in connection with animals is an offence under this Act unless it was for the purpose of destroying insects or vermin and providing that sufficient care was taken to prevent access to the poison by dogs, cats, fowls and other domestic animals. Authorised officers are empowered to slaughter without the owner’s consent, any animal considered to be so diseased or injured that to move it would be cruel. If an animal can be moved without suffering then the Act deems it to be a legal responsibility of the owner to move it with as little suffering as possible and without delay. The Act also provides for the recovery of Court costs from animal owners convicted under this Act.The Sudan regulates the export and import of animals via the Animals Export and Import Act71 which prohibits the export from Sudan of camels, horses, mules, donkeys, cattle, sheep or goats unless they have been purchased in a public market and a market certificate of sale obtained or unless an export permit has been obtained. Quarantine prior to export is provided for under the Act. The responsibility of owners to provide water and forage for animals in quarantine is also established.Somalian ships and vehicles carrying animals must possess a special permit from the Secretary of State for livestock, stating the maximum numbers of animals that may be carried by that particular vehicle, etc. The provision for this is made under the Veterinary Code.72 Import and export of animals is also controlled under this Code although the provisions are generally in connection with animal health and consequently this aspect will be discussed elsewhere in this review. However special provisions are made for ensuring the Welfare of Animals during transport and the code generally prohibits the export of female animals.North African animal legislation: a review

15August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareLegislation relating to animals in agricultureSeveral countries permit the killing of protected animals in defence of crops.8,24,25,50 The Statutory Instrument passed in the Ivory Coast,73 specifies the conditions under which harmful animals may be destroyed. It requires the weight and length of the tusks of an animal which is killed in defence of crops to be reported.Egypt specifically protects birds used in agriculture under the Protection of Birds Beneficial to Agriculture and Protection of Wild Fauna Act.74 Open seasons for Quail are established in a subsequent S.I., which also protects animals beneficial to agriculture.39The Liberian National Livestock Artificial Insemination Act 1956,75 defines Artificial Insemination (AI) and lists the aims of the Act as an establishment of a National Livestock Bureau which is to establish contact with Institutions already familiar with AI practices in tropical countries. The Act provides for the development of regional AI programmes, the enforcement of sanitary measures and the provision of training in AI techniques and practices.Included in the remit of the Nigerian National Animal Production Research Institute established under the Agricultural Research Institute’s Decree 1973,76 is the genetic and nutritional improvement of livestock of economic importance, the improvement of livestock management and husbandry practices, the economies of meat production, the integration of livestock in farming systems and the effects on rural populations and any other problems related to animal production. Also established under this Act is the National Veterinary Research Institute. The remit of this establishment includes conducting research into all aspects of animal disease, all aspects of animal nutrition, the introduction of exotic stock to improve meat, milk and egg production and the standardisation and quality control of manufactured animal feeds.Somalian legislation,77 recognises the growing importance of livestock in the country’s economy and establishes a national livestock development agency. The original I.S. legislation passed in 1966 was superseded by Law number 34 in 1970.78 The agency’s function cover’s all aspects of the development of livestock production and marketing. In addition to disease control measures which will be discussed in the section of this review concerning animal health, the agency’s function is to establish a regular flow of graded cattle for the national canning factories, the enforcement of quality control and the rationalisation of the marketing of livestock.Agricultural animals in Uganda are protected via the Cattle Grazing Act, 1918.79 The Act defines cattle as bulls, cows, oxen, calves, sheep and goats. It establishes the powers of Veterinary Officers to declare areas as controlled and Veterinary Officers are empowered to set the number allowed to graze. The impounding of animals found grazing in contravention of the Act is permitted. Animals found straying may be seized under the provisions of the Animals (Straying) Act 1922.80 The eventual disposal of such animals is provided under the Act and lays down penalties for owners convicted of allowing the animals to stray. Zebra are included in the provisions of the Branding of Stock Act 1918 and a multitude of terms, including brand, cattle, etc., are defined.81 The Act provides for a National Brand Register and makes provision for the transfer of brands. It establishes the offence of brand defacement and lays down penalties for the same. The Trout Protection Act 1936,82 would appear to be more concerned with the protection of trout as a food source. This Act establishes that this is unlawful to fish for, capture or kill any trout in any water unless authorised by licence.Whilst not relating specifically to animals in agriculture it is appropriate to include legislation concerning the destruction of locust in this section. Both Sierra Leone,83 and the Sudan,84 have legislation which provides for the declaration of locust areas and to provide for their destruction.Many African countries have legislation connected with animals, such as Hide and Skin Acts but as these are generally related to the marketing or the production of animal products, they have been excluded from this review.Animal health legislation Under Gambian legislation85 the import or export of cattle from the Gambia without the permission of the principal Veterinary Officer is prohibited. Measures for the control of Rabies in the Gambia are established in the Dogs (Prevention of Hydrophobia) Order 1975.86 This declares various areas as rabies infected areas and empowers the police to destroy all dogs found at large, other than those found on private premises. Provisions for the control of rabies are contained in The Animal Health Regulations of the Ivory Coast.87 Special measures for quarantine periods of animals are established as is the wearing of muzzles by dogs. These regulations also contain provisions for the control of other contagious diseases including Cattle Plague or Rinderpest, Tuberculosis, Glanders, Brucellosis, Epizootic Lymphangitis, Plague, Swine Salmonellosis and Swine Pasteurellosis.Liberia via its Plant and Animal Quarantine Act 1956,88 provides for the prevention of entry into Liberia of injurious plant and animal pests and diseases. Provision under the Act is made for the prevention of the spread of North African animal legislation: a review

16Animal Technology and Welfare August 2020such pests and diseases should they become established.Nigeria has several acts concerned with animal health enacted during colonial rule which appear to have been adopted into Federal Law on independence. The Animals (Diseases) Act 1917,89 makes provisions for preventing the introduction and spread of infectious or contagious diseases amongst animals. Animals for the purposes of the Act are defined as stock, camels, dogs and ostriches and includes any animals or birds which the Governor may by notice in the North Nigerian Gazette, declares to be included in the term animal for the purpose of this Act. (Sic. ‘Gazette’ does not mean the local newspaper. The ‘Gazette’ was an official publication containing official notices, orders, etc, which was posted in the District Offices during the colonial era.) The Act also defines the term disease and includes such infections as Rinderpest, Foot and Mouth Disease, Rabies and Swine Fever together with 14 others by notice from the Governor in the North Nigerian Gazette. The Act provides for examination, testing, isolation, inoculation. Slaughter of animals, etc. and, regulates the movement of infected or suspected infected animals. Methods of disposal of carcasses are established. The Diseases of Animals Act 1962,90 regulates the importation and exportation by air of animals, animal products and ‘things’ which may cause disease. It provides for restrictions of the import and export of animals including the importation and exportation, by any means, of animal vaccines and similar substances and animal semen. A Veterinary Council was established by Nigeria for the purpose of determining standards for registration for veterinary surgeons. The Council maintains a register of vets and publishes periodical lists of registered veterinary surgeons. The Veterinary Surgeons decree, 1969,91 by which The Veterinary Council was established, empowers the Council to approve courses and to supervise instruction and examinations leading to approved qualifications. Sierra Leone makes provision against dangerous dogs in legislation, The Dogs Act enacted in 1900.92 The Act defines dangerous to mean savage, vicious or likely to spread rabies, distemper, mange or any kind of disease. The licensing of dogs is ordered and penalties for keeping a dog without license are established. The police are empowered to seize dangerous dogs and to destroy them. The Act defines the term animal to mean all stock and ruminating animals, cats and dogs but it does not include any other animals except as may be declared in the ‘Gazette’. Diseases are listed and provision is made for additions to the list to be made via the ‘Gazette’. It establishes steps which are to be taken for checking the spread of disease e.g. compulsory slaughter, isolation, disinfection, etc., and provides for compensation in the event of compulsory slaughter.Somalia enacted a Veterinary Code72 which governs matters such as the prevention of notifiable diseases of animals, the transport of animals, exportation and importation of animals and the production of animal products. The code casts a duty on owners, veterinarians and police officers to report if there is any outbreak of notifiable disease. The Secretary of State for Livestock may declare any area as infected by new notifiable disease and movement and restriction, etc., may be imposed. Slaughter of infected animals and the disposal of carcasses may be ordered by the Director of Livestock. Inspection of buildings, vehicles, ships or aircraft for infected animals may be ordered. If a notifiable disease is prevalent, the importation of any specified type of animal may be prohibited. Preventative vaccination against infectious and contagious diseases of livestock is compulsory and free of charge. Provision is made for the establishment of regulated markets for animals and animal products exported from the Republic. No camel, cattle, sheep, goats or any other kind of animal may be exported unless they have previously been brought into a quarantine station or holding ground and detained there for a specified period for the purpose of vaccination or treatment. If free of disease on inspection by a Veterinary Officer a Certificate of Health will be issued. The Code also restricts the movement of animals released from quarantine until they are exported and prescribes that animals for export must only travel to quarantine stations via certain authorised routes and must be checked at certain check points. For both animals and humans, protection from rabies is made through the Sudanese Rabies Act 1948.94 It establishes a duty to notify cases of suspected animals and the need for confinement or destruction of any suspected rabid animal. Destruction of in-contact animals is also provided for under the Act. Measures for carcass destruction are established by the Act, as is the provision for the declaration of infected areas. The Act also established penalties for failure to comply with the provision against rabies. The Diseases of Animals Act 1901,95 was enacted to deal with Cattle Plague and other diseases of animals. It establishes the duty of an owner or carer to declare any incidents or suspected case of Cattle Plague or other contagious disease. Isolation, disposal and prevention of movement of infected, suspected or in-contact animals is provided for in this Act. The practice of Veterinary Surgery in the Sudan is controlled through the Veterinary Council Act of 1954.96 The responsibility of owners or carers in terms of notification of disease incidence and for isolation of an affected animal is established in Uganda by the Animal Disease Act 1918.97 This is an enabling act (section 2) and defines animals to mean all stock, camels and other ruminating animals, cats and dogs but does not include any other animal, except those as may be declared by the Minister by Statutory Instrument to be included in the term animals for the purpose of this Act. It empowers Veterinary Officers, etc., to direct burial, destruction of carcasses of infected animals, etc., and prohibits the exhibition of animals. The Act also gives officers power North African animal legislation: a review

17August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfareof entry into premises etc., suspected of containing animals. It establishes the right to compensation for compulsory slaughter and empowers the Minister to declare ‘infected areas’ and lays down rules for such areas. The Rabies Act 1935,98 establishes the power of Veterinary Officers, police, etc., to seize and detain any stray dog in a proclaimed district with the term ‘stray‘ being defined. Veterinary Officers may dispose of any stray dog after three days if unclaimed. The Act sets down the duties of owners in the event of a dog becoming rabid or suspected of being infected and imposes penalties for breaches of these duties. Provision for the restriction of imports of animals from countries where rabies is known to exist is also made.Conclusions It is often imagined by some, that animals are given little attention in less developed countries and even less protection in law. However it can be seen from the legislation reviewed here that in fact comprehensive legislation exists in several states, particularly in respect to the conservation of animals, although how much is still valid in 2023 has been impossible to ascertain. The rapidly changing political situation in several states adds to the confusion with legislation changing each time the Government changes.12,13,18Understandably perhaps there appears to be little concern for the protection of animals used in science and one would imagine that this is due to scientific use being of little economic importance. The exception to this rule is to be found in Uganda,1 which dates back to colonial administration but appears to have still been valid in 1964 which was post-independence (1962). The issuance of scientific capture permits is a feature of much of the conservation legislation and this would appear to be the only other reference to scientific use made in any of the legislation.7,10,17,19 etc Although most states have conservation legislation the degree of enforcement of these Acts is also unknown. It should be noted that much of the conservation legislation provides for the issuance of scientific permits and hunting licences in return for fees or royalties and several Acts rest ownership of wild animals with the State.8,9,29 The majority of conservation legislation affords special protection to the females and the young of the species.It is interesting to note that the legislation of many countries surveyed have an extremely realistic approach to animals, many Acts removing protection afforded to animals if they are deemed to pose a threat to human life, either in a direct form or an indirect way through damage to crops.8,15,46,72 This practical approach to animals is also reflected in the legislation relating to animals in agriculture, appearing to be aimed more at protecting important economic reserves than protecting animals in their own right. Several Acts,8,17,22,48 excludes rats, mice, rats and other vermin from protection and usually the only reptiles that are listed in schedules of protected animals are monitor lizards and pythons, often receiving special mention in the body of the legislation,8,9,22,65 and only The Gambia appears to protect amphibians.46The special needs of native hunters are recognised in much of the conservation legislation and special provision is often made.27,32,42 Traditional hunting methods are often regulated and the use of fire is specifically prohibited by some Acts.19,42 The control of pest species is recognised,46,49,53 with Morocco uniquely prohibiting bacteriological methods for this purpose.52 Understandably colonial administration has had a great deal of influence on legislation relating to animals and several former English colonies demonstrate this for example Uganda1 and Nigeria.90There is little legislation specifically aimed at preventing cruelty to animals the exception to this, being Ugandan law.1 Where regulations regarding the transport or importation/exportation of animals exist these appear to be more as a means of protecting valuable economic resource or for the prevention of the entry of disease into a country than for the protection of animals in their own right.Where animal health legislation exists it is often aimed as much as protecting human health as it is the health of the animals. Rabies is of particular concern in many states,86,94,98 and comprehensive measures are on the statute book in several countries. Economically, diseases for example, Rinderpest and Swine Fever, are often subject to control measures and are specifically mentioned in several of the Acts.86,90,93Many of the Acts reviewed here were in enacted at an early date,59,84,92 although according to the most recent sources available they are still valid, where particular doubt exists this has been indicated. The scarcity of up-to-date information regarding legislation pertaining to animals in the African states of north of the Equator has been particularly frustrating. References1 Slade, G.B. (1964). The Laws of Uganda 1964. The Animals (Prevention of Cruelty) Act 1957. https://books.google.com/books/about/Laws_of_Uganda.html?id=RsQ0AQAAIAAJ2 Protection of Animals Act 1911, United Kingdom https://www.legislation.gov.uk/ukpga/Geo5/1-2/273 Cruelty to Animals Act 1876, United Kingdom https://www.legislation.gov.uk/ukpga/Vict/39-40/ 77/enactedNorth African animal legislation: a review

18Animal Technology and Welfare August 20204 Algeria – Hunting Act document date 1982/08/21 valid as of 1985-02-5. 00 (Algeria) Updated in August 2014. http://datazone.birdlife.org/userfiles/ file/hunting/HuntingRegulations_Algeria.pdf5 Algeria – Decree concerning the protection of wild animal species (S.I.). Ref: National Legislation: Enabling/Implementation texts: 801550 (I-9830282100) LE /025402 Document date: 1983/08/20. Valid as of: 1985/ O2/00. https://cites.org/eng/parties/country-profiles/dz6 Algeria – Environment Protection Act Document date: 1983/08/20 Valid as of: 1985/02/00. https://core.ac.uk/download/pdf/277480513.pdf7 Benin – Decree concerning hunting and capturing licence, bag limits and professional hunters. (S.I.). Ref. National Legislation: Enabling/Implementation.textx: 829410(H-98002100) L.E/02/o27080 Document date: 1980/02/11. Valid as of: 1984/12/12 Status of Legislative Progress for Implementing CITES | CITES8 Benin – ordinance concerning protection of nature and hunting. Document date: 1980/02/11. Valid as of: 1984/12/12.9 Burkina Faso – Wildlife Conservation and Hunting Act. Document Date: 1968/12/31. Valid as of: 1983/12/12.10 Burkina Faso – Decree implementing the Wildlife Conservation and Hunting Act (S.I.). Ref: National Legislation:Enabled/Implenetastion. Texts 943360(1-968123100) L.E/000839 Documents Date: 1968/12/31. Valid as of: 1983/10/17. 11 Burkina Faso – ordinance concerning: 1. Categories of hunting and capturing licences. 2. Modifications of licence fee. 3. Changes in bag limits. Ref: National Legislation: CITES:943360 (I-96812310000) L.E./000839 Document date: 1977/11/11. Valid as of: 1983/10/17. 12 Burkina Faso – Act re-opening hunting in Burkina Faso. Ref. National Legislation: CITES:943360 (I-9681231000) LE/000839. Document Date: 1985/12/15. 13 Burkina Faso – Hunting Regulations (S.I.). Ref: National Legislation: Enabling/Implementation texts: 9433609 (I- 968123100) LE/000839 Document Date: 1985/12/00.14 Cameroon – Forests, Wildlife and Fisheries Act. Document Date: 1981/11/27. Valid as of: 1983/07/22. https://www.observatoire-comifac.net/ file/eyJtb2RlbCI6IkFwcFxcTW9kZWxzXFxDYXRhbG9 ndWVcXE115 Cameroon – Order listing classes A,B,C animals and listing game species which may be hunted under the various types of hunting licences and corresponding bag limits (S.I.). Ref: National Legislation: Enablling/Implementation texts: 815690 (U-9811127000 le/021541. Document Date: 1983/07/29. 16 Central African Republic – Decree concerning the issuance of licences and the collection of fees for the capture of live wild animals (S.I.). Ref. National Legislation: Enabling/Implementation texts: 817380(H-984100900)LE/0209015 Document date: 1984/10/09. Valid as of: 1985/04/25.17 Ethiopia – Wildlife Conservation regulations 1972 (S.I.). Ref. National Legislation: Enabling/Implementation texts: 833360 (I-944052900)LE/000249. Document date: 1972/01/19. Valid as of: 1985/03/06. 18 Ethiopia – Forest and Wildlife Conservation and Development Proclamation. Document date: 1972/01/05. Valid as of: 1985/03/06. https://www.fao.org/faolex/results/details/en/c/LEX-FAOC00290119 Ghana – The Wild Animal Preservation Act. Document Date: 1961/03/22. Valid as of 1985/01/30. https://www.clientearth.org/latest/documents/act-1961-wild-animals-preservation-act-act-4320 Guinea Bissau – Hunting regulations (S.I.). Document Date: 1980/05/12. Valid as of 1985/06/28. https://www.fao.org/faolex/results/details/en/c/LEX-FAOC01671721 Ivory Coast – Decree regulating the trade in and the transport, import and export of the trophies and skins of protected and spectacular animals. (S.I.). Ref. National Legislation: Enabling/implementation texts: 865040(H-965080400) LE/007338 Document Date: 196/09/15 22 Ivory Coast – Wildlife and Hunting Act. Document Date: 1965/08/04. 23 Ivory Coast – Decree regulating the issuance and validity of game licences in the ivory Coat (S.I.). Ref. National Legislation. Enabling/Implementation texts 865040(I-966091501) LE/007341. Document Date: 1966/09/15.24 Nigeria (Anambra, Imo Rivers, Cross Rivers) – The Wild Animals Law 1965. Document date: 1965/05/20 Wild Animals Law, 1965. | FAOLEX25 Nigeria (Bauchi, Borno, Gongola) – The Wild Animals Law 1965. Document date: 1965/00/00 Valid as of: 1968/04/15 (Bauchi) 1968/04/22 (Gongola). 26 Nigeria (Bendel) – The Wild Animals Preservation Act. Document date: 1916/05/30. Valid as of: 1985/01/16 27 Nigeria (Benue, Kaduna, Kwara, Niger, Plateau, Sokoto) – The Wild Animals Law 1963. Document date: 1963/04/13 Valid as of: 1985/04/29 (Plateau), 1985/05/02 (Kwara), 1985/01/10 (Niger). 28 Nigeria (Kano) – The Wild Animals Law 1963. Document date: 1963/04/13 Valid as of: 1985/04/29 (Plateau), 1985/03/26. 29 Senegal – Game and Wildlife protection Regulations (S.I.). Ref. National Legislation. Enabling/ Implementation texts 911600 (I-967052300) LE/007341. Document Date: 1967/05/09.North African animal legislation: a review

19August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfare30 Benin – Ordinance establishing royalties for the taking of game species. Ref. National Legislation. CITES:8294100 LE /027080. Document Date: 1980/02/11. Valid as of: 1984/12/12.31 Burkina Faso – Fishing Ordinance (S.I.). Document date: 1955/07/27. Valid as of 1983/07/17. 32 Cameroon – Wildlife Decree (Statutory Instrument). Document date: 1983/04/12. Valid as of: 1983/07/22.33 Cape Verde – Decree establishing rules for the protection of soil, flora and fauna (Statutory instrument). Document Date: 1955/01/20. Valid as of: 1985/05/15. 34 Central African Republic – Ordinance concerning the protection of wildlife and regulated hunting (Statutory Instrument). Document date 1984/07/27. Valid as of: 1985/04/25.35 Central African Republic – Ordinance concerning the capturing and export of live wild animals (Act). Ref. National Legislation. CITES:817380 (H-984072700) LE /0279016. Document Date: 1984/07/27. Valid as of: 1985/04/25. 36 Central African Republic – Ordinance establishing a ban on elephant hunting (Act). Ref. National Legislation. CITES:817380 (H-984072700) LE / 029016. Document Date: 1985/01/30. Valid as of: 1985/04/25. 37 Egypt – Decree forbidding to hunt various kinds of birds and mammals in Sinai Goverances (S.I.). Ref. National Legislation. Enabling/Implementation texts:*Law 43of 1979. Document Date: 1982/04/26. 38 Egypt (Matrouh) – Decree forbidding hunting of wild animals in Matrouh Governate (S.I.). Ref. National Legislation. Enabling/Implementation texts: *Law 43of 1979. Document Date: 1982/06/02. 39 Egypt (North Sinai) – Decree forbidding hunting of birds useful agriculture and of wild animals (S.I.). Ref. National Legislation. Enabling/Implementation texts:931520(1966000000) LE /000780. Document Date: 1980/07/17. 40 Egypt (South Sinai) – Decree concerning the preservation of wild animals (S.I.). Ref. National Legislation. Enabling/Implementation texts:* Law 43/79 Law 84/78. Document Date: 1980/02/18. 41 Egypt (South Sinai) – Decree prohibiting the hunting of coloured fish and the removal of specimens from coral reefs in the Ras Muhammed areas (S.I.). Ref. National Legislation. Enabling/Implementation texts: *Law 43/79 Law 84/78 Document Date: 1980/02/18.42 Equatroial Guinea – Hunting regulations for the Spanish Territories of the Gulf of Guinea (S.I.). Document date: 1981/04/0343 Gabon – Decree concerning wildlife protection (S.I.). Ref. National Legislation: Enabling/Implementation texts:837900 (I-960060800). L.E. 003454. Document Date: 1981/10/03. 44 Gabon – Decree establishing an open season for small game (S.I). Ref. National Legislation: Enabling/Implementation texts: 837900 (I-960060800) L.E. 003454 Document Date: 1982/04/09 Valid as 0f:1985/01/08 45 Gabon – Waters and Forest Act. Document Date: 1982/07/22. Valid as of: 1985/08/07. 46 Gambia – Wildlife Conservation Act 1977. Ref. National Legislation. 10 of 1977 Gambia Gazette No.18 1977/03/09 Supplement C, Document Date: 1977/02/14. Valid as of: 1985/11/21. 47 Gambia – Wildlife Conservation (Sale) Regulations 1977 (S.I). Ref. L.N.. 10 of 1977. Gambia Gazette No. 34 1977/05/27 Supplement A. Document date: 197/09/01. Valid as of: 1985/11/21. 48 Gambia – Wildlife Conservation Regulations 1978 (S.I). Ref: Nat legisl: Enabl/implem. text 838640(H- 9770211400) L.E./001451 Reference: 1 of 1977. Gambia Gazette NO. 18 1977/O3/09 Supplement C Document date 1978/12/ 21. Valid as of: 1985/11/21.49 Ghana – Wildlife Conservation Regulations 1971 (S.I). Ref. Nat Legisl. :Ebabl./Implemt. Texts 841340(H-961032200)LE/000388 Document Date: 1971/03/04. Valid as of: 1985/01/30. 50 Mauritania – Hunting and Wildlife Protection Act. Document date: 1975/01/15.51 Morocco – Hunting Act 1923. Document date: 1923/07/21. Valid as of: 1985/11/12. http://datazone.birdlife.org/userfiles/file/hunting/HuntingRegulations_Morocco.pdf52 Morocco – Standing Hunting Regulations (S.I.). Ref Nat Legisl.: Enabl./Implem. Texts 881879 (I-923072100)LE/007407) Document date: 1985/08/14. 53 Morocco – Order establishing open and close seasons and special rule for the season 1985- 1986. (S.I.). Ref Nat legisl: Enabl./Implem. Texts: 8818799I-923072100)le/007407. Document Date: 1985/08/14. Valid as of: 1985/11/12. HuntingRegulations_Morocco.pdf (birdlife.org)54 Niger – Hunting Act. Document Date: 1962/04 Valid as of: 1985/09/30. 55 Niger – Decree (S.I.). prohibiting trade in and export of wild animal products (S.l.). Document date: 1964/06/17.56 Niger – Decree prohibiting Hunting (S.I.). Document date: 1972/07/20. Valid as of: 1985/09/30.57 Niger – Decree concerning traffic in wild animals and animal products (S.I.). Document date: 1974/09/06. Niger – Animal Protection Index 58 Nigeria (Federal) – Endangered species (Control of International Trade and Traffic) Decree 1985. Ref treaties: Enabl/Impm. Texts 800000/aH – 973030300/TR/002343 CITES Document date: 1985/04/20. https://www.animallaw.info/statute/nigeria-endangered-species-endangered-species-act-englishNorth African animal legislation: a review

20Animal Technology and Welfare August 202059 Nigeria (Lagos) – Wild Animals Preservation Act. Document date: 1916/05/30. Valid as of: 1983/11/21. https://globalhistorydialogues.org/ projects/a-history-of-wildlife-preservation-in-southwestern-nigeria60 Senegal – Hunting and Wildlife Protection Code (Statutory Part) Act. Document date: 1967/05/23.61 Sierra Leone – Wild Animals (Export) Rules (Statutory instrument). Ref. Nat legisl: Enabl./Implemt full. Texts: 912340 (H – 901 00000) LE/005918. Document date: 1950/00/00. Valid as of: 1981/12/11. Sierra Leone CONSERVATION AND WILDLIFE POLICY 2010 – LSE https://www.lse.ac.uk/GranthamInstitute/wp-content/uploads/laws/4754.pdf62 Sierra Leone – The Wildlife Conservation Act 1972. Ref. number: Act 27 of 1972 Reference treaties: Enabl./Implementing texts: 800000/Q – 933110800/uppercase TR001834. Convention relative to the preservation of and flora in their natural state. London 1933/1108. Document date 1972/00/00. Valid as of: 1981/12/11. https://faolex.fao.org/docs/pdf/sie41659.pdf63 Somalia – Law on Fauna (Hunting) and Forest Act. Document date: one 1969/01/25. http://www.somalilandlaw.com/Law_on_Fauna_Hunting_and_Forest_Conservation_1969.pdf64 Sudan – Preservation of Wild Animals Act. Ref: 919751231. Laws of Sudan Volume 2 chapter 36. Document date 1935/00/00. Valid as of: 1983. https://www.fao.org/faolex/results/details/en/c/LEX-FAOC01006865 Togo – Ordinance on wildlife protection and hunting in Togo (Act). Ref. Treaties CITES: 800000/U – 933110800/TR001834. Document date: 1968/010/16 Valid as of: 1985/04/05.66 Togo – Decree prohibiting night hunting (S.I.). Reference Nat Legisl: Enabl./Implement. texts: 926080 (I – 968011601) LE/001240. Document date: 1968/01/16 Valid as of: 1985/04/05.67 Togo – Decree implementing Ordinance No. 4 of 1968/01/16 regulating protection of fauna and Hunting (S.I.). Document Date: 1973/04/18. Valid as of: 1985/04/05. Wildlife Law (No.439). | FAOLEX Feb 16, 2023. https://www.fao.org/faolex/results/details/en/c/LEX-FAOC07485668 Tunisia – Forest Act Government of Tunisia. 1966. Code forestier tunisien. In Journal Officiel de la République Tunisienne. https://www.fao.org/3/x1880e/x1880e04.htm Document date: 1966/07/04.69 Uganda – The Fish and Crocodiles Act. Ref. Number: ordinance 47 of 1950. Document date: 1950/00/00. Valid as of: 1985/04/00 https://core.ac.uk/download/pdf/33720701.pdf70 Uganda – Game (Preservation and Control) Act. Ref. Number ordinance for of 1959. Document date: 1959/00/00. CHAPTER 198 · CHAPTER 198 · THE GAME (PRESERVATION AND CONTROL) ACT. Valid as of: 1984/04/02 https://old.ulii.org/ug/legislation/consolidated-act/19871 Sudan – The Animals Export and Import Act 1913. https://swm-programme.info/web/guest/legal-hub/sudan/legal-framework?swmlegalsector=[%22FD%72 Somalia – Veterinary Code. Ref. Legal systems of Africa series – Democratic Republic of Somalia. Valid as of: 1972. https://leap.unep.org/countries/ so/national-legislation/veterinary-law-code-201673 Ivory Coast – Order concerning conditions upon which noxious animals may be destroyed. (S.I.). Ref: Nat Legisl: EnAbl./Implem. texts 865040 (H – 9650800400) LE/007338. Document date: 1961/12/29.74 Egypt – Protection of Birds Beneficial to Agriculture and Protection of Wild Fauna Act 1966. Ref. Number: 53 of 1966 Document date: 1966/00/00. Valid as of: 1986/01000. https://api.worldanimalprotection.org/country/egypt75 Liberia – National Artificial Insemination Act 1956. https://pflanzengesundheit.julius-kuehn.de/dokumente/upload/lr-1973pqact_en.pdf76 Nigeria – Agricultural Research Institutes Decree 1973. Dec 11, 1975 ... _ LN. 107 of 1975. AGRICULTURAL RESEARCH INSTITUTES DECREE 1973. |. (1973 No. 35). ResearchInstitutes (Establishment, etc.) Order 1975.77 Somalia – Livestock Development Agency Law (Act). Decree law No. 2 of 1966/02/16 converted into law by Law No. 3 of 1966/03/21. Ref: Legal Systems of Africa series – Democratic Republic Somalia valid as of: 1972. https://leap.unep.org/countries/so/national-legislation/livestock-development-agency-law-no-34-197078 Somalia – Livestock Development Agency Law (Act). Law No.34 of 1970/06/26. Reference: Legal systems of Africa series – Democratic Republic Somalia.79 Uganda – Cattle Grazing Act 1918. https://faolex.fao.org/docs/pdf/uga96015.pdfreference80 Uganda – Animals (Straying) Act 1922. https://faolex.fao.org/docs/pdf/uga95979.pdf81 Uganda – Branding of Stock Act 1918. https://www.fao.org/faolex/results/details/en/c/LEX-FAOC096014r82 Uganda – Trout Protection Act 1936. https://faolex.fao.org/docs/pdf/uga132679.pdf83 Sierra Leone – Locusts (Destruction) Act 1931. Ref the laws of Sierra Leone 1960, volume VII Subsidiary Legislation, Chapter 191. Cap. 188 – Sierra Leone Web http://www.sierra-leone.org/Laws/Cap%20188.pdf84 Sudan – Locusts Destruction Act 1907. https://leap.unep.org/countries/sd/national-legislation/locust-destruction-act-1907North African animal legislation: a review

21August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfare85 Gambia – Diseases of Animals (Restrictions of Import and Export) Regulations 1968. https://www.trade.gov/country-commercial-guides/gambia-import-requirements-and-documentation86 Gambia – Dogs (Prevention of Hydrophobia Order 1975. Ref: The Constitutional Law of the Gambia: 1965 – 2010 by Ousman A.S. Jammeh… ISBN 13. 9781467007405. ISBN 10 1467007404.87 Ivory Coast – Animal Health Regulations 1985. https://www.aphis.usda.gov/aphis/ourfocus/animalhealth/export/international-standard-setting-activ88 Liberia – Plant and Quarantine Act 1956. https://www.wto.org/english/thewto_e/acc_e/lbr_e/WTACCLBR5_LEG_10.pdf89 Nigeria – Animals (Diseases) Act 1960. Ref: laws of the Federation of Nigeria and Lagos 1958 https://lawcat.berkeley.edu/record/525470?ln=en90 Nigeria – Diseases of Animals Act 1962. Ref: Laws of the Federation of Nigeria. https://laws.lawnigeria.com/2018/05/02/laws-of-the-federation-of-nigeria-291 Nigeria – Veterinary Surgeons Decree 1969. THE VETERINARY SURGEONS DECREE 1969 17. 20. ment https://gazettes.africa/archive/ng/1969/ng-government-gazette-supplement-dated-1969-09-18-no-48-part-a.pdf92 Sierra Leone – The Dogs Act 1900. http://www.sierra-leone.org/Laws/Cap%20193.pdf Ref: the laws of Sierra Leone 1960: volume 7 subsidiary legislation.93 Sierra Leone – Animal Diseases Act 1944. The laws of Sierra Leone 1960, volume 7 subsidiary legislation. https://books.google.com/books/about/The_Laws_of_Sierra_Leone_in_Force_on_the.html?id=7ug-AQ94 Sudan – The Rabies Act 1948. https://swm-programme.info/web/guest/legal-hub/ sudan/legal-framework?swmlegalsector=[%22AN95 Sudan – The Diseases of Animals Act 1901. https://www.woah.org/app/uploads/2021/03/ 2020-vetlegislation-identificationreport-sudan-en-.pdf96 Sudan – Veterinary Council Act 1954 repealed 1995. sud189208E.pdf (fao.org)97 Uganda – The Animals Diseases Act 1918. The laws of Uganda CHAPTER 38 THE ANIMAL DISEASES ACT.C ommencement https://faolex.fao.org/docs/pdf/uga95975.pdf98 Uganda – Rabies Act 1935. Ref: Rabies Act 1935 | Uganda Legal Information Institute (ulii.org)North African animal legislation: a review

22Animal Technology and Welfare August 2020Animal Technology and Welfare April 2023PAPER SUMMARY TRANSLATIONSCONTENU DE LA REVUELégislation nord-africaine sur les animaux : examenA.N. OTHERInstitut de technologie animaleCorrespondance: atweditor@iat.org.comRésumé Cet examen a été mené par un étudiant dans le cadre du cours de l’Institut de technologie animale menant au diplôme d’études supérieures (PGDip) en technologie et sciences animales de laboratoire, créé en 1992 en collaboration avec l’Université du Middlesex. Les étudiants ayant réussi à obtenir leur PGDip ont pu poursuivre leurs études pour obtenir un Master en technologie et sciences animales de laboratoire, par la rédaction d’une thèse. Dans le cadre du module de droit, les étudiants étaient amenés à examiner la législation relative aux animaux dans différentes régions du monde. La portée initiale de ce projet était d’étudier la législation relative aux animaux dans les États africains situés au nord du désert du Sahara. Malheureusement, aucune autorité ne semble s’entendre sur les limites du Sahara, par conséquent, une décision arbitraire a été prise pour étudier les États africains où la majorité de la masse terrestre se trouve au nord de l’équateur. Dans les pays où la masse terrestre se trouve également de part et d’autre de l’équateur, la position de la capitale a été utilisée comme facteur décisif, d’où le fait que le Gabon ait été inclus dans cet examen, alors que le Kenya et le Congo ont été classés comme appartenant à l’Afrique du Sud. Sur la base de cette justification, 30 pays ont été identifiés comme relevant de la classification « Afrique du Nord ». 19 ambassades ou consulats britanniques des pays concernés ont été approchés et 10 autres États ont été contactés par l’intermédiaire des Nations unies. Ce n’est que dans le cas du Cap-Vert que le contact n’a pas été établi, parce que l’auteur n’a pas pu déterminer si les îles étaient un État indépendant ou une dépendance d’une autre puissance nationale (il a été constaté depuis qu’il s’agissait désormais d’un État indépendant, autrefois une colonie du Portugal). La plupart des demandes de contact auprès des ambassades du Royaume-Uni appropriées se sont avérées infructueuses et une grande partie de l’information utilisée dans cet examen a donc été tirée de sources de bibliothèque. Les références ont été mises à jour dans la mesure du possible en 2023. Mots-clés: Législation sur les animaux. Afrique du Nord ★ ★ ★

23August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsValidation de l’utilisation de l’entraînement à la boîte pour raffiner la manipulation des lapinsALICIA KINALLY, LUCY ONIONS ET SARAH GLENNUniversité de LeicesterCorrespondance: ak716@leicester.ac.uk Basé sur la candidature au prix Andrew Blake Tribute AwardRésumé Dans leur habitat naturel, les lapins sont des animaux de proie, c’est pourquoi le fait d’être pris en main et manipulé peut s’avérer pour eux extrêmement stressant. Des études antérieures menées chez des lapins utilisés comme animaux de compagnie ont montré que lorsqu’ils sont pris en main, les lapins montrent des signes de lutte et d’agression due à la peur ; il est donc conseillé aux propriétaires de ces animaux de compagnie d’éviter de soulever directement les animaux si possible (Bradbury et Dickens, 2016).1 La méthode la plus couramment utilisée pour manipuler le lapin de laboratoire consiste à le saisir par la peau du cou, tout en plaçant l’autre main sous l’animal pour soutenir les pattes arrière, en le soulevant de la cage/de l’enclos. L’animal est alors tenu contre soi, son poids étant supporté par le bras (Norecopa, 2015). Nous voulions introduire une nouvelle méthode de prise en main et de transport du lapin qui permettrait de réduire son stress, semblable à la méthode déjà reconnue et largement utilisée de manipulation des souris dans des tunnels (Gouevia et Hurst, 2013).Dans notre établissement, les lapins sont manipulés plusieurs fois par semaine, car ils passent du temps dans des parcs spécialement conçus pour faire de l’exercice ou subir des pesées hebdomadaires et des contrôles de santé. Les lapins que nous hébergeons sont utilisés pour les modèles cardiovasculaires d’infarctus du myocarde, et il est donc important de leur éviter du stress pour assurer la validité des données produites. Nous pensons qu’une manipulation moins invasive affectera également le tempérament général des lapins et qu’ils seront, par la suite, plus susceptibles de répondre positivement à l’interaction humaine plutôt que de chercher à l’éviter comme lorsqu’ils étaient attrapés par le cou précédemment.Mots-clés: Lapins, manipulation, entraînement à la boîte, raffinement, réponse positive★ ★ ★

24Animal Technology and Welfare August 2020Paper Summary TranslationsComment faire se reproduire et élever des grenouilles africaines à griffes (Xenopus laevis)THERESE JONES-GREENUniversité de CambridgeCorrespondance: tj222@cam.ac.ukRésumé Il s’agit d’un article d’instruction sur la façon de faire se reproduire des grenouilles Xenopus laevis en laboratoire autorisé en vertu de la loi sur les procédures scientifiques impliquant des animaux de 1986.Les nombreux défis de l’année 2020 ont eu un impact négatif sur les fournisseurs de grenouilles Xenopus, principalement la fermeture du plus grand fournisseur mondial NASCO. Bien qu’il existe d’autres fournisseurs en raison de l’augmentation de la demande sur toutes les entreprises, une reproduction à l’interne peut s’avérer nécessaire pour compléter le stock.L’élevage en interne constitue également un excellent raffinement du bien-être des animaux car il élimine le stress causé par les voyages et les animaux sont acclimatés à l’installation.Avantages et inconvénients des différents organismes modèles. Les Xenopusare sont largement utilisés pour étudier l’embryologie et le développement des vertébrés, la biologie cellulaire et moléculaire de base, la génomique, la neurobiologie et la toxicologie, et pour modéliser les maladies humaines (Xenbase.org, 2022). Mots-clés: Xenopus laevis, reproduction, raffinement ★ ★ ★Maladie de la grenouille africaine à griffes Xenopus laevis et signes cliniques visibles sur des photographiesTHERESE JONES-GREENUniversité de Cambridge Correspondance: tj222@cam.ac.ukRésumé De nombreuses maladies touchant les grenouilles Xenopus laevis ont été décrites dans la littérature. Il est toutefois souvent difficile de trouver des images qui illustrent clairement les signes cliniques qui leur sont associés. Cet article a été conçu comme un raffinement pour combler ce fossé en utilisant des photographies de grenouilles présentant des signes cliniques afin de partager mes expériences, par des images, pour aider à améliorer le bien-être et l’élevage de cette espèce. J’espère également stimuler un dialogue qui permettra un partage plus approfondi des idées et des informations. Mots-clés: Xenopus laevis, maladies, signes cliniques

25August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePaper Summary TranslationsINHALTVERZEICHNISEine kritische Betrachtung der Tierschutzgesetzgebung in NordafrikaA.N. OTHERInstitute of Animal TechnologyKorrespondenz: atweditor@iat.org.comAbstract Diese kritische Betrachtung war eine Studienaufgabe im Rahmen des Postgraduierten-Diplomkurses (PGDip) des Institute of Animal Technology in Laboratory Animal Technology and Science, der 1992 zusammen mit der University of Middlesex eingerichtet wurde. Studenten, die ihren PGDip erfolgreich absolvierten, konnten ihr Studium fortsetzen und mit einer Abschlussarbeit einen Master of Science in Versuchstiertechnologie erwerben.Im Rahmen des Moduls „Recht“ erhielten die Studenten die Aufgabe, die Tierschutzgesetzgebung in verschiedenen Regionen der Welt zu untersuchen. Der ursprüngliche Gegenstand dieses Projekts war die Untersuchung der Tierschutzgesetze in den afrikanischen Staaten nördlich der Sahara. Angesichts der anscheinend zwischen den Behörden bestehenden Uneinigkeit über die Grenzen der Sahara wurde beschlossen, jene afrikanischen Staaten zu erfassen, deren Großteil der Landmasse nördlich des Äquators liegt. Bei Ländern, in denen die Landmasse zu gleichen Teilen auf beiden Seiten des Äquators liegt, gab die Lage der Hauptstadt den Ausschlag, weshalb Gabun in diese Untersuchung einbezogen wurde, Kenia und der Kongo hingegen als Südafrika eingestuft wurden. Auf dieser Grundlage wurden 30 Länder als „Nordafrika“ klassifiziert. 19 britische Botschaften oder Konsulate in den betreffenden Ländern wurden kontaktiert, weitere 10 Staaten wurden über die Vereinten Nationen angesprochen. Lediglich im Fall von Kap Verde wurde kein Kontakt hergestellt, da der Verfasser nicht klären konnte, ob die Inseln ein unabhängiger Staat oder eine Dependance einer anderen Staatsmacht sind (inzwischen wurde geklärt, dass die ehemalige Kolonie Portugals heute ein unabhängiger Staat ist). Die meisten Anfragen bei der zuständigen britischen Botschaft waren ergebnislos, und ein Großteil der in diesem Bericht verwendeten Informationen stammt aus Bibliotheksquellen. Die Quellenangaben wurden, soweit möglich, im Jahr 2023 aktualisiert. Schlagwörter: Tierschutzgesetzgebung, Nordafrika ★ ★ ★

26Animal Technology and Welfare August 2020Validierung des Boxentrainings als Verbesserung des Umgangs mit KaninchenALICIA KINALLY, LUCY ONIONS UND SARAH GLENNUniversity of LeicesterKorrespondenz: ak716@leicester.ac.ukBasierend auf dem Andrew Blake Tribute Award-SiegerbeitragAbstract In freier Wildbahn sind Kaninchen Flucht- und Beutetiere, weswegen Hochheben und Handling für sie extrem belastend sein können. In früheren Studien an Hauskaninchen zeigten sich beim Hochheben durch Angst ausgelöste Anzeichen von Abwehr und Aggression – daher wird Haustierhaltern empfohlen, ein direktes Hochheben der Tiere nach Möglichkeit zu vermeiden (Bradbury and Dickens, 2016).1 Die am häufigsten angewandte Handling-Methode bei Versuchskaninchen besteht darin, das Tier am Nackenfell zu fassen und mit der anderen Hand das Hinterteil zu stützen, um das Tier so aus dem Käfig/der Box herauszuheben. Es wird dann eng am Körper festgehalten, wobei das Körpergewicht auf dem Arm lastet (Norecopa, 2015). Wir wollten eine neue Methode zum Heben und Tragen von Kaninchen einführen, die Stress reduziert, ähnlich wie die bereits anerkannte und verbreitete Methode des Tunnel-Handlings von Mäusen (Gouevia and Hurst, 2013).In unserer Einrichtung werden die Kaninchen mehrmals pro Woche gehandhabt, da sie in speziell dafür vorgesehenen Gehegen wöchentlichen Auslauf haben oder wöchentlich gewogen und gesundheitlich untersucht werden. Die bei uns untergebrachten Kaninchen werden für kardiovaskuläre Modelle des Myokardinfarkts verwendet, und daher ist es wichtig, sie nicht zu sehr zu stressen, um die Validität der gewonnenen Daten zu gewährleisten. Wir glauben, dass ein weniger invasives Handling sich auch auf das allgemeine Temperament der Kaninchen auswirkt, sodass sie dann eher positiv auf menschliche Interaktion reagieren dürften und nicht mehr so sehr ausweichen wie beim Hochheben am Nackenfell.Schlagwörter: Kaninchen, Handling, Boxentraining, Verbesserung, positive Reaktion ★ ★ ★Paper Summary Translations

27August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAnweisungen zu Züchtung und Aufzucht afrikanischer Krallenfrösche (Xenopus laevis)THERESE JONES-GREENUniversity of CambridgeKorrespondenz: tj222@cam.ac.ukAbstract Dies ist ein Artikel mit Anweisungen zur Züchtung von Xenopus laevis im Labor, die nach dem britischen Tierschutzgesetz (Animals [Scientific Procedures] Act) von 1986 zugelassen ist. Die zahlreichen Herausforderungen des Jahres 2020 haben sich negativ auf die Lieferanten von Xenopus-Fröschen ausgewirkt, vor allem die Schließung des weltweit größten Lieferanten NASCO. Es gibt zwar aufgrund der gestiegenen Nachfrage bei allen Unternehmen andere Lieferanten, eine eigene Zucht kann jedoch zur Bestandsergänzung erforderlich sein.Die einrichtungseigene Zucht stellt auch eine erhebliche Verbesserung des Tierwohls dar, da der durch Transport verursachte Stress entfällt und die Tiere mit der Einrichtung vertraut sind.Vor- und Nachteile verschiedener Modellorganismen. Xenopus werden häufig zur Erforschung der Embryologie und Entwicklung von Wirbeltieren, der grundlegenden Zell- und Molekularbiologie, der Genomik, der Neurobiologie und der Toxikologie sowie zur Modellierung menschlicher Krankheiten verwendet (Xenbase.org, 2022).Schlagwörter: Xenopus laevis, Züchtung, Verbesserung ★ ★ ★Afrikanischer Krallenfrosch Xenopus laevis – Krankheiten und klinische Symptome auf FotosTHERESE JONES-GREENUniversity of CambridgeKorrespondenz: tj222@cam.ac.ukAbstract In der Literatur werden viele Krankheiten von Xenopus laevis-Fröschen beschrieben, doch oft ist es schwierig, Bildmaterial zu finden, das damit verbundene klinische Symptome klar veranschaulicht. Mit diesem Artikel möchte ich diese Lücke anhand von Fotos von Fröschen mit klinischen Symptomen schließen und meine Erfahrungen mittels Bildmaterial teilen, um Wohlergehen und Haltung dieser Art zu verbessern. Ich hoffe auch, einen Dialog anstoßen zu können, der zu einem weiteren Austausch von Ideen und Informationen führt.Schlagwörter: Xenopus laevis, Krankheiten, klinische SymptomePaper Summary Translations

28Animal Technology and Welfare August 2020INDICE DELLA REVISTARassegna della legislazione sugli animali del Nord AfricaA.N. OTHER Corrispondenza: atweditor@iat.org.comAbstract Questa rassegna è scaturita da un compito per studenti assegnato nell’ambito del Diploma post laurea (PGDip) in Scienze e Tecnologie degli Animali da Laboratorio dell’Institute of Animal Technology, istituito nel 1992 in collaborazione con l’Università del Middlesex. Gli studenti che hanno ottenuto il PGDip hanno proseguito gli studi per conseguire un Master in Scienze, su presentazione di una tesi su Scienze e Tecnologie degli Animali da Laboratorio. Come parte del modulo di Diritto, agli studenti è stato richiesto di esaminare la legislazione sugli animali di vari territori del mondo. Il compito originario di questo progetto era l’analisi della legislazione relativa agli animali degli Stati africani a nord del Deserto del Sahara. Dal momento che non esiste un consenso unanime sui confini del Sahara, si è deciso arbitrariamente di valutare gli Stati africani la cui massa terrestre giace principalmente a nord dell’Equatore. Nei Paesi cui la massa di terra si trova in pari misura sia a nord che a sud dell’Equatore, il fattore determinante è stato la posizione della capitale, per cui il Gabon è stato incluso nella rassegna, mentre il Kenya e il Congo sono stati classificati come Africa meridionale. Basandosi su questa logica, 30 Paesi sono rientrati nella classificazione di “Africa settentrionale”. Sono stati contattati 19 ambasciate o consolati britannici dei Paesi in questione e ci si è rivolti ad altri 10 Stati attraverso le Nazioni Unite. Solo nel caso di Capo Verde non è stato stabilito alcun contatto, poiché l’autore non è riuscito a verificare se le isole fossero Stati indipendenti o territori dipendenti di un’altra potenza nazionale (sin d’allora, si è determinato che l’arcipelago era un’ex colonia del Portogallo, ma è ora uno Stato indipendente). Gran parte dei contatti con l’ambasciata britannica competente sono stati improduttivi e molte delle informazioni condivise nella presente rassegna sono state ottenute da biblioteche. Ove possibile, la bibliografia è stata aggiornata nel 2023. Parole chiave: Legislazione sugli animali, Nord Africa ★ ★ ★Paper Summary Translations

29August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareValidazione dell’allenamento in box per il miglioramento della manipolazione dei conigliALICIA KINALLY, LUCY ONIONS E SARAH GLENNUniversità di LeicesterCorrispondenza: Ak716@leicester.ac.ukArticolo basato sul progetto vincitore del premio Andrew Blake Tribute AwardAbstract In natura, i conigli sono animali da preda e, pertanto, l’atto di essere afferrati e maneggiati può essere fonte di forte stress per questi mammiferi. Studi precedenti condotti su conigli domestici hanno mostrato che, durante il sollevamento, questi animali mostravano segni di difesa e aggressione scatenati da un senso di paura; di conseguenza, si consiglia ai proprietari di evitare un sollevamento per le orecchie ove possibile (Bradbury e Dickens, 2016).1 Il metodo di manipolazione dei conigli da laboratorio più comunemente usato prevede che l’animale venga afferrato dalla collottola, sostenendo il dorso con l’altra mano e sollevando l’animale dalla gabbia/dal recinto. Il coniglio deve essere poi avvicinato al proprio corpo, sostenendone il peso sul braccio (Norecopa, 2015). Il nostro intento era quello di introdurre un nuovo metodo di sollevamento e trasporto dei conigli in grado di ridurre lo stress provato dagli animali, simile ai tunnel di movimentazione dei topi già ampiamente riconosciuti e utilizzati (Gouevia e Hurst, 2013).Presso la nostra struttura, i conigli vengono maneggiati varie volte a settimana, in quanto vengono trasferiti in box appositamente progettati a fini di allenamento o per essere sottoposti a pesatura e controlli di salute settimanali. I conigli da noi ospitati vengono utilizzati per generare modelli cardiovascolari dell’infarto miocardico e, pertanto, è importante che non manifestino segni di stress in modo da garantire la validità dei dati ottenuti. Siamo convinti che una manipolazione meno invasiva possa influire anche sul temperamento generale dei conigli che, di conseguenza, risponderanno molto probabilmente in maniera positiva all’interazione con l’uomo piuttosto che evitarla, come si è riscontrato nel caso di regolari sollevamenti per la nuca.Parole chiave: Conigli, manipolazione, allenamento in box, miglioramento, risposta positiva ★ ★ ★Paper Summary Translations

30Animal Technology and Welfare August 2020Allevamento di rane artigliate africane (Xenopus laevis)THERESE JONES-GREENUniversità di Cambridge Corrispondenza: tj222@cam.ac.ukAbstract Questo articolo si propone di approfondire l’allevamento di Xenopus laevis in laboratorio autorizzato ai sensi della legge Animals (Scientific Procedures) Act del 1986.Le numerose sfide emerse nel 2020 hanno avuto un impatto negativo sui fornitori di rane Xenopus, che ha portato preminentemente alla chiusura di NASCO, il fornitore più grande al mondo. Benché esistano altri fornitori per via della crescente domanda, potrebbe essere necessario ricorrere anche all’allevamento interno per integrare le scorte.L’allevamento interno è, inoltre, un’ottima soluzione di perfezionamento del benessere animale, in quanto elimina lo stress causato dagli spostamenti e gli animali sono già acclimatati alla struttura.Vantaggi e svantaggi di organismi modello diversi. Le Xenopus sono ampiamente utilizzate per studiare l’embriologia e lo sviluppo dei vertebrati, i concetti fondamentali della biologia cellulare e molecolare, la genomica, la neurobiologia e la tossicologia e per creare modelli di malattie umane (Xenbase.org, 2022).Parole chiave: Xenopus laevis, allevamento, perfezionamento ★ ★ ★Malattie delle rane artigliate africane (Xenopus laevis) e segni clinici visibili nelle fotografieTHERESE JONES-GREENUniversità di Cambridge Corrispondenza: tj222@cam.ac.ukAbstract La letteratura annovera vari esempi di malattie delle Xenopus laevis, ma spesso è difficile trovare immagini che illustrino chiaramente i segni clinici associati. Questo articolo è stato concepito come strumento di perfezionamento per colmare tale divario utilizzando fotografie di rane con segni clinici e per condividere le mie esperienze personali, attraverso l’uso di immagini, al fine di migliorare il benessere e l’allevamento di questa specie. Mi auguro, inoltre, di poter stimolare un dialogo che promuova l’ulteriore condivisione di idee e informazioni.Parole chiave: Xenopus laevis, malattie, segni clinici Paper Summary Translations

31August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareINDICE DE LA REVISTALegislación norteafricana sobre animales: análisisA.N. OTHERInstituto de Tecnología AnimalContacto: atweditor@iat.org.com Resumen Este análisis fue un trabajo realizado como parte del Diploma de Postgrado en y Ciencia y Tecnología de Animales de Laboratorio del Instituto de Tecnología Animal, creado en 1992 junto con la Universidad de Middlesex. Los estudiantes que hubieron superado el Diploma de Postgrado con éxito pudieron continuar sus estudios para obtener el título de Máster en Tecnología de Animales de Laboratorio después de realizar un trabajo final.Como parte del módulo de Derecho, se encargó a los estudiantes la tarea de revisar la legislación relativa a los animales en distintas zonas del mundo.El objetivo inicial de este proyecto era estudiar la legislación relativa a los animales en los estados africanos situados al norte del desierto del Sáhara. Sin embargo, ninguna autoridad ha llegado a un acuerdo sobre los límites del Sáhara, por lo que se tomó la decisión arbitraria de estudiar los estados africanos cuya mayor parte de masa terrestre se encuentra al norte del ecuador. En el caso de los países en los que la masa terrestre se distribuye de igual manera a ambos lados del ecuador, se empleó la posición de la capital como factor decisivo, por lo que Gabón se ha incluido en este estudio, mientras que Kenia y el Congo se consideraron como parte de África del Sur. Según este criterio, se incluyeron 30 países dentro de la clasificación «África del Norte». Se contactó con 19 embajadas o consulados británicos de los países en cuestión y con otros 10 estados a través de las Naciones Unidas. Únicamente en el caso de Cabo Verde no se estableció contacto, debido a que el autor no pudo determinar si las islas eran un estado independiente o dependían de otra nación (desde entonces se ha verificado que, tras haber sido colonia de Portugal, en la actualidad se trata de un estado independiente). La mayoría de los contactos realizados ante las embajadas del Reino Unido correspondientes resultaron infructuosos y gran parte de la información utilizada en este estudio se obtuvo de fuentes bibliográficas. En la medida de lo posible, las referencias se actualizaron en 2023.Palabras clave: Legislación animal. África del Norte★ ★ ★Paper Summary Translations

32Animal Technology and Welfare August 2020Validación del uso del adiestramiento en caja como perfeccionamiento de la manipulación de conejosALICIA KINALLY, LUCY ONIONS Y SARAH GLENNUniversidad de LeicesterCorrespóndanse: Ak716@leicester.ac.ukBasado en la obra ganadora del premio Andrew Blake Tribute AwardResumen En estado salvaje, los conejos son animales de presa, por lo que el acto de ser cogidos y manipulados puede resultarles muy estresante. Estudios anteriores en conejos domésticos han demostrado que, a la hora de levantarlos, los conejos muestran signos de lucha y agresividad debido al miedo; por consiguiente, se recomienda a los propietarios de mascotas que eviten levantar directamente a los animales siempre que sea posible (Bradbury y Dickens, 2016).1 El método más común para manipular conejos de laboratorio consiste en agarrar al animal por la piel de la nuca y, mientras se sostiene la parte trasera con la otra mano, levantarlo y sacarlo de la jaula o el recinto. A continuación, se sujeta al animal contra el cuerpo, sosteniendo el peso corporal en el brazo (Norecopa, 2015). La intención era introducir un nuevo método para levantar y transportar conejos que redujera el estrés, de modo similar al método ya reconocido y ampliamente utilizado de manipulación de ratones con túneles (Gouevia y Hurst, 2013).En nuestras instalaciones, los conejos se manipulan varias veces a la semana, ya que pasan tiempo en recintos específicamente diseñados para hacer ejercicio o se someten a pesajes y controles sanitarios de manera semanal. Los conejos que albergamos se utilizan para modelos cardiovasculares de infarto de miocardio, por lo que es importante que no sufran demasiado estrés a fin de garantizar la validez de los datos obtenidos. Creemos que manipular de forma menos invasiva también afectará el temperamento general de los conejos y, como resultado, es más probable que respondan de forma positiva a la interacción humana en lugar de evitarla, tal y como hacían cuando se les sujetaba del cuello con regularidad.Palabras clave: Conejos, manipulación, adiestramiento en caja, refinamiento, respuesta positiva★ ★ ★Paper Summary Translations

33August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareCómo criar ranas de uñas africanas (Xenopus laevis) THERESE JONES-GREENUniversidad de Cambridge Correspóndanse: tj222@cam.ac.ukResumen Este es un artículo instructivo sobre cómo criar Xenopus laevis en laboratorios autorizados en virtud de la Ley de Animales (Procedimientos Científicos) de 1986.Las numerosas dificultades de 2020 tuvieron un impacto negativo en los proveedores de ranas Xenopus, sobre todo el cierre del mayor proveedor mundial, NASCO (North American Science Associates, Inc.). Aunque existen otros proveedores, la cría interna puede ser necesaria para complementar las existencias debido al aumento de la demanda en todas las empresas.La cría interna es también un excelente perfeccionamiento del bienestar animal, ya que evita el estrés causado por los viajes, además del hecho de que los animales ya están aclimatados a las instalaciones.Ventajas y desventajas de los diferentes organismos modelo. Las Xenopus se utilizan generalmente para estudiar la embriología y el desarrollo de los vertebrados, la biología celular y molecular básica, la genómica, la neurobiología y la toxicología, así como para modelar enfermedades humanas (Xenbase.org, 2022).Parole chiave: Xenopus laevis, cría, perfeccionamiento ★ ★ ★Enfermedades de la rana de uñas africana Xenopus laevis y signos clínicos mediante fotografíasTHERESE JONES-GREENUniversidad de Cambridge Correspóndanse: tj222@cam.ac.ukAbstract Muchas enfermedades de las ranas Xenopus laevis se han descrito en múltiples publicaciones, sin embargo, a menudo es difícil encontrar imágenes que ilustren claramente los signos clínicos asociados. Este artículo ha sido diseñado como un refinamiento para llenar este vacío utilizando fotografías tomadas de ranas con signos clínicos, así como para compartir mis experiencias, a través de imágenes, a fin de ayudar a mejorar el bienestar y la cría de esta especie. También espero estimular un diálogo que resulte en un mayor intercambio de ideas e información.Parole chiave: Xenopus laevis, enfermedades, signos clínicos.Paper Summary Translations

The College of Laboratory Animal Science and Technology (CLAST) and Advance at MRC Harwell are proud to announce a new partnership for the provision of Levels 4, 5 and 6 Institute of Animal Technology (IAT) HE Diplomas for animal technologists and professionals in the field of laboratory animal science and technology.New Curriculum for the 2023/24 yearWe are due to launch teaching of the new CLAST curriculum in Autumn 2023.Progression: ◦ Level 4 – 6 IAT HE Diplomas in Laboratory Animal Science and Technology◦ Complete a diploma Level and start applying for the next Level◦ Or take a break between Levels, returning to study when the time is right for youFlexibility: ◦ Smaller, bite-size units, ideal for CPD◦ More choice of courses◦ Select the courses that suit your career pathwayQualications for:◦ Animal Technologists◦ Management◦ Specialist Roles

35August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareIntroductionIn the wild, rabbits are prey animals and so the act of being picked up and handled can be extremely stressful for them. Previous studies in pet rabbits have shown that during the act of lifting, rabbits show signs of struggling and aggression due to fear; consequently, petowners are recommended to avoid directly lifting animalswhere possible.1 The most commonly used method ofhandling laboratory rabbits is to grasp the animal bythe scruff, while placingyour other hand to support therear, lifting the animal free of the cage/pen. The animalis then held against the body, with the bodyweight being supported on the arm.2 We wanted to introduce a new method of lifting and carrying the rabbit that would reduce stress, similar to the already recognised and widely used method of tunnel handling mice.3In our facility the rabbits are handled multiple times a week, due to them spending time in specifi cally designed playpens for weekly exercise or undergoing weekly weighing and health checks. The rabbits we house are used for cardiovascular models of Myocardial Infarction and so it is important they are not experiencing a lot of stress to ensure validity in data produced. We believe less invasive handling will also affect the overall temperament of the rabbits and subsequently they will be more likely to positively respond to human interaction rather than be avoidant as they have previously been when being scruffed regularly.Validating the use of box training as a refi nement to rabbit handlingALICIA KINALLY, LUCY ONIONS and SARAH GLENNUniversity of Leicester Correspondence: ak716@leicester.ac.uk Based on the Winning Entry of the Andrew Blake Award 2023 Figure 1.Standard rabbit housing at the University of Leicester.The Andrew Blake Tribute Award commemorates the work and life of Andrew Blake, who suffered from Friedrich’s ataxia, a hereditary condition described as one of the “worst of neurological diseases”. Andrew died in May 2002 aged 39. Andrew was passionate about the need to support scientists in their work and his commitment to speaking out against animal rights activists took up much of the last ten years of his life. He died shortly before he was to collect his MBE.ANDREW BLAKETRIBUTE AWARDSPONSORED BY THE ABPIANDREW BLAKETRIBUTE AWARDDON’T KEEP YOUR GOOD IDEA TO YOURSELF!WE WANT TO HEAR ABOUT IT FOR THE 2023 AWARDDETAILS OF THE AWARD This Award is given annually, where sponsorship allows, to the Animal Technician/Technologist judged to have made the most significant contribution to improving standards in laboratory animal welfare over the previous twelve months. All qualified Animal Technologists are guided in their work by the Institute of Animal Technology’s Ethical Statement: In the conduct of their Professional duties Animal Technologists have a moral and legal obligation, 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 good laboratory animal technology and science. The Institute recognises and supports the application of the principles of the 3Rs (Replacement, Reduction, Refinement) in all areas of animal research. The Award is made to acknowledge the professional and personal commitment of Animal Technologists to improving standards in all aspects of laboratory animal care and welfare. THE PRIZE CONSISTS - CONGRESS 2023 FREE ATTENDANCEnext March WHICH WILL INCLUDE DISPLAYING YOUR POSTER(WITH THE OPTION TO ALSO GIVE AN ORAL PRESENTATION)- AN ENGRAVED GLASS PLAQUE - AND £250 CASH AWARDCLOSING DATE FRIDAY 30TH SEPTEMBER 2022 Need advice – or you wish to discuss anything regarding a possible entry? Then please email the IAT Administrator admin@iat.org.uk with your contact details and one of the organisers will respond and give you all the support you need.ARE YOU AN ANIMAL TECH?HAVE YOU BEEN PART OF A TEAM OR HAVE YOU REFINED ANIMAL CARE AND WELFARE IN YOUR FACILITY?ALL ANIMAL TECHNICIANS AND TECHNOLOGISTS, QUALIFIED AT ANY LEVEL AND PRIMARILY WORKING IN THE UK CAN ENTERCRITERIA – The topic of work that you describe in your application may be undertaken as part of a project and PRESENTED AS A POSTER.YOUR POSTER SUBMISSION SHOULD CONTAIN THE FOLLOWING HEADINGS: TITLE, AIM, METHOD, RESULTS, DISCUSSION, CONCLUSION, REFERENCES and ACKNOWLEDGEMENTS The Poster should also contain the content below:- Why did you undertake this work? (what was the potential problem you were trying to improve?) - How did you undertake it? (species, numbers, sex, materials used) - Describe in a comprehensive and concise manner that allows a complete understanding facilitating reproducibility. - Explain if the work contributes to one of the 3Rs. - Explain how the welfare of the animals was improved. - Describe the results you obtained including data generated with assessment. - Were there any statistics undertaken? Please provide this information. Include a brief CV outlining your overall contribution to the work. Please also list your supervisors or PPL holder if applicable for the work. Submit your Poster online via this link https://www.iat.org.uk/abta where you will see the Submission form for completion.To allow others to be able to replicate the work, please consult the ARRIVE guidelines: https://www.nc3rs.org.uk/arrive-guidelinesOur rabbit modelsHousing and acclimatisation ANDREW BLAKE TRIBUTE AWARDThe Andrew Blake Tribute Award commemorates the work and life of Andrew Blake, who suffered from Friedrich’s ataxia, a hereditary condition described as one of the “worst of neurological diseases”. Andrew died in May 2002 aged 39. Andrew was passionate about the need to support scientists in their work and his commitment to speaking out against animal rights activists took up much of the last ten years of his life. He died shortly before he was to collect his MBE.ANDREW BLAKETRIBUTE AWARDSPONSORED BY THE ABPIANDREW BLAKETRIBUTE AWARDDON’T KEEP YOUR GOOD IDEA TO YOURSELF!WE WANT TO HEAR ABOUT IT FOR THE 2023 AWARDDETAILS OF THE AWARD This Award is given annually, where sponsorship allows, to the Animal Technician/Technologist judged to have made the most significant contribution to improving standards in laboratory animal welfare over the previous twelve months. All qualified Animal Technologists are guided in their work by the Institute of Animal Technology’s Ethical Statement: In the conduct of their Professional duties Animal Technologists have a moral and legal obligation, 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 good laboratory animal technology and science. The Institute recognises and supports the application of the principles of the 3Rs (Replacement, Reduction, Refinement) in all areas of animal research. The Award is made to acknowledge the professional and personal commitment of Animal Technologists to improving standards in all aspects of laboratory animal care and welfare. THE PRIZE CONSISTS - CONGRESS 2023 FREE ATTENDANCEnext March WHICH WILL INCLUDE DISPLAYING YOUR POSTER(WITH THE OPTION TO ALSO GIVE AN ORAL PRESENTATION)- AN ENGRAVED GLASS PLAQUE - AND £250 CASH AWARDCLOSING DATE FRIDAY 30TH SEPTEMBER 2022 Need advice – or you wish to discuss anything regarding a possible entry? Then please email the IAT Administrator admin@iat.org.uk with your contact details and one of the organisers will respond and give you all the support you need.ARE YOU AN ANIMAL TECH?HAVE YOU BEEN PART OF A TEAM OR HAVE YOU REFINED ANIMAL CARE AND WELFARE IN YOUR FACILITY?ALL ANIMAL TECHNICIANS AND TECHNOLOGISTS, QUALIFIED AT ANY LEVEL AND PRIMARILY WORKING IN THE UK CAN ENTERCRITERIA – The topic of work that you describe in your application may be undertaken as part of a project and PRESENTED AS A POSTER.YOUR POSTER SUBMISSION SHOULD CONTAIN THE FOLLOWING HEADINGS: TITLE, AIM, METHOD, RESULTS, DISCUSSION, CONCLUSION, REFERENCES and ACKNOWLEDGEMENTS The Poster should also contain the content below:- Why did you undertake this work? (what was the potential problem you were trying to improve?) - How did you undertake it? (species, numbers, sex, materials used) - Describe in a comprehensive and concise manner that allows a complete understanding facilitating reproducibility. - Explain if the work contributes to one of the 3Rs. - Explain how the welfare of the animals was improved. - Describe the results you obtained including data generated with assessment. - Were there any statistics undertaken? Please provide this information. Include a brief CV outlining your overall contribution to the work. Please also list your supervisors or PPL holder if applicable for the work. Submit your Poster online via this link https://www.iat.org.uk/abta where you will see the Submission form for completion.To allow others to be able to replicate the work, please consult the ARRIVE guidelines: https://www.nc3rs.org.uk/arrive-guidelinesApril 2023 Animal Technology and Welfare

36Animal Technology and Welfare August 2020Once our intact male New Zealand white rabbits (all 2.25-2.5kg on arrival) came into the unit from Envigo, they were singularly housed in an Allentown double rabbitrack (see fi gure 1). One side of the rack trays were lined with a paper tray liner and fi lled with a thick layer of aspen chips to allow natural behaviour such as digging. The rabbits also received boxes, tubes, chew blocks and play balls as environmental enrichment. They were acclimatised here for one week; The rabbit holding room is on a 12/12-hour light cycle with lights gradually fading to off at 7pm and gradually turning back on at 7am. They are housed at an average temperature of approximately 19°C which is monitored daily and they are fed the Teklad™ Global Rabbit Diet 2030 in their hopper. Their housing always had a water bottle on both sides of their cage due to the rabbits having access to both sides of their accommodation.During this time, no handling took place and they were allocated into either of two groups – the fi rst group were to be trained with a carrying box containing a shared piece of Vetbed and the second group were given their own Vetbed. Because the size of the batches of rabbits arriving at one time was so small, the fi rst batch was allocated randomly to either of the two groups. A month later when the next group arrived, they were allocated to the remaining group. We wanted to use rabbits that were going to be staying with us for an extended period of time and so we chose to use our MI/SHAM rabbits as they would be with us for around 10 weeks (2 weeks before and 8 weeks post-surgery). This gave us plenty of time to collect data for the training and allowed us to explore whether the surgery affected the training in any way. The control rabbits we house typically are only with us for two weeks and so whilst it is possible to box train them in that time, we would not have been able to collect much data from them.As the rabbits were already here and being used for surgeryand the handling study was not an intrusive project, we did not require any ethical approval to undertake it.Incentives Whilst trialling the carrying box training, we wanted to see if there was something that would be a good incentive for the rabbits that would encourage them into the box quite quickly. We had previously tried treats and clicker training to help encourage them into the boxes but our rabbits did not seem very interested. We took the idea of the Vetbed from the fact that in their playpen already, the carrying box was being used as a ‘hiding house’ and it contained a piece of Vetbed that everyone shared whilst in there (see fi gure 2) so with this, we created 2 groups. Shared Vetbed rabbits (2 rabbits) – A piece of Vetbed was placed in the carrying box that was not changed Figure 2. Rabbit carrying box with Vetbed in situ. or washed in between every rabbit and therefore picked up all of the rabbits’ scent (washed only when very dirty, typically once a week). This was the same as the routine used in the playpen – the rabbits are used to smelling each other only in the playpen and otherwise do not have access to other rabbits’ scent. The thought process was that the shared Vetbed may smell interesting to the rabbit due to everyone else’s scent being on there and therefore he may want to jump in and investigate the scent.Own Vetbed rabbits (3 rabbits) – A piece of Vetbed that was only for the individual rabbit was provided for these rabbits. It was not shared among the group. The Vetbed would only smell like the rabbit who it was assigned to and was only washed if really dirty. The idea behind this was that the rabbit might have felt more comfortable to jump into the box if the box smelled of his own scent.Introduction of the box After a week the rabbits arrived into the unit (usually the following Monday) the carrier box was introduced to the rabbits at the front of the rabbit cage where we were to hold it steadily at the opening waiting for the rabbit to fully jump in (see fi gure 3). They were timed for 3 minutes using a timer and after 3 minutes, if they had not fully jumped into the box (all 4 legs and body) they were scooped up and were carried to the playpen by hand. If they did jump into the box, the entrance to the box was covered using our arms to ensure the rabbit could not fall or jump out and the rabbit was carried to either the playpen or scales where the box was placed down and the rabbit could jump out on his own (see fi gures 4 and 5).Validating the use of box training as a refi nement to rabbit handling

37August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareValidating the use of box training as a refi nement to rabbit handlingFigure 3.8 Figure 4. Figure 5 Figure 4.Figure 5.Figure 6.The timings were recorded for each rabbit on a spreadsheetso we could monitor how long the rabbits took to jump in.After the rabbits had their allocated time in the playpens or had been weighed on the scales, they were gently encouraged back into the box by lightly guiding them in from behind. They were then taken back to their cage the same way as before, with the entrance covered. Once they arrived back at their cage, the box was placed back in the same position in front of the cage as before and the rabbits were able to hop back into the cage in their own time (see fi gure 6). They were not forced out of the box and most rabbits jumped back into their cage straight away.Positive associationWhen training the rabbits, we wanted to ensure they associated the box as being a positive activity that they would voluntarily do. This again would reduce stress. The way in which we did this was by making sure not to rush the rabbits into the box; whilst waiting for them to jump in, they were not pushed, touched or made to feel like they had to jump in. The same goes for jumping out at their end destination; the rabbits are not forced out of the box or picked up out of the box as it could cause the rabbit to form a negative association with the box and effect the end result of becoming a box trained rabbit.We also wanted the box training to be repeatable and a positive experience with whoever was in the rabbit room at the time, due to there being different people actively working in the rabbit room during staff annual leave, training, post op care and weekend cover. Each person who was going to be handling rabbits in the room at various time points regularly came into the room to

38Animal Technology and Welfare August 2020familiarise themselves with the rabbits and vice versa. It was important that the rabbits were comfortable with a range of people so that they felt confi dent jumping into the box regardless of who would be handling them. This involved giving them some extra attention during the day or at end of day sign-offs such as encouraging them to play, talking to and stroking them. We also ensured everyone knew exactly how to undertakethe box training, how to time it and how to record the data. We did this by carrying out training sessions with everyone who was involved. For it to have successfully worked, the handling method needed to be carried out in the same way by everyone so that the rabbits were confi dent enough to jump in regardless of who was handling them.ResultsDuring the fi rst week of box training, the rabbits took a while to jump into the box or did not jump in at all after 3 minutes of timing them. The box was new to them at this point and so they were hesitant to jump in, this was shown by them either running to the other side of their cage or sitting looking at the box not wanting to interact with it at all. After the fi rst minute however, most showed interest in the box either sniffi ng it or rubbing their chins on the front of the box. If they did not jump in the box, they were picked up and carried by hand and the box was left in the playpen with them as normal. After one week of exposure to the box, the rabbits overall became quicker jumping into the box and they seemed to be more comfortable with it and the effect of the different Vetbed also started to show (see fi gure 7).The shared Vetbed was increasingly preferred during the second week and the rabbits were more interested to jump in due to the mixed scent. We found they were sniffi ng and rubbing their chins on it a lot more in the initial few seconds of being presented with the box and were much less wary of its presence. They also did jump in a lot quicker, in almost half the time than in the fi rst week. It looked as though they knew what to do and were excited to explore the smells on the Vetbed.Although the own Vetbed group did jump into the box quicker than the fi rst week, they did not continue to improve and slightly lost interest in the box after the fi rst 2 weeks. There was less chin rubbing and sniffi ng when the rabbits were presented with the box and this was probably due to the Vetbed not being covered in the strong scent of the other rabbits, so it was not exciting enough for them. They did however jump in eventually.These results seemed to be quite consistent through the 6 week period except for week 3 and 5 of the ownVetbed group and week 5 of the shared Vetbed group where the rabbits had myocardial infarction surgery. During the 3 day postoperative period, they were presentedwith the box as normal as they needed to be weighed and although we did record their data, it was however, discounted. The rabbits did not really show interest in thebox during this time as they were recovering from surgery.We have found that since though, some rabbits are still happy to jump into the box during the post op period however, it is usually on days 2 and 3 of the checks.By the end of the study, we found that the shared Vetbed rabbits were still very interested in the box every time the box was presented to them and they got quicker overall jumping in (an average of 14.6 seconds by theend of the study compared to an average of 42 seconds the fi rst week). It seemed to become routine for them when they saw the box – they were coming to the front ofthe cage as soon as they saw us; perhaps in anticipationthat they would get to come out into the box.Figure 7.12Results Fig 7.During the first week of box training, the rabbits took a while to jump into the box or did not jump in at all after 3 minutes of timing them. The box was new to them at this point and so they were hesitant to jump in, this was shown by them either running to the other side of their cage or sitting looking at the box not wanting to interact with it at all. After the first minute however, most showed interest in the box either sniffing it or rubbing their chins on the front of the box. If they did not jump in the box, they were picked up and carried by hand and the box was left in the playpen with them as normal. Shared vetbed Average time: 28.9sShared vetbed STDEV: 18.52Own vetbed Average time: 99.5Own vetbed STDEV: 59.08Shared Vetbed Average time: 28.9sShared Vetbed STDEV: 18.52Own Vetbed Average time: 99.5Own Vetbed STDEV: 59.08Validating the use of box training as a refi nement to rabbit handling

39August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareValidating the use of box training as a refinement to rabbit handlingThe own Vetbed group still showed interest; however, it was lost after a minute or so, most likely due to the rabbits only smelling their own scent and therefore not as interesting. The rabbits would just walk away from the box and no longer interact with it after spending some time sniffing it. This meant the majority of the times they had passed the 3-minute time limit and the rabbits were carried by hand to their destination. At the start of the study the average time it took for them to jump into the box was 118 seconds, at the end it was 133 seconds.Conclusions Overall, we now know that rabbits can successfully accept the box training as a less invasive and more positive method of handling. The rabbits seem to really enjoy it and are often sat ready and waiting when they can see the box is coming towards them. We also know that the shared Vetbed helps to encourage the rabbits to jump into the carrier and it is a positive incentive for them whilst training. We found that after a couple of weeks of training the rabbits with the shared Vetbed, when we offered freshly washed ones at the start of the week, still jumped in and were interested in scenting the box themselves. The rabbits appeared to have learned to jump into the box regardless of whether it was scented or not after getting used to the shared one for a while.The rabbits typically will not jump into the box the first time but after two or three attempts during the first week will usually have learned what to do. This completely removes the need to physically pick up the rabbit as once the rabbit has jumped into the box, the rabbit can then be carried to the destination e.g., scales, playpen, etc., and back. It was easy to implement into the facility as a primary method of handling and typically, the rabbits will jump into the box, once they have learned the method, regardless of who is handling the box. In our facility, we have multiple members of staff working in each room as well as routine visits by our vet and so it is an important point that they are able to jump in regardless of the member of staff handling them. A good example of successful box handling with unfamiliar people was when a group of visitors from a different facility came in to view the box training in action and were able to box the rabbits themselves with no problems. The rabbits jumped in almost straight away. Similarly, we have had our Named Veterinary Surgeon (NVS) come in recently to check on one of our rabbits and she commented on how easy it was to get the rabbit out of the cage to be taken to the scales and how she was impressed how the rabbit jumped into the box for her swiftly when she had never met that rabbit before.Resource: Quantity Cost – each (£)Animal carrier 4 27.11Vetbed 2 40Total: 67.11BudgetWe have broken down the budget of the box training. Both the Vetbed and carriers were relatively cheap and have lasted us a long time.Box training female rabbitsOver the last few weeks, we have had the opportunity to trial the box handling with a few female rabbits we were housing. We were interested to see if they were as responsive as the males to the Vetbed/box with the same scent-based incentive.We undertook all the same steps from acclimatisation to the box handling, however we were only able to trial a shared Vetbed due to the limited number of rabbits we had housed. The trial also only lasted 2 weeks as the females were all experimental controls. The training took place every day and as there was no playpen to be taken to in the box, the rabbits were walked around the room in the box for a couple of minutes.We found that the female rabbits actually were just as responsive to the box and followed the same pattern during the training. At first, they were quite wary of the box but eventually came out from the back of their cage to explore the box. During the first 2 days, the box was placed inside of the cage rather than held at the front as the females were a lot more timid than the males and we wanted to let them explore in their cage where they felt safe. After those first days though, the box was held as normal at the front of the cage and the rabbits started to jump in relatively quickly after it was presented to them, all jumping in before the 3-minute time limit. We noticed they did not show the same sniffing, chin rubbing and scenting behaviours as the males did however it seemed as though they were jumping in to find a nice comfy place to hide. We hope to continue to collect data from female rabbits in the future if we have the chance to house them again but it is encouraging to see that so far, they have taken well to the box handling.

40Animal Technology and Welfare August 2020AcknowledgementsWith thanks to Professor Andre Ng, Dr Reshma Chauhan and Dr Emily Allen of Cardiovascular Sciences for their support and permission to trial the box handling method on their experimental rabbits. And thanks to colleagues at the PRF for continuing the implementation of box training as a standard within the facility.References1 Bradbury, A.G. & Dickens, G.J.E. (2016). Appropriate handling of pet rabbits: a literature review. [Online]. PubMed. Available at: https://pubmed.ncbi.nlm.nih.gov/27558778/2 NC3RS. (2013). Handling and restraint: General principles. [Online]. https://nc3rs.org.uk/ Last Updated: 20 February 2022. https://nc3rs.org.uk/3rs-resources/ handling-and-restraint3 Kelly Gouveia and Jane L Hurst. (2013). Reducing Mouse Anxiety during Handling: Effect of Experience with Handling Tunne. [Online]. PLOS ONE. Available at: https://journals.plos.org/plosone/article?id=10.1371/ journal.pone.00664014 Meadow Lane Vets. (2021). Health and welfare information about your rabbit from Vetlexicon Lapis. [Online]. Meadow Lane Vets. Last Updated: 2021. Available at: https://meadowlanevets.co.uk/ wp-content/ plugins/vetstream_swagger/pet-health/ pet/pet_info_print.php? Validating the use of box training as a refinement to rabbit handling49Haven’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 Centr e for the 3RsApril 2020 Animal Technology and WelfareTECH-2-TECHBackgroundEmma was invited to write an article as a 3Rschampion in NC3Rs ‘Tech 3Rs’ Issue 5, November2019.Here is her response describing how she has used anautomated system to reduce how frequently mousecage bedding ischanged without compromisingcleanliness.IntroductionOur unit opened in 2017, during the procurement ofnew equipment we had the opportunity to purchase adigital ventilated rack system from Tecniplast UK. Thecages are referred to as the Digitally Ventilated Cage orDVC. This system uses the data collected by sensorsbelow the cage to flag when to clean out based on thechange in an electromagnetic signal. To have thisfunctionality we first needed to create an algorithmduring a learning phase.The learning phase: devising analgorithmWe held a meeting to agree what warranted a cage basechange based on pictures to avoid being subjective. Wereferred to the Home Office Codes of Practice for thehousing and care of animals bred, supplied or used forscientific purposes (HOCoP) for advice on husbandrypractices to set our criteria, balancing hygiene and theimportance of olfactory cues to rodents and their needfor control over their environment.1We started the trial, noting when the cage reached thepoint it required a base change. We assessed airquality, what proportion of the cage base was wet andwhether the animals still had choice over theirenvironment and their ability to show spatial separationof different behaviours such as nesting and excretion,for example their nest was free of faeces. During the‘learning phase’ we a sked our Named VeterinarySurgeon (NVS) and Home Office inspector (HOI) tocheck that they agreed with our assessment.APRIL_1-628207435_4-628196990.e$S:Animal Technology and Welfare 24/9/20 06:51 Page 49

41August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2023 Animal Technology and Welfare49Haven’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 Centr e for the 3RsApril 2020 Animal Technology and WelfareTECH-2-TECHBackgroundEmma was invited to write an article as a 3Rschampion in NC3Rs ‘Tech 3Rs’ Issue 5, November2019.Here is her response describing how she has used anautomated system to reduce how frequently mousecage bedding is changed without compromisingcleanliness.IntroductionOur unit opened in 2017, during the procurement ofnew equipment we had the opportunity to purchase adigital ventilated rack system from Tecniplast UK. Thecages are referred to as the Digitally Ventilated Cage orDVC. This system uses the data collected by sensorsbelow the cage to flag when to clean out based on thechange in an electromagnetic signal. To have thisfunctionality we first needed to create an algorithmduring a learning phase.The learning phase: devising analgorithmWe held a meeting to agree what warranted a cage basechange based on pictures to avoid being subjective. Wereferred to the Home Office Codes of Practice for thehousing and care of animals bred, supplied or used forscientific purposes (HOCoP) for advice on husbandrypractices to set our criteria, balancing hygiene and theimportance of olfactory cues to rodents and their needfor control over their environment.1We started the trial, noting when the cage reached thepoint it required a base change. We assessed airquality, what proportion of the cage base was wet andwhether the animals still had choice over theirenvironment and their ability to show spatial separationof different behaviours such as nesting and excretion,for example their nest was free of faeces. During the‘learning phase’ we a sked 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 49Improving the welfare of mice used in the study of ageing mice: Collated Workshop Feedback LINDA HORAN and MICHAEL WILKINSONUniversity of Strathclyde Correspondence: Linda.horan@strathclyde.ac.uk Based on presentations and feedback from IAT Congress 21 and Congress 22TECH-2-TECHIntroductionHuman lifespan is increasing globally whereas healthspan – the period of life free from age-related diseases – is not increasing at the same rate. One consequence of this is that we are witnessing a concerted research effort into the causes of ageing and the control of its consequences and this effort has resulted in an increase in the number of research groups that use ageing rodents – particularly mice – in their scientific studies.The workshops presented at the 2021 and 2022 Institute of Animal Technology Congresses, aimed to explore the more common clinical signs of ageing and identify husbandry, care measures and humane endpoints. Groups also looked at two example protocols using mice aged for scientific purposes combined with the study of a disease process; the examples were for cancer and arthritis.

42Animal Technology and Welfare August 2020Planning, preparation and considerations for the teamThe preparation and planning of the project are considered important processes. It is essential to have initial discussions to ensure everyone had the same picture/idea of what is expected:1. Include technicians, Veterinary Surgeons, scientists Project (PPL) holders and Personal Licence holders (PIL) NIO and NTCO as appropriate.2. Discuss baselines; weight, body condition and activity levels if possible. Ensure agreement regarding interpretation of endpoints, e.g. what constitutes the weight used as the baseline for any weight loss, agreement on body conditions score, etc. For the arthritis study example, you would need to consider what baseline will be taken for gait and activity/behaviour and consider how this can be measured.3. Clarify (i) intervention points and (ii) endpoints.4. Discuss how interventions and clinical signs accumulate in terms of severity – this should also be reflected in endpoints.5. Training of personnel in the light of discussions, if required; to give consistency/in any new techniques/ monitoring practices.6. Need to consider the impact of any mitigating steps on the scientific outcomes.7. Provide separate score sheets for ageing and the signs associated with the inter-current disease of interest.8. Understanding of the differences between normal ageing and pathology, especially in concurrent models and in the strain being used.9. Comparison can be made with an ageing animal without an inter-current disease.10. A considered approach so that animals are not (inadvertently) killed prior to completion of study for welfare issues associated with normal ageing.11. This is a team process.Mitigations to consider for ageingThe list below emerged as new potential considerations to take into account when monitoring ageing animals in general. – Repeated anaesthesia is an issue for older mice. Gaseous anaesthesia should always be used and there must be steps taken to keep the animals warm. Ensure prolonged body temperature maintenance. – Warmer ambient room temperatures for ageing animals, increase slowly week by week from 15 months to a minimum of 24˚C (check with Home Office (HO) if exceeding Housing Code of Practice). – House ageing studies in a dedicated room – ideally in a quieter part of the animal unit. – Increased aggression may be minimised by improved/ longer acclimatisation period.– Can enrichment items be used to encourage activity to prevent obesity and behavioural problems, e.g. access to running wheel, use of foraging, increased level of complexity in cage, larger floor area, more items for climbing?– Ageing mice ‘bought in’ have a high tendency to fight even if transported in an already established group, offer a larger area with enrichment and foraging material.– Have food pellets specially made that are smaller – does not have the same problems of incisor teeth overgrowth that can be associated with softened foods and smaller pellets easier to handle by older mice. – Be aware of risk of tarsal joint injury (destabilisation of the joint and subsequent dislocation) this is particularly a risk with ageing males after 6 months and it can often be mistaken for a fight injury. – In strains prone to ulcerative dermatitis, consider a scoring system to determine end-point. – Visually, muscle loss can be seen in ageing mice; it is a continual, slow weight loss separate from acute, pathology-related weight loss. – Cage-side monitoring for behavioural changes. – Consider the use of a (non-invasive) frailty index to monitor ageing and welfare and to identify abrupt changes in frailty indicative of pathology. – Use of automated behavioural monitoring of cage (especially in dark phase) to detect changes in behaviour. – Allow the animals to take control of their own environment, give males more space; if they are allowed to make their own hut from cardboard, they will often create just one entry and exit point. – Cages with more height appeared to reduce aggression.– All listed on the common signs of ageing document (see below). In order of prevalence: i.e. skin/hair; musculoskeletal; respiratory; cardiovascular; neurological/behavioural.Points specific to the arthritis model1. Score sheet to assess the level of arthritis (multiple joints, degree of swelling) – however the current ‘standard’ measures (in brackets) were NOT considered a good indicator of disease progression or Animal Welfare. 2. General welfare score (how ‘well’ the animal is) – lack of grooming, etc.3. Evidence of pain – GRIMACE SCALE (not validated for this model and prone to issues with variability e.g. when just woken up).4. An endpoint BEFORE non-weight bearing should be identified.5. Monitoring of gait/activity/behaviour – spontaneous versus evoked. 6. Could there be an ‘analgesic test’ – i.e. animals suspected of arthritis receive dose(s) of analgesic to see if activity increased/lameness decreased? Could this be used as an endpoint to demonstrate the presence of pathology?Improving the welfare of mice used in the study of ageing mice

43August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareImproving the welfare of mice used in the study of ageing miceMitigations to consider for arthritis– Deep bedding/particle size of bedding.– Easy access to food and water with food on floor of cage (to limit the need for climbing).– Pre-exposure to soft food to encourage use later if needed.– Analgesics: what, when, how much, how long? If not, why not? – Discuss possible adjustments for ageing animals within the team – Animal Technicians, Veterinary Surgeons or researchers with experience of the model or species may have ideas on possible solutions. Ask Named Information Officer (NIO) to look at published information to see whether they can identify any other species-specific adjustments that could be made, e.g. ‘easier’ ways to enable climbing. – Ideally monitor in cage via automated technology – decrease in activity likely to flag onset of arthritis.Points specific to the colon cancer modelThe following three clinical signs will be impacted by ageing but not necessarily mean there is an imminent welfare issue: weight loss; body condition score; piloerection; and the animal may be considered as in ‘severe’ when in fact it is just normal ageing.1. Consider weight of tumour vs natural weight loss with ageing vs weight loss due to health issue. Tumour burden assessment should be done to help with this, visually the muscle loss can be seen in ageing mice and a continual, slow weight loss should be expected with ageing animals. 2. Monitoring of stool consistency. 3. Tumour burden assessment – Tumour may not be visible, so rapid weight gain or loss may be an indication of tumour growth. Body scoring may be more useful for this model, especially abdominal distension.Mitigations to consider for colon cancer– Easy access to food and water. – More absorbent bedding. ‘Common’ signs of ageing in mice grouped by organ/system (Reproduced from Progressing the care, husbandry and management of ageing mice used in scientific studies. Michael JA Wilkinson et al).1Organ/System Possible Clinical Signs/Pathology’Skin and hairHair thinning, hair loss, greyingLoss of vibrissaeDry, flaky skin; unkempt coatDelayed wound healing, greater propensity to post-op infections/abscessationIncreased risk/incidence of cutaneous or subcutaneous tumoursSpecial sensesOcular opacities, loss of vision, ocular or periocular infectionsDry, sunken eye/sHearing lossLoss of vibrissaeCardiovascularGeneral slowing down; exercise intoleranceIncreased risk/incidence of:Strokes (neurological signs; sudden death)Heart attacks/fatal arrhythmias (extreme collapse; sudden death)Poorer post-anaesthetic recovery/anaesthetic-related deathRespiratoryNasal discharge, sneezing, coughing/chattering, rapid/shallow breathing, dyspnoea, aerophagiaIncreased risk/incidence of tumoursDigestive Malocclusions, dental abscessesGastrointestinal dysfunction (diarrhoea; constipation; changed Body Condition Score31 [BCS])Rectal prolapseIncreased risk/incidence of tumoursMusculoskeletalArthritis/arthrosis/loss of muscle tone (swollen/painful joints; reluctance to move general slowing down; exercise intolerance; gait abnormalities; decreased grip strength)Hunched postureIncreased risk/incidence of bone fractures/dislocations (pain)Metabolic/endocrineLess able to thermoregulate (shivering; cold stress; piloerection; heat intolerance; decreased core body temperature)Less able to keep energy balance (weight loss; weight gain)Less able to process/clear drugs (increased risk of toxicity)Increased risk/incidence of tumoursReferences1 Wilkinson, M.J.A. et al. (2020). Laboratory Animals. 54.3 pp 225-238.

44Animal Technology and Welfare August 2020Abstract Common marmosets (Callithrix jacchus) are New World primates that are found within the coastal rainforests of North East (NE) Brazil. They are a multisensory species, relying on all their senses to survive. It is the duty of all animal carers to fulfi l the 5 needs of captive animals,1 one of which being the need to express natural behaviours. Apart from the environmental enrichment that is placed in a primate’s home cage, the majority of enrichment ideas used usually involve hiding foods and sweet treats encouraging the sense of taste to keep the animals engaged for longer. Within the marmoset breeding colony,at the Defence Science and Technology Laboratory (Dstl),a form of enrichment that would encourage the primates to use sense of smell was trialled. To do this thehammock within the marmoset’s home cage was soakedin other scents to encourage a use of the sense of smell. This poster describes the trial designed to assess 2 different scents on one of the breeding families.HypothesisThe marmosets would smell the hammocks morefrequently with different scents more compared to the scentless hammock. The marmosets would prefer the scent of oregano compared to cinnamon as cinnamon has a sharper scent than oregano. The hammock scent would reduce the time spent by the marmosets in the hammock owing to the scent being unfamiliar, the hammock scent would also affect the playing, relaxing and grooming activities whilst in the hammock.Sensory enrichment for the common marmoset (Callithrix jacchus)MILLIE FULLER CBR; Defence Science and Technology Laboratory (Dstl) Correspondence: melfuller@dstl.gov.uk Based on a poster displayed at IAT Congress 22Trialling ideasWhen it came to looking for a scent it was decided to use something that is similar to that which a marmoset might come across in the wild but also easy to get hold of here in the UK. Looking into the natural habitat of the common marmoset (NE Brazil) there are a number of different herbs and spices that grow there. The end decision was to use oregano (Origanum vulgare) and cinnamon (genus Cinnamomum) allowing the comparison of a herb and a spice.4 After trialling a couple methods of getting the scent to attach to the wooden perch, such as soaking the perch in a bucket of scented water or applying a paste of the desired scent to the perch, it was realised that using wooden perches was not feasible. Tea towel hammocks were chosen as a better option. Figure 1. Top: Cinnamon Hammock, Bottom: Plain Hammock Figure 1.Top: Cinnamon Hammock. Bottom: Plain Hammock.4 After trialling a couple methods of getting the scent to attach to the wooden perch, such as soaking the perch in a bucket of scented water or applying a paste of the desired scent to the perch, it was realised that using wooden perches was not feasible. Tea towel hammocks were chosen as a better option. Figure 1. Top: Cinnamon Hammock, Bottom: Plain Hammock Animal Technology and Welfare April 2023

45August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareThe next stage was to fi nd a way to present the scent to the family group in the home cage. To ensure that the marmosets were interested in a scent, not a novel item, a piece of regularly used cage furniture was assessed. The object had to be easily replicated, cleaned and placed back into the cage. The items of choice were wooden perches or tea towel hammock.After trialling a couple of methods of getting the scent to attach to the wooden perch, such as soaking the perch in a bucket of scented water or applying a paste of the desired scent to the perch, it was realised that using wooden perches was not feasible. Tea towel hammocks were chosen as the better option.MethodOne breeding family was chosen to record during the whole scent hammock trial. This family consisted of 6 individuals. Dam and sire, 2 juveniles 6+ months and 2 infants under 6 months. Each day before fi lming, the camera was placed on top of the family cage to allow the marmosets to acclimatise to the camera. This then ensured focus was on the enrichment given and not the camera. Over the space of a week, the family group were given the three types of hammock, one at a time, and recorded for an hour between 1100-1200. All the essential husbandry duties were completed in the room for the day before any recording took place and access was restricted to the room to prevent any other distractions. This was repeated for three consecutive weeks.The two scented hammocks were created by soaking each in a mixture of the chosen scent (either cinnamon or oregano) and water for an hour before being hung up to dry overnight. The following day they were ready to be placed in the cage for monitoring.Once all the footage was recorded it was then analysed to see how many times specifi c behaviours were noticed.These behaviours were:– Smelling, seeing how many instances of smelling the hammock were recorded. – Licking/chewing, either chewing the hammock material itself or the individual spices/herbs.– Grooming, the marmosets sitting on the hammock grooming one another. – Relaxing, The marmosets either sitting or lying on the hammock for a certain period of time without getting off the hammock or performing other behaviours.– Playing, The marmosets chasing or play fi ghting with one another on the hammock.Results/Discussion On average the marmoset family spent 117 seconds (1 minutes 57 seconds) smelling the cinnamon hammock.This is 35 seconds more than the average time spent smelling the oregano hammock. However the hammock that had the least time spent smelling was the plain hammock which had only an average of 33 seconds. This shows the fi rst statement of the hypothesis, the 6- Smelling, seeing how many instances of smelling the hammock were recorded. - Licking/chewing, either chewing the hammock material itself or the individual spices/herbs.- Grooming, the marmosets sitting on the hammock grooming one another. - Relaxing, The marmosets either sitting or lying on the hammock for a certain period of time without getting off the hammock or performing other behaviours.- Playing, The marmosets chasing or play fighting with one another on the hammock.Sensory enrichment for the common marmoset (Callithrix jacchus)

46Animal Technology and Welfare August 2020marmosets would smell the hammocks with different infused scents more, compared to plain hammocks was proved correct.The second part of the hypothesis had to be rejected as the data shows the opposite of what was predicted. Overall the family group spent more time on the cinnamon hammock, a total average of 842 seconds (14 minutes) compared to a total average of 429 seconds (7 minutes) on the oregano hammock.The data collected shows time spent playing, relaxing and grooming was reduced on the oregano hammock, a total average of 90 seconds (1 minute 30 seconds), compared to the plain hammock, a total average of 503 seconds (8 minutes 23 seconds). However the longest time spent playing, grooming and relaxing was the cinnamon hammock, a total average of 577 seconds (9 minutes 37 seconds) This was surprising as this scent was the strongest scent. This meant the final statement of the hypothesis ‘with the other behaviours that will be monitored the marmosets would be more inclined to play, relax and groom on the hammock with no extra scent as it is more familiar to the family group’ was rejected. ConclusionIn conclusion the trial was a success in showing that the use of scented enrichment works to stimulate the marmosets and keep them occupied. This is the first time within the colony’s history that scents were used to stimulate our marmosets in other ways than by taste and even though attempts were made to lick/chew the scented hammocks, whether it was picking at each individual oregano herb/grain of cinnamon on the hammock, the marmosets still spent more time smelling the material compared to licking/chewing it. Cinnamon was the favoured scent of the two scented hammocks in all areas apart from licking/chewing which is most likely down to the ease of picking at individual pieces of oregano compared to grains of cinnamon.Since this trial was completed we have now implemented the use of scent hammocks into our enrichment rotation. With the success of this trial there are also plans in place for further investigations into other scents as well as looking into other senses and how we can encourage marmosets to use other senses, such as sound and touch.Acknowledgements I would firstly like to thank Phillipa Gent for helping me with the data collecting for this project. I would also like to thank the rest of my colleagues for being patient and supportive throughout.References1 https://wikipedia/wiki/fivefreedoms2 WWF (2021). ‘Ten products and ingredients that come from tropical rainforests’ [online] Available at https://www.wwf.org.uk/updates/ten-products-and-ingredients-come-tropical-forests (Accessed 16/12/2021).3 Scielo (2013). ‘Photodegradation of essential oil from marjoram’ [online]. Available at http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S0370-59432013000200001 (Accessed 16/12/2021).Sensory enrichment for the common marmoset (Callithrix jacchus)

47August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAbstract/IntroductionThis is an instructional poster on how to breed Xenopus laevis in the licenced laboratory.The many challenges of 2020 had a negative impact on xenopus suppliers, predominantly the closure of the world’s largest supplier NASCO.However other suppliers include, Portsmouth UK (EXRC), Xenopus1 USA and Xenopus Express. Due to the increase in demand on all companies, in-house breeding may be needed to supplement stock.In-house breeding is also an excellent refinement in Animal Welfare as it eliminates the stress caused by travel and the animals are acclimatised to the facility. Plus it is cost-effective. Advantages and disadvantages of different model organisms. Xenopus are widely used to study vertebrate embryology and development, basic cell and molecular biology, genomics, neurobiology and toxicology and to model human diseases (Xenbase.org, 2022).Superovulation equipment– MS222 (Tricaine methanesulfonate) – aquatic species anaesthetic. – 1ml syringe and 27 or 26 gauge needle.– L15 (Leibrovitz without Glutamine) Sigma L5520.– Penicillin and streptomycin.– Adult male xenopus (testes).– Forceps, scissors.How to breed and raise African Clawed-Toed frogs (Xenopus laevis)THERESE JONES-GREEN University of CambridgeCorrespondence: tj222@cam.ac.uk Based on a poster displayed at IAT Congress 22 Category:C. elegansDrosophila Zebrafish Xenopus Chicken MouseBroodsize250-300 80-100 100-200 500-3000+ 1 5-8Cost per embryolow low low low mediumhighHigh-throughput multiwell-format screeninggood good good goodpoor poorAccess to embryosgood good good goodpoor poorMicro-manipulation of embryoslimited limited fairgood goodpoorGenomeknown known known known known knownGeneticsgood good goodfair nonegoodKnockdowns (RNAi, morpholinos)good good good goodlimited limitedTransgenesisgood good good goodpoorgoodEvolutionary distance to humanvery distant very distant distant intermediate intermediatecloseColour code: green, best in category; red, worst in category.Taken from Xenbase.org, adapted from Wheeler & Brändli 2009 Dev Dyn 238:1287-1308April 2023 Animal Technology and Welfare

48Animal Technology and Welfare August 2020– Human Chorionic GonadotropinhCG (Sigma). – Adult female xenopus.– 1XMMR (Marc’s Modified Ringer solution.– 4ml pipette, turkey baster, 5ml petri dishes.– Holding tanks, 1L beakers, tadpole rearing tanks.– Spirulina (Sera micron, ZM Spirulina etc) protein powder, small bite froglet food. Superovulation eggs– Inject 150ul of PMSG (Intervet) into the female frog’s dorsal lymph sac a week before eggs are needed.– The day before eggs are required inject 400ul of hCGinto the dorsal lymph sac. Frogs will begin laying 8 hours later.– The next day dispose of any laid eggs as these will not be viable.– Place the female into a small holding tank containing 5L of 1XMMR (High salt/egg laying solution).3 Superovulation Equipment • MS222 (Tricaine methanesulfonate) -aquatic species anaesthetic. • 1ml syringe and 27 or 26 gauge needle – Allow the frog a few hours to lay the desired number of eggs you wish to fertilise.– Collect eggs using a turkey baster and place into 5ml petri dishes ready for fertilisation.– Return frog to fresh system water once finished. Eggs to avoid– Try to select eggs that are laid singular and round. Colour is not important and depends on parent’s pigmentation.– Eggs laid in strings, clumps, or as larger white spheres are no good.– Sometimes the frogs eat their eggs and then vomit. You will know if this has happened if there are large clumps of eggs. A barrier can be placed in the tank to discourage this behaviour.6 - Place the female into a small holding tank containing 5L of 1XMMR (High salt/egg laying solution) - Allow the frog a few hours to lay the desired number of eggs you wish to fertilise. - Collect eggs using a turkey baster and place into 5ml petri dishes ready for fertilisation - Return frog to fresh system water once finished. How to breed and raise African Clawed-Toed frogs (Xenopus laevis)Figure 1. Superovulation equipment. Figure 2. Female frog in holding tank. Figure 3. Unsuitable eggs.

49August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareHow to breed and raise African Clawed-Toed frogs (Xenopus laevis)Testes: non-regulated procedure– Select an adult male with well-defined black nuptial pads.– Euthanise the xenopus using a Schedule 1 method.– Make an incision across the frog’s waist. With forceps, pull out the yellow fat strands. The testes will be attached to the fat towards the back of the frog. Carefully cut the testes away from the connective tissues.– Place testes into a petri-dish with L15 and 20ul of penicillin and streptomycin. This is to stop the sperm activating and prevent unwanted growth of bacteria. (Figure 5)– Store in the fridge for up to a week.9 In--vitro fertilisation– Cut a small amount of teste off (2 mm³).– Using a pipette remove as much 1XMMR as possible.– Using the forceps pull the fragment of cut teste apart on the dish containing the dry eggs.– Pick up the testes with forceps and waft the teste over each individual egg, then leave the fragments of testes sitting with the eggs.– Wait 5 minutes. 10 11 In--vitro Fertilisation - Cut a small amount of teste off (2 mm³) - Using a pipette remove as much 1XMMR as possible. - Using the forceps pull the fragment of cut teste apart on the dish containing the dry eggs - Pick up the testes with forceps and waft the teste over each individual egg, then leave the fragments of testes sitting with the eggs - Wait 5 minutes 12 Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water a). Do not use de-mineralised water, as a low salt concentration is required. - The eggs and sperm will now fertilise. This processtakes10-20minutes. You know if it’s successful as the eggs will rotate so that the animal pole is facing upwards (b) Eggs can be left on the bench at 22°Celsius or put in an incubator to either slow down or speed up development. 12 Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water a). Do not use de-mineralised water, as a low salt concentration is required. - The eggs and sperm will now fertilise. This processtakes10-20minutes. You know if it’s successful as the eggs will rotate so that the animal pole is facing upwards (b) Eggs can be left on the bench at 22°Celsius or put in an incubator to either slow down or speed up development. 12 Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water a). Do not use de-mineralised water, as a low salt concentration is required. - The eggs and sperm will now fertilise. This processtakes10-20minutes. You know if it’s successful as the eggs will rotate so that the animal pole is facing upwards (b) Eggs can be left on the bench at 22°Celsius or put in an incubator to either slow down or speed up development. 11 In--vitro Fertilisation - Cut a small amount of teste off (2 mm³) - Using a pipette remove as much 1XMMR as possible. - Using the forceps pull the fragment of cut teste apart on the dish containing the dry eggs - Pick up the testes with forceps and waft the teste over each individual egg, then leave the fragments of testes sitting with the eggs - Wait 5 minutes Figure 5. Testes in petri dish. Figure 4. Male anatomy. Figure 6. In-vitro fertilisation process.

50Animal Technology and Welfare August 2020Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water – a) Do not use de-mineralised water, as a low salt concentration is required. – The eggs and sperm will now fertilise. This processtakes 10-20 minutes. You know if it is successful as the eggs will rotate so that the animal pole is facing upwards. b) Eggs can be left on the bench at 22°C or put in anincubator to either slow down or speed up development.13 A) B) Natural Mating - To preserve your male stock, inject a male with 50μl of Human Chorionic Gonadotropin (hCG) two days before mating. Use the same steps for superovulation for the female. - Put the male and female together in a holding tank in a quiet dark place for two days (make sure they still get a discrete daily health check). - After the two days, many eggs will have been laid and fertilised. These will be at different stages of development. - Remove the male and female.•Drain off excess water and add fresh system water. - Keep the water clean, pipette out any dead eggs. - After 3 or 4 days the eggs will hatch, and feeding can begin. (A) (B)Natural mating –To preserve your male stock, inject a male with 50µlof Human Chorionic Gonadotropin (hCG) two days before mating. Use the same steps for superovulation for the female.– Put the male and female together in a holding tank in a quiet dark place for two days (make sure they still get a discrete daily health check).– After the two days, many eggs will have been laid and fertilised. These will be at different stages of development.– Remove the male and female.– Drain off excess water and add fresh system water.– Keep the water clean, pipette out any dead eggs.– After 3 or 4 days the eggs will hatch and feeding can begin.Caring for fertilised eggs– Remove dead eggs/embryos daily and change the media if it gets cloudy (a).– Top up water if evaporation has occurred.–Make sure the eggs are not too crowded, as dead eggswill infect healthy eggs. – (b) 30 eggs in a 70mm petri dish works well.–Eggs should hatch 2/3 days post fertilisation at roomtemperature (22°C), development can be sped up orslowed down dependent on the temperature they arekept at.Abstract/IntroduconAdvantages and disadvantages of different model organisms. Xenopusare widely used to study vertebrate embryology and development, basic cell and molecular biology, genomics, neurobiology and toxicology and to model human diseases (Xenbase.org, 2022).Category: C. elegans Drosophila Zebrafish Xenopus Chicken MouseBroodsize 250-300 80-100 100-200 500-3000+ 1 5-8Cost per embryo low low low low medium highHigh-throughput multiwell-format screening good good good good poor poorAccess to embryos good good good good poor poorMicro-manipulation of embryos limited limited fair good good poorGenome known known known known known knownGenetics good good good fair none goodKnockdowns (RNAi, morpholinos) good good good good limited limitedTransgenesis good good good good poor goodEvolutionary distance to human very distant very distant distant intermediate intermediate closeColour code: green, best in category; red, worst in category. Taken from Xenbase.org, adapted fromWheeler & Brändli 2009 Dev Dyn 238:1287-1308Superovulation Equipment• MS222• 1ml syringe and 27 or 26 gauge needle• L15 (leibrovits without Glutamine) Sigma L5520• Penicillin and streptomycin• Adult male xenopus (testes)• Forceps, scissors• Human Chorionic Gonadotropin hCG (Sigma)• Adult female xenopus• 1XMMR (Marc's Modified Ringer solution• 4ml pipette, turkey baster, 5ml petri dishes• Holding tanks, 1L beakers, tadpole rearing tanks• Spirulina( Sera micron, ZM Spirulina etc) protein powder, small bite froglet foodTherese Jones-Green, Chief Aquatic Technician/Named Animal Care and Welfare Ocer, University of CambridgeHow to Breed and Raise African Clawed-Toed Frogs (Xenopus laevis)Superovulation Eggs• Inject 150ul of PMSG (Intervet) into the female frog’s dorsal lymph sac a week before eggs are needed• The day before eggs are required inject 400ul of hCG into the dorsal lymph sac. Frogs will begin laying 8 hours later.• The next day dispose of any laid eggs as these will not be viable.• Place the female into a small holding tank containing 5L of 1XMMR (High salt/egg laying solution)• Allow the frog a few hours to lay the desired number of eggs you wish to fertilise.• Collect eggs using a turkey baster and place into 5ml petri dishes ready for fertilisation• Return frog to fresh system water once finishedEggs to avoid• Try to select eggs that are laid singular and round. Colour is not important and depends on parent's pigmentation.• Eggs laid in strings, clumps, or as larger white spheres are no good.• Sometimes the frogs eat their eggs and then vomit, you will know if this has happened if there are large clumps of eggs. A barrier can be placed in the tank to discourage this behaviour.Testes: Non-RegulatedProcedure• Select an adult male with well-defined black nuptial pads.• Euthanise the xenopus using a schedule 1 method.• Make an incision across the frog's waist. With forceps, pull out the yellow fat strands. The testes will be attached to the fat towards the back of the frog. Carefully cut the testes away from the connective tissues.• Place testes into a petri- dish with L15 and 20ul of penicillin and streptomycin. This is to stop the sperm activating and prevent unwanted growth of bacteria.• Store in the fridge for up to a week.In-vitro Fertilisation• Cut a small amount of teste off (2 mm³)• Using a pipette remove as much 1XMMR as possible.• Using the forceps pull the fragment of cut teste apart on the dish containing the dry eggs• Pick up the testes with forceps and waft the teste over each individual egg, then leave the fragments of testes sitting with the eggs• Wait 5 minutesIn-vitroFertilisation cont..ab• Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water (a). Do not usede-mineralised water, as a low salt concentration is required.• The eggs and sperm will now fertilise. This process takes 10-20minutes. You know if it's successful as the eggs will rotate so that the animal pole is facing upwards (b)• Eggs can be left on the bench at 22° Celsius or put in an incubator to either slow down or speed up development.Natural Mating• To preserve your male stock, inject a male with 50µl of hCG two days before mating. Use the same steps for superovulation for the female.• Put the male and female together in a holding tank of their system’s water and leave them in a quiet dark place for two days (make sure they still get a discrete daily health check).• After the two days, many eggs will have been laid and fertilised. These will be at different stages of development.• Remove the male and female.• Drain off excess water and add fresh system water.• Keep the water clean, pipette out any dead eggs.• After 3 or 4 days the eggs will hatch, and feeding can begin.Caring forfertilised eggs• Remove dead eggs/embryos daily and change the media if it gets cloudy (a).• Top up water if evaporation has occurred.• Make sure the eggs are not too crowded, as dead eggs will infect healthy eggs (b). 30 eggs in a 70mm petri dish works well.• Eggs should hatch 2/3 days post fertilisation at room temperature (22C), development can be sped up or slowed down dependant on the temperature they are kept at.abCaringfor tadpoles• The first couple of weeks are the most sensitive.• Remove dead tadpoles using a pipette.• As the tadpoles increase in size, feed more food and increase the size of their holding container.• The water should have a slightly sweet aroma, if it starts to smell foul, change 75% of the water. Sometimes a whole water change will be needed. Knowing when and how much water to change is a fine balance.Tadpolefeeding• Add small amounts of spirulina or Sera Micron suspended in demineralised water to the tanks, so the water turns a very light green colour.• If the water becomes cloudy refrain from feeding until it is clear. Change 50% of the water.• As the tadpoles grow more food should be added. Protein (egg powder) can also be used to supplement their diet. Sera Micron contains extra proteins.Aeration?Aeration (using a pump and air stones) helps maintain water quality, but it often requires a difficult and elaborate set up to make sure the bubbling is neither too fast nor too weak.Natural Mating versus SuperovulationNatural Mating SuperovulationPreserve male stockPreciseAllow animals to exhibit natural behaviour Can use left-over testes and eggs from experimentsTime efficient Easier to keep clean post fertilisationGet many embryos at different stages of developmentBatches can unexpectantly crash (die)No consumables cost Hormones needed to induce ovulationCan be difficult to know if fertilisation was successfulEasier to know if fertilisation has been successfulSick tadpoles and thingsto look out for• Some tadpoles have crooked tails, this is usually Okay. Feed them more spirulina.• Some tadpoles have developmental defects (left). They should be euthanised with a Schedule 1 method.• If a tadpole is corkscrewing (spinning) remove and euthanise as above.• If many tadpoles are floating, change their water, they may need more oxygen.• If a single tadpole is floating or sitting on bottom of the tank, and unresponsive (but alive) remove and euthanise Sometimes after a water change some tadpoles turn upside down. Don't panic, they usually sort themselves out after a couple of hours.Healthy tadpolesUnhealthy tadpolesUnhealthy tadpolesConclusionThis poster is aimed at animal technicians as a tool to support in-house breeding. Some parts may need adapting depending on local water conditions, type of aquatic system used, and availability of resources. Please feel free to contact me with any ideas or questions related to in-house breeding: tj222@cam.ac.ukAcknowledgementsThe Gurdon InstituteCambridge University Biomedical ServicesBackground to the Home Office designated bio-facility• Opened in 2005, the bio-facility can hold up to 900 female and 240 male frogs in a Marine Biotech (now serviced by MBK Installations Ltd.) recirculating aquatic system.• The system monitors parameters such as: temperature, pH, conductivity, and Total Gas Pressure. • Frogs are maintained on mains fed water with a 20% water exchange. • There are several filtration steps, including: biological, mechanical, activated carbon, and UV sterilization.• Frogs are held under a 12-hour light/dark cycle.• Each tank has a PVC enrichment tube (30cm x 10cm), with smoothed cut edges.• Frogs are fed three times a week with a commercial trout pellet. • The focus of the research undertaken in the bio-facility is embryonic and developmental biology. All procedures are conducted under Home Office licence authority.• Frogs are euthanized humanely using a Schedule 1 methodFroglet care• Xenopus can take anywhere between 6 weeks to 6 months to metamorphize. It may be an evolutionary advantage to the species to have it spaced out this way.• You must remove froglets from the tank, as they will eat smaller tadpoles.• Froglets can be fed on 2mm trout pellets (manufactured by Skretting), Baby Reptomin, or large bite Zebrafish food• Many froglets can be kept together but make sure they are matched in size as larger frogs will attack smaller during feeding• Froglets like plastic plants and small tubes• Very easy to raise• Reach maturity at around 2 years of age and can live to over 30 years!References• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownReferences• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownReferences• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownUniversity Biomedical Services14 Caring for fertilised eggs - Remove dead eggs/embryos daily and change the media if it gets cloudy (a). - Top up water if evaporation has occurred. - Make sure the eggs are not too crowded, as dead eggs will infect healthy eggs - (b). 30 eggs in a 70mm petri dish works well. - Eggs should hatch 2/3 days post fertilisation at room temperature(22C), development can be sped up or slowed down dependant on the temperature they are kept at. Caring for tadpoles– The fi rst couple of weeks are the most sensitive.– Remove dead tadpoles using a pipette.– As the tadpoles increase in size, feed more food and increase the size of their holding container.– The water should have a slightly sweet aroma, if it starts to smell foul, change 75% of the water. Sometimes a whole water change will be needed. Knowing when and how much water to change is a fi ne balance. Abstract/IntroduconAdvantages and disadvantages of different model organisms. Xenopusare widely used to study vertebrate embryology and development, basic cell and molecular biology, genomics, neurobiology and toxicology and to model human diseases (Xenbase.org, 2022).Category: C. elegans Drosophila Zebrafish Xenopus Chicken MouseBroodsize 250-300 80-100 100-200 500-3000+ 1 5-8Cost per embryo low low low low medium highHigh-throughput multiwell-format screening good good good good poor poorAccess to embryos good good good good poor poorMicro-manipulation of embryos limited limited fair good good poorGenome known known known known known knownGenetics good good good fair none goodKnockdowns (RNAi, morpholinos) good good good good limited limitedTransgenesis good good good good poor goodEvolutionary distance to human very distant very distant distant intermediate intermediate closeColour code: green, best in category; red, worst in category. Taken from Xenbase.org, adapted fromWheeler & Brändli 2009 Dev Dyn 238:1287-1308Superovulation Equipment• MS222• 1ml syringe and 27 or 26 gauge needle• L15 (leibrovits without Glutamine) Sigma L5520• Penicillin and streptomycin• Adult male xenopus (testes)• Forceps, scissors• Human Chorionic Gonadotropin hCG (Sigma)• Adult female xenopus• 1XMMR (Marc's Modified Ringer solution• 4ml pipette, turkey baster, 5ml petri dishes• Holding tanks, 1L beakers, tadpole rearing tanks• Spirulina( Sera micron, ZM Spirulina etc) protein powder, small bite froglet foodTherese Jones-Green, Chief Aquatic Technician/Named Animal Care and Welfare Ocer, University of CambridgeHow to Breed and Raise African Clawed-Toed Frogs (Xenopus laevis)Superovulation Eggs• Inject 150ul of PMSG (Intervet) into the female frog’s dorsal lymph sac a week before eggs are needed• The day before eggs are required inject 400ul of hCG into the dorsal lymph sac. Frogs will begin laying 8 hours later.• The next day dispose of any laid eggs as these will not be viable.• Place the female into a small holding tank containing 5L of 1XMMR (High salt/egg laying solution)• Allow the frog a few hours to lay the desired number of eggs you wish to fertilise.• Collect eggs using a turkey baster and place into 5ml petri dishes ready for fertilisation• Return frog to fresh system water once finishedEggs to avoid• Try to select eggs that are laid singular and round. Colour is not important and depends on parent's pigmentation.• Eggs laid in strings, clumps, or as larger white spheres are no good.• Sometimes the frogs eat their eggs and then vomit, you will know if this has happened if there are large clumps of eggs. A barrier can be placed in the tank to discourage this behaviour.Testes: Non-RegulatedProcedure• Select an adult male with well-defined black nuptial pads.• Euthanise the xenopus using a schedule 1 method.• Make an incision across the frog's waist. With forceps, pull out the yellow fat strands. The testes will be attached to the fat towards the back of the frog. Carefully cut the testes away from the connective tissues.• Place testes into a petri- dish with L15 and 20ul of penicillin and streptomycin. This is to stop the sperm activating and prevent unwanted growth of bacteria.• Store in the fridge for up to a week.In-vitro Fertilisation• Cut a small amount of teste off (2 mm³)• Using a pipette remove as much 1XMMR as possible.• Using the forceps pull the fragment of cut teste apart on the dish containing the dry eggs• Pick up the testes with forceps and waft the teste over each individual egg, then leave the fragments of testes sitting with the eggs• Wait 5 minutesIn-vitroFertilisation cont..ab• Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water (a). Do not usede-mineralised water, as a low salt concentration is required.• The eggs and sperm will now fertilise. This process takes 10-20minutes. You know if it's successful as the eggs will rotate so that the animal pole is facing upwards (b)• Eggs can be left on the bench at 22° Celsius or put in an incubator to either slow down or speed up development.Natural Mating• To preserve your male stock, inject a male with 50µl of hCG two days before mating. Use the same steps for superovulation for the female.• Put the male and female together in a holding tank of their system’s water and leave them in a quiet dark place for two days (make sure they still get a discrete daily health check).• After the two days, many eggs will have been laid and fertilised. These will be at different stages of development.• Remove the male and female.• Drain off excess water and add fresh system water.• Keep the water clean, pipette out any dead eggs.• After 3 or 4 days the eggs will hatch, and feeding can begin.Caring forfertilised eggs• Remove dead eggs/embryos daily and change the media if it gets cloudy (a).• Top up water if evaporation has occurred.• Make sure the eggs are not too crowded, as dead eggs will infect healthy eggs (b). 30 eggs in a 70mm petri dish works well.• Eggs should hatch 2/3 days post fertilisation at room temperature (22C), development can be sped up or slowed down dependant on the temperature they are kept at.abCaringfor tadpoles• The first couple of weeks are the most sensitive.• Remove dead tadpoles using a pipette.• As the tadpoles increase in size, feed more food and increase the size of their holding container.• The water should have a slightly sweet aroma, if it starts to smell foul, change 75% of the water. Sometimes a whole water change will be needed. Knowing when and how much water to change is a fine balance.Tadpolefeeding• Add small amounts of spirulina or Sera Micron suspended in demineralised water to the tanks, so the water turns a very light green colour.• If the water becomes cloudy refrain from feeding until it is clear. Change 50% of the water.• As the tadpoles grow more food should be added. Protein (egg powder) can also be used to supplement their diet. Sera Micron contains extra proteins.Aeration?Aeration (using a pump and air stones) helps maintain water quality, but it often requires a difficult and elaborate set up to make sure the bubbling is neither too fast nor too weak.Natural Mating versus SuperovulationNatural Mating SuperovulationPreserve male stockPreciseAllow animals to exhibit natural behaviour Can use left-over testes and eggs from experimentsTime efficient Easier to keep clean post fertilisationGet many embryos at different stages of developmentBatches can unexpectantly crash (die)No consumables cost Hormones needed to induce ovulationCan be difficult to know if fertilisation was successfulEasier to know if fertilisation has been successfulSick tadpoles and thingsto look out for• Some tadpoles have crooked tails, this is usually Okay. Feed them more spirulina.• Some tadpoles have developmental defects (left). They should be euthanised with a Schedule 1 method.• If a tadpole is corkscrewing (spinning) remove and euthanise as above.• If many tadpoles are floating, change their water, they may need more oxygen.• If a single tadpole is floating or sitting on bottom of the tank, and unresponsive (but alive) remove and euthanise Sometimes after a water change some tadpoles turn upside down. Don't panic, they usually sort themselves out after a couple of hours.Healthy tadpolesUnhealthy tadpolesUnhealthy tadpolesConclusionThis poster is aimed at animal technicians as a tool to support in-house breeding. Some parts may need adapting depending on local water conditions, type of aquatic system used, and availability of resources. Please feel free to contact me with any ideas or questions related to in-house breeding: tj222@cam.ac.ukAcknowledgementsThe Gurdon InstituteCambridge University Biomedical ServicesBackground to the Home Office designated bio-facility• Opened in 2005, the bio-facility can hold up to 900 female and 240 male frogs in a Marine Biotech (now serviced by MBK Installations Ltd.) recirculating aquatic system.• The system monitors parameters such as: temperature, pH, conductivity, and Total Gas Pressure. • Frogs are maintained on mains fed water with a 20% water exchange. • There are several filtration steps, including: biological, mechanical, activated carbon, and UV sterilization.• Frogs are held under a 12-hour light/dark cycle.• Each tank has a PVC enrichment tube (30cm x 10cm), with smoothed cut edges.• Frogs are fed three times a week with a commercial trout pellet. • The focus of the research undertaken in the bio-facility is embryonic and developmental biology. All procedures are conducted under Home Office licence authority.• Frogs are euthanized humanely using a Schedule 1 methodFroglet care• Xenopus can take anywhere between 6 weeks to 6 months to metamorphize. It may be an evolutionary advantage to the species to have it spaced out this way.• You must remove froglets from the tank, as they will eat smaller tadpoles.• Froglets can be fed on 2mm trout pellets (manufactured by Skretting), Baby Reptomin, or large bite Zebrafish food• Many froglets can be kept together but make sure they are matched in size as larger frogs will attack smaller during feeding• Froglets like plastic plants and small tubes• Very easy to raise• Reach maturity at around 2 years of age and can live to over 30 years!References• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownReferences• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownReferences• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownUniversity Biomedical ServicesHow to breed and raise African Clawed-Toed frogs (Xenopus laevis)Figure 7. Fertilised eggs. Figure 8. Frogs in amplexus. Figure 10. Developing tadpoles. Figure 9. Developing eggs.

51August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareHow to breed and raise African Clawed-Toed frogs (Xenopus laevis)Tadpole feeding– Add small amounts of spirulina or sera Micron suspended in demineralised water to the tanks, so the water turns a very light green colour.– If the water becomes cloudy refrain from feeding until it is clear. Change 50% of the water.– As the tadpoles grow more food should be added. Protein (egg powder) can also be used to supplement their diet (sera Micron contains extra proteins).16 Tadpole feeding - Add small amounts of spirulina or Sera Micron suspended in demineralised water to the tanks, so the water turns a very light green colour. - If the water becomes cloudy refrain from feeding until it is clear. Change 50% of the water. - As the tadpoles grow more food should be added. Protein (egg powder) can also be used to supplement their diet. Sera Micron contains extra proteins. Aeration?? Aeration (using a pump and air stones) helps maintain water quality, but it often requires a difficult and elaborate set up to make sure the bubbling is neither too fast nor too weak. Abstract/IntroduconAdvantages and disadvantages of different model organisms. Xenopusare widely used to study vertebrate embryology and development, basic cell and molecular biology, genomics, neurobiology and toxicology and to model human diseases (Xenbase.org, 2022).Category: C. elegans Drosophila Zebrafish Xenopus Chicken MouseBroodsize 250-300 80-100 100-200 500-3000+ 1 5-8Cost per embryo low low low low medium highHigh-throughput multiwell-format screening good good good good poor poorAccess to embryos good good good good poor poorMicro-manipulation of embryos limited limited fair good good poorGenome known known known known known knownGenetics good good good fair none goodKnockdowns (RNAi, morpholinos) good good good good limited limitedTransgenesis good good good good poor goodEvolutionary distance to human very distant very distant distant intermediate intermediate closeColour code: green, best in category; red, worst in category. Taken from Xenbase.org, adapted fromWheeler & Brändli 2009 Dev Dyn 238:1287-1308Superovulation Equipment• MS222• 1ml syringe and 27 or 26 gauge needle• L15 (leibrovits without Glutamine) Sigma L5520• Penicillin and streptomycin• Adult male xenopus (testes)• Forceps, scissors• Human Chorionic Gonadotropin hCG (Sigma)• Adult female xenopus• 1XMMR (Marc's Modified Ringer solution• 4ml pipette, turkey baster, 5ml petri dishes• Holding tanks, 1L beakers, tadpole rearing tanks• Spirulina( Sera micron, ZM Spirulina etc) protein powder, small bite froglet foodTherese Jones-Green, Chief Aquatic Technician/Named Animal Care and Welfare Ocer, University of CambridgeHow to Breed and Raise African Clawed-Toed Frogs(Xenopus laevis)Superovulation Eggs• Inject 150ul of PMSG (Intervet) into the female frog’s dorsal lymph sac a week before eggs are needed• The day before eggs are required inject 400ul of hCG into the dorsal lymph sac. Frogs will begin laying 8 hours later.• The next day dispose of any laid eggs as these will not be viable.• Place the female into a small holding tank containing 5L of 1XMMR (High salt/egg laying solution)• Allow the frog a few hours to lay the desired number of eggs you wish to fertilise.• Collect eggs using a turkey baster and place into 5ml petri dishes ready for fertilisation• Return frog to fresh system water once finishedEggs to avoid• Try to select eggs that are laid singular and round. Colour is not important and depends on parent's pigmentation.• Eggs laid in strings, clumps, or as larger white spheres are no good.• Sometimes the frogs eat their eggs and then vomit, you will know if this has happened if there are large clumps of eggs. A barrier can be placed in the tank to discourage this behaviour.Testes: Non-RegulatedProcedure• Select an adult male with well-defined black nuptial pads.• Euthanise the xenopus using a schedule 1 method.• Make an incision across the frog's waist. With forceps, pull out the yellow fat strands. The testes will be attached to the fat towards the back of the frog. Carefully cut the testes away from the connective tissues.• Place testes into a petri- dish with L15 and 20ul of penicillin and streptomycin. This is to stop the sperm activating and prevent unwanted growth of bacteria.• Store in the fridge for up to a week.In-vitro Fertilisation• Cut a small amount of teste off (2 mm³)• Using a pipette remove as much 1XMMR as possible.• Using the forceps pull the fragment of cut teste apart on the dish containing the dry eggs• Pick up the testes with forceps and waft the teste over each individual egg, then leave the fragments of testes sitting with the eggs• Wait 5 minutesIn-vitroFertilisation cont..ab• Flood the eggs and testes with a low salt solution such as 0.1XMMR or mineral water (a). Do not usede-mineralised water, as a low salt concentration is required.• The eggs and sperm will now fertilise. This process takes 10-20minutes. You know if it's successful as the eggs will rotate so that the animal pole is facing upwards (b)• Eggs can be left on the bench at 22° Celsius or put in an incubator to either slow down or speed up development.Natural Mating• To preserve your male stock, inject a male with 50µl of hCG two days before mating. Use the same steps for superovulation for the female.• Put the male and female together in a holding tank of their system’s water and leave them in a quiet dark place for two days (make sure they still get a discrete daily health check).• After the two days, many eggs will have been laid and fertilised. These will be at different stages of development.• Remove the male and female.• Drain off excess water and add fresh system water.• Keep the water clean, pipette out any dead eggs.• After 3 or 4 days the eggs will hatch, and feeding can begin.Caring forfertilised eggs• Remove dead eggs/embryos daily and change the media if it gets cloudy (a).• Top up water if evaporation has occurred.• Make sure the eggs are not too crowded, as dead eggs will infect healthy eggs (b). 30 eggs in a 70mm petri dish works well.• Eggs should hatch 2/3 days post fertilisation at room temperature (22C), development can be sped up or slowed down dependant on the temperature they are kept at.abCaringfor tadpoles• The first couple of weeks are the most sensitive.• Remove dead tadpoles using a pipette.• As the tadpoles increase in size, feed more food and increase the size of their holding container.• The water should have a slightly sweet aroma, if it starts to smell foul, change 75% of the water. Sometimes a whole water change will be needed. Knowing when and how much water to change is a fine balance.Tadpolefeeding• Add small amounts of spirulina or Sera Micron suspended in demineralised water to the tanks, so the water turns a very light green colour.• If the water becomes cloudy refrain from feeding until it is clear. Change 50% of the water.• As the tadpoles grow more food should be added. Protein (egg powder) can also be used to supplement their diet. Sera Micron contains extra proteins.Aeration?Aeration (using a pump and air stones) helps maintain water quality, but it often requires a difficult and elaborate set up to make sure the bubbling is neither too fast nor too weak.Natural Mating versus SuperovulationNatural Mating SuperovulationPreserve male stockPreciseAllow animals to exhibit natural behaviour Can use left-over testes and eggs from experimentsTime efficient Easier to keep clean post fertilisationGet many embryos at different stages of developmentBatches can unexpectantly crash (die)No consumables cost Hormones needed to induce ovulationCan be difficult to know if fertilisation was successfulEasier to know if fertilisation has been successfulSick tadpoles and thingsto look out for• Some tadpoles have crooked tails, this is usually Okay. Feed them more spirulina.• Some tadpoles have developmental defects (left). They should be euthanised with a Schedule 1 method.• If a tadpole is corkscrewing (spinning) remove and euthanise as above.• If many tadpoles are floating, change their water, they may need more oxygen.• If a single tadpole is floating or sitting on bottom of the tank, and unresponsive (but alive) remove and euthanise Sometimes after a water change some tadpoles turn upside down. Don't panic, they usually sort themselves out after a couple of hours.Healthy tadpolesUnhealthy tadpolesUnhealthy tadpolesConclusionThis poster is aimed at animal technicians as a tool to support in-house breeding. Some parts may need adapting depending on local water conditions, type of aquatic system used, and availability of resources. Please feel free to contact me with any ideas or questions related to in-house breeding: tj222@cam.ac.ukAcknowledgementsThe Gurdon InstituteCambridge University Biomedical ServicesBackground to the Home Office designated bio-facility• Opened in 2005, the bio-facility can hold up to 900 female and 240 male frogs in a Marine Biotech (now serviced by MBK Installations Ltd.) recirculating aquatic system.• The system monitors parameters such as: temperature, pH, conductivity, and Total Gas Pressure. • Frogs are maintained on mains fed water with a 20% water exchange. • There are several filtration steps, including: biological, mechanical, activated carbon, and UV sterilization.• Frogs are held under a 12-hour light/dark cycle.• Each tank has a PVC enrichment tube (30cm x 10cm), with smoothed cut edges.• Frogs are fed three times a week with a commercial trout pellet. • The focus of the research undertaken in the bio-facility is embryonic and developmental biology. All procedures are conducted under Home Office licence authority.• Frogs are euthanized humanely using a Schedule 1 methodFroglet care• Xenopus can take anywhere between 6 weeks to 6 months to metamorphize. It may be an evolutionary advantage to the species to have it spaced out this way.• You must remove froglets from the tank, as they will eat smaller tadpoles.• Froglets can be fed on 2mm trout pellets (manufactured by Skretting), Baby Reptomin, or large bite Zebrafish food• Many froglets can be kept together but make sure they are matched in size as larger frogs will attack smaller during feeding• Froglets like plastic plants and small tubes• Very easy to raise• Reach maturity at around 2 years of age and can live to over 30 years!References• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownReferences• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownReferences• Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it• Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308• https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022• Photography- My ownUniversity Biomedical ServicesAerationAeration (using a pump and air stones) helps maintain water quality but it often requires a diffi cult and elaborateset up to make sure the bubbling is neither too fast nor too weak.Sick tadpoles and things to look out for–Some tadpoles have crooked tails, this is usually not detrimental to them. Feed them more spirulina. – Some tadpoles have developmental defects (below). They should be euthanised with a Schedule 1 method.– If a tadpole is corkscrewing (spinning) remove and euthanise as above.– If many tadpoles are fl oating, change their water, they may need more oxygen.– If a single tadpole is fl oating or sitting on bottom ofthe tank and unresponsive (but alive) remove and euthanise. Sometimes after a water change some tadpoles turn upside down. Do not panic, they usuallysort themselves out after a couple of hours.17 Sick tadpoles and things too look out for - Some tadpoles have crooked tails, this is usually okay. Feed them more spirulina. - Some tadpoles have developmental defects (left).They should be euthanised with a Schedule 1 method. - If a tadpole is corkscrewing (spinning) remove and euthanise as above. - If many tadpoles are floating, change their water, they may need more oxygen. - If a single tadpole is floating or sitting on bottom of the tank, and unresponsive (but alive) remove and euthanise. Sometimes after a water change some tadpoles turn upside down. Do not panic, they usually sort themselves out after a couple of hours. 18 Froglet care - Xenopus can take anywhere between 6 weeks to 6 months to metamorphise. It may be an evolutionary advantage to the species to have it spaced out this way. - You must remove froglets from the tank, as they will eat smaller tadpoles. 18 Froglet care - Xenopus can take anywhere between 6 weeks to 6 months to metamorphise. It may be an evolutionary advantage to the species to have it spaced out this way. - You must remove froglets from the tank, as they will eat smaller tadpoles. Figure 11. Tadpoles being fed. Figure 12. Aeration system. Figure 13. Tadpoles showing signs of stress, etc.

52Animal Technology and Welfare August 2020Froglet care– Xenopus can take anywhere between 6 weeks to 6 months to metamorphise. It may be an evolutionary advantage to the species to have it spaced out this way.– You must remove froglets from the tank, as they will eat smaller tadpoles.– Froglets can be fed on 2mm trout pellets (manufactured by Skretting), Baby Reptomin, or large bite Zebrafish food.– Many froglets can be kept together but make sure they are matched in size as larger frogs will attack smaller during feeding.– Froglets like plastic plants and small tubes.– Very easy to raise.– Reach maturity at around 2 years of age and can live to over 30 years!Natural mating versus Superovulation Natural mating SuperovulationPreserve male stock PreciseAllow animals to exhibit natural behaviourCan use left over testes and eggs from experimentsTime efficient Easier to keep clean post fertilisationGet many embryos at different stages of developmentBatches can unexpectantly crash (die)No consumables cost Hormones needed to induce ovulationCan be difficult to know if fertilisation was successfulEasier to know if fertilisation has been successfulBackground to the Home Office designated bio-facility– Opened in 2005, the bio-facility can hold up to 900 female and 240 male frogs in a Marine Biotech (now serviced by MBK Installations Ltd) recirculating aquatic system.– The system monitors parameters such as: temperature, pH, conductivity and Total Gas Pressure. – Frogs are maintained on mains fed water with a 20% water exchange. – There are several filtration steps, including: biological, mechanical, activated carbon and UV sterilisation.– Frogs are held under a 12-hour light/dark cycle.– Each tank has a PVC enrichment tube (30cm x 10cm) with smoothed cut edges.– Frogs are fed three times a week with commercial trout pellets. – The focus of the research undertaken in the bio-facility is embryonic and developmental biology. All procedures are conducted under Home Office licence authority.– Frogs are euthanised humanely using a Schedule 1 method.Conclusion This poster is aimed at Animal Technicians as a tool to support in-house breeding. Some parts may need adapting depending on local water conditions, type of aquatic system used and availability of resources. Please feel free to contact me with any ideas or questions related to in-house breeding: tj222@cam.ac.ukAcknowledgements The Gurdon Institute Cambridge University Biomedical Services. References1 Xenbase: Facilitating the use of Xenopus to model human disease. www.frontiersin.org/articles/ 10.3389/fphys.20/92 Xenopus laevis Anatomy Poster. Gheorghe M. Constantinescu, www.aalas.org and www.tecniplast.it3 Wheeler and Brändli 2009, Developmental Dynamics, 238:1287-1308.4 https://www.xenbase.org/anatomy/intro.do accessed on 16/2/2022. Photography – author’s own.How to breed and raise African Clawed-Toed frogs (Xenopus laevis)

53August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareApril 2023 Animal Technology and WelfareAbstractMany diseases of Xenopus laevis frogs have been described in the literature however, it is often difficult to find images which clearly illustrate associated clinical signs. This poster has been designed as a refinement to bridge this gap using photographs taken of frogs with clinical signs.My aims are:– To share my experiences, via images, to help improve the welfare and husbandry of this species.– To stimulate a dialogue which results in further sharing of ideas and information.IntroductionBetween January 2016 and October 2018, cases of ill health and disease in a colony of Xenopus laevis frogs housed by the University Biological Services (UBS) at Cambridge University, have been documented via paper records and photographs. Over this 2-year and 10 months period, there were over 2000 adult frogs housed in this facility, of which 1940 (97%) have shown no clinical signs of disease or other abnormality. However, a number of incidents such as: Gas Bubble Disease, an infestation with nematodes and a suspected adverse reaction to a batch of HCG hormone, have all contributed to our usual low number of clinical cases.Background to the Home Office designated Bio-facility– Opened in 2005, the Bio-facility can hold up to 900 female and 240 male frogs in a Marine Biotech (now serviced by MBK Installations Ltd) recirculating aquatic system. – The system monitors parameters such as: temperature, pH, conductivity and Total Gas Pressure. – Frogs are maintained on mains fed water with a 20% water exchange. – There are several filtration steps, including: biological, mechanical, activated carbon and UV sterilisation. – Frogs are held under a 12-hour light/dark cycle.– Each tank has a PVC enrichment tube (30cm x 10cm) with smoothed cut edges.– Frogs are fed three times a week with a commercial trout pellet. – The focus of the research undertaken in the Bio-facility is embryonic and developmental biology. All procedures are conducted under Animal (Scientific Procedures) 1986 (ASPA) licence authority, issued by the UK Home Office. – Frogs are euthanised humanely using an ASPA prescribed Schedule 1 method.African Clawed Frog (Xenopus laevis ) diseases and clinical signs in photographsTHERESE JONES-GREEN University of CambridgeCorrespondence: tj222@cam.ac.uk Based on a poster displayed at IAT Congress 22

54Animal Technology and Welfare August 2020Reported clinical signs associated with ill health and disease● swimming in circles, tilting or loss of righting refl ex●buoyancy problems●changes in activity e.g. lethargy or easily startled●unusual amount of skin shedding●weight loss or failure to feed correctly●coelomic (body cavity) distention ●whole body bloat●fungal strands/tufts●sores or tremors at extremities (forearms and toes)●redness or red spots●excessive slime/mucous production or not enough (dry dull skin)●bite wounds, open cuts, lesions, abrasions of the skinor ulcerations●cloudy corneas●prolapsed cloaca(Reed 2005, Green 2010)1,201020304050607080Oedema Red leg Red leg (badhormone)Red leg(nematodes)Gas bubblediseaseSores on forearm Opaque cornea SkeletaldeformityGrowth (cartilageunder skin)Scar Cabletie Missing claws EggboundNumber of casesClinical signsClinical signs found between 2016 to 2018201820172016Attributes of a healthy Xenopus frogAlert and calmClear eyesMuscular hind legsAll claws intactPear shaped with smooth, clear, slimy skinThe dorsal pattern should contine over the frog’s fore legs and tops of the toes. The abdomen should be a pearly solid paler colour, although freckles and spots are common5 Clinical signs found between 2016 and 2018 5 Clinical signs found between 2016 and 2018 Gas bubble disease6 Gas bubble disease Figure 1. Xenopus frog exhibiting signs of Gas Bubble disease Description:: Frogs float horizontally and develop redness in the webbing of their feet which may extent over the toes, hind legs and back. Cause: Water that has been supersaturated with gasses. Figure 1.Xenopus frog exhibiting signs of Gas Bubble disease. Description:Frogs fl oat horizontally and develop redness in the webbing of their feet which may extend over the toes, hind legs and back. Cause:Water that has been super-saturated with gasses. Treatment:Shut down the system. Find andrepair crack(s) and allow time forthe gasses to disperse. Frogs that show clinical signs should be removed and closely monitored until the system is repaired and the gasses dispersed. Frogs that have severe clinical signs (unresponsive and/or redness over more that 25% of their body) should be euthanised.African Clawed frog (Xenopus laevis) diseases and clinical signs in photographsClinical signs found between 2016 and 2018

55August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAfrican Clawed frog (Xenopus laevis) diseases and clinical signs in photographsPersonal experienceThis happened at my facility when a small crack developedin the pipework attached to the pump allowing air to be drawn in and then pressurised. The fi rst clinical sign noticed was frogs fl oating abnormally at the top of their tanks (approximately 10% of the colony). On closer inspection redness could be seen in the webbing of their feet. The system was immediately shut down. From this point onwards, 99% of the frogs made a rapid and excellent recovery. Those that did not were euthanised using a Schedule 1 method. Recovering frogs that displayed clinical signs such as redness on their hind legs, feet or abdomens were taken out of the re-circulating system and housed in a static tank and closely monitored. Water was changed daily. Most frogs made a full recovery within days and were returned to their home tanks once the crack in the pipe was repaired. Recovery in photographs The photographs below clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs signifi cantly improved.8 Recovery in photographs The photographs below clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved.3 days after exposure7 days after exposure14 days after exposure. Figure 2. 3 days after exposure Figure 2.3 days after exposure.Red Leg disease (general)Description:Clinical signs include: redness of the hind legs, abdomen and/or red spotting that may extend to the frog’s back, thickening of the skin and/or swelling of limbs.Cause: There are many types of Red Leg, the way it presents can be attributed to different types of infectious organisms (Aeromonas, Pseudomonas, Mycobacterium, Chryseobacterium, etc). They may be carried concurrently(Hill et al 2010) and usually lie dormant in the frog but can cause disease when frogs are exposed to a stressful event.3 If secondary fungal infection occurs this can confound the clinical signs.Treatment: Red Leg can be a complicated condition to treat especially where there is an underlying systemic disease (Green 2010).2 Salt baths have been found to help milder, earlycases.9 Figure 3. 7 days after exposure Figure 4 . 14 days after exposure. Figure 3.7 days after exposure.9 Figure 3. 7 days after exposure Figure 4 . 14 days after exposure. Figure 4.14 days after exposure.11 Figures 5 and 6. Xenopus afflicted with Red Leg. Figure 5.Xenopus affl icted with Red Leg.

56Animal Technology and Welfare August 2020Red Leg: Aeromonas hydrophilaDescription: The photographs clearly demonstrate red lesions on the underside of affected frogs. Diagnosis: Skin swabs taken from these lesions confi rmed the causal agent was the bacterium Aeromonas hydrophila. In addition the bacterial cultures identifi ed Aeromonas hydrophilia and Citrobacter freundiiand Comamonas spp.Treatment: These frogs were euthanised. Their tanks were thoroughly cleaned and left to completely dry.Red Leg: after use of new hormone batchDescription: Red patches developed on super ovulated frog’s fl anks and at the tops of their hind legs either during laying eggs or a few days later. Diagnosis:These frogs were swabbed and bacterial cultures isolated both Aeromonas spp. and Pseudomonas spp. Treatment: After ruling out husbandry practices, it was thought thatthe change to a new veterinary approved brand of hormonewas the likely stressor. Upon returning to the old brand, no more cases were found. Frogs showing clinical signswere monitored where most made an excellent recovery.11 Figures 5 and 6. Xenopus afflicted with Red Leg. Figure 6.Xenopus affl icted with Red Leg.12 Red Leg: Aeromonas hydrophila Description: The photographs clearly demonstrate red lesions on the underside of affected frogs. Diagnosis: Skin swabs taken from these lesions confirmed the causal agent was the bacterium Aeromonas hydrophila. In addition the bacterial cultures identified Aeromonas hydrophilia and Citrobacter freundiiand Comamonas spp. Treatment: These frogs were euthanised. Their tanks were thoroughly cleaned and left to completely dry. Figure 7 and 8. Xenopus infected with Aeromonas hydrophila.13 Figure 7 and 8. Xenopus infected with Aeromonas hydrophila. Red Leg: after use of new hormone batch Description: Red patches developed on super ovulated frog’s flanks and at the tops of their hind legs either during laying eggs or a few days later. Diagnosis: These frogs were swabbed and bacterial cultures isolated both Aeromonasspp. and Pseudomonasspp. Treatment: After ruling out husbandry practices, it was thought that the change to a new veterinary approved brand of hormone was the likely stressor. Upon returning to the old brand, no more cases were found. Frogs showing clinical signs were monitored where most made an excellent recovery. 14 Figure 9 Figures 9 and 10. Xenopus showing signs of Red Leg following use of a different brand of hormone. Figure 9 and 10. Xenopus showing signs of Red Leg following use of a different brand of hormone.14 Figure 9 Figures 9 and 10. Xenopus showing signs of Red Leg following use of a different brand of hormone. Foreleg clinical signs: Grey armsDescription:Forearms that normally show a continuation of the dorsalpattern are faded and appear ‘grey’ sometimes with white marks or open sores.African Clawed frog (Xenopus laevis) diseases and clinical signs in photographs

57August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAfrican Clawed frog (Xenopus laevis) diseases and clinical signs in photographsCause: Unknown but thought to be stress related and/or associated with bites occurring during feeding. Histopathology is often inconclusive.Treatment: Provide extra food. Separate (rehouse with another frog) and salt bath for those with wounds (sodium chloride 5g/L bath for 72 h). Also check/change UV bulbs.15 Foreleg clinical signs: Grey arms Description: Forearms that normally show a continuation of the dorsal pattern are faded and appears ‘grey’ sometimes with white marks or open sores. Cause: Unknown but thought to be stress related and/or associated with bites occurring during feeding. Histopathology is often inconclusive. Treatment: Provide extra food. Separate (rehouse with another frog) and salt bath for those with wounds (sodium chloride 5g/L bath for 72 h). Also check/change UV bulbs. Figure 11 and 12. Forelimbs showing greying and bite wounds.16 Figures 11 and 12. Forelimbs showing greying and bite wounds. Opaque corneas Description: One or both eyes appear cloudy. Cause: Unknown, but could be a sign of ill health (Green 2010), stress, nematodes, or a developmental abnormality. Treatment: None. The eye may decrease in size leaving a scar. This frog was still able to forage and behaved normally. Opaque corneasDescription: One or both eyes appear cloudy.Cause: Unknown but could be a sign of ill health (Green 2010),2stress, nematodes or a developmental abnormality.Treatment: None. The eye may decrease in size leaving a scar. This frog was still able to forage and behaved normally.17 Figure 13. Exenopus frog showing opaqueness of one cornea. Fluid filled cyst Description: Bulge on the underside of the foot, filled with a clear fluid. Figure 13. Xenopus frog showing opaqueness of one cornea.Fluid fi lled cystDescription: Bulge on the underside of the foot, fi lled with a clear fl uid.Cause: Unknown, although the cyst appeared benign. The frog was euthanised as the cyst appeared to increase in sizeduring the monitoring period. Granular scarringDescription: Strange irregular granular scarring was observed on a frog’s fl ank which did not heal.Cause: Unknown, perhaps a chemical burn caused by Chlorhexidinewhich is found in many cosmetic products (Philips et al2015).18 Cause: Unknown, although the cyst appeared benign. The frog was euthanised as the cyst appeared to increase in size during the monitoring period. Figure 14. Frog exhibiting a fluid filled cyst. Figure 14. Frog exhibiting a fl uid fi lled cyst.19 Granular scarring Description: Strange irregular granular scarring was observed on a frog’s flank which did not heal. Cause: Unknown, perhaps a chemical burn caused by Chlorhexidine which is found in many cosmetic products (Philips et al 2015). Treatment: None, the frog was euthanized as there was no improvement after monitoring. Histopathology was inconclusive. Figure 15. Signs of granular scarring Figure 15. Signs of granular scarring.

58Animal Technology and Welfare August 2020Treatment: None, the frog was euthanised as there was no improvement after monitoring. Histopathology was inconclusive.Frogs will eat anythingDescription: A frog was found with a sore on her back that would notheal.Cause: The frog had eaten a cable tie (we do not know when thishappened and we do not use this type of tie). The cable tie had passed through her digestive system down to her colon. The end of the tie had pierced through the colon and through the skin on her back.Treatment: Euthanise frog and record. Be extremely careful and do not leave debris around tanks. Keep small inedible items away from tanks and procedure areas. Keep track of everything you use.Nematodes: CapillariaxenopodisDescription: The nematodes inhabit the epidermis (skin) typically on the frog’s back that can then result in skin lesions (Green 2010, Reed 2006)2.1. Figure 17 shows lesions on the underside of this frog.Cause: Frogs show clinical signs when stressed. This frog was found positive for nematodes following laboratory tests.Treatment: Anthelmintic (Levamisole or Ivermectin). Thorough cleaningand drying of aquatic system.21 Figure 16. Ingested cable tie eaten by a frog. Nematodes: Capillariaxenopodis Description: The nematodes inhabit the epidermis (skin) typically on the frog’s back that can then result in skin lesions (Green 2010, Reed 2006)2.1. This photo shows lesions on the underside of this frog. Figure 16. Ingested cable tie eaten by a frog.Loss of hind clawsDescription: One or more claws are absent from the hind feet. Once afrog loses a claw the toe turns red or white over subsequent days. Claws do not regrow. Cause: Unknown, histology is often inconclusive. Perhaps viral orautoimmune.Treatment: Rehouse affected frogs and monitor. Drain tanks that occupied affected frogs and allow to air dry before re-fi lling. 22 Cause: Frogs show clinical signs when stressed. This frog was found positive for nematodes following laboratory tests. Treatment: Anthelmintic (Levamisole or Ivermectin). Thorough cleaning and drying of aquatic system. Figure 17. Clinical signs of nematode infections in a Xenopus frog. Figure 17. Clinical signs of nematode infections in a Xenopus frog.23 Loss of hind claws Description: One or more claws are absent from the hind feet. Once a frog loses a claw the toe turns red or white over subsequent days. Claws do not regrow. Cause: Unknown, histology is often inconclusive. Perhaps viral or autoimmune. Treatment: Rehouse affected frogs and monitor. Drain tanks that occupied affected frogs and allow to air dry before re-filling. Figure 18. Frog with missing claw. Figure 18. Frog with missing claw.African Clawed frog (Xenopus laevis) diseases and clinical signs in photographs

59August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAfrican Clawed frog (Xenopus laevis) diseases and clinical signs in photographsSkeletal deformityDescription: As froglets grow to adult size a small percentage (< 1%) often have skeletal deformities. The spine curves either upwards (as illustrated in Figure 19) or may bend to the left or right. Cause: Unknown but could be a genetic mutation or damage caused from the frog’s environment when they were a small froglet or tadpole.Treatment: None. If the deformity is impacting on the frog’s welfare, causing diffi culty in swimming, feeding or any other behaviours, then the frog should be euthanised. Frogs with mild skeletal deformities still produce good oocytes. 24 Skeletal deformity Description: As froglets grow to adult size a small percentage (< 1%) often have skeletal deformities. The spine curves either upwards (as illustrated) or may bend to the left or right. Cause: Unknown but could be a genetic mutation or damage caused from the frog’s environment when they were a small froglet or tadpole. Treatment: None. If the deformity is impacting on the frog’s welfare, causing difficulty in swimming, feeding or any other behaviours, then the frog should be euthanised. Frogs with mild skeletal deformities still produce good oocytes. Figure 19. Frog with skeletal deformity – hind leg absent. Figure 19. Frog with skeletal deformity – hind leg absent. Egg boundDescription: The frog may lay few or no eggs after superovulation. The next 1-2 days, the ‘non-layer’ has a bloated appearance and feels fi rm. The frog may be less responsive.Cause: Eggs get blocked in the oviduct which can lead to a build up of bacteria inside the frog which, if left untreated, will result in the death of the frog.Treatment: Close monitoring of ‘non layers’, then immediate euthanasia as soon as egg retention is recognised. Some frogs fail to lay due to other reasons. Orange blotchesDescription: Irregular orange/red marks appear on the abdomen.Cause: Change in water quality. In this case the cause was dueto a sudden switch from mains water to Reverse Osmosis(RO) water.Treatment: Put source back to mains water supply. Note – when changing water quality/source, this should be done slowly25 Egg bound Description: The frog may lay few or no eggs after superovulation. The next 1-2 days, the ‘non-layer’ has a bloated appearance and feels firm. The frog may be less responsive. Cause: Eggs get blocked in the oviduct which can lead to a build up of bacteria inside the frog which, if left untreated, will result in the death of the frog. Treatment: Close monitoring of ‘non layers’, then immediate euthanasia as soon as egg retention is recognised. Some frogs fail to lay due to other reasons. Figure 20. Egg bound xenopus frog. Figure 20.Egg bound Xenopus frog. 26 Orange blotches Description: Irregular orange/red marks appear on the abdomen. Cause: Change in water quality. In this case the cause was due to a sudden switch from mains water to Reverse Osmosis (RO) water. Treatment: Put source back to mains water supply. Note, when changing water quality/source, this should be done slowly with the frogs closely monitored. Most facilities in the UK use mains water as it is well suited to xenopusdue due to its ‘hardness’ (Water hardness is defined as the measured content of Calcium (Ca++) and magnesium (Mg++). Figure 21. Frog showing orange blotches on underside. Figure 21.Frog showing orange blotches on underside.

60Animal Technology and Welfare August 2020with the frogs closely monitored. Most facilities in the UKuse mains water as it is well suited to xenopus due toits ‘hardness’ (water hardness is defi ned as the measuredcontent of Calcium (Ca++) and Magnesium (Mg++).Wounds affecting males Description: Red sores may occur on abdomen of male frogs.Cause: Male gripping onto another male during amplexus (mating).This is common in male frogs housed together.Treatment: Sores should heal in a couple of days (lesions should change from red to white, then decrease in size until they are healed). Separate and isolate affected animal with a different male. Salt treatment if required, althoughthis can add further stress to the frog.Wound affecting males (2)Description: This male frog was found to have lines all over his body (Note: The redness on the right-hand side was due to MS222 anaesthetic agent).Cause: This frog was probably trying to mate with another male who defensively kicked with his hind claws to get away or out of amplexus.Treatment: Lines healed quickly. Isolate with another male and salt treatment if required, although this can add further stress to the frog.27 Wounds affecting males Description: Red sores may occur on abdomen of male frogs. Cause: Male gripping onto another male during amplexus (mating). This is common in male frogs housed together. Treatment: Sores should heal in a couple of days (lesions should change from red to white, then decrease in size until they are healed). Separate and isolate affected animal with a different male. Salt treatment if required, although this can add further stress to the frog. Figure 22. Male frog exhibiting wounds. Figure 22.Male frog exhibiting wounds.Conclusion By sharing this photography I hope to expand and refi ne the material available to those working with Xenopusfrogs. These cases were recorded over a period of 2 years and 10 months and in some the cause was anecdotal, based on our experience at that time.If you work with Xenopus spp. and have experienced similar or different cases, or have any ideas or opinions on the cause and treatment, then please get in touch: tj222@cam.ac.uk it would be great to hear from you!Acknowledgements Cambridge University Biomedical Services and The GurdonInstitute.References1Reed. B.T. (2005). R.S.P.C.A Guidance on the housingand care of African clawed frog Xenopus laevis. Research Animals Department-RSPCA.2Green, S.L. (2010). ‘The Laboratory Xenopus sp.’ CRS Press. The Laboratory Xenopus sp., 2010 - Book – Xenbase. XB-BOOK-222. 2010; Boca Raton, FL USA:Taylor & Francis Group. The Laboratory Xenopus sp. Green, Sherill L (Author). ISBN: 1420091093.28 Wound affecting males (2) Description: This male frog was found to have lines all over his body (Note: The redness on the right-hand side was due to MS222 anaesthetic agent). Cause: This frog was probably trying to mate with another male who defensively kicked with his hind claws to get away or out of amplexus. Treatment: Lines healed quickly. Isolation with another male and salt treatment if required, although this can add further stress to the frog. Figure 24. Further of examples incurred by male frogs. Figure 23.Further examples incurred by male frogs.African Clawed frog (Xenopus laevis) diseases and clinical signs in photographs

61August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAfrican Clawed frog (Xenopus laevis) diseases and clinical signs in photographs3Hill, A.H., Newman, S.J., Craig, L., Carter, C., Czarra, J.,Brown, J.P. (2010). Diagnosis of Aeromonas hydrophila,Mycobacterium species, and Batrachochytrium dendrobatidis in an African Clawed Frog (Xenopus laevis). J.Am.Assoc.Lab.Anim.Sci. 49(2): 215–220.Additonal sources Densmore, C.L., Green, D.E. (2007). Diseases ofAmphibia. ILAR Journal, 48(3): 235–254. Philips, B.H., Crim, M.J., Hankenson, F.C., Steffan, E.K.,Klein, P.S., Brice, A.K., Carty, A.J. (2015). Evaluationof Presurgical Skin Preparation Agent in African Clawed Frogs (Xenopus laevis). J.Am.Assoc.Lab.Anim.Sci.54(6): 788-798.

It’s OK … NOT TO BE OKAY …IAT – Virtual Network Support ForumHas it not been your day, your week, your month or even your year? Or just fancy a catch up with friendly people?Then drop in to say a quick ‘hello’ or for longer, if you have time for a chat the IAT will be there for you! Every Thursday at 12.30 join us on zoom: CLICK TO JOINhttps://tinyurl.com/5ax86u9tMental Health Awarenesswww.iat.org.ukInstitute of Animal TechnologyCOUNCILEDI GroupEquity, Diversity and Inclusion

63August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareValidating in-cage mouse enrichmentCHELSEA BURBIDGE, ZOE BERESFORD and VALENTINA SERRANO-GALLEG University of BirminghamCorrespondence: Z.beresford@bham.ac.uk, c.burbidge@bham.ac.ukPOSTER PRESENTATIONSOriginally presented at:IAT Congress 2022151IntroductionA 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, ankle or footpad width isa commonly used surrogate marke r 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 measure 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 151Abstract There is a wide variety of different types of mouse enrichment currently available, some items of which are more practical than others. Validating items used for providing enrichment can be time consuming and costly, nevertheless it is important to ensure that anyobjects being provided for this purpose are truly offering enrichment opportunities for the animals. The purpose of this study was to demonstrate how a quick and simplecage side assessment can be undertaken to identify which items are most effective as enrichments and whether they offer short- or long-term solutions. The studycompared four different types of enrichment, all of whichare easily available, in fi ve cages of pair housed 6-month-old CD1 mice. Through this approach, we found that the mice preferred items suspended from the cage lid. Using this information, we have identifi ed a practical and cost-effective solution and confi rmed that it maintains novelty over longer periods. This validated enrichment is now being used routinely in the Biomedical Services Unit at the University of Birmingham.Hol-ee Roller BallsValidating in-cage mouse enrichmentAbstract: There is a wide variety of different types of mouse enrichment currently available, some items of which are more practical than others. Validating items as providing enrichment can be time consuming and costly, neverthelessit is important to ensure that any objects being provided for this purpose are truly offering enrichment opportunities for the animals. The purpose of this study was to demonstrate how a quick and simple cage side assessment can beundertaken to identify which items are most effective as enrichments and whether they offer short- or long-term solutions. The study compared four different types of enrichment, all of which are easily available, in five cages of pairhoused 6-month-old CD1 mice. Through this approach, we found that the mice preferred items suspended from the cage lid. Using this information, we have identified a practical and cost effective solution, and confirmed that itmaintains novelty over longer periods. This validated enrichment is now being used routinely in the Biomedical Services Unit at the University of Birmingham.Chelsea Burbidge, Zoe Beresford, Valentina Serrano-Galleg, Biomedical Services Unit, University of BirminghamResources:• Time: It takes a minimum of 30 minutes to set it up, including sanitising, drying, stuffing the ball with new nesting material and then putting it into the cage.• Costs: balls are relatively expensive, require frequent replacement, nesting inside is cheap and easy to replace.• Maintenance: sanitise before and after use, no autoclave due to material. It requires someone to take out the little pieces of chewed material from the cages daily to reduce possible chokage.Observations: Instant attraction, did not lose interest. Showed interest in pulling enrichment out of it as well as the ball itself. The mice excessively chewed the blue and green balls for unknown reasons. Provides different enrichment of interest outside of this study.Pros:• Very high degree of interest seen in all animals.• Prolonged enrichment.• Can use them in different ways.Cons: • Time consuming to put the nesting in.• Expensive as balls need replacing frequently.• Cleaning takes time as has to be done by hand.• Enrichment ends once nesting removed.Hol-ee Roller BallsScenario: We stuffed each ball with nesting material, in addition to that provided within the cage and suspended the ball from the cage lid. Each morning we scored use of the balls from 0 to 3, 0 being the nesting material was notdisturbed, 1 being the nesting material was disturbed but not removed, 2 being that some nesting material was removed but not entirely, and 3 being that all nesting material was removed from the ball. Each cage was scored eachand the nesting replenished.Resources:• Time: Easy and fast to put into cage after autoclaving by clicking into place on the wire lid. • Costs: Relatively expensive, do not require frequent replacement.• Maintenance: Sanitise in autoclave. Replaced during routine cage clean, or more often if dirty. Observations: Interest varied between animals, with some becoming more interested over time, and some less. No animal was routienly observed using the wheels, generally use was assumed based on movement of the sticker. Where wheels have been provided to other animals outside of the study, young animals appear to have a preference for them.Pros: • Provides an opportunity for exercise.• Easily cleaned and can be autoclaved.• Cost effective over the longer term.Cons:• High degree of variability between animals, so not a universal enrichment for all.• Hard to monitor use to confirm if providing enrichment for individual animals.• Wheel uses a lot of cage space.WheelsScenario: We attached a wheel to the cage lid and applied a sticker to the top of the wheel itself. We observed the cage each morning for 15 minutes and then checked the wheel for the position of the sticker. We scored use of thewheels from 0 to 2, 0 being no evidence of use, 1 being we hadn’t observed use but the sticker had moved indicating interaction, and 2 being they had used it during the observation period.Resources: • Time: Easy and fast to add to each cage.• Costs: Very cheap, need replacing often• Maintenance: Sanitise in autoclave before use. Need to regularly remove broken pieces of chew sticks from cage. Replace weekly in some cages. Observations: Immediate interest but this lessens over time. Follow-up observations indicate that novelty restored after removed for a week. Interest varied between individual animals. Pros: • Helps prevent overgrown teeth.• Low maintenance.• Easily autoclaved and stored.• Do not take up much room in the cage.• Easy to judge if providing enrichment on a case-by-case basis.Cons:• Need to remove splinters and monitor for blocks becoming too small.• Enrichment lost as item consumed.• Interest decreases over time so needs to be rotated with other enrichment.Aspen chew blocksScenario: At the start of each week we removed any old chew blocks and replaced with new (one per mouse in the cage). We weighed each chew stick before and after it was added to the cage to determine how much they had beenused over time. The graph shows the average amount of chew stick (grams) consumed each week per cage.Resources:• Time: Easy and fast to add to each cage.• Costs: Very cheap, need replacing frequently• Maintenance: Sanitise in autoclave before use. Need to replace at least at cage change, if not more regularly to provide a constant source of enrichment. Observations: Immediate and maintained interest. Some variation between individuals but,overall, use was high, and has also been observed in other strains outside of the study. All animals incorporated the majority of the material into the nest.Pros:• Enrichment from both destroying the CellPad and incorporating into nest. • Novelty maintained over time.• Universal interest from all animals trialled.• Easily autoclaved and stored.• Provides additional insulation for litters.Cons:• Enrichment lost as item is consumed.• Have to limit number of CellPads provided between cage cleans as material accumulates and blocks view of animals.CellPadScenario: We attached a new cell pad to the cage lid at the start of the week and scored use each morning. CellPad was provided ina ddition to normal nesting material. We scored on two levels; percentage of CellPad destroyed, and ofthat, the percentage that had been incorportated into a nest. These percentage are plotted on the graph.Summary: From the study, we have come to the conclusion that all of the items trialled have their pros and cons, and it is important to validate items as providing enrichment, rather than assuming this.Whilst the mice showed a prolonged interest in the balls, this was not sustainable due to the high level of resources required. Based on this, we identified the CellPads as a possible alternative that offered a similar type ofenrichment. Both the cell pads and chew sticks were the most effective, due to their low resource requirements, ease of use, and interest shown by the animals. They were also the most universally suitable for use in differentscenarios.Further studies are required to determine whether novelty is retained over longer periods, or can be reinstated after removal for a period of time, or if there are certain sceanrios where wheels consistentlyroutinely offer aenrichment opportunity. However, based on the data from this study, as a facility we are now confident in our routine use of chew sticks as an enrichment item, and also now additionally use the CellPads where we requireadditional enrichment such as mice that are showing signs of aggression or are unavoidably singularly housed.Figure 1.Hole-ee ball. April 2023 Animal Technology and Welfare

64Animal Technology and Welfare August 2020Scenario: We fi lled each ball with nesting material, in addition to that provided within the cage and suspended the ball from the cage lid. Each morning we scored use of the balls from 0 to 3, 0 being the nesting material was not disturbed, 1 being the nesting material was disturbed but not removed, 2 being that some nesting material was removed but not entirely and, 3 being that all nesting material was removed from the ball. Each cage was scored each morning and the nesting replenished.Resources:– Time: It takes a minimum of 30 minutes to set it up, including sanitising, drying, stuffi ng the ball with new nesting material and then putting it into the cage.– Costs: balls are relatively expensive, require frequentreplacement, nesting inside is cheap and easy to replace.–Maintenance: sanitise before and after use, autoclavenot used due to manufacturing material. It requires someone to take out the little pieces of chewed material from the cages daily to reduce possible choking.Observations: Instant attraction, did not lose interest. Showed interest in pulling enrichment out of it as well as the ball itself. The mice excessively chewed the blue and green balls for unknown reasons. Provides different enrichment opportunities as can change the contents and position in the cage (fl oor vs lid). C57Bl6/J animals also showed high level of interest outside of this study.Pros:– Very high degree of interest seen in all animals.– Prolonged enrichment.– Can use them in different ways.Cons: – Time consuming to put the nesting in.– Expensive as balls need replacing frequently.– Cleaning takes time as has to be done by hand.Enrichment ends once nesting removed.Validating in-cage mouse enrichmentAbstract: There is a wide variety of different types of mouse enrichment currently available, some items of which are more practical than others. Validating items as providing enrichment can be time consuming and costly, neverthelessit is important to ensure that any objects being provided for this purpose are truly offering enrichment opportunities for the animals. The purpose of this study was to demonstrate how a quick and simple cage side assessment can beundertaken to identify which items are most effective as enrichments and whether they offer short- or long-term solutions. The study compared four different types of enrichment, all of which are easily available, in five cages of pairhoused 6-month-old CD1 mice. Through this approach, we found that the mice preferred items suspended from the cage lid. Using this information, we have identified a practical and cost effective solution, and confirmed that itmaintains novelty over longer periods. This validated enrichment is now being used routinely in the Biomedical Services Unit at the University of Birmingham.Chelsea Burbidge, Zoe Beresford, Valentina Serrano-Galleg, Biomedical Services Unit, University of BirminghamResources:• Time: It takes a minimum of 30 minutes to set it up, including sanitising, drying, stuffing the ball with new nesting material and then putting it into the cage.• Costs: balls are relatively expensive, require frequent replacement, nesting inside is cheap and easy to replace.• Maintenance: sanitise before and after use, no autoclave due to material. It requires someone to take out the little pieces of chewed material from the cages daily to reduce possible chokage.Observations: Instant attraction, did not lose interest. Showed interest in pulling enrichment out of it as well as the ball itself. The mice excessively chewed the blue and green balls for unknown reasons. Provides different enrichment of interest outside of this study.Pros:• Very high degree of interest seen in all animals.• Prolonged enrichment.• Can use them in different ways.Cons: • Time consuming to put the nesting in.• Expensive as balls need replacing frequently.• Cleaning takes time as has to be done by hand.• Enrichment ends once nesting removed.Hol-ee Roller BallsScenario: We stuffed each ball with nesting material, in addition to that provided within the cage and suspended the ball from the cage lid. Each morning we scored use of the balls from 0 to 3, 0 being the nesting material was notdisturbed, 1 being the nesting material was disturbed but not removed, 2 being that some nesting material was removed but not entirely, and 3 being that all nesting material was removed from the ball. Each cage was scored eachand the nesting replenished.Resources:• Time: Easy and fast to put into cage after autoclaving by clicking into place on the wire lid. • Costs: Relatively expensive, do not require frequent replacement.• Maintenance: Sanitise in autoclave. Replaced during routine cage clean, or more often if dirty. Observations: Interest varied between animals, with some becoming more interested over time, and some less. No animal was routienly observed using the wheels, generally use was assumed based on movement of the sticker. Where wheels have been provided to other animals outside of the study, young animals appear to have a preference for them.Pros: • Provides an opportunity for exercise.• Easily cleaned and can be autoclaved.• Cost effective over the longer term.Cons:• High degree of variability between animals, so not a universal enrichment for all.• Hard to monitor use to confirm if providing enrichment for individual animals.• Wheel uses a lot of cage space.WheelsScenario: We attached a wheel to the cage lid and applied a sticker to the top of the wheel itself. We observed the cage each morning for 15 minutes and then checked the wheel for the position of the sticker. We scored use of thewheels from 0 to 2, 0 being no evidence of use, 1 being we hadn’t observed use but the sticker had moved indicating interaction, and 2 being they had used it during the observation period.Resources: • Time: Easy and fast to add to each cage.• Costs: Very cheap, need replacing often• Maintenance: Sanitise in autoclave before use. Need to regularly remove broken pieces of chew sticks from cage. Replace weekly in some cages. Observations: Immediate interest but this lessens over time. Follow-up observations indicate that novelty restored after removed for a week. Interest varied between individual animals. Pros: • Helps prevent overgrown teeth.• Low maintenance.• Easily autoclaved and stored.• Do not take up much room in the cage.• Easy to judge if providing enrichment on a case-by-case basis.Cons:• Need to remove splinters and monitor for blocks becoming too small.• Enrichment lost as item consumed.• Interest decreases over time so needs to be rotated with other enrichment.Aspen chew blocksScenario: At the start of each week we removed any old chew blocks and replaced with new (one per mouse in the cage). We weighed each chew stick before and after it was added to the cage to determine how much they had beenused over time. The graph shows the average amount of chew stick (grams) consumed each week per cage.Resources:• Time: Easy and fast to add to each cage.• Costs: Very cheap, need replacing frequently• Maintenance: Sanitise in autoclave before use. Need to replace at least at cage change, if not more regularly to provide a constant source of enrichment. Observations: Immediate and maintained interest. Some variation between individuals but,overall, use was high, and has also been observed in other strains outside of the study. All animals incorporated the majority of the material into the nest.Pros:• Enrichment from both destroying the CellPad and incorporating into nest. • Novelty maintained over time.• Universal interest from all animals trialled.• Easily autoclaved and stored.• Provides additional insulation for litters.Cons:• Enrichment lost as item is consumed.• Have to limit number of CellPads provided between cage cleans as material accumulates and blocks view of animals.CellPadScenario: We attached a new cell pad to the cage lid at the start of the week and scored use each morning. CellPad was provided ina ddition to normal nesting material. We scored on two levels; percentage of CellPad destroyed, and ofthat, the percentage that had been incorportated into a nest. These percentage are plotted on the graph.Summary: From the study, we have come to the conclusion that all of the items trialled have their pros and cons, and it is important to validate items as providing enrichment, rather than assuming this.Whilst the mice showed a prolonged interest in the balls, this was not sustainable due to the high level of resources required. Based on this, we identified the CellPads as a possible alternative that offered a similar type ofenrichment. Both the cell pads and chew sticks were the most effective, due to their low resource requirements, ease of use, and interest shown by the animals. They were also the most universally suitable for use in differentscenarios.Further studies are required to determine whether novelty is retained over longer periods, or can be reinstated after removal for a period of time, or if there are certain sceanrios where wheels consistentlyroutinely offer aenrichment opportunity. However, based on the data from this study, as a facility we are now confident in our routine use of chew sticks as an enrichment item, and also now additionally use the CellPads where we requireadditional enrichment such as mice that are showing signs of aggression or are unavoidably singularly housed.Figure 2.Mouse in cage interacting with Hole-ee ball. 6 Wheels Figure 3. Running Wheel including attachment to the cage. Mouse 1 Mouse 2 Mouse 3 Mouse 4 Mouse 5Averaged per weekWheelsScenario: We attached a wheel to the cage lid and applied a sticker to the top of the wheel itself. We observed the cage each morning for 15 minutes and then checked the wheel for the position of the sticker. We scored use of the wheels from 0 to 2, 0 being no evidence of use, 1 being we had not observed use but the sticker had moved indicating interaction and 2 being they had used it during the observation period.6 Wheels Figure 3. Running Wheel including attachment to the cage. Figure 3. Running Wheel including attachment to the cage.Poster Presentations

65August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsResources:– Time: Easy and fast to put into cage after autoclaving by clicking into place on the wire lid. –Costs: Relatively expensive but do not require frequentreplacement.–Maintenance: Sanitised in autoclave. Replaced duringroutine cage clean, or more often if dirty. Observations: Interest varied between animals, with some becoming more interested over time and some less. No animal was routinely observed using the wheels, generally use was assumed based on movement of the sticker. Where wheels have been provided to other animals outside of the study, young animals appear to have a preference for them.Pros: – Provides an opportunity for exercise.– Easily cleaned and can be autoclaved.– Cost effective over the longer term.Cons:– High degree of variability between animals, so not a universal enrichment for all.– Hard to monitor use to confi rm if providing enrichment for individual animals.Wheel uses a lot of cage space.Aspen chew blocks8 Pros: • Provides an opportunity for exercise. • Easily cleaned and can be autoclaved. • Cost effective over the longer term. Cons: • High degree of variability between animals, so not a universal enrichment for all. • Hard to monitor use to confirm if providing enrichment for individual animals. Wheel uses a lot of cage space. Figure 4. Mouse in cage with wheel.9 Figure 4. Mouse in cage with wheel. Aspen chew blocks Figure 5. Aspen blocks – available sizes. Mouse 1 Mouse 2 Mouse 3 Mouse 4 Mouse 5Wheels averageScenario: At the start of each week, we removed any old chew blocks and replaced with new (one per mouse in the cage). We weighed each chew stick before and after it was added to the cage to determine how much they had been used over time (fi gure 7). The graph shows the average amount of chew stick (grams) consumed each week per cage. Resources: – Time: Easy and fast to add to each cage.– Costs: Very cheap, need replacing frequently.– Maintenance: Sanitise in autoclave before use. Needto regularly remove broken pieces of chew sticks fromcage. Replace weekly in some cages. Observations: Immediate interest but this lessens overtime. Follow-up observations indicate that novelty restoredafter removed for a week. Interest varied betweenindividual animals. 9 Figure 4. Mouse in cage with wheel. Aspen chew blocks Figure 5. Aspen blocks – available sizes. Figure 5. Aspen blocks – available sizes. 12 Figure 6. Mouse ‘enjoying’ an Aspen block. Figure 6. Mouse ‘enjoying’ an Aspen block.

66Animal Technology and Welfare August 2020Pros: – Helps prevent overgrown teeth.– Low maintenance.– Easily autoclaved and stored.– Do not take up much room in the cage.– Easy to judge if providing enrichment on a case-by-case basis.Cons:– Need to remove splinters and monitor for blocks becoming too small.– Enrichment lost as item consumed.Interest decreases over time so needs to be rotated withother enrichment.CellPadScenario: We attached a new cell pad to the cage lid at the start of the week and scored use each morning. CellPad was provided in addition to normal nesting material. We scored on two levels; percentage of CellPaddestroyed and, of that, the percentage that had been incorporated into a nest. These percentages are plotted on the graph.Resources:– Time: Easy and fast to add to each cage.– Costs: Very cheap, need replacing frequently.–Maintenance: Sanitise in autoclave before use. Needto replace at least at cage change, if not more regularlyto provide a constant source of enrichment. Observations: Immediate and maintained interest. Somevariation between individuals but overall, use was high and has also been observed in other strains outside of the study. All animals incorporated most of the material into the nest.Pros:– Enrichment from both destroying the CellPad and incorporating into nest. 13 CellPad Scenario: We attached a new cell pad to the cage lid at the start of the week and scored use each morning. CellPad was provided in addition to normal nesting material. We scored on two levels; percentage of CellPad destroyed, and of that, the percentage that had been incorporated into a nest. These percentages are plotted on the graph. Resources: • Time: Easy and fast to add to each cage. • Costs: Very cheap, need replacing frequently Mouse 1 Mouse 2 Mouse 3 Mouse 4 Mouse 5Chew sticks average13 CellPad Scenario: We attached a new cell pad to the cage lid at the start of the week and scored use each morning. CellPad was provided in addition to normal nesting material. We scored on two levels; percentage of CellPad destroyed, and of that, the percentage that had been incorporated into a nest. These percentages are plotted on the graph. Resources: • Time: Easy and fast to add to each cage. • Costs: Very cheap, need replacing frequently – Novelty maintained over time.– Universal interest from all animals trialled.– Easily autoclaved and stored.– Provides additional insulation for litters.Cons:– Enrichment lost as item is consumed.Have to limit number of CellPads provided between cagecleans as material accumulates and blocks view of the animals. 15Figure 7. Mouse breaking down the CellPad. Figure 8. Shows incorporation of CellPad into next structure. Figure 8. Mouse breaking down the CellPad. 15Figure 7. Mouse breaking down the CellPad. Figure 8. Shows incorporation of CellPad into next structure. Figure 9. Shows incorporation of CellPad into next structure. Poster PresentationsFigure 7. Amounts of chew stick utilised by mice.

67August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsSummary From the study, we have concluded that all of the items trialled have their pros and cons and that it is important to validate items as providing enrichment, rather than assuming this. Whilst the mice showed a prolonged interest in the balls, this was not sustainable due to the high level of resources required. Based on this, we identifi ed the CellPads as a possible alternative that offered a similar type of enrichment. Both the Cellpads and chew sticks were the most effective, due to their low resource requirements, ease of use and interest shown by the animals. They were also the most universally suitable for use in different scenarios. Further studies are required to determine whether novelty is retained over longer periods or can be revivedafter removal for a period of time or if there are certainsceanrios where wheels consistently offer an enrichment opportunity. However, based on the data from this study, as a facility we are now confi dent in our routine use of chew sticks as an enrichment item and also use the CellPads where we require additional enrichment such as mice that are showing signs of aggression or are unavoidably singularly housed. 16 Summary From the study, we have concluded that all of the items trialed have their pros and cons and that it is important to validate items as providing enrichment, rather than assuming this. Whilst the mice showed a prolonged interest in the balls, this was not sustainable due to the high level of resources required. Based on this, we identified the CellPads as a possible alternative that offered a similar type of enrichment. Both the cell pads and chew sticks were the most effective, due to their low resource requirements, ease of Mouse 1CellPads16 Summary From the study, we have concluded that all of the items trialed have their pros and cons and that it is important to validate items as providing enrichment, rather than assuming this. Whilst the mice showed a prolonged interest in the balls, this was not sustainable due to the high level of resources required. Based on this, we identified the CellPads as a possible alternative that offered a similar type of enrichment. Both the cell pads and chew sticks were the most effective, due to their low resource requirements, ease of Mouse 2 Mouse 3 Mouse 4 Mouse 5Figure 10. Analysis of CellPad use.

68Animal Technology and Welfare August 2020Animal Technology and Welfare April 2023AbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identification aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent refinement in their welfare and care.Introduction The need to identify xenopus frogs individually has grown in recent years. In 2016 photo- graphy was trialled over the other methods of identification. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet and, implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were fixed and, the refinements found in health monitoring of clinical signs.When identification of animals is necessary then it is of paramount importance that only the most humane methods are used (Reed 2005).1 Digital photography and the care and welfare of African Clawed frogs (Xenopus laevis ) THERESE JONES-GREEN University of CambridgeCorrespondence: tj222@cam.ac.uk Variance in Xenopus laevis dorsal patternsThere is a huge variance in the colour, size and pattern on a Xenopus laevis frog’s back. Technicians and researchers quickly develop a skill for discriminating one from the other. (Figure 1)3 In this poster our photography method is described, how we got thebest photographs, some of the issues that arose, how they werefixed, and the refinements found in health monitoring of clinical signs. When identification of animals is necessary then it is of paramount importance that only the most humane methods are used1 (Reed 2005).1 Variance in Xenopus laevis dorsal patterns 3 In this poster our photography method is described, how we got thebest photographs, some of the issues that arose, how they werefixed, and the refinements found in health monitoring of clinical signs. When identification of animals is necessary then it is of paramount importance that only the most humane methods are used1 (Reed 2005).1 Variance in Xenopus laevis dorsal patterns 3 In this poster our photography method is described, how we got thebest photographs, some of the issues that arose, how they werefixed, and the refinements found in health monitoring of clinical signs. When identification of animals is necessary then it is of paramount importance that only the most humane methods are used1 (Reed 2005).1 Variance in Xenopus laevis dorsal patterns Figure 1. Xenopus frog dorsal patterns.

69August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsMethod Example of a good photographEquipmentYou will need a suitable holding tank, digital camera, a suitable computer programme, laminate pouches, permanent marker pen and a laminator.ActionPut frog to be identifi ed into a holding tub. Wait for the frog to be static.PhotographEnsure lighting is correct (no shadows/refl ections) and the whole frog is in the frame. Take the photograph.UploadUpload the photograph onto a computer for printing and fi ling.LabelLabel the photograph with any desirable permanent information (spawn date, type, ID number etc).LaminateLaminate the card.● whole of the frog is visible● water is clean● frog is stationary● photograph is in focus● no refl ections● not too dark or too light5 Example of a good photograph. • Whole of the frog is visible• • Water is clean• • Frog is stationary• • Photograph is in focus• • No reflections • Not too dark or too light Example of a badly taken photograph 1. Hind limbs are missing: this photo would not be useful for comparing health status. Example of a badly taken photograph 6 2. Frog is moving Figure 4. Frog is moving.6 2. Frog is moving Figure 5. Image is too dark: once printed marking will be diffi cult to see. 7 3. Image is too dark: once printed marking will be difficult to see. 4. Reflections should be avoided Figure 3.Hind limbs are missing: this photo would not be useful for comparing health status.Figure 2.

70Animal Technology and Welfare August 2020Figure 6. Refl ections should be avoided.Albinos are diffi cult to differentiate between 7 3. Image is too dark: once printed marking will be difficult to see. 4. Reflections should be avoided 8 Albinos are difficult to differentiate between. 8 Albinos are difficult to differentiate between. Example of Xenopus IDs9 Example of Xenopus IDs Identification used to compare clinical signs Identifi cation used to compare clinical signsVariance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesFigure 10. The original ID was used to measure the extentof this frogs unusual clinical signs (Figure 11).Variance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesPoster PresentationsFigure 8. Figure 7. Figure 11. Figure 9.

71August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareVariance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesComparison of Xenopus markings with a 54-month interval Figure 12-17. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have changed little over a period of at least 4 years. Mrozek et al (1994)2 claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Variance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesVariance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesVariance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesVariance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesVariance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesPoster Presentations

72Animal Technology and Welfare August 2020Variance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesXenopus markings rarely changeDiscrete changes (affecting <0.1% of frogs) are easily nullifi ed by either updating the photo or marking prominentfeatures. Recovery in photographs The photographs below clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs signifi cantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Figure 18. July 2017. Variance in Xenopus laevis dorsal patterns.There is a huge variance in the colour, size and pattern on a Xenopus laevis frogs’ back. Technicians and researchers quickly develop a skill for discriminating one from the other.MethodYou will need a suitable holding tank, digital camera, computer program such as iphoto, laminate pouches, permanent marker pen, and a laminator.EquipmentPut frog to be identified into a holding tub. Wait for the frog to be static.ActionEnsure lighting is correct (no shadows/reflections) and the whole frog is in the frame. Take the photograph.PhotographUpload the photograph onto a computer for printing and filing.UploadLabel the photograph with any desirable permanent information (spawn date, type, ID number etc.)LabelLaminate the card.LaminateExample of Xenopus IDsExample of a good photograph.• Whole of the frog is visible• Water is clean• Frog is stationary• Photograph is in focus• No reflections• Not too dark or too lightExample of badly taken photographs.1) Hind limbs are missing: this photo would not be useful for comparing health status2) Frog is moving3) Image is too dark: once printed marking will be difficult to see4) Reflections should be avoided.1324Some Xenopus look very alikeHowever, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Albinos are difficult to discriminate.Comparison of Xenopus markings with a 54-month interval. Photos on the left were taken in August 2016. The photos on the right were taken in January 2020. These frogs (3 out of 100) were chosen using a random number generator to show that the markings have little change over a period of at least 4 years. Mrozek et al (1994) claimed that the patterning on xenopus was disruptive and inconspicuous, we have found this to not be the case.Xenopus markings rarely changeJuly 2017 October 2018. Arrows indicate changesDiscrete changes (affecting <0.1% of frogs) are easily nullified by either updating the photo or marking prominent features.ID used to compare clinical signsThe original ID (left), was used to measure the extent of this frogs unusual clinical signs (right).Recovery in photographs3 days after exposure 7 days after exposure14 days after exposure.The below photographs clearly illustrate the progress of a frog after exposure of Gas Bubble Disease. After two weeks this frog’s clinical signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. Digital Photography and the Care and Welfare of African Clawed Frogs (Xenopus Laevis)Therese Jones-Green, Chief Aquatic Technician/NACWO, University of CambridgeAbstractDigital photography has been used at The Gurdon Institute, Cambridge University since 2016, primarily as an identication aid. We have found it to be: non-invasive, quick, cheap, reliable and accessible. We also discovered it to be advantageous in monitoring the health and body condition of our large colony of over 600 Xenopus laevis frogs. This has been an excellent renement in their welfare and care.IntroductionThe need to identify xenopus frogs individually has grown in recent years. In 2016 photography was trialled over the other methods of identication. Those methods include: freeze branding, tattooing, toe clipping, threading tags in the hind feet, and implanting microchips in the dorsal lymph sac. All are invasive, causing pain and stress to the animal. In this poster our photography method is described, how we got the best photographs, some of the issues that arose, how they were xed, and the renements found in health monitoring of clinical signs.“When identication of animals is necessary then it is of paramount importance that only the most humane methods are used” (Reed 2005).DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we’re able to check back to the reference point to monitor their condition. This helps us to easily make informed decisions of the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identication (ID) card for quick reference.The few issues we found only aected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changed over time. Frog’s with similar markings can usually be swapped with another from a dierent tank. Both issues can easily be negated by marking out dierences/prominent features on the ID cards.Other methods of Identication such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M., (2011) Housing and Husbandry of Xenopus laevis Aect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science : JAALAS. 50. 46-53.Green, S.L. (2010) The Laboratory Xenopus Sp.CRC Press, Taylor & Francis Group.Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system used for animal identication in laboratory rabbits, guineapigs, woodchucks and in amphibians’ Laboratory Animals 29.Schultz TW, Dawson DA. Housing and husbandry of Xenopus for oocyte production.Lab Anim (NY). 2003;32(2):34–39. doi:10.1038/laban0203-34Reed, B.T. (2005) Guidance on the housing and care of the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.Wolfensohn, S. & Lloyd, M. (2010) ‘Handbook of laboratory animal management and welfare - 3rd edition’ Blackwell Science, Oxford.ReferencesUniversity Biomedical ServicesFigure 19. October 2018. Arrows indicate changes.13 signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. 3 days after exposure 7 days after exposure 13 signs significantly improved. These visual records enabled us to make better informed decisions on the frog’s progress. 3 days after exposure 7 days after exposure Figure 20. 3 days after exposure.Figure 21. 7 days after exposure.14 14 days after exposure. Some Xenopus look very alike However, this is unusual. In order to expedite identification put similar looking frogs in different tanks. Figure 22. 14 days after exposure.Poster Presentations

73August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareSome Xenopus look very alike However, this is unusual. In order to expedite identifi cationput similar looking frogs in different tanks. 14 14 days after exposure. Some Xenopus look very alike However, this is unusual. In order to expedite identification put similar looking frogs in different tanks. 14 14 days after exposure. Some Xenopus look very alike However, this is unusual. In order to expedite identification put similar looking frogs in different tanks. DiscussionPhotographing our female wild type frogs, provides us with a reference point for each frog at their healthiest stage in life. Dependent on the reason; we are able to refer back to the reference point to monitor their condition. This helps us to easily make informed decisionsof the fate of that animal, and what treatment to provide. It can help to detect subtle changes, such as weight loss/gain or changes in dorsal colour, all of which could indicate an underlying heath issue. All information can be recorded on each frog’s identifi cation (ID) card for quick reference.The few issues we found only affected a small minority of frogs. Less than 0.1% of frog’s dorsal pattern changedover time. Frogs with similar markings can usually be swapped with another from a different tank. Both issues can easily be negated by marking out differences/prominent features on the ID cards.Other methods of identifi cation such as microchips and tags, may cause pain, stress and are an infection risk and would be more appropriate for identifying albino frogs.References1 Reed, B.T. (2005). Guidance on the housing and careof the African Clawed Frog Xenopus Laevis, Research Animal Department, RSPCA.2 Mrozek, M., Fischer, R., Trendelenburg, M. & Zillmann, U. (1995) ‘Microchip implant system usedfor animal identifi cation in laboratory rabbits, guinea-pigs, woodchucks and in amphibians’ Laboratory Animals. Jul;29(3):339-44.Additional bibliographyGreen, S.L. (2010). The Laboratory Xenopus sp. CRC Press. ISBN 9781138437296.Delpire E., Gagnon K.B., Ledford J.J., Wallace J.M.(2011). Housing and Husbandry of Xenopus laevisAffect the Quality of Oocytes for Heterologous Expression Studies. Journal of the American Association for Laboratory Animal Science: JAALAS. 50. 46-53Schultz, T.W., Dawson, D.A. Housing and husbandry of Xenopus for oocyte production. Lab Anim (NY). 2003;32(2): 34–39. doi:10.1038/laban0203-34Wolfensohn, S. & Lloyd, M. (2010). ‘Handbook of laboratory animal management and welfare – 3rd edition’ Blackwell Science, Oxford.Poster PresentationsFigure 23-24. Similarly marked Xenopus frogs.

74Animal Technology and Welfare August 2020IntroductionCage-mate aggression is a well-known problem that is more prevalent in certain strains of mice and is one of the most common reasons for single housing, increased stress levels, premature death and euthanasia. It is known in the Animal Technology fi eld that there is a higher-prevalence of aggression in strains with the following lineage; C3H, CBA and CD-1 (Lidster, K et al., 2019).1From a retrospective review of health conditions between April 2020-April 2021 approximately one third of male mice on a CD-1 background were single housed due toaggression and fi ght wounds (Figure 1). This was observedboth in CD-1 background males generated in the facility and those ordered in from an external supplier.Numerous methods can be implemented to ameliorate and reduce aggression, including; reducing cage density Minimising aggression in CD-1 and CD-1 background male mice with different enrichment typesAMY VENESS, CHIS COYLE, SIAN MURPHY, JAMIE REDDEN, TINA O’MAHONY and ELENI M AMANITISainsbury Wellcome CentreCorrespondence: a.veness@ucl.ac.ukBased on a poster displayed at IAT Congress 22MINIMISING AGGRESSION IN CD-1 AND CD-1 BACKGROUND MALE MICE WITH DIFFERENT ENRICHMENT TYPESCage-mate aggression is a well-known problem that is more prevalent in certain strains of mice and is one of the most common reasons for single housing, increased stress levels, premature death and euthanasia. It is known in the animal technology field that there is a higher-prevalence of aggression in strains with the following lineage; C3H, CBA and CD-1 (Lidster, K et al., 2019).From a retrospective review of health conditions between April 2020-April 2021 approximately one third of male mice on a CD-1 background where single housed due to aggression and fight wounds (Figure 1). This was observed both in CD-1 background males generated in the facility and ordered in from an external supplier.Numerous methods can be implemented to ameliorate and reduce aggression, including; reducing cage density (number of mice per cage), altering cage cleaning routines, and changing environmental enrichment. It is accepted throughout the animal technology industry that single housing should be the last resort. Several combinations of environmental enrichment can be trialled, including tunnel clips, mezzanines, mouse loft/balconies, and different nesting materials such as sizzle nesting and nestlets (Figure 2).❑ Tunnel clips, mezzanines and mouse loft/balconies had minimal effect on reducing observed aggression rates, therefore the use of these enrichment types were discontinued.❑ Adding sizzle nest and a nestlet to cages with CD-1 and CD-1 background males considerably reduced the aggression rates and the amount of single housed males (Figure 3). ❑ The decrease of 27.51% in the aggression rates, observed in CD-1 and CD-1 background males between the two years, was statistically significant (P < 0.0001) (Figure 4).Amy Veness1, Chis Coyle1, Sian Murphy1, Jamie Redden1, Tina O'Mahony1, Eleni M Amaniti11 Sainsbury Wellcome Centre, 25 Howland Street, London, W1T 4JGAcknowledgements: Yoh Isogai and NRF Team (Aaron Donaldson, Amy Lynch, Ben Foster Georgie Spibey, Molly Pettengell, Olivia Adams, Rosie Keenan, Sandra Stojkovska)References: Lidster, K., Owen, K., Browne, W.J. and Prescott, M.J. (2019). Cage aggression in group-housed laboratory male mice: an international data crowdsourcing projec.t Scientific Reports, [online] 9(1), pp.1–12. doi:https://doi.org/10.1038/s41598-019-51674-z. ; Weber, E.M., Zidar, J., Ewaldsson, B., Askevik, K., Udén, E., Svensk, E. and Törnqvist, E. (2022). Aggression in Group-Housed Male Mice: A Systematic Review. Animals, 13(1), p.143. ; Jirkof, P., Bratcher, N., Medina, L., Strasburg, D., Ebert, P. and Gaskill, B.N., (2020). The Effect of Group Size, Age and Handling Frequency on Inter-male Aggression in CD-1 Mice. scientific reports, 10(1), pp.1-13.: Howerton, C.L., Garner, J.P. and Mench, J.A. (2008). Effects of a running wheel-igloo enrichment on aggression, hierarchy linearity, and stereotypy in group-housed male CD-1 (ICR) mice. Applied Animal Behaviour Science, 115(1-2), pp.90–10330.57%***3.06%0.00%5.00%10.00%15.00%20.00%25.00%30.00%35.00%April 2020-2021 April 2021-2022Aggression Rates of CD-1 and CD-1 Background Male Mice Between April 2020-2021 and April 2021-2022Background/ Introduction Results❑ A data review between April 2020-2021 and April 2021-2022 was conducted using the animal facility software PyRAT. The criteria used were: 1) CD-1 (supplied by Charles River) and CD-1 background males 2) Having the condition of ‘fight wounds’ and/or ‘aggressive’ 3) Single housed males. The total number of CD-1 and CD-1 background males included in this study was 507 (April 2020-2021) and 458 (April 2021- 2022). ❑ Between February 2021 and April 2021 numerous environmental enrichments (Figure 2) were trialled/added to cages of CD-1 and CD-1 background males. In April 2021, 10g of sizzle nest and 1 nestlet were added as part of standard enrichment to all cages housing CD-1 and CD-1 background males.❑ Fisher’s exact test was used to determine if there was a significant difference between rates of aggression during April 2020-2021 and April 2021-22. MethodFigure 2; Examples of enrichment: a) Mezzanine b) Mouse balcony c) Tunnel clips d) Cardboard house e) Nestlet f) Sizzle nesting g) Standard enrichment used at the Sainsbury Wellcome Centreb)c)a)d)f)e)g)❑ Previous literature has suggested that certain enrichment such as wheel-igloos may increases aggression in CD-1 males due to them becoming territorial over it, however when used in conjunction with nesting material this aggression was reduced (Howerton, C.L et al., 2008). This may explain why there were higher levels of aggression seen between April 2020-2021, but the aggression was reduced after adding sizzle nest and nestlets.❑ These results support current literature that shows manipulatable enrichment reduces aggression seen in CD-1 and CD-1 background mice (Weber E. M., 2022: Jirkof, P, et al., 2020).❑ This study accomplished its aim to reduce the aggression rates in group housed CD-1 and CD-1 background males and reduce the number of males single housed. In turn, animal welfare has increased as well as reducing the requirement for resources in the facility. Due to the results of this study, adding sizzle nest and nestlets to cages and strains showing aggression has become standard practice.DiscussionFigure 1; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Examples of fight woundsa)b)Figure 3; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Example of a cage with sizzle nest and a nestlet and our current enrichment for CD-1 malesa)b)Figure 4; Bar chart showing the rates of aggression, Fisher exact test (***P < 0.0001)Figure 1. a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Examples of fi ght wounds.Animal Technology and Welfare April 2023MINIMISING AGGRESSION IN CD-1 AND CD-1 BACKGROUND MALE MICE WITH DIFFERENT ENRICHMENT TYPESCage-mate aggression is a well-known problem that is more prevalent in certain strains of mice and is one of the most common reasons for single housing, increased stress levels, premature death and euthanasia. It is known in the animal technology field that there is a higher-prevalence of aggression in strains with the following lineage; C3H, CBA and CD-1 (Lidster, K et al., 2019).From a retrospective review of health conditions between April 2020-April 2021 approximately one third of male mice on a CD-1 background where single housed due to aggression and fight wounds (Figure 1). This was observed both in CD-1 background males generated in the facility and ordered in from an external supplier.Numerous methods can be implemented to ameliorate and reduce aggression, including; reducing cage density (number of mice per cage), altering cage cleaning routines, and changing environmental enrichment. It is accepted throughout the animal technology industry that single housing should be the last resort. Several combinations of environmental enrichment can be trialled, including tunnel clips, mezzanines, mouse loft/balconies, and different nesting materials such as sizzle nesting and nestlets (Figure 2).❑ Tunnel clips, mezzanines and mouse loft/balconies had minimal effect on reducing observed aggression rates, therefore the use of these enrichment types were discontinued.❑ Adding sizzle nest and a nestlet to cages with CD-1 and CD-1 background males considerably reduced the aggression rates and the amount of single housed males (Figure 3). ❑ The decrease of 27.51% in the aggression rates, observed in CD-1 and CD-1 background males between the two years, was statistically significant (P < 0.0001) (Figure 4).Amy Veness1, Chis Coyle1, Sian Murphy1, Jamie Redden1, Tina O'Mahony1, Eleni M Amaniti11 Sainsbury Wellcome Centre, 25 Howland Street, London, W1T 4JGAcknowledgements: Yoh Isogai and NRF Team (Aaron Donaldson, Amy Lynch, Ben Foster Georgie Spibey, Molly Pettengell, Olivia Adams, Rosie Keenan, Sandra Stojkovska)References: Lidster, K., Owen, K., Browne, W.J. and Prescott, M.J. (2019). Cage aggression in group-housed laboratory male mice: an international data crowdsourcing projec.t Scientific Reports, [online] 9(1), pp.1–12. doi:https://doi.org/10.1038/s41598-019-51674-z. ; Weber, E.M., Zidar, J., Ewaldsson, B., Askevik, K., Udén, E., Svensk, E. and Törnqvist, E. (2022). Aggression in Group-Housed Male Mice: A Systematic Review. Animals, 13(1), p.143. ; Jirkof, P., Bratcher, N., Medina, L., Strasburg, D., Ebert, P. and Gaskill, B.N., (2020). The Effect of Group Size, Age and Handling Frequency on Inter-male Aggression in CD-1 Mice. scientific reports, 10(1), pp.1-13.: Howerton, C.L., Garner, J.P. and Mench, J.A. (2008). Effects of a running wheel-igloo enrichment on aggression, hierarchy linearity, and stereotypy in group-housed male CD-1 (ICR) mice. Applied Animal Behaviour Science, 115(1-2), pp.90–10330.57%***3.06%0.00%5.00%10.00%15.00%20.00%25.00%30.00%35.00%April 2020-2021 April 2021-2022Aggression Rates of CD-1 and CD-1 Background Male Mice Between April 2020-2021 and April 2021-2022Background/ Introduction Results❑ A data review between April 2020-2021 and April 2021-2022 was conducted using the animal facility software PyRAT. The criteria used were: 1) CD-1 (supplied by Charles River) and CD-1 background males 2) Having the condition of ‘fight wounds’ and/or ‘aggressive’ 3) Single housed males. The total number of CD-1 and CD-1 background males included in this study was 507 (April 2020-2021) and 458 (April 2021- 2022). ❑ Between February 2021 and April 2021 numerous environmental enrichments (Figure 2) were trialled/added to cages of CD-1 and CD-1 background males. In April 2021, 10g of sizzle nest and 1 nestlet were added as part of standard enrichment to all cages housing CD-1 and CD-1 background males.❑ Fisher’s exact test was used to determine if there was a significant difference between rates of aggression during April 2020-2021 and April 2021-22. MethodFigure 2; Examples of enrichment: a) Mezzanine b) Mouse balcony c) Tunnel clips d) Cardboard house e) Nestlet f) Sizzle nesting g) Standard enrichment used at the Sainsbury Wellcome Centreb)c)a)d)f)e)g)❑ Previous literature has suggested that certain enrichment such as wheel-igloos may increases aggression in CD-1 males due to them becoming territorial over it, however when used in conjunction with nesting material this aggression was reduced (Howerton, C.L et al., 2008). This may explain why there were higher levels of aggression seen between April 2020-2021, but the aggression was reduced after adding sizzle nest and nestlets.❑ These results support current literature that shows manipulatable enrichment reduces aggression seen in CD-1 and CD-1 background mice (Weber E. M., 2022: Jirkof, P, et al., 2020).❑ This study accomplished its aim to reduce the aggression rates in group housed CD-1 and CD-1 background males and reduce the number of males single housed. In turn, animal welfare has increased as well as reducing the requirement for resources in the facility. Due to the results of this study, adding sizzle nest and nestlets to cages and strains showing aggression has become standard practice.DiscussionFigure 1; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Examples of fight woundsa)b)Figure 3; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Example of a cage with sizzle nest and a nestlet and our current enrichment for CD-1 malesa)b)Figure 4; Bar chart showing the rates of aggression, Fisher exact test (***P < 0.0001)

75August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and Welfare(number of mice per cage), altering cage cleaning routinesand changing environmental enrichment. It is considered throughout the Animal Technology industry that single housing should be the last resort. Several combinations of environmental enrichment can be trialled, including tunnel clips, mezzanines, mouse loft/balconies and different nesting materials such as sizzle nesting and nestlets (Figure 2). Method – A data review between April 2020-2021 and April 2021-2022 was conducted using the animal facility software PyRAT. The criteria used were: 1) CD-1 (supplied by Charles River) and CD-1 background males, 2) Having the condition of ‘fi ght wounds’ and/or ‘aggressive’, 3) Single housed males. The total number of CD-1 and CD-1 background males included in this study was 507 (April 2020-2021) and 458 (April 2021-2022). – Between February 2021 and April 2021 numerous environmental enrichments (Figure 2) were trialled/added to cages of CD-1 and CD-1 background males. In April 2021, 10g of sizzle nest and 1 nestlet were added as part of standard enrichment to all cages housing CD-1 and CD-1 background males.–Fisher’s exact test was used to determine if there was a signifi cant difference between rates of aggression during April 2020-2021 and April 2021-22. Results– Tunnel clips, mezzanines and mouse loft/balconies had minimal effect on reducing observed aggression rates, therefore the use of these enrichment types was discontinued.– Adding sizzle nest and a nestlet to cages with CD-1 and CD-1 background males considerably reduced the aggression rates and the amount of single housed males (Figure 3). – The decrease of 27.51% in the aggression rates, observed in CD-1 and CD-1 background males betweenthe two years, was statistically signifi cant (P < 0.0001)(Figure 4).MINIMISING AGGRESSION IN CD-1 AND CD-1 BACKGROUND MALE MICE WITH DIFFERENT ENRICHMENT TYPESCage-mate aggression is a well-known problem that is more prevalent in certain strains of mice and is one of the most common reasons for single housing, increased stress levels, premature death and euthanasia. It is known in the animal technology field that there is a higher-prevalence of aggression in strains with the following lineage; C3H, CBA and CD-1 (Lidster, K et al., 2019).From a retrospective review of health conditions between April 2020-April 2021 approximately one third of male mice on a CD-1 background where single housed due to aggression and fight wounds (Figure 1). This was observed both in CD-1 background males generated in the facility and ordered in from an external supplier.Numerous methods can be implemented to ameliorate and reduce aggression, including; reducing cage density (number of mice per cage), altering cage cleaning routines, and changing environmental enrichment. It is accepted throughout the animal technology industry that single housing should be the last resort. Several combinations of environmental enrichment can be trialled, including tunnel clips, mezzanines, mouse loft/balconies, and different nesting materials such as sizzle nesting and nestlets (Figure 2).❑ Tunnel clips, mezzanines and mouse loft/balconies had minimal effect on reducing observed aggression rates, therefore the use of these enrichment types were discontinued.❑ Adding sizzle nest and a nestlet to cages with CD-1 and CD-1 background males considerably reduced the aggression rates and the amount of single housed males (Figure 3). ❑ The decrease of 27.51% in the aggression rates, observed in CD-1 and CD-1 background males between the two years, was statistically significant (P < 0.0001) (Figure 4).Amy Veness1, Chis Coyle1, Sian Murphy1, Jamie Redden1, Tina O'Mahony1, Eleni M Amaniti11 Sainsbury Wellcome Centre, 25 Howland Street, London, W1T 4JGAcknowledgements: Yoh Isogai and NRF Team (Aaron Donaldson, Amy Lynch, Ben Foster Georgie Spibey, Molly Pettengell, Olivia Adams, Rosie Keenan, Sandra Stojkovska)References: Lidster, K., Owen, K., Browne, W.J. and Prescott, M.J. (2019). Cage aggression in group-housed laboratory male mice: an international data crowdsourcing projec.t Scientific Reports, [online] 9(1), pp.1–12. doi:https://doi.org/10.1038/s41598-019-51674-z. ; Weber, E.M., Zidar, J., Ewaldsson, B., Askevik, K., Udén, E., Svensk, E. and Törnqvist, E. (2022). Aggression in Group-Housed Male Mice: A Systematic Review. Animals, 13(1), p.143. ; Jirkof, P., Bratcher, N., Medina, L., Strasburg, D., Ebert, P. and Gaskill, B.N., (2020). The Effect of Group Size, Age and Handling Frequency on Inter-male Aggression in CD-1 Mice. scientific reports, 10(1), pp.1-13.: Howerton, C.L., Garner, J.P. and Mench, J.A. (2008). Effects of a running wheel-igloo enrichment on aggression, hierarchy linearity, and stereotypy in group-housed male CD-1 (ICR) mice. Applied Animal Behaviour Science, 115(1-2), pp.90–10330.57%***3.06%0.00%5.00%10.00%15.00%20.00%25.00%30.00%35.00%April 2020-2021 April 2021-2022Aggression Rates of CD-1 and CD-1 Background Male Mice Between April 2020-2021 and April 2021-2022Background/ Introduction Results❑ A data review between April 2020-2021 and April 2021-2022 was conducted using the animal facility software PyRAT. The criteria used were: 1) CD-1 (supplied by Charles River) and CD-1 background males 2) Having the condition of ‘fight wounds’ and/or ‘aggressive’ 3) Single housed males. The total number of CD-1 and CD-1 background males included in this study was 507 (April 2020-2021) and 458 (April 2021- 2022). ❑ Between February 2021 and April 2021 numerous environmental enrichments (Figure 2) were trialled/added to cages of CD-1 and CD-1 background males. In April 2021, 10g of sizzle nest and 1 nestlet were added as part of standard enrichment to all cages housing CD-1 and CD-1 background males.❑ Fisher’s exact test was used to determine if there was a significant difference between rates of aggression during April 2020-2021 and April 2021-22. MethodFigure 2; Examples of enrichment: a) Mezzanine b) Mouse balcony c) Tunnel clips d) Cardboard house e) Nestlet f) Sizzle nesting g) Standard enrichment used at the Sainsbury Wellcome Centreb)c)a)d)f)e)g)❑ Previous literature has suggested that certain enrichment such as wheel-igloos may increases aggression in CD-1 males due to them becoming territorial over it, however when used in conjunction with nesting material this aggression was reduced (Howerton, C.L et al., 2008). This may explain why there were higher levels of aggression seen between April 2020-2021, but the aggression was reduced after adding sizzle nest and nestlets.❑ These results support current literature that shows manipulatable enrichment reduces aggression seen in CD-1 and CD-1 background mice (Weber E. M., 2022: Jirkof, P, et al., 2020).❑ This study accomplished its aim to reduce the aggression rates in group housed CD-1 and CD-1 background males and reduce the number of males single housed. In turn, animal welfare has increased as well as reducing the requirement for resources in the facility. Due to the results of this study, adding sizzle nest and nestlets to cages and strains showing aggression has become standard practice.DiscussionFigure 1; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Examples of fight woundsa)b)Figure 3; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Example of a cage with sizzle nest and a nestlet and our current enrichment for CD-1 malesa)b)Figure 4; Bar chart showing the rates of aggression, Fisher exact test (***P < 0.0001)Figure 2. Examples of enrichment: a) Mezzanine. b) Mouse balcony. c) Tunnel clips. d) Cardboard house. e) Nestlet. f) Sizzle nesting. g) Standard enrichment used at the Sainsbury Wellcome Centre.MINIMISING AGGRESSION IN CD-1 AND CD-1 BACKGROUND MALE MICE WITH DIFFERENT ENRICHMENT TYPESCage-mate aggression is a well-known problem that is more prevalent in certain strains of mice and is one of the most common reasons for single housing, increased stress levels, premature death and euthanasia. It is known in the animal technology field that there is a higher-prevalence of aggression in strains with the following lineage; C3H, CBA and CD-1 (Lidster, K et al., 2019).From a retrospective review of health conditions between April 2020-April 2021 approximately one third of male mice on a CD-1 background where single housed due to aggression and fight wounds (Figure 1). This was observed both in CD-1 background males generated in the facility and ordered in from an external supplier.Numerous methods can be implemented to ameliorate and reduce aggression, including; reducing cage density (number of mice per cage), altering cage cleaning routines, and changing environmental enrichment. It is accepted throughout the animal technology industry that single housing should be the last resort. Several combinations of environmental enrichment can be trialled, including tunnel clips, mezzanines, mouse loft/balconies, and different nesting materials such as sizzle nesting and nestlets (Figure 2).❑ Tunnel clips, mezzanines and mouse loft/balconies had minimal effect on reducing observed aggression rates, therefore the use of these enrichment types were discontinued.❑ Adding sizzle nest and a nestlet to cages with CD-1 and CD-1 background males considerably reduced the aggression rates and the amount of single housed males (Figure 3). ❑ The decrease of 27.51% in the aggression rates, observed in CD-1 and CD-1 background males between the two years, was statistically significant (P < 0.0001) (Figure 4).Amy Veness1, Chis Coyle1, Sian Murphy1, Jamie Redden1, Tina O'Mahony1, Eleni M Amaniti11 Sainsbury Wellcome Centre, 25 Howland Street, London, W1T 4JGAcknowledgements: Yoh Isogai and NRF Team (Aaron Donaldson, Amy Lynch, Ben Foster Georgie Spibey, Molly Pettengell, Olivia Adams, Rosie Keenan, Sandra Stojkovska)References: Lidster, K., Owen, K., Browne, W.J. and Prescott, M.J. (2019). Cage aggression in group-housed laboratory male mice: an international data crowdsourcing projec.t Scientific Reports, [online] 9(1), pp.1–12. doi:https://doi.org/10.1038/s41598-019-51674-z. ; Weber, E.M., Zidar, J., Ewaldsson, B., Askevik, K., Udén, E., Svensk, E. and Törnqvist, E. (2022). Aggression in Group-Housed Male Mice: A Systematic Review. Animals, 13(1), p.143. ; Jirkof, P., Bratcher, N., Medina, L., Strasburg, D., Ebert, P. and Gaskill, B.N., (2020). The Effect of Group Size, Age and Handling Frequency on Inter-male Aggression in CD-1 Mice. scientific reports, 10(1), pp.1-13.: Howerton, C.L., Garner, J.P. and Mench, J.A. (2008). Effects of a running wheel-igloo enrichment on aggression, hierarchy linearity, and stereotypy in group-housed male CD-1 (ICR) mice. Applied Animal Behaviour Science, 115(1-2), pp.90–10330.57%***3.06%0.00%5.00%10.00%15.00%20.00%25.00%30.00%35.00%April 2020-2021 April 2021-2022Aggression Rates of CD-1 and CD-1 Background Male Mice Between April 2020-2021 and April 2021-2022Background/ Introduction Results❑ A data review between April 2020-2021 and April 2021-2022 was conducted using the animal facility software PyRAT. The criteria used were: 1) CD-1 (supplied by Charles River) and CD-1 background males 2) Having the condition of ‘fight wounds’ and/or ‘aggressive’ 3) Single housed males. The total number of CD-1 and CD-1 background males included in this study was 507 (April 2020-2021) and 458 (April 2021- 2022). ❑ Between February 2021 and April 2021 numerous environmental enrichments (Figure 2) were trialled/added to cages of CD-1 and CD-1 background males. In April 2021, 10g of sizzle nest and 1 nestlet were added as part of standard enrichment to all cages housing CD-1 and CD-1 background males.❑ Fisher’s exact test was used to determine if there was a significant difference between rates of aggression during April 2020-2021 and April 2021-22. MethodFigure 2; Examples of enrichment: a) Mezzanine b) Mouse balcony c) Tunnel clips d) Cardboard house e) Nestlet f) Sizzle nesting g) Standard enrichment used at the Sainsbury Wellcome Centreb)c)a)d)f)e)g)❑ Previous literature has suggested that certain enrichment such as wheel-igloos may increases aggression in CD-1 males due to them becoming territorial over it, however when used in conjunction with nesting material this aggression was reduced (Howerton, C.L et al., 2008). This may explain why there were higher levels of aggression seen between April 2020-2021, but the aggression was reduced after adding sizzle nest and nestlets.❑ These results support current literature that shows manipulatable enrichment reduces aggression seen in CD-1 and CD-1 background mice (Weber E. M., 2022: Jirkof, P, et al., 2020).❑ This study accomplished its aim to reduce the aggression rates in group housed CD-1 and CD-1 background males and reduce the number of males single housed. In turn, animal welfare has increased as well as reducing the requirement for resources in the facility. Due to the results of this study, adding sizzle nest and nestlets to cages and strains showing aggression has become standard practice.DiscussionFigure 1; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Examples of fight woundsa)b)Figure 3; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Example of a cage with sizzle nest and a nestlet and our current enrichment for CD-1 malesa)b)Figure 4; Bar chart showing the rates of aggression, Fisher exact test (***P < 0.0001)Figure 3. a) A pie chart of the amount of singly housed CD-1 and CD-1 background males. b) Example of a cage with sizzle nest and a nestlet and our current enrichment for CD-1 males.Poster Presentations

76Animal Technology and Welfare August 2020Discussion Previous literature has suggested that certain enrichmentsuch as wheel-igloos may increase aggression in CD-1males due to them becoming territorial over it, however when used in conjunction with nesting material this aggression was reduced (Howerton, C.L et al., 2008).2This may explain why there were higher levels of aggression seen between April 2020-2021, but the aggression was reduced after adding sizzle nest and nestlets. These results support current literature that shows manipulatable enrichment reduces aggression seen in CD-1 and CD-1 background mice (Weber E.M., 2017: Jirkof, P, et al., 2020).3,4This study accomplished its aim to reduce the aggression rates in group housed CD-1 and CD-1 background males and reduce the number of males single housed. In turn, Animal Welfare has increased as well as reducing the requirement for resources in the facility. Due to the results of this study, adding sizzle nest and nestlets to cages and strains showing aggression has become standard practice.MINIMISING AGGRESSION IN CD-1 AND CD-1 BACKGROUND MALE MICE WITH DIFFERENT ENRICHMENT TYPESCage-mate aggression is a well-known problem that is more prevalent in certain strains of mice and is one of the most common reasons for single housing, increased stress levels, premature death and euthanasia. It is known in the animal technology field that there is a higher-prevalence of aggression in strains with the following lineage; C3H, CBA and CD-1 (Lidster, K et al., 2019).From a retrospective review of health conditions between April 2020-April 2021 approximately one third of male mice on a CD-1 background where single housed due to aggression and fight wounds (Figure 1). This was observed both in CD-1 background males generated in the facility and ordered in from an external supplier.Numerous methods can be implemented to ameliorate and reduce aggression, including; reducing cage density (number of mice per cage), altering cage cleaning routines, and changing environmental enrichment. It is accepted throughout the animal technology industry that single housing should be the last resort. Several combinations of environmental enrichment can be trialled, including tunnel clips, mezzanines, mouse loft/balconies, and different nesting materials such as sizzle nesting and nestlets (Figure 2).❑ Tunnel clips, mezzanines and mouse loft/balconies had minimal effect on reducing observed aggression rates, therefore the use of these enrichment types were discontinued.❑ Adding sizzle nest and a nestlet to cages with CD-1 and CD-1 background males considerably reduced the aggression rates and the amount of single housed males (Figure 3). ❑ The decrease of 27.51% in the aggression rates, observed in CD-1 and CD-1 background males between the two years, was statistically significant (P < 0.0001) (Figure 4).Amy Veness1, Chis Coyle1, Sian Murphy1, Jamie Redden1, Tina O'Mahony1, Eleni M Amaniti11 Sainsbury Wellcome Centre, 25 Howland Street, London, W1T 4JGAcknowledgements: Yoh Isogai and NRF Team (Aaron Donaldson, Amy Lynch, Ben Foster Georgie Spibey, Molly Pettengell, Olivia Adams, Rosie Keenan, Sandra Stojkovska)References: Lidster, K., Owen, K., Browne, W.J. and Prescott, M.J. (2019). Cage aggression in group-housed laboratory male mice: an international data crowdsourcing projec.t Scientific Reports, [online] 9(1), pp.1–12. doi:https://doi.org/10.1038/s41598-019-51674-z. ; Weber, E.M., Zidar, J., Ewaldsson, B., Askevik, K., Udén, E., Svensk, E. and Törnqvist, E. (2022). Aggression in Group-Housed Male Mice: A Systematic Review. Animals, 13(1), p.143. ; Jirkof, P., Bratcher, N., Medina, L., Strasburg, D., Ebert, P. and Gaskill, B.N., (2020). The Effect of Group Size, Age and Handling Frequency on Inter-male Aggression in CD-1 Mice. scientific reports, 10(1), pp.1-13.: Howerton, C.L., Garner, J.P. and Mench, J.A. (2008). Effects of a running wheel-igloo enrichment on aggression, hierarchy linearity, and stereotypy in group-housed male CD-1 (ICR) mice. Applied Animal Behaviour Science, 115(1-2), pp.90–10330.57%***3.06%0.00%5.00%10.00%15.00%20.00%25.00%30.00%35.00%April 2020-2021 April 2021-2022Aggression Rates of CD-1 and CD-1 Background Male Mice Between April 2020-2021 and April 2021-2022Background/ Introduction Results❑ A data review between April 2020-2021 and April 2021-2022 was conducted using the animal facility software PyRAT. The criteria used were: 1) CD-1 (supplied by Charles River) and CD-1 background males 2) Having the condition of ‘fight wounds’ and/or ‘aggressive’ 3) Single housed males. The total number of CD-1 and CD-1 background males included in this study was 507 (April 2020-2021) and 458 (April 2021- 2022). ❑ Between February 2021 and April 2021 numerous environmental enrichments (Figure 2) were trialled/added to cages of CD-1 and CD-1 background males. In April 2021, 10g of sizzle nest and 1 nestlet were added as part of standard enrichment to all cages housing CD-1 and CD-1 background males.❑ Fisher’s exact test was used to determine if there was a significant difference between rates of aggression during April 2020-2021 and April 2021-22. MethodFigure 2; Examples of enrichment: a) Mezzanine b) Mouse balcony c) Tunnel clips d) Cardboard house e) Nestlet f) Sizzle nesting g) Standard enrichment used at the Sainsbury Wellcome Centreb)c)a)d)f)e)g)❑ Previous literature has suggested that certain enrichment such as wheel-igloos may increases aggression in CD-1 males due to them becoming territorial over it, however when used in conjunction with nesting material this aggression was reduced (Howerton, C.L et al., 2008). This may explain why there were higher levels of aggression seen between April 2020-2021, but the aggression was reduced after adding sizzle nest and nestlets.❑ These results support current literature that shows manipulatable enrichment reduces aggression seen in CD-1 and CD-1 background mice (Weber E. M., 2022: Jirkof, P, et al., 2020).❑ This study accomplished its aim to reduce the aggression rates in group housed CD-1 and CD-1 background males and reduce the number of males single housed. In turn, animal welfare has increased as well as reducing the requirement for resources in the facility. Due to the results of this study, adding sizzle nest and nestlets to cages and strains showing aggression has become standard practice.DiscussionFigure 1; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Examples of fight woundsa)b)Figure 3; a) A pie chart of the amount of singly housed CD-1 and CD-1 background males b) Example of a cage with sizzle nest and a nestlet and our current enrichment for CD-1 malesa)b)Figure 4; Bar chart showing the rates of aggression, Fisher exact test (***P < 0.0001)Figure 4. Bar chart showing the rates of aggression, Fisher exact test (***P < 0.0001).Acknowledgements Yoh Isogai and NRF Team – Aaron Donaldson, Amy Lynch,Ben Foster, Georgie Spibey, Molly Pettengell, Olivia Adams,Rosie Keenan, Sandra Stojkovska. References1Lidster, K., et al., (2019). Scientifi c reports 9:152. DOI: 10/1038/s41598-0 51874-2.2Howerton, C.L., Garner, J.P., Mench, J.A. Applied Animal Behaviour Science. 115: 90-130. J. Applarim. 2008.05.04.3Weber, E.M. (2017). Lab Anim (NY) 46, 157-161.4Jirkof, P. et al., (2020). Laboratory Animals. 54.2.Poster Presentations

77August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfareAbstractAs a commercial breeder, Envigo had a requirement to relocate a hamster breeding colony from the United States of America (USA) to the UK Contract Breeding Services (CBS) Facility. Due to Animals (Scientifi c Procedures) Act 1986 (ASPA) Code of Practice for the Housing and Care of Animals Bred, Supplied or Used for Scientifi c Purposes specifi cations,1 there was a necessity to amend the established mating regime from trios (2 males, 1 female) to monogamous pairs. On implementing this regime, the number of successful matings was inconsistent and litter sizes were below the expected output. Consequently, the current breeding programme required review. Due to the lack of relevant literature, we had to establish a novel colony plan based on our existing data and expertise as breeders. This poster demonstrates an alternative scheme to produce a sustainable colony with a reduction in numbers of breeding animals used and increased productivity.IntroductionThe Golden Syrian Hamster, Mesocricetus auratus, is adesert rodent belonging to the Cricetidae family. Due to its unique anatomical and physiological features, it is widely used across a multitude of research areas. The Envigo HsdHan®:AURA colony was fi rst established in 1994 with stock originating from Zentralinstitut fur Versuchstiere in Hannover, Germany. Envigo acquired theline along with the Sprague-Dawley Company in 1973.In 2016, Envigo was approached by Public Health England(PHE), an executive agency of the Government of the United Kingdom Department of Health and Social Care, tobreed and supply cohorts of hamsters for a 4-yearexperimental programme of work, focussing on theresearch and development of new treatments forClostridium diffi cilerelated illnesses. At this time, theHsdHan®:AURA colony was based in Envigo’s NorthAmerican barrier facility. Due to the duration of thisprogramme of work and the considerable ethical burdenassociated with transatlantic travel it was deemed more appropriate to have this model readily available in the UK where the experimental work was to be performed.On relocation of the line from the USA to the UK, due to ASPA Code of Practice specifi cations, there was a need to revise the established breeding strategy used in the USA of 2 males to 1 female. On review of the literature, of which there was very little relating to hamster breeding, monogamous pairings of hamsters from weaning were deemed the most productive and labour effi cient method of breeding. The colony was fi rst housed in isolators before being relocated to Individually Ventilated Cages (IVCs) in November 2019. A refi nement in hamster breeding: creating a sustainable colony for vaccine research K. ANDERSON, M. DAVENPORT, J. FINNEY and S. MACCUISHENVIGO EnvigoCorrespondence: melanie.davenport@envigo.comBased on a poster displayed at IAT Congress 22April 2023 Animal Technology and Welfare

78Animal Technology and Welfare August 2020MethodContinuous mating in isolators–15 male and 15 female hamsters were imported fromEnvigo RMS USA to establish monogamous breeding pairs for continuous mating.– 15 monogamous pairs were fi rst mated on 18thNovember 2017 with an expected output of 135 pupsborn based on the average litter size of 9 pups per female at Envigo’s USA barrier. Results Continuous mating in isolatorsA refinement in hamster breeding – creating a sustainable colony for vaccine researchAbstractAs a commercial breeder, Envigo had a requirement to relocate a hamsterbreeding colony from the US to the UK Contract Breeding Services (CBS) Facility.Due to Code of Practice specifications, there was a necessity to amend theestablished mating regime from trios (2 males, 1 female) to monogamous pairs. Onimplementing this regime, the number of successful matings was inconsistent andlitter sizes were below the expected output. Consequently, the current breedingprogram required review. Due to the lack of relevant literature, we had to establisha novel colony plan based on our existing data and expertise as breeders. Thisposter demonstrates an alternative scheme to produce a sustainable colony, with areduction in numbers of breeders used and increased productivity.K Anderson, M Davenport, J Finney, S MacCuishEnvigo, Hillcrest, Dodgeford Lane, Belton, Loughborough, Leicestershire, LE12 9TEIntroductionThe Golden Syrian Hamster, Mesocricetus auratus, is a desert rodent belonging tothe Cricetidae family. Due to its unique anatomical and physiological features, it iswidely used across a multitude of research areas. The Envigo HsdHan®:AURAcolony was first established in 1994 with stock originating from Zentralinstitut furVersuchstiere in Hannover, Germany. Envigo acquired the line along with theSprague-Dawley Company in 1973.In 2016, Envigo was approached by Public Health England, an executive agencyof the Government of the United Kingdom Department of Health and Social Care,to breed and supply cohorts of hamsters for a 4 year experimental program ofwork, focusing on the research and development of new treatments for ClostridiumDifficile related illnesses. At this time, the HsdHan®:AURA colony was based inEnvigo’s North American barrier facility. Due to the duration of this program ofwork and the considerable ethical burden associated with transatlantic travel it wasdeemed more appropriate to have this model readily available in the UK where theexperimental work was to be performed.On relocation of the line from the US to the UK, due to Code of Practicespecifications, there was a need to revise the established breeding strategy usedin the US of 2 males to 1 female. On review of the literature, of which there wasvery little relating to hamster breeding, monogamous pairings of hamsters fromweaning was deemed the most productive and labour efficient method ofbreeding. The colony was first housed in isolators before being relocated toindividually ventilated cages IVCs) in November 2019.0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected Actual0204060801001201401601 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected ActualMethod: Continuous Mating in Isolators+ 15 male and 15 female hamsters were imported from Envigo RMS US toestablish monogamous breeding pairs for continuous mating+ 15 monogamous pairs were first mated on 18thNovember 2017 with anexpected output of 135 pups born based on the average litter size of 9pups per female at Envigo’s US barrier+ Whilst the first mating was 88% successful expected versus actual output, anticipated subsequent matings were sporadic with low productivity and high pre-wean loss+ Subsequently, it was deemed critical to review the continuous mating schemeDiscussionIn addition to the measurable benefits published in this poster on the use of a peakmating scheme versus a continuous mating scheme, it was generally noted thatfemale hamsters showed reduced aggression when being handled. Wehypothesise the increased productivity observed in IVCs versus isolators to be dueto the females only being able to smell the one male scented to her, rather thanmany animals, as is the case within an isolator where animals are held in open topcages. In optimising our hamster breeding strategy Envigo has been able to readilysupport PHE’s cohort requirements which has proved critical during the COVID-19pandemic. The peak mating scheme has also been used with transgenic mousemodels and has proved equally successful. As a refinement, we plan to trial aprocess that reduces the requirement to single house males for an extended periodof time.Following the transfer of a hamster colony from the US to the UK, we set out tobreed cohorts of hamsters for PHE experimental studies. On review of theliterature, a monogamous continuous breeding scheme was established. Whilst theoutput from the first mating was not as successful as the US production colonyoutput, it was in line with expectations. Anticipated subsequent matings weresporadic with low productivity generating cohorts of animals that were unusable forthe customer and therefore went to waste. Consequently, it was necessary toreview the established breeding scheme. A peak mating scheme was selected.Pre-assigned monogamous pairs were housed separately for a period of 10 daysafter which the scent of the male hamster was introduced to the females cage bythe transfer of nesting material. Mating was predominantly observed immediatelyon introduction of the female into the male cage 48 hours after scenting. After 10days, the female was removed from the male cage and returned to her untouchedhome cage. On average, the three peak matings performed generated 20% morepups born than anticipated, based on the US production colony output, with anaverage litter size of 11. There was a concern that the larger litter sizes would bean increased burden on the female resulting in a higher pre-wean loss. Our datademonstrates that this is not the case. On average pre-wean loss was reduced to11%. This scheme enabled the production of large cohorts of animals born withinon average an 8 day window as per the customers specifications utilising fewernumbers of breeders to achieve the required output, therefore refining our breedingscheme and eliminating animal wastage.Method: Peak Mating in Isolators and IVCs+ All breeding pairs were separated for a period of 10 days+ Future monogamous pairs (9 males and 9 females) were selected+ 48 hours prior to mating, nesting material was removed from the female cage and placed into the corresponding male cage+ Simultaneously, nesting material was removed from the male cage and placed into the corresponding female cage+ On day 0 the female was transferred into the male cage for mating+ The female home cage was not cleaned or modified in any way+ On day 9 the female was removed from the male cage and transferred back to her home cageResults: Peak Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 100/81 123% 11 15% 9Average Weeks 1-27334/279 120% 11 11% 8Since transferring the colony to IVCs, the number of breeders successfullyproducing a litter has increased from 120% to 129% over a comparative period andthe average age range per cohort of pups born has been further refined to 1 day.Results: Continuous Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born(days)Weeks 1-5 119/135 88% 9 19% 4 daysAverage Weeks 1-27656/1881 35% 6 49%Highly variable and largely unpredictableEnvigo would like to say a very special thank you to PHE for their continued support without which this poster would not have been possible. MatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 26/18 144% 13 0% 1Average Weeks 1-27116/90 129% 11 11% 1Results: Peak Mating in IVCs* Colony was relocated to IVCs in Nov 2019ancompany0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27Expected ActualWEEK NUMBERNUMBER OF PUPS BORNFigure 1. Golden Syrian Hamster (Mesocricetus auratus). MatingActual pups born versus expected pups bornSuccess%Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5119/135 88% 9 19% 4 daysAverage weeks 1-27656/1881 35% 6 49% Highly variable end largely unpredict-able– Whilst the fi rst mating was 88% successful expected versus actual output, anticipated subsequent matingswere sporadic with low productivity and high pre-weanloss.– Subsequently, it was deemed critical to review the continuous mating scheme.MethodPeak mating in isolators and IVCs– All breeding pairs were separated for a period of 10 days.– Future monogamous pairs (9 males and 9 females) were selected.– 48 hours prior to mating, nesting material was removed from the female cage and placed into the corresponding male cage.– Simultaneously, nesting material was removed from the male cage and placed into the corresponding female cage.– On day 0 the female was transferred into the male cage for mating.– The female home cage was not cleaned or modifi ed in any way.– On day 9 the female was removed from the male cage and transferred back to her home cage.ResultsPeak mating in isolatorsMatingActual pups born versus expected pups bornSuccess%Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5100/81 123% 11 15% 9Average weeks 1-27334/279 120% 11 11% 8Peak mating in IVCs Colony was relocated to IVCs in Nov 2019MatingActual pups born versus expected pups bornSuccess%Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-526/18 144% 13 0% 1Average weeks 1-27116/90 129% 11 11% 1Poster Presentations

79August 2020 Animal Technology and WelfareAugust 2020 Animal Technology and WelfarePoster PresentationsConclusionsFollowing the transfer of a hamster colony from the USA to the UK, we set out to breed cohorts of hamsters for PHE experimental studies. On review of the literature, a monogamous continuous breeding scheme was established. Whilst the output from the fi rst mating was not as successful as the US production colony output, it was in line with expectations. Anticipated subsequent matings were sporadic with low productivity generating cohorts of animals that were unusable for the customer and therefore went to waste. Consequently, it was necessary to review the established breeding scheme. A peak mating scheme was selected. Pre-assigned monogamous pairs were housed separately for a period of 10 days after which the scent of the male hamster was introduced to the female’s cage by the transfer of nesting material. Mating was predominantly observed immediately on introduction of the female into the male cage 48 hours after scenting. After 10 days, the female was removed from the male cage and returned to her untouched home cage. On average, the three peak matings performed generated 20% more pups born than anticipated, based on the USA production colony output, with an average litter size of 11. There was a concern that the larger litter sizes would be an increased burden on the female resulting in a higher pre-wean loss. Our data demonstrates that this is not the case. On average pre-wean loss was reduced to 11%. This scheme enabled the production of large cohorts of animals born within on average an 8-day window as per the customer’s specifi cations utilising fewer numbers of breeders to achieve the required output, therefore refi ning our breeding scheme and eliminating animal wastage. Since transferring the colony to IVCs, the number of breeders successfully producing a litter has increased from 120% to 129% over a comparative period and the average age range per cohort of pups born has been further refi ned to 1 day. A refinement in hamster breeding – creating a sustainable colony for vaccine researchAbstractAs a commercial breeder, Envigo had a requirement to relocate a hamsterbreeding colony from the US to the UK Contract Breeding Services (CBS) Facility.Due to Code of Practice specifications, there was a necessity to amend theestablished mating regime from trios (2 males, 1 female) to monogamous pairs. Onimplementing this regime, the number of successful matings was inconsistent andlitter sizes were below the expected output. Consequently, the current breedingprogram required review. Due to the lack of relevant literature, we had to establisha novel colony plan based on our existing data and expertise as breeders. Thisposter demonstrates an alternative scheme to produce a sustainable colony, with areduction in numbers of breeders used and increased productivity.K Anderson, M Davenport, J Finney, S MacCuishEnvigo, Hillcrest, Dodgeford Lane, Belton, Loughborough, Leicestershire, LE12 9TEIntroductionThe Golden Syrian Hamster, Mesocricetus auratus, is a desert rodent belonging tothe Cricetidae family. Due to its unique anatomical and physiological features, it iswidely used across a multitude of research areas. The Envigo HsdHan®:AURAcolony was first established in 1994 with stock originating from Zentralinstitut furVersuchstiere in Hannover, Germany. Envigo acquired the line along with theSprague-Dawley Company in 1973.In 2016, Envigo was approached by Public Health England, an executive agencyof the Government of the United Kingdom Department of Health and Social Care,to breed and supply cohorts of hamsters for a 4 year experimental program ofwork, focusing on the research and development of new treatments for ClostridiumDifficile related illnesses. At this time, the HsdHan®:AURA colony was based inEnvigo’s North American barrier facility. Due to the duration of this program ofwork and the considerable ethical burden associated with transatlantic travel it wasdeemed more appropriate to have this model readily available in the UK where theexperimental work was to be performed.On relocation of the line from the US to the UK, due to Code of Practicespecifications, there was a need to revise the established breeding strategy usedin the US of 2 males to 1 female. On review of the literature, of which there wasvery little relating to hamster breeding, monogamous pairings of hamsters fromweaning was deemed the most productive and labour efficient method ofbreeding. The colony was first housed in isolators before being relocated toindividually ventilated cages IVCs) in November 2019.0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected Actual0204060801001201401601 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected ActualMethod: Continuous Mating in Isolators+ 15 male and 15 female hamsters were imported from Envigo RMS US toestablish monogamous breeding pairs for continuous mating+ 15 monogamous pairs were first mated on 18thNovember 2017 with anexpected output of 135 pups born based on the average litter size of 9pups per female at Envigo’s US barrier+ Whilst the first mating was 88% successful expected versus actual output, anticipated subsequent matings were sporadic with low productivity and high pre-wean loss+ Subsequently, it was deemed critical to review the continuous mating schemeDiscussionIn addition to the measurable benefits published in this poster on the use of a peakmating scheme versus a continuous mating scheme, it was generally noted thatfemale hamsters showed reduced aggression when being handled. Wehypothesise the increased productivity observed in IVCs versus isolators to be dueto the females only being able to smell the one male scented to her, rather thanmany animals, as is the case within an isolator where animals are held in open topcages. In optimising our hamster breeding strategy Envigo has been able to readilysupport PHE’s cohort requirements which has proved critical during the COVID-19pandemic. The peak mating scheme has also been used with transgenic mousemodels and has proved equally successful. As a refinement, we plan to trial aprocess that reduces the requirement to single house males for an extended periodof time.Following the transfer of a hamster colony from the US to the UK, we set out tobreed cohorts of hamsters for PHE experimental studies. On review of theliterature, a monogamous continuous breeding scheme was established. Whilst theoutput from the first mating was not as successful as the US production colonyoutput, it was in line with expectations. Anticipated subsequent matings weresporadic with low productivity generating cohorts of animals that were unusable forthe customer and therefore went to waste. Consequently, it was necessary toreview the established breeding scheme. A peak mating scheme was selected.Pre-assigned monogamous pairs were housed separately for a period of 10 daysafter which the scent of the male hamster was introduced to the females cage bythe transfer of nesting material. Mating was predominantly observed immediatelyon introduction of the female into the male cage 48 hours after scenting. After 10days, the female was removed from the male cage and returned to her untouchedhome cage. On average, the three peak matings performed generated 20% morepups born than anticipated, based on the US production colony output, with anaverage litter size of 11. There was a concern that the larger litter sizes would bean increased burden on the female resulting in a higher pre-wean loss. Our datademonstrates that this is not the case. On average pre-wean loss was reduced to11%. This scheme enabled the production of large cohorts of animals born withinon average an 8 day window as per the customers specifications utilising fewernumbers of breeders to achieve the required output, therefore refining our breedingscheme and eliminating animal wastage.Method: Peak Mating in Isolators and IVCs+ All breeding pairs were separated for a period of 10 days+ Future monogamous pairs (9 males and 9 females) were selected+ 48 hours prior to mating, nesting material was removed from the female cage and placed into the corresponding male cage+ Simultaneously, nesting material was removed from the male cage and placed into the corresponding female cage+ On day 0 the female was transferred into the male cage for mating+ The female home cage was not cleaned or modified in any way+ On day 9 the female was removed from the male cage and transferred back to her home cageResults: Peak Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 100/81 123% 11 15% 9Average Weeks 1-27334/279 120% 11 11% 8Since transferring the colony to IVCs, the number of breeders successfullyproducing a litter has increased from 120% to 129% over a comparative period andthe average age range per cohort of pups born has been further refined to 1 day.Results: Continuous Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born(days)Weeks 1-5 119/135 88% 9 19% 4 daysAverage Weeks 1-27656/1881 35% 6 49%Highly variable and largely unpredictableEnvigo would like to say a very special thank you to PHE for their continued support without which this poster would not have been possible. MatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 26/18 144% 13 0% 1Average Weeks 1-27116/90 129% 11 11% 1Results: Peak Mating in IVCs* Colony was relocated to IVCs in Nov 2019ancompany0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27Expected ActualWEEK NUMBERNUMBER OF PUPS BORNA refinement in hamster breeding – creating a sustainable colony for vaccine researchAbstractAs a commercial breeder, Envigo had a requirement to relocate a hamsterbreeding colony from the US to the UK Contract Breeding Services (CBS) Facility.Due to Code of Practice specifications, there was a necessity to amend theestablished mating regime from trios (2 males, 1 female) to monogamous pairs. Onimplementing this regime, the number of successful matings was inconsistent andlitter sizes were below the expected output. Consequently, the current breedingprogram required review. Due to the lack of relevant literature, we had to establisha novel colony plan based on our existing data and expertise as breeders. Thisposter demonstrates an alternative scheme to produce a sustainable colony, with areduction in numbers of breeders used and increased productivity.K Anderson, M Davenport, J Finney, S MacCuishEnvigo, Hillcrest, Dodgeford Lane, Belton, Loughborough, Leicestershire, LE12 9TEIntroductionThe Golden Syrian Hamster, Mesocricetus auratus, is a desert rodent belonging tothe Cricetidae family. Due to its unique anatomical and physiological features, it iswidely used across a multitude of research areas. The Envigo HsdHan®:AURAcolony was first established in 1994 with stock originating from Zentralinstitut furVersuchstiere in Hannover, Germany. Envigo acquired the line along with theSprague-Dawley Company in 1973.In 2016, Envigo was approached by Public Health England, an executive agencyof the Government of the United Kingdom Department of Health and Social Care,to breed and supply cohorts of hamsters for a 4 year experimental program ofwork, focusing on the research and development of new treatments for ClostridiumDifficile related illnesses. At this time, the HsdHan®:AURA colony was based inEnvigo’s North American barrier facility. Due to the duration of this program ofwork and the considerable ethical burden associated with transatlantic travel it wasdeemed more appropriate to have this model readily available in the UK where theexperimental work was to be performed.On relocation of the line from the US to the UK, due to Code of Practicespecifications, there was a need to revise the established breeding strategy usedin the US of 2 males to 1 female. On review of the literature, of which there wasvery little relating to hamster breeding, monogamous pairings of hamsters fromweaning was deemed the most productive and labour efficient method ofbreeding. The colony was first housed in isolators before being relocated toindividually ventilated cages IVCs) in November 2019.0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected Actual0204060801001201401601 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected ActualMethod: Continuous Mating in Isolators+ 15 male and 15 female hamsters were imported from Envigo RMS US toestablish monogamous breeding pairs for continuous mating+ 15 monogamous pairs were first mated on 18thNovember 2017 with anexpected output of 135 pups born based on the average litter size of 9pups per female at Envigo’s US barrier+ Whilst the first mating was 88% successful expected versus actual output, anticipated subsequent matings were sporadic with low productivity and high pre-wean loss+ Subsequently, it was deemed critical to review the continuous mating schemeDiscussionIn addition to the measurable benefits published in this poster on the use of a peakmating scheme versus a continuous mating scheme, it was generally noted thatfemale hamsters showed reduced aggression when being handled. Wehypothesise the increased productivity observed in IVCs versus isolators to be dueto the females only being able to smell the one male scented to her, rather thanmany animals, as is the case within an isolator where animals are held in open topcages. In optimising our hamster breeding strategy Envigo has been able to readilysupport PHE’s cohort requirements which has proved critical during the COVID-19pandemic. The peak mating scheme has also been used with transgenic mousemodels and has proved equally successful. As a refinement, we plan to trial aprocess that reduces the requirement to single house males for an extended periodof time.Following the transfer of a hamster colony from the US to the UK, we set out tobreed cohorts of hamsters for PHE experimental studies. On review of theliterature, a monogamous continuous breeding scheme was established. Whilst theoutput from the first mating was not as successful as the US production colonyoutput, it was in line with expectations. Anticipated subsequent matings weresporadic with low productivity generating cohorts of animals that were unusable forthe customer and therefore went to waste. Consequently, it was necessary toreview the established breeding scheme. A peak mating scheme was selected.Pre-assigned monogamous pairs were housed separately for a period of 10 daysafter which the scent of the male hamster was introduced to the females cage bythe transfer of nesting material. Mating was predominantly observed immediatelyon introduction of the female into the male cage 48 hours after scenting. After 10days, the female was removed from the male cage and returned to her untouchedhome cage. On average, the three peak matings performed generated 20% morepups born than anticipated, based on the US production colony output, with anaverage litter size of 11. There was a concern that the larger litter sizes would bean increased burden on the female resulting in a higher pre-wean loss. Our datademonstrates that this is not the case. On average pre-wean loss was reduced to11%. This scheme enabled the production of large cohorts of animals born withinon average an 8 day window as per the customers specifications utilising fewernumbers of breeders to achieve the required output, therefore refining our breedingscheme and eliminating animal wastage.Method: Peak Mating in Isolators and IVCs+ All breeding pairs were separated for a period of 10 days+ Future monogamous pairs (9 males and 9 females) were selected+ 48 hours prior to mating, nesting material was removed from the female cage and placed into the corresponding male cage+ Simultaneously, nesting material was removed from the male cage and placed into the corresponding female cage+ On day 0 the female was transferred into the male cage for mating+ The female home cage was not cleaned or modified in any way+ On day 9 the female was removed from the male cage and transferred back to her home cageResults: Peak Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 100/81 123% 11 15% 9Average Weeks 1-27334/279 120% 11 11% 8Since transferring the colony to IVCs, the number of breeders successfullyproducing a litter has increased from 120% to 129% over a comparative period andthe average age range per cohort of pups born has been further refined to 1 day.Results: Continuous Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born(days)Weeks 1-5 119/135 88% 9 19% 4 daysAverage Weeks 1-27656/1881 35% 6 49%Highly variable and largely unpredictableEnvigo would like to say a very special thank you to PHE for their continued support without which this poster would not have been possible. MatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 26/18 144% 13 0% 1Average Weeks 1-27116/90 129% 11 11% 1Results: Peak Mating in IVCs* Colony was relocated to IVCs in Nov 2019ancompany0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27Expected ActualWEEK NUMBERNUMBER OF PUPS BORNA refinement in hamster breeding – creating a sustainable colony for vaccine researchAbstractAs a commercial breeder, Envigo had a requirement to relocate a hamsterbreeding colony from the US to the UK Contract Breeding Services (CBS) Facility.Due to Code of Practice specifications, there was a necessity to amend theestablished mating regime from trios (2 males, 1 female) to monogamous pairs. Onimplementing this regime, the number of successful matings was inconsistent andlitter sizes were below the expected output. Consequently, the current breedingprogram required review. Due to the lack of relevant literature, we had to establisha novel colony plan based on our existing data and expertise as breeders. Thisposter demonstrates an alternative scheme to produce a sustainable colony, with areduction in numbers of breeders used and increased productivity.K Anderson, M Davenport, J Finney, S MacCuishEnvigo, Hillcrest, Dodgeford Lane, Belton, Loughborough, Leicestershire, LE12 9TEIntroductionThe Golden Syrian Hamster, Mesocricetus auratus, is a desert rodent belonging tothe Cricetidae family. Due to its unique anatomical and physiological features, it iswidely used across a multitude of research areas. The Envigo HsdHan®:AURAcolony was first established in 1994 with stock originating from Zentralinstitut furVersuchstiere in Hannover, Germany. Envigo acquired the line along with theSprague-Dawley Company in 1973.In 2016, Envigo was approached by Public Health England, an executive agencyof the Government of the United Kingdom Department of Health and Social Care,to breed and supply cohorts of hamsters for a 4 year experimental program ofwork, focusing on the research and development of new treatments for ClostridiumDifficile related illnesses. At this time, the HsdHan®:AURA colony was based inEnvigo’s North American barrier facility. Due to the duration of this program ofwork and the considerable ethical burden associated with transatlantic travel it wasdeemed more appropriate to have this model readily available in the UK where theexperimental work was to be performed.On relocation of the line from the US to the UK, due to Code of Practicespecifications, there was a need to revise the established breeding strategy usedin the US of 2 males to 1 female. On review of the literature, of which there wasvery little relating to hamster breeding, monogamous pairings of hamsters fromweaning was deemed the most productive and labour efficient method ofbreeding. The colony was first housed in isolators before being relocated toindividually ventilated cages IVCs) in November 2019.0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected Actual0204060801001201401601 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27NUMBER OF PUPS BORNWEEK NUMBERExpected ActualMethod: Continuous Mating in Isolators+ 15 male and 15 female hamsters were imported from Envigo RMS US toestablish monogamous breeding pairs for continuous mating+ 15 monogamous pairs were first mated on 18thNovember 2017 with anexpected output of 135 pups born based on the average litter size of 9pups per female at Envigo’s US barrier+ Whilst the first mating was 88% successful expected versus actual output, anticipated subsequent matings were sporadic with low productivity and high pre-wean loss+ Subsequently, it was deemed critical to review the continuous mating schemeDiscussionIn addition to the measurable benefits published in this poster on the use of a peakmating scheme versus a continuous mating scheme, it was generally noted thatfemale hamsters showed reduced aggression when being handled. Wehypothesise the increased productivity observed in IVCs versus isolators to be dueto the females only being able to smell the one male scented to her, rather thanmany animals, as is the case within an isolator where animals are held in open topcages. In optimising our hamster breeding strategy Envigo has been able to readilysupport PHE’s cohort requirements which has proved critical during the COVID-19pandemic. The peak mating scheme has also been used with transgenic mousemodels and has proved equally successful. As a refinement, we plan to trial aprocess that reduces the requirement to single house males for an extended periodof time.Following the transfer of a hamster colony from the US to the UK, we set out tobreed cohorts of hamsters for PHE experimental studies. On review of theliterature, a monogamous continuous breeding scheme was established. Whilst theoutput from the first mating was not as successful as the US production colonyoutput, it was in line with expectations. Anticipated subsequent matings weresporadic with low productivity generating cohorts of animals that were unusable forthe customer and therefore went to waste. Consequently, it was necessary toreview the established breeding scheme. A peak mating scheme was selected.Pre-assigned monogamous pairs were housed separately for a period of 10 daysafter which the scent of the male hamster was introduced to the females cage bythe transfer of nesting material. Mating was predominantly observed immediatelyon introduction of the female into the male cage 48 hours after scenting. After 10days, the female was removed from the male cage and returned to her untouchedhome cage. On average, the three peak matings performed generated 20% morepups born than anticipated, based on the US production colony output, with anaverage litter size of 11. There was a concern that the larger litter sizes would bean increased burden on the female resulting in a higher pre-wean loss. Our datademonstrates that this is not the case. On average pre-wean loss was reduced to11%. This scheme enabled the production of large cohorts of animals born withinon average an 8 day window as per the customers specifications utilising fewernumbers of breeders to achieve the required output, therefore refining our breedingscheme and eliminating animal wastage.Method: Peak Mating in Isolators and IVCs+ All breeding pairs were separated for a period of 10 days+ Future monogamous pairs (9 males and 9 females) were selected+ 48 hours prior to mating, nesting material was removed from the female cage and placed into the corresponding male cage+ Simultaneously, nesting material was removed from the male cage and placed into the corresponding female cage+ On day 0 the female was transferred into the male cage for mating+ The female home cage was not cleaned or modified in any way+ On day 9 the female was removed from the male cage and transferred back to her home cageResults: Peak Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 100/81 123% 11 15% 9Average Weeks 1-27334/279 120% 11 11% 8Since transferring the colony to IVCs, the number of breeders successfullyproducing a litter has increased from 120% to 129% over a comparative period andthe average age range per cohort of pups born has been further refined to 1 day.Results: Continuous Mating in IsolatorsMatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born(days)Weeks 1-5 119/135 88% 9 19% 4 daysAverage Weeks 1-27656/1881 35% 6 49%Highly variable and largely unpredictableEnvigo would like to say a very special thank you to PHE for their continued support without which this poster would not have been possible. MatingActual pups born versus expected pups bornSuccess %Average litter sizePre-wean lossAge range per cohort of pups born (days)Weeks 1-5 26/18 144% 13 0% 1Average Weeks 1-27116/90 129% 11 11% 1Results: Peak Mating in IVCs* Colony was relocated to IVCs in Nov 2019ancompany0204060801001201 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27Expected ActualWEEK NUMBERNUMBER OF PUPS BORNContinuous MatingPeak Mating – IsolatorsPeak Mating – IVCs

80Animal Technology and Welfare August 2020DiscussionIn addition to the measurable benefits published in this poster on the use of a peak mating scheme versus a continuous mating scheme, it was generally noted that female hamsters showed reduced aggression when being handled. We hypothesise the increased productivity observed in IVCs versus isolators to be due to the females only being able to smell the one male scented to her, rather than many animals, as is the case within an isolator where animals are held in open top cages. In optimising our hamster breeding strategy Envigo has been able to readily support PHE’s cohort requirements which has proved critical during the COVID-19 pandemic. The peak mating scheme has also been used with transgenic mouse models and has proved equally successful. As a refinement, we plan to trial a process that reduces the requirement to single housed males for an extended period of time. AcknowledgementsEnvigo would like to say a very special thank you to PHE for their continued support without which this poster would not have been possible. References1 Code of Practice for the Housing and Care of Animals Bred, Supplied or Used for Scientific Purposes. Print ISBN 9781474112390. Web ISBN 978147411240. https://assets.publishing.service.gov.uk/Poster Presentations

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