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ATW April 2019

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IAT JournalAnimal Technologyand WelfareOfficial Journal of the Institute of Animal Technologyand European Federation of Ani mal TechnologistsVol 18 No 1 April 20 19ISSN 1742-0385G Education, Education EducationG Intravenous Training ProjectG Animal Technology Exchange ProgrammeG Congress 18 Posters – Part 3G Edinburgh 3Rs symposium posters

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Editorial ixJas Barley, Chair of the Editorial BoardAn introduction to the Higher Education Programme Level 6 1Intravenous Injection Training Project: A Level 6 Higher Education project 3Carmen AbelaAnimal Technologist Exchange Programme: A Level 6 Higher Education project 19Matthew BiltonPAPER SUMMARY TRANSLATIONS 31TECH-2-TECHUsing software to tackle the 3Rs – A scientist’s perspective 39Chris W.D. ArmstrongANDREW BLAKE TRIBUTE AWARDGive us a break! 45Emma FilbyPOSTER PRESENTATIONSImpact of water hardness on Zebrafish early stage development 49Carole Wilson, Paul Barwood, Visilia Moiche, Jenna Hakkesteeg and Karen DunfordComparing Zebrafish embryo production methods 53Heather Callaway and Carole WilsonComparison of feeding regimes for Zebrafish (Danio rerio) 56Paul Barwood and Carole WilsonUsing novel equipment to aid reduction and replacement of the ferret model in influenza transmission 59Teresa BorehamOptimising health & welfare tracking using The Sanger Mouse Database 62Joshua Dench, Robbie McLaren-Jones, Valerie Vancollie, Emma Siragher, David Tino Lafont, Lauren Anthony,Simon Maguire, Mark Griffiths, Hannah Wardle-Jones, Carl Shannon, James Bussell and Chris LelliottThe use of Hyperova®to produce oocytes from aged WT mice and small groups of transgenic 65mice for IVF rederivationJulie Thomson, Ailsa Travers, Jacek Mendr ychowski and Emma AllanAllele conversion using cell-permeable Cre recombinase 68Ailsa Travers, Julie Thomson and Matt SharpThe use of a par-vaginal impedance checker to improve rat plugging efficiency 70Julie Thomson and William MungallRefining mouse re-derivation by using IVF with fresh or frozen sperm as opposed to embryos 73Ailsa Travers, Julie Thomson and Matthew SharpUsing subcuticular stitching in rats to replace skin closure clips as a refinement 75Julie Thomson and William MungallIAT recognised in South Africa 77Jabulane S. Magagula and Bert J. MohrInstructions to Authors 78CONTENTSVol 18 No 1 April 2019iApr LATEST:Animal Technology and Welfare 22/3/19 12:27 Page i

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iiIAT REPRESENTATIVESOFFICERSPresidentDr Robin Lovell-Badge CBE FRSImmediate Past PresidentProfessor Sir Richard Gardner MA PhD FRSB FIAT (Hon) FRSVice-PresidentsDavid Anderson MRCVS, Stephen Barnett BA MSc FIAT (Hon)CBiol FRSB RAnTech, Miles Carroll PhD, Brian Cass CBE,Gerald Clough BSc PhD EurBiol CBiol MRSB SFZSL,Paul Flecknell MA Vet MB PhD DLAS DipLECVA MRCVS, FIAT (Hon)Penny Hawkins PhD BSc, Wendy Jarrett MA,Judy MacArthur-Clark CBE BVMS DLAS FRSB DVMS (h.c.), DipECLAMFRAgS DipACLAM MRCVS, Fiona McEwen BSc BVM&S MSc MRCVS,Tim Morris BVetMed PhD DipACLAM DipECLAM CBiol FRSB CertLASMRCVS, José Orellana BVSc MSc, Clive Page OBE PhD BSc,Jan-Bas Prins PhD MSc, Vicky Robinson CBE BSc PhD,Gail Thompson RLATG, Robert Weichbrod PhD RLATGLife MembersCharlie Chambers MIAT RAnTech, Roger Francis MSC FIAT RAnTech,Pete Gerson MSc FIAT RAnTech, Cathy Godfrey FIAT RAnTech,John Gregory BSc (Hons) FIAT CBiol FRSB RAnTech, Patrick HayesFIAT DipBA RAnTech, Robert Kemp FIAT (Hon) RAnTech,Phil Ruddock MIAT RAnTech, Ted Wills HonFIAT RAnTech,Honorary MembersMark Gardiner MIAT RAnTech, Andy Jackson MIAT, Sarah Lane MScFIAT, Brian Lowe MSc FIAT RAnTech, Sue McHugh BSc FIAT,Norman Mortell BA (Hons) MIAT RAnTech, Terry Priest MBE FIATRAnTech, Trevor Richards BEM MIAT, David Spillane FIAT,Wendy Steel, Pete Willan DMS FInstLM MIATMembers of CouncilKen Applebee OBE, Matthew Bilton, Kally Booth, Charlie Chambers,Steven Cubitt, Simon Cumming, Haley Daniels, Glyn Fisher,Nicky Gent, Cathy Godfrey, Alan Graham, Nathan Hill, Linda Horan,Sam Jameson, Elaine Kirkum, Adele Kitching, Theresa Langford,Sylvie Mehigan, Steve Owen, Alan Palmer, Allan Thornhill,John Waters, Lynda Westall, Carole Wilson, Adrian WoodhouseCouncil OfficersChair: Ken Applebee OBE FIAT CBiol FRSB RAnTechVice Chair: Linda Horan BSc (Hons) MIAT RAnTechHonorary Secretary: Linda Horan BSc (Hons) MIAT RAnTechHonorary Treasurer: Charlie Chambers MIAT RAnTechAssistant Treasurer: Glyn Fisher FIAT RAnTechChair Board of Educational Policy: Glyn Fisher FIAT RAnTechChair Board of Moderators: Haley Daniels MBA MSc MIAT RAnTechCIPDChair Registration & Accreditation Board: Ken Applebee OBE FIATCBiol FRSB RAnTechATW Editor: Jas Barley MSc FIAT RAnTechBulletin Editor: Carole Wilson BSc MIATBranch Liaison Officer: Lynda Westall BSc (Hons) FIAT DMS RAnTechEFAT Representative: Charlie Chambers MIAT RAnTechWebsite Coordinator: Allan Thornhill FIAT RAnTechAnimal Welfare Officers and LABA Representatives:Matthew Bilton, Kally Booth, Simon Cumming, Nicky Gent,Sylvie Mehigan, John WatersATW/Bulletin Editorial Board: Jas Barley (Chair), Matthew Bilton,Nicky Gent, Patrick Hayes, Elaine Kirkum, Carole Wilson,Lynda WestallBoard of Educational Policy: Glyn Fisher (Chair), Steven Cubitt(Secretary), Adele KitchingBoard of Moderators: Haley Daniels (Chair), Simon Cumming,Cathy GodfreyModerators: Anthony Iglesias, Theresa Langford, Jenny Parks,Sarah ReedCommunications Group: Adrian Woodhouse (Chair), Nathan Hill,Elaine Kirkum, Teresa Langford, Sylvie Mehigan, Allan Thornhill,Lynda WestallCPD Officer: Charlie ChambersRegistration and Accreditation Board: Ken Applebee (Chair),Charlie Chambers, John Gregory, Cathy Godfrey, Gerald Clough,Kathy Ryder (Home Office), Stuart StevensonObserver: Ngaire Dennison (LAVA)Congress Committee: Alan Graham (Chair), Haley Daniels,Linda Horan, Adele Kitching, Allan Thornhill, John WatersDiversity Officer: Haley Daniels MBA MSc MIAT RAnTech CIPDUK Biosciences ASG Representative/Home Office: Steve Owen,Charlie Chambers, Alan PalmerIAT OFFICERS MAY BE CONTACTED VIA:IAT Administrator: OR VIA THE IAT WEBSITE VIA THE REGISTERED OFFICE:5 South Parade, Summertown, Oxford OX2 7JLAdvertisement Managers: PRC Associates LtdEmail: every effort is made to ensure that no inaccurate or misleading data,opinion or statement appear in the journal, the Institute of Animal Technologywish to expound that the data and opinions appearing in the articles, posterpresentations and advertisements in ATW are the responsibility of thecontributor and advertiser concerned. Accordingly the IAT, Editor and theiragents, accept no liability whatsoever for the consequences of any suchinaccurate or misleading data, opinion, statement or advertisement beingpublished. Furthermore the opinions expressed in the journal do notnecessarily reflect those of the Editor or the Institute of Animal Technology.© 2019 Institute of Animal TechnologyAll rights reserved. No part of this publication may be reproducedwithout permission from the publisher.BRANCH SECRETARIES 2019Cambridge: Sarah Shorne Janice Young & Essex: Joanna Cruden, Suffolk & Norfolk: Jo Martin Lisa Watson Rebecca Towns Ian Fielding East England: Rachel Sandy and Joanne Bland West: Nicky Windows April Shipton, Hampshire & Sussex: Francesca Whitmore Middlesex: Josefine Woodley of Scotland: Linda Horan LATEST:Animal Technology and Welfare 22/3/19 12:27 Page ii

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April 2019 Animal Technology and WelfareEditorialJas BarleyChair of the Editorial BoardAs you know we frequently have themed issues, sometimes they are planned and sometimes they just happen as is thecase with this issue. Planning for each issue starts at least three months in advance. In the case of the April issuebecause of the Christmas break interrupting things and as we like to have it available at IAT Congress each year, it oftenstarts even earlier – I started this issue in November last year. At the time there was no plan but over the weeks theedition has developed an education theme.For over 60 years the main raison d’etre of the IAT has been education. Although Tony Blair may have used the sloganeducation, education, education, I like to think he got it from us! Our motto curando docemus – through learning we care,to my mind says it all. Animal Technology qualifications have been available almost from the start of the Institute and oureducation programme is expanding to meet the demands of an increasingly technical based industry. Gone are the dayswhen we were just cage scrapers and the growing reliance on technology demands technologists with more complex skills.Fortunately, the Institute has been able to meet this need and is now able to offer Higher Education (HE) qualifications.The main reason we have the Journal of Animal Technology and Welfare is of course education. By disseminating the workof Animal Technologists and our colleagues we can improve the welfare of the animals we care for. This is still importantfor all Animal Technologists but especially those who are unable to attend meetings and courses.I am delighted to be able to include an introduction to the Level 6 Higher Education programme by one of the leadingadvocates of the programme Steven Cubitt as well as two articles based on Level 6 projects. Both these projects lookat training and expanding technicians’ skills. Carmen Abela’s project discusses an approach for training researchers andtechnologists in intravenous tail vein injections, one of the more difficult procedures to master. Matt Bilton, alongside hisrole as Chair of the IAT Animal Welfare Group, has looked at the development of an Animal Technologists’ ExchangeProgramme. Both papers on the projects are lengthy but are well worth reading. As one of the participants of the firstPost Graduate Animal Technology courses which eventually led to my gaining my MSc, I know that the students willremember the days spent on the course with fondness for not only do you learn about your profession but you establishfriendships that will stay with you the rest of your life. I look forward to seeing the latest students receive their degreesat this year’s graduation.However, it is not all good news as it appears that some major employers and, in particular, a few of the Universities, arenot financially supporting technologists on courses above Level 3. Continuing Professional Development (CPD) is arequirement both here in the UK under ASPA and in Europe via the 2010 Directive so theoretically we should be continuingour education until at least the day we retire. Thankfully the charity Animals in Science Education Trust) (AS-ET), can helpby providing bursaries to attend both relevant courses and meetings for those working with animals in science.More concise offerings are the last few posters from IAT Congress 2018 and some from the University of Edinburgh, 3Rs’symposium. Congress posters include another education-based offering with the poster by Jabulane Magagula and BertMohr from the University of Cape Town. This outlines how IAT qualifications are now available in South Africa.Although the material for this issue of ATW is at the printers, the work is not over and I must now start thinking aboutAugust’s issue, it is a bit like education – there is always something new!THE INSTITUTE OF ANIMAL TECHNOLOGYETHICAL STATEMENTOur purpose is to advance knowledge and promote excellence in the care and welfare ofanimals in science and to enhance the standards and status of those professionallyengaged in the care, welfare and use of animals in science.ixApr LATEST:Animal Technology and Welfare 22/3/19 12:27 Page ix

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April 2019 Animal Technology and Welfare1An introduction to the Higher EducationProgramme Level 6The course is part of the prospectus of the College ofLaboratory Animal Science and Technology (CLAST).CLAST is a non-profit course provider that deliversHigher Education (HE) qualifications designed for thoseworking with animals in research, including AnimalTechnologists, Registered Veterinary Professionals andSenior Researchers.The Higher Education provides Levels 4-6 IAT HEDiplomas and CPD units in a range of subjects specificto Laboratory Animal Science and Technology.Qualifications are awarded by the IAT are designed to prepareAnimal Technologists and related professionals for therole of senior and middle management, or specialistroles within the animal facility.Animal Facility Design andManagement Level 6 ProjectManagement – ProjectIntroductionLevel 6 HE units can be taken as stand-alone CPDcourses. On completion an IAT CPD Certificate for theunit studied will be awarded. These certificates can beused at any time to contribute towards a full HEqualification; there is no requirement to repeat the unit.Animal Facility Design and ManagementThis unit is designed to develop knowledge ofbiomedical facility design but equips students with arange of vital practical skills, as their careers progressinto a more senior role. The unit covers order ofmagnitude budget setting, financial management andhow to manage risk accountably, introducing a range ofuseful management tools. Students develop theirunderstanding of all aspects of briefing, specification,design and construction. This enables the student tounderstand and engage with both the project team andto represent the institutional requirements effectively.The unit also covers legislative aspects and developsskills on the translation of these into the brief.This issue of Animal Technology and Welfare contains two papers based on project carriedout as part of the Level 6 Animal Facility Design and Management Module: ProjectManagement. It is intended that these will be the first in a series of papers from Level 6students.Students learning about stages of design and buildprojects from an architect on a visit to an architecturalpractice.Because change and development are continual in theindustry, the central focus of the management part ofthe unit is on managing change. Students are taughtthe leadership skills required to manage change withconfidence, bringing staff on board in commitment tocontinuous improvement where the goal is the veryhighest standard in science, animal welfare, staffwelfare and dignity at work.Besides written assignments, students visit a sitewhere a successful change in the workplace culturehas been achieved, ask questions, discuss freely andengage in practical exercises and role play to addressa range of workplace problems. Students also designand deliver a presentation and answer stakeholderquestions, building their communication skills andconfidence to interact confidently with stakeholders atall levels.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 1

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Project ManagementTheory and PracticeThis unit is ideally geared towards people who are nowprogressing their career in a management position.Students get to grips with relevant managementtheories underpinning their practical experience atwork. So often, difficulties arise in the workplace wheninsufficient consideration is given to effectivecommunication strategies. Misunderstandings anddisagreements arise which can impede the progress ofimportant work objectives.Students are encouraged to identify the keystakeholders they will be working with, whoseperspectives and concerns may be very different fromthose of the Animal Technologist; senior academics,representatives and managers of various otherdepartments within their organisation andprofessionals such as engineers, architects andsurveyors, to name but a few. The unit is deliveredthrough practical taught sessions, distance learningand visits to sites of interest.Students develop effective collaborative working andcommunication strategies and get to hone theirinterviewing, negotiating and information gatheringskills in a range of course activities designed to buildconfidence in understanding the ‘bigger picture’ whilststaying on track in the world of management. There isa balance of academic study, management, soft skillsand learning through shared experience.ProjectBuilding on learning acquired during the ProjectManagement; Theory and Practice unit. This unit givesthe student the opportunity to complete a writtenaccount on their own project and present the projectbenefits and achievements to key stakeholders.Students are invited to outline, plan and evaluate aproject which could fall into any of the followingcategories:1. Scientific/Animal Welfare research.2. Biomedical facility project.3. Biomedical business or training project.The unit is taught through a flexible tutorial modelwhich can be accessed by email, telephone or skypedepending on individual need and is ideal for peoplecombining study with work and home responsibilities.Support is tailored to meet the needs of the individual.The unit is academically robust, requiring submissionof a project proposal to agreed guidelines, twoextended written reports and a presentation on theproject objectives, achievements and benefits.Students are encouraged to evaluate the lessonslearnt and demonstrate a commitment to both thehighest standards of animal and staff welfare and theirown continuous professional development.An introduction to the Higher Education Programme Level 62Students visit a Building Information Management(BIM) room.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 2

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3April 2019 Animal Technology and WelfareIntravenous Injection Training Project:A Level 6 Higher Education projectCARMEN ABELABiological Services Facility, Faculty of Infectious and Tropical Diseases, London School ofHygiene and Tropical Medicine, Keppel Street, London WC1E 7HTCorrespondence: (i.v.) injection via the lateral tail vein is aroute of administration when using animals in medicalresearch. It is the preferred route delivery ofsubstances, especially where avoidance of the first-pass effect through hepatic metabolism is desired. It isalso used to mimic the proposed route of finaladministration. However, it is technically difficult andrequires specialist training. Inaccuracies duringinjection are common which deter researchers fromusing the route, preferring instead to use morecommon routes of dosing such as intraperitonealinjection or oral gavage.The use of live animals to gain technical skills isethically and legally prohibited and usually technicalcompetence is gained by practising on cadavers. Thisproject aims to establish whether there is a need touse animal cadavers – dead mice for training orwhether an alternative training tool is as effective. Thedesign of a training project is integral to the outcomeand in order for it to be successful, it must be on time,within budget and to specification.Key words: training, intravenous injections, mouse,cadavers, technical skills.Part 1 – Planning and scopingSection 1: Statement of the contextThe London School of Hygiene and Tropical Medicine(LSHTM) is a world-leading centre for research andpostgraduate education in public and global health.Most research carried out using animals relates toneglected or tropical disease and it is renowned for itsresearch as one of the highest rated institutes in thecountry. The school’s mission is to improve healthequity in the UK and worldwide with an aim to createpolicy, excellence in research, education andtranslation of knowledge.It has two buildings based in Bloomsbury and twoMedical Research Council (MRC) units in the Gambiaand Uganda and over 1300 staff working in over 100different countries. LSHTM deploy research in real timecrises and deployed staff during the Ebola crisis inLiberia and Sierra Leone in 2014.1A world leader in infectious tropical disease research,some carried out in the biological services facilitywhere scientists are working on vaccines, treatmentsand cures, to understand the basic biology andprogression of disease. Currently 5 full-time and 2 part-time Animal Technologists cover all animal work. Thefacility holds mice, hamsters, Zebrafish and all work isunderpinned by the 3Rs principles – Reduction,Replacement and Refinement. A member of theConcordat of Openness on Animal Research, animportant objective is the publication of statisticallyvalid data of the highest scientific merit, which ispublicly available and peer reviewed. Researchinfections include tuberculosis, malaria, leishmaniosisand Trypanosome cruzi.2Intravenous (i.v.) injection through the lateral tail vein ofmice in medical research is a common route ofadministration with benefits including rapid distributionthrough the bloodstream,3absorption buffering anddilution.4Experimental design researches which is theoptimum route of administration for the substancesbeing used, with intravenous often used to avoid the‘first-pass’ effect through hepatic metabolism.Additionally, i.v. is used to mimic the proposed route offinal administration.Intravenous tail vein injections into the lateral tail veinsof mice require specialist training as injectioninaccuracy is commonplace. Injection into the lateraltail vein of mice is a procedure used by a smallernumber of researchers than other, more commonroutes of dosing, such as intraperitoneal injection (ip)or oral gavage often due to technical difficulty.Intraperitoneal administration results in slowerabsorption into the vasculature. It may be used to injectlarger volumes, although is similar to parenteralApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 3

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Intravenous Injection Training Project: A Level 6 Higher Education project4administration (oral gavage or insertion of liquidstraight into the stomach). With the i.p. routesubstances pass through the mesenteric vessels whichdrain into the portal vein and pass through to the liver.5Data was analysed from the BSF in 2017 and therewas a high failure rate for i.v. administration of 42%. Incomparison, researchers assessed for intraperitonealinjection in 2017 had a success rate of 100%.Most literature available explains how to carry out thisprocedure on live animals. Legally and ethically, atrainee should not carry out this or any procedure on alive animal until they are at least competent on acadaver.6This study will establish whether there is a need to useanimals (cadavers – dead mice) for training or whetheran alternative training tool is as effective. According toMesser5apart from the skill of the experimenter, usingthe right technical aids and material is critical forsuccessful intravenous injections in mice.The design of a training project is integral to theoutcome and in order for it to be successful, it must beon time, within budget and to specification. Essential tothe project are the participants, trainer and assessor orstakeholders.7The use of cadavers for training in biomedical settingsis a controversial one, particularly concerning ethicsdepending on where or how animals are sourced.8TheNational Centre for the Replacement, Refinement andReduction of Animals in Research (NC3Rs) launch in2004 was in response to a House of Lords’ selectcommittee report, Animals in Scientific Proceduresreport, which recommended a National Centre.9TheNC3Rs aim is to reduce, refine and replace animals inscientific studies and provide e-learning resources,which help toward training.The Biological Services Facility (BSF) at LSHTM is aContainment Level 3 (CL3) facility with aerosolcapacity. A small facility with only 56 Personal Licenceholders (PILs) of whom the majority have nointravenous lateral tail vein experience.This study aims to compare training with an aid, whichif successful, could potentially remove the need forcadavers and eliminate the problem of unethical animaluse in practical training. A minimum of 20 participantswill be required for relevant statistical analysis.Enrolment of participants will take place throughdiscussion with the Named Training and CompetenceOfficer (NTCO) to take part in a research project with asurety that should they fail their competenceassessment in the trial, training would continue untilcompetence success. All participants expressed a wishto learn this skill for future experimental purposes,some explaining that it would have been a first-choicemethod but difficulty with this method made themchoose an alternative route.A trainer who could teach this technique in bothcadaver and the tool and an assessor ‘blinded’ to theteaching method is essential for the project as well asparticipants. The trainer will randomly train in onemethod or another until considered competent at thatlevel. Number of training sessions and a formalassessment of competence on a live animal by anassessor will take place and result recorded.Context has great bearing on a project and opens upstrategic issues that the project will need to address.Tonnquist (2010)10has four criteria suggested as thebasis for a projectG determined and defined goal – unique taskG determined time frame – set scheduleG determined resources – own budgetG special work model – temporary organisationThese points will be addressed in order for projectsuccess.Project work is often used in the short term tocomplete a task, especially innovative techniques.Learning within a project does not always transfer tothe day-to- day organisational setting. Project work isseen as useful in allowing precise autonomy anddiscretion with an allowance for flexibility in order tomove the project along. Usually they are employed in acurrent or forward thinking setting and respond to rapidchanges in the market.Organisational learning, particularly from projectsshould be successful but quite often fails due to a lackof capturing or translating newly learned routines orpractices. Organisational learning tends to happen insub-groups and may not be shared or transferable on alarger scale. Projects tend to be in isolation from thewider organisational context.Another difference between project individuals andorganisations is that project members do notnecessarily see themselves as part of a team as theymay enter and leave the project at different points.Organisation tends to have a strategy or vision aimedat the whole team working towards a shared practice.11Project based learning is closely related to problem-based learning. Learning is driven by challenging open-ended problems and can be interchangeable.12Saveryand Duffy12suggest true knowledge lies in ourinteractions with our environment.Teamwork emphasis on learning for a project is anApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 4

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Intravenous Injection Training Project: A Level 6 Higher Education project5important factor, this project could not run without it.Distributive cognition or the study of knowledge beingpassed on and growing is also an important aspect ofproject success.13Distributive cognition is concerned with structure andfinding a stable design which can be widely applicable.In the normal working environment, teamwork is alsoan important factor with distribution of knowledge andskills. This tends to happen over a longer period incomparison to a project where adhering to timeframesare essential for success.The defined goal or unique task in this project will bethe practical task of teaching intravenous tail veininjections to participants (a training method) with therecognised defined goal being the non-use of animals(a successful training method).The determined time frame will be realised through aGantt chart as a realistic set schedule. A budget will benecessary to purchase equipment for the project. Agrant proposal must be put together to apply forfunding.The special work model is a temporary re-organisationof staff. The project will need use of staff resources forsuccess.Section 2: SynopsisIn this section, I will review previous research and theproblems this project will address. The purpose of thisstudy is comparative to identify whether two alternativemethods of training in intravenous administration intolaboratory mice via the lateral tail veins are equal orbetter regarding competence outcome.The physical characteristics of these models includesize, composition and ease of administration withconsideration of ethical acceptance.This study will use wild type outbred cadavers versus atraining tool (Mimicky®mouse model)14to comparecompetence outcome.15,16A learning and developmentstrategy is employed at LSHTM in order to developskills and competencies as well as maintainingContinued Professional Development (CPD). The NTCOis tasked with identifying gaps in skills or education andensuring training is available. SMART (specific,measurable, achievable, realistic, time-bound)objectives are useful as a measure and outline ofachievable goals. Procurement is also important for acost-effective approach and understanding.There appears to be a problem in animal researchwhere researchers cannot train in a practicable manneron their chosen animal model because:G it is a live animalG the law prohibits thisThis however, poses a huge potential gap or absence intraining, as the route that must be followed tends to beobservational and on either cadavers or simulators.There seems to be disparity among researchinstitutions and the methods they employ with varyingdegrees of confidence.17For this study participants will undertake training inparallel groups in a randomised controlled trial. Datawill be analysed by the number of training sessionsrequired and attempts to achieve competence.The result of this study will determine whethercadavers are necessary as training tools to achievecompetence in i.v. administration into the lateral tailveins of mice.The anticipated outcome would be that the training toolis equally suitable removing the need for real mice.These findings may be useful as a 3Rs (Replacement,Refinement, Reduction) improvement to eliminate theneed for real mice.18Intravenous administration has a high failure rate. Outof 1054 people on death row in the United States ofAmerica awaiting execution from 1890 to 2010 bylethal injection, 75 attempts or 7.12% of instancesinitially failed for various reasons. Failure varied fromdifficulty in finding a suitable vein (often due toprevious drug abuse in the prisoners), collapsed veins,a blowout where the syringe comes out of the vein,allergic reaction to the drugs, equipment failure, needleinserted incorrectly and flowing away from the heart orpuncturing through the vein resulting in administrationinto the soft tissue. The main cause of failure washuman error with questions raised regarding theexperience and training of prison staff in i.v.administration.19Tail vein injections require skill and expertise, onaverage 14% of material injected by this method sits inthe tail tissue.20An Automated Vascular System (A-VAS) system hasbeen developed to overcome this by Chang et al20but itis not readily available on the market and requiresanaesthesia, system set up and considerably moretime for successful injection.Qualitative and quantitative methods can be used,qualitative by the researcher and level of resistancewhen injecting and quantitative by Positron EmissionTomography (PET) scans with the disadvantage beingtime and cost.21There are three veins and one artery in the tail of aApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 5

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Intravenous Injection Training Project: A Level 6 Higher Education project6mouse. Two lateral veins and one dorsal caudal vein aswell as a ventral caudal artery. The lateral veins arethick enough to allow for intravenous administration aswell as blood collection.22Failure to inject successfully can occur for a number ofreasons such as:Incorrect placement of the needle: Needle inserted intothe artery and not one of the veins. The mouse has twolateral tail veins and injection initiated nearer the tip ofthe tail, if failure results there is an opportunity toattempt again half way along the tail and further nearthe base of the tail. This technique allows for sixattempts if required in order to achieve successfulinjection in the mouse model or cadaver. Four attemptsmaximum in a live mouse as any more may causedistress.Incorrect bevel position: The bevel should always faceupwards towards you for insertion. This allows for lessresistance when puncturing the skin. If the bevel isincorrectly positioned it may cause occlusion to thelumen.Incorrect needle size: There are various guidelinesregarding recommended needle gauge size for thisprocedure, the smallest chosen, as a refinement. 27gauge is standard size. Using too large a needle mayresult in tissue trauma and too small may result inpressure to the tail from prolonged plunger depression.Length is equally important as too small may result insubcutaneous injection and too long may be difficult todepress resulting in puncture to the vein.Subcutaneous placement: This results in a bleb orbulge occurring on the tail usually seen by the nakedeye.Needle pierced through vein: Needle inserted too farexiting through the vein resulting in a subcutaneousinjection.Movement when aspirating syringe: If restraint is notfirm enough and allows movement the needle candislodge from the original position.Blunt needle: Each time a needle is used it becomesdulled therefore should not be used for any more than4 times for competence assessment beforereplacement.Whilst there are alternative methods of measuringsuccess or failure of the administration of material intothe lateral tail vein of mice there is no previousresearch or studies investigating the ethical context ofthe use of cadavers.Henri Fayol’s management theory is useful in keepingmanagement targets for the project on track.23Fayolhas 14 principles of management but these can bebroken down into 5 main components.– Plan – A good plan needs to be flexible in itsapproach. It must be appropriate for the businessand the staff involved needs to be competent tocarry out tasks.– Organise – Includes communication, responsibility,authority and ensuring resources are available.– Co-ordinate – For the project to run smoothly eachpart must be completed in time for the next part ofthe project to start. Many parts may need to runconcurrently.– Command – The manager of the project should havea good understanding of each aspect of the project.Understand how the staff involved work and removestaff who are incompetent or slowing the projectdown unnecessarily.22Project goals and objectivesSMARTThere is no previous research in the field of comparinglateral tail vein injection training methods in a researchsetting using mouse cadavers and a simulator.A study to compare the two methods to determine ifsimulator training is as effective is appropriate asethically to use cadavers is a controversial topic.Training will take place over the course of a year inorder to encapsulate all researchers. The trainer willdetermine how many sessions are necessary beforeadvising a competence assessment.A member of staff whom is ‘blinded’ to the trainingmethod will undertake the competence assessmentsto determine a pass or fail result. i.v. injection outcomecan be easily assessed as the solution (Sodiumpentobarbitone) will either flow through the veinresulting in schedule 1 euthanasia or the plunger willnot depress correctly. If unsuccessful it has no effecton the animal. All mice in this study are excessbreeding stock and scheduled for euthanasia.ObjectiveTo reduce the use of cadavers for training in i.v. lateraltail vein procedure by using a simulator, through acomparison research study and a consultativeapproach with AWERB for implementation by 2019.Specific– To reduce/remove the use of mouse cadavers fortraining in i.v. lateral tail vein injections.– To compare training method outcome through use ofa simulator and cadavers.– To implement a new training format using asimulator with endorsement through AWERB by2019.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 6

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Intravenous Injection Training Project: A Level 6 Higher Education project7Measurable– To implement a new training format for i.v. lateraltail vein injections by 2019.– In order to implement the new training format by2019 the research comparison study must beconcluded with a positive outcome (higher successpercentage).– Submission to AWERB as an agenda item for thenext meeting in January 2019.Achievable– The objective to reduce the need for mice supportsthe 3Rs vision and part of the AWERB agenda.Realistic– By decreasing the use of cadavers, we will supportthe 3Rs’ vision. There is the opportunity to drive thisalternative method to other research institutes,particularly those who have no cadaver availability.Time-bound– To complete the research comparison study by theend of 2018.– To consult with AWERB for a change to the internaltraining policy by January 2019.– To implement a new training programme in i.v.lateral tail vein procedures by February 2019.Peter Drucker was an innovative thinker and studied hiswork using ideas to create management theory.Drucker placed business ethics and morals high on hisagenda, which mirrors this study.In 1954 Drucker published a book which looked atmanagement by objectives.24The belief is that ifemployees manage their own objectives, they willadhere to set standards and are more likely to fulfilthem. Using the SMART method helps to ensure anobjective is valid. Delegation of tasks empowersemployees allowing more control and interest.24According to the International Network for HumaneEducation (InterNICHE) there is a need for change intraining methods due to the ethical nature of how andwhat we train with. (See Figure 1.)Figure 1. Flowchart to assess the sourceof an animal cadaver, organ or tissueaccording to the InterNICHE policy.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 7

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Intravenous Injection Training Project: A Level 6 Higher Education project8Organisational contextIdentification of stakeholders is realised by determiningwho is interested in the project, who has influence, whocan help to achieve it and equally who can obstruct. Inevery project, there will be strengths, weaknesses andrecognition of these as near to the beginning aspossible is crucial. Staff expertise sourced internallyand the use of equipment and facilities can speed theproject up but will add a cost and burden to it.PESTLE analysis (Political, Economic, Social,Technological, Legal and Environmental) is relevant tothis project as research, development, marketing,image, sales and reputation all potentially have a role.Stakeholder map!""#$%&'(%)("*$+#,'"-'(&.$/0&-1"$&1"-'%2 !""#$%&'(%)("*#$%&'()!"3$#.&3"4%$+ !""#$'0"5$%&'(%)("*2 5 &-&1"$/.,%".3*+%+,'(-./01(-*())"4"-'$+ 5(-(5&.$")),4'$-""*"*2$5,-(',423+34)3454+%!""#$'0"5$(-),45"*6(+4%'(-))'))7(8+(34549+%3)0678$(9:;<=9>=.?@$(9:;<=9>=.?@$(9A=B=CA0678$(9A=B=CAFigure 2. Stakeholder map. (Bryson, 1995)25G Animal Welfare and Ethics Review Board (AWERB)members – Chair, Statistician, Scientist (AnimalResearch), Scientist (Non-animal research),Layperson, Quality and Governance manager,NACWO x 2 (Named Animal Care and Welfareofficer), BSF Manager, NTCO (Named Training andCompetence Officer), NVS (Named VeterinarySurgeon)G managerG trainerG assessorG research participantsG fundersStakeholders are important to manage as part of aproject as their level of influence and interest canimpact heavily. They may be individuals or groups andshould be included in strategy when planning a project.The project could be successful in delivery and cost butstakeholders should be satisfied in order for it to besuccessful.AWERB Committee members(Group)High influence, high interestThe AWERB can be seen as a whole group rather thanindividuals. There are various levels of influence withinthe group members but there is high influence andinterest overall. As the project involves ethical andwelfare issues surrounding training using animals thereis potential for conflict between members. In order tosatisfy members’ specific expectations, a presentationcovering statistics, animal welfare, possible publicationor where the information will be going. The presentationwill also have to show the advantages with a guaranteeof completion of paperwork such as risk assessmentsand standard operating procedures, General DataProtection Regulation (GDPR) and that it is following thelaw. Time management and training issues will alsohave to be presented and how these will be dealt with.A presentation to the AWERB seeking permission withregular updates at their quarterly meetings to keepthem informed. Good communication is necessary forthis group. Without their permission the project will notgo ahead.ManagerHigh influence, high interestHigh influence because without support the projectcannot go ahead. High interest because of timeconstraints on staff taking them away from normalduties. They will also have interest regarding welfareimplications and will expect timeframes and legalobligations to be met. Updates should be at leastweekly, keeping them well informed. This would be bestdone face-to-face in a meeting.TrainerHigh interest, low influenceAdequate information about the project and regularcommunication should apply to ensure there are noproblems arising from the project. They can be helpfulwhen designing fine detail about the method ofprocedures they will be training. As trainers they willhave a positive welfare interest in the project.Motivation will come from the potential outcome. Detailof the project and paperwork must be in place and dailycommunication in person when trials are running.AssessorHigh interest, low influenceAssessor has the same criteria as the trainer.Research participantsHigh interest, low influenceResearch participants will gain skill in a difficultprocedure so have interest but little influence on theApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 8

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Intravenous Injection Training Project: A Level 6 Higher Education project9project overall. They will require ongoing informationregarding dates, paperwork, etc., which will be providedby the trainer as this is a blinded study. Email will bethe best communication method.FunderHigh influence, low interestFunding will be sought through a public engagementgrant for the cost of the simulator therefore influence ishigh as budget is required but interest low in thisproject as funding is primarily for an alternative project.If funding was aimed primarily at this project only,influence and interest would be high. Feedback formsto complete and specific deadlines for each step of theproject are necessary. Email is the best format to use.StatisticianLow influence, low interestStatistician will be asked to randomise the participantsfor it to be blinded to the assessor.Risk RegisterRefer to Excel spreadsheet (available on request fromthe author).A special work model or temporary organisation of staffis necessary as described by Tonnquist in his 4strategic issues for the project to succeed.10It would benaïve to assume a project of this scale could runwithout reorganisation of staff for a period of time.BudgetRefer to Excel spreadsheet. (available on request fromthe author).Determined resources – Own budget.The amount of resources available can influence aproject. Stakeholders are important in order to supportthe project in several ways. A budget is essential.Statistician1. Standard operating procedureSOP 64: i.v. Study – Intravenous injection via lateral tailvein training for cadaver and mouse simulator study.SOP 100: i.v. Study – Competence assessment forintravenous injection via the lateral tail vein for mouse(cadaver/mimicky®mouse) comparison study.The SOPs explain step by step how each process in thisstudy should be carried out. This guides theparticipants and trainer/assessor. It also reduces driftand increases the likelihood that each step is carriedout similarly for each participant encouragingstandardisation in techniques, training and expectedstandards.2. Stakeholder engagementAWERB – High influence, high interest.FUNDERS – High influence, low interest.TRAINER – High interest, low influence.STATISTICIAN – Low interest, low influence.Stakeholder theory states that the interests of allstakeholders should be taken into consideration.AWERBR. Edward Freeman uses the term of business ethicswhich appears to be an oxymoron.26He explains thatpeople have more than just a financial expectation orinterest. He also suggests that too much time is spentidentifying primary and secondar y stakeholders.(Primary such as employees and secondary such aslegal authorities) This is a prime example in the case ofAWERB members where stakeholders come from bothprimary and secondary backgrounds. The task ofcommunication needs to be very clear to this group bydefining each member’s stake or interest ensuringeach is communicated in as positive way as possible.AWERB members are high interest, high influencestakeholders as without permission from them for theproject it would not be able to proceed. The primaryagenda is welfare and ethics and so these valuesshould be communicated within the project. AWERBmeets every three months.Communication– A document will need to be put together andsubmitted in advance of the AWERB meeting. Ashort presentation with a Q & A session should beincluded on the agenda.– Updates should be presented as a written piece ofinformation (report) or as a short presentation atAWERB dependent upon their preference.FundersThe funders are willing to give small grant awards forpublic engagement. These are high influence, highinterest in terms of public engagement but in thisinstance after Public engagement (PE) work with thesimulator we would go on to use the model for thestudy. Therefore, interest is lowered.Communication would occur as requested by thefunders and needs to be on time and meetingdeadlines. In this case my starting point is working outhow I can add value to this project which meets theneeds of the funders.Communication– Through an application form by email.– Through feedback form after the project.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 9

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Intravenous Injection Training Project: A Level 6 Higher Education project10TrainerLow influence but extremely important for the project,this stakeholder must work together with various othermembers of the stakeholders for success. Littleinfluence politically but important for interaction withvarious groups in order to move the project forward.Communication– Face-to-face meetings to be clear and share ideasand criticisms to improve the process.– Email in order to communicate SOP’s, riskassessments and other important information.StatisticianStatistician does not politically have an effect and haslow interest as they are being asked to carry out partof a procedure. Stakeholders in these positions can beuseful at looking at a project from an outsideperspective and may offer ideas or innovations.26Risk Register: Refer to risk register documentappendix (available from author on request).Gantt Chart: Refer to Gantt chart document appendix(available from author on request).Budget: Refer to budget document appendix. (availablefrom author on request).Ethics Committee approvalThe AWERB at LSHTM reviews all work undertaken at orassociated with the institute. Initially an AWERB formwill need to be submitted to the chair of the committee,this form will request details such as:G plan of workG objectivesG number of animalsG use of 3RsG severity bandingG endpoints, etcThe chair will then decide if the project is acceptable toput forward to the committee which is made up of avaried group which include:Lay people, Non-animal scientists, Scientist, NTCO,NACWO, NVS, etc.The AWERB meets quarterly and so unless anapplication is especially urgent and needs to have ameeting convened, they will review every three months.At the meeting the proposer of the project will beinvited to give a presentation and will then bescrutinised by the board who will ask scientific andwelfare questions and attempt to refine, reduce andreplace the project as much as possible.They will be asked to make any changes necessaryafter suggestions as the project will be permitted to goahead after changes via email or they will be calledback to the next meeting if it requires furtherclarification.AWERB will require regular updates and a retrospectivereview completed in due course.Part 2 – Implementing andevaluationDelivery Methodology – Alternative training methodusing a mouse simulator in i.v. lateral tail veinprocedures reducing unethical cadaver usage.Objective: To reduce the use of cadavers for trainingin i.v. lateral tail vein procedure using a simulatorthrough a comparison research study and aconsultative approach with AWERB for implementationby 2019.Investigative Searches on Comparisons of lateral tailvein injection training methods and ethics regardingthe use of cadavers revealed that this subject matterwas scarce. Relevant information was difficult to findand it appeared, not to be investigated in depth. Thiswas important for the overall design of the projectbecause animals were involved and in order toeliminate repetition, a systematic style approach wasnecessary.27Investigating data from 2017 regarding pass/failurerates for competence assessments of procedures onmice within the BSF at LSHTM it was noted that someprocedures had a higher success rate than others.Intravenous injections into the lateral tail veins of miceclearly exhibited as a more difficult technique with ahigher failure rate. (Data obtained and collated frommandatory training files within the facility.)Training is completed through a training process withcadavers (dead mice) initially before moving on to thenext step with live animals which may take days,weeks or months dependent upon the skill of thetrainee. Cadavers utilised at the facility for trainingpurposes are breeding stock surplus to requirementand scheduled for culling. This is not an ethical sourceof cadavers and this study will determine whether atraining tool (mimicky®mouse simulator)14would beequally effective for successful assessment incompetence for this procedure. The simulator has twolateral tail veins, mimicking a real mouse and is thesame size as a small to medium sized laboratory wildtype mouse. Liquid injected straight into the vein,leads to a reservoir within the main body. (Emptied forre-use) Participants selected had no previousexperience in i.v. injection technique.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 10

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Intravenous Injection Training Project: A Level 6 Higher Education project11The project required an enormous amount of planningand communication and the best way to see anoverview was through a Gantt chart.A budget was necessary in order to purchase therelevant equipment and so funds had to be sourced.Health and Safety implications needed to be addressedas participants would be working with live animals,sodium pentobarbitone and sharps.The study had to be blinded and randomised forauthenticity as statistical analysis would determine theoutcome.Animal Welfare and Ethical Review Board (AWERB) andthe Manager were highly important stakeholders in thisproject and without their support the project would nothave worked. Delays occurred throughout the projectbut most got completed within the necessar ytimeframe. Some issues came about from externalinfluence such as the delay for the simulator and thesetypes of issues were unavoidable.Project deliveryOn timeWithin budgetTo specificationCompare standardtraining processwith a simulatorStatistical analysisEnrolment ofparticipantsTrainingCompetenceImplementationAdhere to Ganttchart specificationsBudget split into 3divisional areasSpecificationincluding riskassessments,standard operatingprocedures in placeBlinded study tocompare a trainingaid to determine abetter, worse orsimilar outcomeA minimum numberof 20 candidatesnecessary for thestudy to havestatisticalsignificanceParticipants enrolledthrough the NTCO(named training andcompetence officer)with no priorexperience in lateraltail vein injectionsTraining delivered asa randomised trial inhouse by the trainerCompetenceassessmentsdelivered through aclear process ofpass/fail criteriaTimingDelayedOn timeDelayedOn scheduleDelayedOn scheduleDelayedDelayedProblemsDelays occurred butmajor milestoneswere achievedBudget increasedslightly throughprocurement butpartly offset asinflation increase inoperating costs in2019 were notnecessarySlight delay ofcompetence SOP forone participantInitially extremelydelayed but studycompleted on targetSome candidatesleft resulting in newrecruits for thestudy. Twentyenrolled in totalInitial difficultysourcing candidateswith no previousexperienceDelays due to staffshortages, intake ofnew species intothe facility, annualleave and sicknessProcess itselfworked well but onecandidate unable toread SOP due todelayContingencyA Gantt chart withspecific timeframesto followRequested furtherbudget for additionaltail from managerStarted study withone participantbefore completeassessment waswritten up in orderto prevent furtherdelaySourced the CurvetRat model initiallyas mouse simulatorwasn’t available butveins too large tobe a comparison sowaitedHad to enrol newparticipants mid-term through thestudyDiscussed thepossibility ofsourcingparticipants fromother local researchinstitutes butlegislation madethis difficultRequested helpfrom other staffmembers to helpcover trainers workSome participantshad to be re-testeddue to incorrectrestraint usedinitiallyStakeholderEngagementManager –communicationManager – deliverysolution negotiatedand agreedHealth & Safetyrepresentative– Risks identifiedManager –StakeholderacceptedManufacturer –Regular consultationParticipants –StakeholderopportunityParticipants –CompromiseStaff members– ProductiveParticipants –Delivery solutionnegotiated andagreedAssessor –Stakeholder acceptedApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 11

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Intravenous Injection Training Project: A Level 6 Higher Education project12Trainer –Experienced in bothmethodsExperiencedassessorNumber of trainingsessionsResults recordedSet scheduleOwn budgetTemporaryorganisationProject successInnovative projectImpact on day today organisationalsettingPrecise autonomyDiscretionFlexibilityCapturing ortranslating newlylearned routines orpracticesAdaptable trainer todevelop a systemfor training with bothcadaver andsimulatorAssessor to followprocess laid out inthe SOP and scoreto competenceassessmentrequirementsResults to berecorded on atemplateTemplates recordingresultsGantt ChartManaged through anExcel spreadsheetProject managedthrough planning,communication andre-shuffling of staffand teamworkSuccessful anddeliverableNew product to themarket to be testedand comparedDiscussion,meetings and staffcommunicationUtilising projectmanagementdeliveryAllowing for thepossibility of sometechnical errorsWorking aroundpotential staffingand participantissuesCaptured throughwrittenassessments, SOPsand re-organisationof staffOn timeOn scheduleOn scheduleSlight delayDelayedOn scheduleDelayedDelayedDelayedDelayedDelayedOn scheduleDelayedDelayedSourced with noproblemsSourced with noproblemsNo problemsNo problemsNot all exacttimeframes weremetMinor adjustmentsIssues not expectedsuch as a sicknessand new specieswhich wasn’t takeninto considerationDelayed in parts butsuccessful overallPromise of thesimulator tooklonger to materialisethan originallyinformedDifficult with a smallstaff teamProject Managementdelivery could havebeen utilised moreefficiently withexperienceCorrect restraint notused initiallyresulting in re-assessmentsStaff were extremelyflexible but issuessuch as a newspecies beingintroduced to thefacility only allowedso muchSome staff resistantto changeNoneNoneNoneNoneNoneNoneOther staff asked tohelp cover work tofree up time fortrainer andassessorSome contingency inplace but shouldhave had moreMore reproduciblestudies necessaryOther staffmembers requiredto step up and helpReferred to Kotter,Tonnquist, etc. forautonomySome assessmentswere re-taken withthe correctequipmentAt certain pointsthroughout theproject when therewas time trainingand competencebecame a muchfocussed areaOnly volunteerparticipants initiallyasked to use thisnew methodTrainer – RegularcommunicationAssessor –RegularcommunicationTrainer –Delivery solutionnegotiated andagreedTrainer –ProductiveProject Manager –StakeholderengagementProject Manager –PlanningStaff members –CompromiseAWERB –StakeholdersacceptedManufacturer –CommunicationAWERB –StakeholdersacceptedStaff members –PlanningProject Manager –PlanningParticipants –Assessor –Delivery solutionnegotiated andagreedManager –Regular consultationStaff members –CommunicationParticipants –Build a stakeholderrelationshipApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 12

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Intravenous Injection Training Project: A Level 6 Higher Education project13Sub GrouplearningWider organisationalcontextStrategy/visionwhole team sharedpracticeTeamwork learningDistributed cognitionor the study ofknowledge beingpassed on andgrowingStable design topass on –Distribution ofknowledge andskillsTraining method –Practical taskteaching injectionsSuccessful? –Non-use of animalsTraining to bedelivered to othermembers of staffusing the newequipment andtechniqueSeek AWERBendorsement,change in culturethrough delivery ofan alternativetraining method andleadership action topromote time andinvestmentInvolvement of thewhole team indifferent aspects ofthe projectPractical sessionsfor the training teamTrainees to becomeadept at utilising thecorrect processesand passing thatinformation on tonew traineesA trainingprogramme ofstable designRight facilities,staff, equipmentand experienceEvaluation throughstatistical and non-statistical analysisDelayedDelayedDelayedDelayedOn scheduleDelayedDelayedOn schedulePhysical training isthe optimal methodbut not all staffhave been trainedAWERB only meet 4times per yearunless essential.Slow processDelay to objectivesthrough increasedworkloadTraining went wellNoneNoneFinding enoughparticipantsResults concludenon-significanceNo recourse forcontingencyNo recourse forcontingency yet asAWERB initiallyendorsed the study.Results yet to bedistributedWhole team supportrequested anddeliveredNo recourse forcontingencyNo recourse forcontingencyNoneChanging some ofthe originalparticipantsLarger sample sizeand repeatableexperiment to beconductedStaff members –ConsultationAWERB –Stakeholder risk andopportunityStaff members –StakeholdersuccessTrainers –ProductiveTrainers – deliverysolution negotiatedand agreedAWERB –StakeholderendorsementParticipants,Manager –Stakeholdercommunication andengagementAWERB –EndorsementAll otherstakeholders –ConsultationProject ResultsThis research was conducted for the reduction ofunethical cadaver usage and refinement by use of asimulator for this procedure. The research outcome of‘as good as’ a comparison between the two methodswould indicate the use of this alternative method withno reduction in the quality of training and competenceassessment outcome.This research is important in order to reduce theunethical use of animals within research institutions.While it is recognised that the mice used in thisexperiment and generally for training purposes areexcess breeding stock they are still sourced accordingto InterNiche28as unethical. There is also arequirement to adhere to the law with regard to trainingstaff in procedures and new methods or unavailabilityof cadavers prioritises this important research.The study took place over a year using CD1 outbredmale and female mice aged between 9-12 weeks ofage and were sourced in-house from the breedingfacility in the Biological Services Facility at the LondonSchool of Hygiene and Tropical Medicine. This strainwas selected as we hold a small breeding colony andbeing albino and a larger strain of mouse, the tail veinsare clearer for training purposes.Participants were a combination of researchers andtechnical staff with mixed experience in differentprocedures such as intraperitoneal injection, oralgavage, etc. All participants had previous experience ininjection technique with a live mouse as an alternativeprocedure. Participants were sourced through theNTCO (Named Training and Competence Officer) andwere chosen according to no previous experience inlateral tail vein injection. Participant names wereApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 13

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passed on to a statistician who randomised them fortraining with either a cadaver or simulator. The trainerwho was experienced in both methods carried out thetraining sessions and requested competenceassessments when necessary.Training involved one or more sessions with the traineron the allocated training tool. Participants were showna short video at the start of the session.29Cadaver orsimulator was placed in the restraint by the trainer andsterile water (0.2mls) was pre-loaded into a 27 guageneedle and 1ml syringe. Participants were given amaximum of four cadavers after CO2administration andcer vical dislocation as secondary method ofconfirmation of death or equivalent opportunities with asimulator. A maximum of three attempts starting at thelower end nearer the tail tip and working up towards thebase on one vein before moving to the other veinchanging the needle after three attempts due topotential blunting.For competence assessment 0.3mls of Sodiumpentobarbitone (Euthatal) was pre-loaded by theassessor ensuring there were no air bubbles presentand presented to the participant. They were informedthat this was a formal assessment of competence andasked to read the accompanying SOP (standardoperating procedure) and competence sheet whichclearly labels pass and fail criteria. Pass/Failassessment is clear because the needle is notinserted into the vein correctly the plunger will notdepress and liquid will not flow resulting in a liveanimal. If successful the mouse will be euthanised.30Intravenous Injection Training Project: A Level 6 Higher Education project14Figure 3.Figure 3 shows there appears to be no differencebetween the simulator and cadaver. The chi square P-value = 0.64. This does not mean that the two methodsare equally as good as each other because thisexperiment was not set up as an equivalence trial. Itdoes mean that there is no evidence of a difference inthe pass rate using the two methods.For this trial to be an equivalence experiment muchlarger groups would be needed to gain sufficientstatistical power. This experiment had power with 10per group to detect a difference in pass rates betweenthe two methods.– In conclusion the pass rate was lower than expected– There was no evidence of a difference in the passrate using the two methods, P = 0.64– The study was not powered for equivalence, or todetect small differences in pass rates.31Stakeholder engagementManufacturerInitially held a meeting with the manufacturer outliningthe study but with a rat model as the mouse model wasnot available at that time. Manufacturer agreed to loanthe rat model for the study. We were informed that amouse model was going to become available whilstpaperwork was being written, weeks into the project.This put the project back considerably but was a bettercomparison model. This delay was not attributed to thecompany supplying the model as they were setback bythe manufacturer. It was an unfortunate problem butattributable to the manufacturer, overseas.G there were some one-to-one meetings but mostlyemail communicationIt may have been prudent to have looked further afieldto see if any other models were available as it delayedthe project but at the time the goal was near but keptbeing delayed. Communication, negotiation and a newproject plan should have been drawn up at an earlierstage.In conclusion as this model was especially new to themarket and unavailable it probably could not have beensourced easily elsewhere. Communication with allstakeholders regarding delay is crucial and particularlyparticipants to ensure numbers were maintained forthe study.Funding – managerThe project started unsuccessfully as funding was notinitially available and alternative funding sources had tobe found. There were three main costs that required abudget for this project.G operational – such as the use of facilities toperform training, etcG labour – employee time used to perform the studyG material procurement – equipment such as thesimulator, syringes, etcStakeholder engagement took place via a meeting withthe manager. The project outline was explained and hewas keen for the project to go ahead. He was happy toallow for the whole project to be costed internally apartApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 14

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Intravenous Injection Training Project: A Level 6 Higher Education project15from the simulator as this was a very expensive pieceof equipment. Persuasion was applied as to the benefitof the project on animal welfare and the application ofthe 3Rs (Replacement, Refinement, Reduction) througha series of one-to-one meetings and it was agreed thathalf the cost would be paid from the budget. This stillleft a problem of how the other half would be sourced.A public engagement Wellcome grant for the fullamount of the simulator was available and so aproposal was put forward to engage schoolchildren inanimal research using the model in a workshop setting.This was a successful bid and after fulfilling publicengagement (PE) duties the simulator was available forthe study. The manager agreed to pay for an additionaltail from the budget and in this instance all workedwell.32It is important to include the manager in all aspects ofinformation regarding the study as in this case he wasvery influential. Without support from the manager thisproject would not have transpired.This situation could have improved if there had beeninitial budget available for this type of project. All staffworking with animals in research should look toimprove, replace, refine or reduce through studies andobservation. Working in a research facility is an ideallocation to carry out research to make improvementswith equipment and staff already in place.A recommendation would be to implement a budgetinternally allowing for innovation in this field.Animal Welfare and Ethics ReviewBoard (AWERB)The AWERB’s role is to improve both animal welfareand the quality of science, endorse the 3Rs andpromote a ‘Culture of Care’.33Made up of a mixedgroup of people representing welfare, science and thegeneral public, projects put forward are usually verywell debated. The project got the green light from thechair of the AWERB but ethical questions were raised:– Excess breeding stock as an unethical source. Thiswas debated as some members regarded excessbreeding stock as an ethical source. There willalways be an excess of breeding stock no matterhow well a colony is bred but refinements will onlygo so far and therefore it would appear fair to usethem as by products.– The use of 3Rs for this project was highlighted andagreed. It is difficult to find a whole cohortagreeable to each part of the project but thedecision was taken to allow the project to go ahead.The main concern was ethics and welfare.– Some concern over how well-trained staff would bewith the simulator on the live animal in acompetence assessment setting. It was explainedthat unsuccessful i.v. injection would not work(depressor will not plunge) and that mice would beunharmed (mild needle stick).In conclusion going forward institutions need tocommunicate and work together to make every animalcount. Collaborative work should potentially be includedin Project Licences to allow for animal tissue to beutilised between research institutes and less secrecyinvolved in results before publishing.ParticipantsDue to the length of time that it took to carry out thewhole project and being a small facility with limitedstaff resources some participants contracts ended ormoved on to other employment. This resulted in lessthan the minimum number of 20 participants neededfor statistical comparison in this study. An influx of newstarters spread out over the whole project alleviatedthis.More communication was needed with participants andin hindsight regular emails should have been provided.Some stakeholders left and this was only discovered afew days before they were due to leave. There wassome discussion into sourcing possible stakeholdersfrom local research institutions but due to the amountof legislation and red tape surrounding local rules whichwould have been substantial it was decided againstthis.Due to the length of time that the whole project took itwas fortunate that new starters were enrolled on to thestudy.These stakeholders were crucial for the actual studyand initially did not need much communication.Recommendation would be more communication withthis group particularly if the project becomes delayed.Some stakeholders thought the project had ended dueto lack of information.33Risk RegisterRefer to Risk Register document.BudgetManaging a budget is a key responsibility as part ofoverseeing a project. The three main budgetarysubsections were:G material procurement – products necessary to carryout the projectG operating costs – room charges and administrationG labour costs – professionals carrying out trainingand competenceFinancial management came in several forms ofoverview.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 15

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Intravenous Injection Training Project: A Level 6 Higher Education project16G sources of cashFull funding was not available internally and so anexternal source had to be sought (Public Engagementgrant)G justification of expenditureThe AWERB played a key part in justifying the ethicalreview and worth of the project with the Manager’ssupport vital for the operating and labour costsincurred.A public engagement (PE) grant of £1000 was awardedwhich was used for the purchase of the simulator andsome other additional items for the PE project. Thepurchase of this piece of equipment was at theforefront of the project initially as without it there wouldnot be a comparison model. VAT was not accounted for,so extra funds had to be sourced as initially I hadoverspent. The PE team agreed to pay the additionaloverspend and I began factoring this into my futurebudget management.Operational and labour costs also hadn’t initially beenconsidered and so it was fortunate that my Managerbacked this project as the following were all extra coststhat needed to be accounted for. This would havemeant applying elsewhere for extra funding such as theNC3Rs and the project would have been delayedfurther.Methodology analysisThe project initially was not delivered on time. Delaysincluded budget issues because there was no provisionavailable for this type of study. This sadly is somethingthat needs to be addressed at managerial level. Thereare more and more innovative opportunities to make orrefine products that benefit animal welfarecommunicated through bulletins and media in industry.Unfortunately, management systems are generally notallocating a budget for innovation. Tied into thisdilemma is time and giving staff the opportunity toproject manage an idea or study. Stakeholderengagement and paperwork are time consuming whencompleted efficiently and effectively.Participant numbers were researched prior to the startof the project and twenty appeared to be a feasiblenumber but with such a small facility and small numberof experienced technical staff this became a challengefor a period. Study design should always be wellthought out and not too adventurous dependent uponthe resources already in place. Only one trainer andassessor were sourced for this project, there was nocontingency available should one of these staffmembers become absent. There should have been aback-up plan to either train or bring an experiencedperson in as contingency.Correct restraint for procedure was unavailable for atleast four candidates and had to be re-tested.Equipment should always be checked prior to anassessment. This resulted in further assessment withthe correct equipment and potentially influencedresults as participants had one more opportunity to, ineffect, train. The study showed that there was nodifference in pass rates but would have to be repeatedwith larger numbers to detect solid differences.Stakeholder feedbackRefer to Questionnaire sheet.Budget analysisRefer to Budget document.ReflectionProfessor D.A. Kolb is quoted saying: ‘Knowledgeresults from the combination of grasping experienceand transforming it’. Kolb’s research displays thatexpertise, reflection, conceptualisation andexperimentation are a continuous cycle and hold arelationship together.34Learning by doing and thenreflecting on new skills or attitude changes.Constructivism states that knowledge is constructedbased on personal experiences and hypotheses of theenvironment. They are both similar concepts and haveplace in the project learning cycle regarding thisproject.35It was surprising to learn that this subject matter wasscarce, when developing a theory, it was important thatthe underlying content was balanced in order thatexperiential learning could occur. There are many morepotential projects to investigate.Experience through delivering this project has taughtme that the unknown risk of making assumptions canmake stakeholders fearful. Impact and risk withcontingency must be created when delivering a projectto alleviate fear.36Learning develops through information gathering. At thebeginning of the project the original hypothesis wasdifferent to the conclusion at the end. It changed anddeveloped, my assumption that excess breeding stockwas a totally ethical source of cadavers changed. Thisinformation further strengthened the project as anadditional criterion was discovered.The project was personally relevant to my role intraining, it had meaning and I was interested to learnthe outcome. Experientially I discovered that thequicker and well organised a project is the better theoutcome. Projects should be completed in as short aspace of time as possible as delay adds to additionalApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 16

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Intravenous Injection Training Project: A Level 6 Higher Education project17cost, both financially and otherwise having a directimpact in this project on candidate placement.It is important to have perspective on a project,completing paperwork such as SOPs and Riskassessments may not seem so important but connectto other relationships within the project. Part ofexperiential learning is connecting and formingrelationships with stakeholders and engaging.The project became immersive at some points andreflective and at a later stage allowing for re-examination of values. Until completely involved in aproject it is hard to understand fully the changes thatoccur on your perspective over time.The project did not have an equivalent level of successand in order to test this, 356 participants withcadavers and 356 participants with simulators wouldbe necessary to obtain enough statistical power. Thisis such a small facility it would take years to completebut could potentially be a collaborative work with otherfacilities. Sometimes one must leave their comfortzone in order to enhance learning and provideinnovative welfare research projects.“If at first you don’t succeed, try, and try again.”38References1 Accessed: 24.09.182ht tps :// www.L SHT M.a c.u k/research/resear ch-governance-and-integrity/animal-researchAccessed: 25.09.183 Accessed:26.09.185Turner, P.V., Brabb, T. Pekow, C. and Vasbinder, M.A.(2011). Administration of substances to laboratoryanimals: Routes of administration and factors to consider.Journal for the American Associates for Laboratory AnimalScience: JAA LAS, 50(5),600-313 Accessed: Accessed: 28.08.187Bannerman, P.L. (2008). Defining the future of projectsuccess: a multilevel framework. Paper presented at PMIResearch conference: Defining the future of projectmanagement, Warsaw, Poland. Newton Square, PA:Project Management Institute. Accessed: 26.09.188Knight, A. A conscientious objection to harmful animaluse within veterinary and other biomedical education.Animals: an open access journal form MDPI.2014;4(1):16-34.doi:10.3390/ani4010016 Accessed: 29.08.189 Accessed: 29.08.1810Tonnquist, B. (2010). projektledning, Stockholm: Bonnierut bildning AB Accessed: 29.08.1811Jacky Swan (Warwick Business School, UK, Sue Newell(Bentley College, US and Warwick Business School, UK)and Harry Scarbrough (Warwick Business School, UK)Why don’t (or do) organisations learn from projects.Accessed: 27.09.1812 Accessed: 26.09.1814 y-supplies/training-simulator/mimicky-mouse-training-simulator.aspxAccessed: 20.08.1717Questionnaire conducted with NTCO’s in LondonInstitutions (Imperial, UCL, Crick, etc.)Accessed: 01.10.1818 Accessed: 19.08.1719 Accessed: 25.09.1720Chang, Y-C., Berry-Pusey, B., Yasin, R. et al. (2015). Anautomated mouse tail vascular access system by visionand pressure feedback. IEEE/ASME transactions onmechatronics: a joint publication of the IEEE industrialelectronics society and the ASME dynamic systems andcontrol division. 2015; 20(4): 1616-1623 doi:10.1109TMECH.2014.2360886 Accessed: 29.08.1821Vines, D.C., Green, D.E., Kudo, G. and Keller, H. (2011).Evaluation of mouse tail-vein injections both Qualitativelyand Quantitatively on small Animal PET tail scans, 2011,J.NMT, vol 39, no 4, 264-270 Accessed: 21.09.1822The Laboratory Mouse, Institute for Laboratory Science,Hannover medical shop, Hanover, Germany, Part 5,Procedures, 2012 Accessed: 16.09.1723 ticles/management-theory-of-henri-fayol/ Accessed: 17.09.1824 ticles/management-theory-of-peter-drucker-key-terms/25Bryson, J. (1995). Strategic planning for public and non-profit organisations. San Francisco, CA. Jossey-BassAccessed: 17.09.1826 10.10.1827 Accessed: 23.12.1830 09.11.1831Andrea Rehman, Statistician, LSHTMAccessed: 20.11.1832 t-writing.html Accessed: 06.12.1833 08.11.18Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 17

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Intravenous Injection Training Project: A Level 6 Higher Education project1834McLeod, S.A. (2017). Kolb – learning styles. Retrievedfrom Accessed: 07.12.1838 10.12.1839Edward Hickson, The Singing Master, 1836Accessed: 13.12.18Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 18

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19April 2019 Animal Technology and WelfareAnimal Technologist Exchange Programme:A Level 6 Higher Education ProjectMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road W12 0NNCorrespondence: on a Level 6 HE project management assignmentAbstractIt was brought to the attention of the Institute of AnimalTechnology that the biomedical industry lacked a formalway in which Animal Technologists can exchange witheach other in other scientific establishments to gainfurther knowledge and skills. The Animal TechnologistExchange Programme (ATEP) has been devised toprovide a structured way for Animal Technologists toexchange.The programme is open to all levels of AnimalTechnologists including apprentices and exchanges canbe of varying durations.Key words: Animal Technologists, exchange programme,skills, training, Continuing Professional DevelopmentIntroductionIt was brought to the attention of the IAT council thatthere is a lack of providing a formal way in which AnimalTechnologists can exchange. There is already a way forNamed Animal Care and Welfare Officers (NACWOs) toexchange. In the light of openness in the industry it wasdeemed strange that animal facilities open their doorsto school children and media but are reluctant to opentheir doors to people in their own profession. If plannedand executed in a professional manner the IAT AnimalTechnologist Exchange Programme (ATEP) would be ofgreat benefit to both Animal Technologists because itwill provide a safe and structured way of working inother facilities which may differ from the facilities theyare currently working in, and for the hosting facility ingaining Animal Technologists whose skill sets may differfrom their own which in turn could provide training.A SWOT analysis is a structured method that evaluatesstrengths, weaknesses, opportunities and threats of aproject. The SWOT analysis below, evaluates the aboveelements for the ATEP.Strengths:– Project will provide a forum for developing/trainingAnimal Technologists. There is currently a lack ofthis type of development.– Project will increase communication between animalfacilities and help build a network.Weaknesses:– Exchanges could take some time to arrange.– Would need support from facilities to be successful.If animal facilities are not supportive of theprogramme it will not work.Opportunities:– Gap in the industry for this kind of programme.– Could be developed further with support from theHome Office, and training and developmentcompanies.– Opportunities for Animal Technologists to upgradekey skills which will improve animal welfare acrossthe industry.– Would identify supportive employers and otherstakeholders.– Could help develop case studies at the end todemonstrate benefits of the programme to others.Threats:– Other organisations could develop a programmethat is similar.– A lack of employer engagement due to resources,costs, barrier concerns and uneasiness over staffbeing headhunted.Project goalsThe aims of the Animal Technologist ExchangeProgramme are:1. To exchange good practice in the care and welfareof animals used in science.2. To develop a network of Animal Technologists for allsectors of the industry.3. To identify designated trainers in skills that arecurrently not widely available.4. To provide Continued Professional Development(CPD) opportunities.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 19

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Animal Technologist Exchange Programme: A Level 6 Higher Education project20It is important to deliver these project goals in a SMARTway. SMART is an acronym for Specific, Measurable,Achievable, Realistic and Timely/time bound (SMART)that has been credited to both Peter Drucker and G.T.Doran, though it is difficult to identify whether either ofthese two was really the first person to use the term.1Key people/key stakeholdersProject lead or project managerChair of the Animal Welfare Group (Matthew Bilton) onbehalf of the Institute of Animal Technology will ensurethat the programme is delivered to the highest ofstandards and ensures the reporting continues atregular intervals.Clients/stakeholdersCareer Animal TechnologistsThese have an interest to participate in the programmeand a major big part to play in upgrading key skills andbuilding a network. They will also have documentationto complete and provide feedback.Animal facilities in academia, pharmaceuticalestablishments and industryWill host participants and ensure their safety while onsite. Hosts will contribute greatly by passing onknowledge and helping participants develop key skills.Hosts will have an interest on how the programme willwork and what their responsibilities are.IAT CouncilWill have overall responsibility for developing andrunning the programme and will deal with anycomplaints regarding the programme. The IAT Councilcan change the remit of the programme.IAT Website CoordinatorWill advertise and develop a webpage. Will also helpdevise online forms and format. Will have an influenceon all these developments.Home OfficeAlthough not a client, any project that will help improveanimal welfare and increase the skills AnimalTechnologists develop will make them a distantstakeholder.IAT membersThe IAT Council acts on behalf of its members and areaccountable to them. Having a successful technologistexchange programme would be a good thing to presentas an achievement for the IAT.Named Training and Competency Officer(NTCO)The Named Training and Competency Officer (NTCO)is responsible for ensuring that all those dealing withanimals are adequately educated, trained andsupervised at the establishment they work, until theyare competent and that they continue to undertakeappropriate fur ther training to maintain theirexpertise.Specific. Outline in a clear statement precisely what isrequired.–––––––––––––––––––––––––––––––––––––––––––––––––––––––All four aims listed above, suggest in clear words whatthe outcomes of everyone involved in the AnimalTechnologist Exchange Programme, from the AnimalTechnologists themselves to the facilities hosting in theprogramme, what the programme will provide.–––––––––––––––––––––––––––––––––––––––––––––––––––––––Measurable. Include a measure to enable monitoring ofprogress and to know when the objective has beenachieved.–––––––––––––––––––––––––––––––––––––––––––––––––––––––1. To exchange good practice in the care and welfare ofanimals used in science.2. To develop a network of highly skilled AnimalTechnologists for all sectors of the industry.Measuring: The amount of technicians taking part inthe programme.Measurement tool: a list of names and facilities andwhich sector they work in. The number of names onthe list will be an indication of how successful thisaim is.3. To identify designated trainers in skills that are notwidely available.Measurement tool: list of skills and trainers.4. To provide Continued Professional Development(CPD) opportunities.Measurement tool: the amount of facilities who usethe programme and include it in their technologists’PRDP.–––––––––––––––––––––––––––––––––––––––––––––––––––––––Achievable. Objectives should be achievable. They canbe stretching but not unachievable.–––––––––––––––––––––––––––––––––––––––––––––––––––––––If developed and communicated in the right way allobjectives can be achieved. The measurement toolsshow accountability and if the outcomes have beenreached.–––––––––––––––––––––––––––––––––––––––––––––––––––––––Realistic. Focus on outcomes rather than the means ofachieving them.–––––––––––––––––––––––––––––––––––––––––––––––––––––––All outcomes are positive and are what all clients wantfrom an Animal Technologist Exchange Programme. It isimportant that the outcomes are viewed as a positive, byemployers and managers. The outcomes of theprogramme will improve the technologists on theprogramme and animal welfare.–––––––––––––––––––––––––––––––––––––––––––––––––––––––Timely or time-bound. Agree the date by which theoutcome must be achieved.–––––––––––––––––––––––––––––––––––––––––––––––––––––––The project should be completed by March 2019 readyfor implementation at IAT Congress 2019.Figure 1. SMART objectives.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 20

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Animal Technologist Exchange Programme: A Level 6 Higher Education project21Core Role: Setting local standards, applying localstandards, Record systems, Supervision andCompetence Assessment Continuing ProfessionalDevelopment (CPD), Review and Communications.Animal Facility ManagersAnimal facility managers will have the skills andqualifications of senior Animal Technologists but will beresponsible for a number of different animal areas/facilities controlled by senior technical staff. They willhave overall responsibility for the day-to-day operations,including budget control, implementation of policy,staffing and appointments and helping to promote aculture of care within their organisation.2Potential change agents: Key players:–––––––––––––––––––––––––––––––––––––––––––––––––––––––––G Animal facilities in G IAT Councilacademia, pharmaceuticalsand industryG Career AnimalTechnologistsG IAT membersG NTCOG IAT Website CoordinatorG Project leadG Animal facility managers–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Indifferent: Back-yarders:G Home Office G Education providers–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Low Interest levelHighName/position Role Reason forcommunication–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Chair of IAT Project lead Oversees theAnimal Welfare running of theGroup programme. Contactwith suggestions forimprovements orcomplaints.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Secretary IAT Point of contact The point of contactAnimal Welfare for programme to arrangeGroup exchanges. Willarrangecommunicationbetween the twointerested facilities.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––IAT Council Overall Project Lead willleadership for report to Council onprogramme progress ofprogramme. Willdeal with anymisconduct issuesor concerns byorganisationsand/or IAT.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Host facility Will host Will be contacted byAnimal Animal WelfareTechnologists Group secretary withinterestedtechnicians.Will contact thesending facility’stechnician withtimes/dates of visit.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Sending facility Will send Will contact Animaltechnologists Welfare Groupsecretary withdetails of technicianto participate inprogramme andpurpose of visit.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––IAT website Advertise the Responsible for thecoordinator programme and content of themaintain the webpage and willwebpage interact withproject lead in thiscontext.In consultation withproject lead/IATCouncil will maintainthe webpage andupdate as required.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Animal Programme Request via theirTechnologists/ participation line managers/IAT members NTCOs to take the programme Will contact the IATAnimal WelfareGroup secretary orIAT Council anyfeedback positive ornegative of theprogramme. Will beinformed on theprogress of theprogramme annually.Table 1. Potential influences leading to change.It is vital for the Animal Technologist ExchangeProgramme (ATEP) to be successful that allstakeholders have an input and are always informed.For stakeholder engagement to be successful severalways of communicating will be needed, this will be face-to-face at IAT Council meetings, online and/or via thephone. When communicating to stakeholders the“POURS” system will be used as this system will avoidany misunderstandings and bring about action pointsthat are relevant.POURS;– Plan – What to tell and ask.– Outline – Understanding, clarify objectives andseek feedback.– Use – Open questions to gain further information.– Reflect – Use closed questions for confirmation.– Summarise – Agree actions.Communication plan:Table 2. Communication plan.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 21

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Animal Technologist Exchange Programme: A Level 6 Higher Education project22In his book Supervisory Management P.W. Bettsdescribes any plan for communicating effectivelyshould conform to a scientific or logical method.3Bettssuggests a general approach as:G clarify the purpose of the communicationG consider the type of communicationG obtain the relevant informationG assess the receiverG determine the best means of communicatingG decide on the channel or channels to useG assess oneselfG arrange the communicationG transmit the messageG check for understandingG follow upThis communication plan and the systems in place willaddress the threats set out in the SWOT analysis ofemployers not engaging in the programme that could bedue to resources, costs and barrier concerns.Risk registerRisk management is critical to the success of anyproject and must be developed during the planningstages of the project management process.The risk register starts, of course, with a riskmanagement plan. The project manager must seekinput from team members as well as stakeholders andpossibly even end users. The risk register or risk logbecomes essential as it records identified risks, theirseverity and the actions to be taken. It can be a simpledocument, spreadsheet or a database system but themost effective format is a table. Tables present a greatdeal of information in just a few pages.Managers should view the risk register as amanagement tool through a review and updatingprocess that identifies, assesses and manages risksdown to acceptable levels. The register provides aframework in which problems that threaten the deliveryof the anticipated benefits are captured. Actions arethen instigated to reduce the probability and thepotential impact of specific risks.4Project planning documentProject Name: Animal Technologist ExchangeProgramme (ATEP).IntroductionPlanning of the project will involve a various number ofstages such as highlighting key issues, generatingideas and opinions, making decisions and assessingresources.Before implementation of the plan the 5 Cs of decisionmaking are a good reminder of the process.1. Consider: Clarify the nature of the project, time andother constrains, ask what information is neededand identify objectives.2. Consult: Gather the most information available, calla meeting of those involved for consultation anddecide when the consultation will stop.3. Crunch: Review all options and make a decision.Write the implementation plan.4. Communicate: Provide briefings on what willhappen, why and who the decision affects, back-upbriefing with written confirmation of decision. Makesure everyone understands when the decision willbe implemented.5. Check: The briefing is carried out. Run spot checksto monitor effectiveness and review the impact ofthe decision and take corrective action.6Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 22

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Animal Technologist Exchange Programme: A Level 6 Higher Education project23Implementation planStakeholder engagementCommunication with stakeholders:Step by step plan Completed Comments–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Identity All stakeholdersstakeholders have been identifiedand included in theproject scopingdocument. Theirimportance andeffect they couldhave on theprogramme isdemonstrated in agrid within thescoping document.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Complete a Scoping documentscoping completed. Thisdocument document containsall relevantinformation relatedto the programme.This includesstatement of theproblem,introduction/background to theproblem, projectgoals which areSMART, identifieskey stakeholdersand has a riskregister andoverview budget ofthe project.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Consult the The Animal WelfareIAT Council Group were taskedon how the by IAT Council toproject will develop andwork implement the plan.Items discussedwere why and howthe programme willwork. It was agreedthe Welfare Groupwill devise adocument anddevelop a way ofadvertising theprogramme.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Develop an Informationinformation document written updocument and is loosely basedrelating to the on the informationAnimal provided for the IATTechnologist NACWO ExchangeExchange Programme. ThisProgramme document identifiesthe aims of theprogramme. Theresponsibilities ofthe hostinginstitution, thevisitor and the IAT.This relates to alldifferent checklistsand reports for theprogramme.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Consult with The IAT Websitethe IAT Coordinator wasWebsite tasked with helpingCoordinator the group and wason how to sent a copy of thedevelop the informationinformation document. This wasdocument into changed with theiran online form input to have aand the professional look.development The forms areof a webpage currently underreview and is hopedthe WebsiteCoordinator willdevelop the forminto a more onlinefriendly format. Awebpage iscompleted for theprogramme.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Draw up a It was suggestedlist of all by the Chair of theinstitutions IAT Council to looksigned up to the at the institutionsConcordat of signed up to theOpenness5concordat5andand develop using the Councila contact list contacts obtainsome emailaddresses anddevelop a standardemail ready to sendout details of theprogramme whenfinalised and askthem to sign up tothe programme.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Send out a To be completedstandard email once programme isto all contacts live.asking initiationsto sign up to theprogrammeTable 4. Implementation plan.Table 5. Methods of communication.Stakeholder Influence Method of Frequencycommunication–––––––––––––––––––––––––––––––––––––––––––––––––––––––––IAT Council Key players Face-to-face At Councilmeetings everytwo months–––––––––––––––––––––––––––––––––––––––––––––––––––––––––IAT Website Potential Face-to-face, At leastCoordinator change agent email monthly–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Career Animal Key player email, When AnimalTechnologists advertisements Technologistin IAT programme isactive–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Home Office Indifferent email As requiredApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 23

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Animal Technologist Exchange Programme: A Level 6 Higher Education project24Betts suggests that any omission of a step may reducethe effectiveness of the communication.3The table above suggests the best way ofcommunication with key stakeholders. Their influence,frequency and method of communication areconsidered. All the steps that Betts suggests as ageneral approach should be taken with all thestakeholders suggested in the table above.3The mainmethods of communication will be either face-to-face orby e-mail. Both ways of communicating will need someconsideration. In the Chartered ManagementInstitute’s checklist series ‘Managing BusinessCommunications’, they describe that email is widelyused, however it could be easy to misuse this tool.They suggest it is advisable to:1. Think about presentation and formality.2. Consider how to open and close the message.3. Use good practice for written businesscommunication. Examples include: be brief andsuccinct, keep things simple and clear, avoid usingjargon, write in full sentences, explain what theemail is about, use good grammar, punctuationand spelling and only communicate things that youwould be willing to commit to paper or would behappy for colleagues or outside contacts to see.4. Structure the message in a logical way.5. Make good use of the subject line.6. Avoid strings of messages, where possible.7. Take care with group messages.8. Use ‘urgent’ markers sparingly.9. Avoid humour, irony or sarcasm.10. Check before sending large attachments.11. Ensure clarity of ownership and legality.All of these recommendations will be used whenemailing the stakeholders. Many stakeholders havelimited time to go through emails and some consist oflarge groups of people, an example being the twenty-sixpeople making up the IAT Council. The steps above area logical way when sending emails.With face-to-face communication the CMI checklistseries suggests:– Consider whether face-to-face communication isneeded.– Clarify the purpose of the communication and itsexpected outcome.– Choose the time and place.– Prepare.– Adopt the right tone.– Set the scene.– Be aware of attitudes, values and expectations.– Develop questioning and listening skills.Most of the face-to-face communication will be with theIAT Council where time and place is alreadydetermined, however the checklist is also a logicalchoice of way/method and will be used in all face-to-face communication.Gantt chart:A Gantt chart is a horizontal bar chart developed as aproduction control tool in 1917 by Henry L. Gantt, anAmerican engineer and social scientist. Frequentlyused in project management, a Gantt chart provides agraphical illustration of a schedule that helps to plan,coordinate and track specific tasks in a project.Gantt charts may be simple versions created on graphpaper or more complex automated versions createdusing project management applications such asMicrosoft Project or Excel.Project management:implementation and evaluationProject Implementation document:IAT Animal Technologist Exchange Programme (ATEP).Delivery methodology:In the project planning document, the following steps inimplementation were identified. Comments in italicsdescribe how the steps were implemented andsuccess:Chart 1. Gantt chart of project activity.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 24

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Animal Technologist Exchange Programme: A Level 6 Higher Education project25Step by step plan Completed Comments–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Identity All stakeholdersstakeholders have been identifiedand included in theproject scopingdocument. Theirimportance andeffect they couldhave on theprogramme isdemonstrated in agrid within thescoping document.Update: This stepwas delivered ontime as identified inthe relevant Ganttchart and includedin the projectscoping document.No newstakeholders havecome forward duringthe implementationprocess.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Complete a Scoping documentscoping completed. Thisdocument document containsall relevantinformation relatedto the programme.This includesstatement of theproblem,introduction/background to theproblem, projectgoals which areSMART. Identifieskey stakeholdersand has a riskregister andoverview budget ofthe project.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Consult the The Animal WelfareIAT Council Group were taskedon how the by IAT Council toproject will develop andwork implement the plan.Items discussedwere why and howthe programme willwork. It was agreedthe Welfare Groupwill devise adocument anddevelop a way ofadvertising theprogramme.Update: This wasdone regularly ateach IAT Councilmeeting as part ofthe Animal WelfareGroup repor ts toCouncil. This is anongoing process asidentified in theGantt chart. Therehave been a fewideas from Councilregarding theTechnologistExchangeProgramme. Thisincludes openingthe exchangeprogramme toapprentices whichwas not included inthe original scopingdocument.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Develop an Informationinformation document written updocument and is loosely basedrelating to the on the informationAnimal provided for the IATTechnologist NACWO ExchangeExchange Programme. ThisProgramme document identifiesthe aims of theprogramme, theresponsibilities ofthe hostinginstitution, thevisitor and the IAT.This relates to alldifferent checklistsand reports for theprogramme.Update: This wasdeveloped followingconsultation withother members ofthe Animal WelfareGroup and the IATWebsiteCoordinator.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Consult with The IAT Websitethe IAT Coordinator wasWebsite tasked with helpingCoordinator the group and wason how to sent a copy of thedevelop the informationinformation document. This wasdocument into changed with theiran online form input to have aand the professional look.development The forms areof a webpage currently underreview and it ishoped the WebsiteCoordinator willdevelop the forminto a more onlinefriendly format. Awebpage iscompleted for theprogramme but notcurrently live.Update: At the timeof writing, this isdelaying the project.The WebsiteCoordinator is busyworking on otherprojects related tothe IAT Congress.There has beenfrequent contactwith the WebsiteCoordinatorregarding this step.Furtherconversationsregarding onlineforms are scheduledin for Congress2019. This hasbeen an unforeseenApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 25

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Animal Technologist Exchange Programme: A Level 6 Higher Education project26Stakeholder engagement report:The following table was included in the project planningdocument and indicated who the stakeholders are andhow they were going to be contacted.IAT CouncilEngagement with the IAT Council was frequent andactivity regarding project included in the Animal WelfareGroup reports. There were always plenty of suggestionsincluding one of using the Concordat on Openness5which proved to be a good starting point in identifyingcontacts and the number of institutes there were totarget. As a key player and Council being responsiblefor delivering the ATEP, the advice was used andfollowed up. The frequency of discussion continues tobe every two months.– IAT Website CoordinatorAs identified in the project planning document theIAT Website Coordinator has been a potentialchange agent. Below are examples of emails sentwith the Coordinator’s suggestions. The originalemail suggested a standard worded email toadvertise the programme.Email 1:I will make a start on getting a webpage set up foryou and let you see a mock up before publishing.Just in relation to the email text below may I makethe suggestion below?Dear Sir/Madam,The IAT Animal Welfare Group are delighted tointroduce the Animal Technologist ExchangeProgramme.This programme will complement the NACWOExchange by providing further opportunities for staffdevelopment.Aims of the programme are:To exchange best practice in the care and welfareof animals used in science.1. To share knowledge between Academia, ContractResearch and Pharmaceutical industries.2. To identify designated trainers in skills that arenot widely available.3. To provide an opportunity to network with othertechnologists.4. To provide Continued Professional Development.Take a look at the attached document and theinformation on the IAT website for more detailsabout the Technologist Exchange Programme.Email 2:We are trying to move many of the forms to onlineforms.Having looked through this report form I am notsure it will work but I will see what I can do.The move to online forms contents of email 2 hasproven to be the current main delay. The delay hasbeen compounded due to the exceptionally heavyworkload caused by his involvement in organising theannual IAT Congress. The identification of the WebsiteCoordinator as a change agent has proven to becorrect.risk which has beenadded to the riskregister.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Draw up a It was suggestedlist of all by the Chair of theinstitutions IAT Council to looksigned at the institutionsup to the signed up to theConcordat of concordat5andOpenness5using the Counciland develop contacts obtaina contact list some emailaddresses anddevelop a standardemail ready to sendout details of theprogramme whenfinalised and askthem to sign up tothe programme.Update: This listwas drawn up usingvarious contactsacross Council andmembers of theAnimal WelfareGroup. However, notall institutes had acontact that anyoneknew. The plan isalso to advertisethe programme onthe animalmanagement andwelfare discussiongroup and in the IATBulletin.–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Send out a To be completedstandard email once programme isto all contacts live.asking initiationsto sign up to theprogrammeTable 6. Project implementation document.Table 7.Stakeholder Influence Method of Frequencycommunication–––––––––––––––––––––––––––––––––––––––––––––––––––––––––IAT Council Key players Face-to-face At Councilmeetings everytwo months–––––––––––––––––––––––––––––––––––––––––––––––––––––––––IAT Website Potential Face-to-face, At leastCoordinator change agent email monthly–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Career Animal Key player email, When AnimalTechnologists advertisements Technologistin IAT Bulletin programme isactive–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Home Office Indifferent email As and whenApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 26

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Animal Technologist Exchange Programme: A Level 6 Higher Education project27On the whole communication regarding the project hasbeen planned and managed well as described in theproject planning document.G Project results:As this is an organisational project the results are inorganisational change rather than to present raw data.However, it is worth noting that as part of the reportingprocess to the IAT Council, the amount of peopleinvolved in the exchange will be reported on an annualbasis.The project has brought together stakeholders fromdifferent backgrounds in the aim of setting theexchange programme up. For example the IAT WebsiteCoordinator’s day to day job is head of BiologicalServices at a large research establishment. While I, asproject lead, am an advanced Animal Technologist andNamed Animal Care and Welfare Officer (NACWO) at aLondon based University. In addition to this the Chairof the IAT who also had various suggestions andquestions is the director of of another major University.With this background of people working closelytogether, this demonstrates the essence of what theATEP is all about. The exchange programme is a forumfor an exchange of ideas for the benefit of staffdevelopment and animal welfare. Which is exactlywhat the originators of the programme havedemonstrated.Personally, the benefits to my organisation in myselfworking on a project on the IAT Council has increasedmy own skillset in terms of developing mycommunication, time management and organisingskills. As this project is different to my workplace job,an opportunity to work on a project such as the ATEPhas allowed the development of a more confident andskilled person and who, as a result, built a network ofpeople that did not exist before the project started.G Budget management:This project has been very difficult to report in a budgetsense as most of the budget planned has not had amonetary value but has been an outlay in time. Theestimate for input time was 40%. This proved an underestimation. A large percentage of personal time hasbeen speaking to people to get the project off theground and running. One issue with dealing with othermembers of the Council is that they are all volunteerswith limited free time which means other prioritiessuch as the involvement with planning the annual IATCongress for example, reduces people’s availability towork on new projects. This has been a frustration anda delay in the project that was unforeseen. Due to thisdelay it has been difficult to assess the transportcosts. However, there will be a yearly overview on allaspects of the ATEP and a more detailed assessmentwill be made then.Project evaluationChallenges and setbacks:During the implementation process a few challengesand setback occurred. One setback was the discussionsurrounding online forms. In the process of developingthe Technologist Exchange Programme webpage theIAT Website Coordinator did email to say all onlineforms were bring reviewed. In his opinion the formsdrawn up in the information document were notsuitable for being online use. This has led to severalemails between the various parties being sent backand forth regarding the necessary action required onto resolving this. A face-to-face meeting with the IATWebsite Coordinator and his deputy allowed us toarrive at a date which suited all of us to discuss thisissue in full. This was an unforeseen risk delay and hasbeen added to the risk register with mitigating actions.A further challenge was during consulting with the IATCouncil during the last stages of resolving the onlineform issue there was a suggestion from the Chair tonot only open up the programme to all AnimalTechnologists and NACWOs but also to include anapprentice element to the ATEP exchange programme.This would add to the programme greatly however, thesuggestion has meant the information has had to beamended again this was not planned for in the originalproject planning document.Positive developmentsOne of the ideas that went well was the drawing up alist of all the institutions that had signed up to theConcordat of Openness5and identifying contacts. Thiswas done by a member of the Animal Welfare Groupwho did a fantastic job in coming up compiling with acontact list.The communication plan set out has been adhered toand frequency was good as well as identifying relevantstakeholders. Throughout the process no newstakeholder have emerged and their lack of influencehave been correctly identified.Communication with all stakeholders have beenpositive and the ambition and willingness on all partiesto have an Animal Technologist Exchange Programmehas made the project run smoother.Overall it has been deemed that the AnimalTechnologist Exchange Programme would benefit ourindustry as a whole, improve staff training anddevelopment, and improve animal welfare. These arethe key elements to the IT mission which is ‘toadvance knowledge and promote excellence in thecare and welfare of animals in science and to enhancethe standards and status of those professionallyengaged in the care, welfare and use of animals inscience’.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 27

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project. This has not been without it faults andfrustrations. The members on IAT Council use a lot oftheir free time on Council business. Due to this, therehas been a delay due to other priorities for Councilmembers. This has meant making decisions on my owninitative to get this project off the ground which hasincreased my confidence in making decisions andstanding by them and then communicating thosedecisions to Council. From a personal view havingnever really worked on a major project and beingunsure of the role I would play on Council, developingthis project that could help a number of technologistsdevelop their careers, has justified my decision injoining Council. This has been useful in improving myconfidence in the workplace which now has someonehow can work and develop complex projects but alsosomeone who is establishing a network of people tocall on for advice.On reflection, things that I would have done differently:– I think I should have consulted the IAT WebsiteCoordinator at an earlier stage in the design of theTechnologist Exchange Programme document. As astakeholder who had been identified as a changeagent, this was an important aspect of the projectand the lack of early consultation meant asignificant delay in the project as his available timewas committed to other projects.– One of the difficulties of this type of project isdeveloping a budget plan. In terms of money therewas none available. A lot of people have invested alot of time on this project. I suggested in theplanning budget that 40% of my time would beinvolved on the project. This was in fact anunderestimation. A lot of my time has been spent onthis project however this is due to wanting to haveand develop the programme correctly and make it aworthwhile project. It must be done in the rightmanner to avoid embarrassment to the IAT andmyself as project manager.The establishment of the Animal Technologist ExchangeProgramme is something that will continue to developand provide oppor tunities for animal welfareimprovements in addition to providing staffdevelopment opportunities which may change theindustry. I feel immensely proud to be giving somethingback for the opportunities that I have been given in mycareer.References1Chartered Management Institute (2013): the checklistseries: Managing others teams and individuals, London,pp85-892Institute of Animal Technology (IAT), Animal Technologist:A caring career brochure. Technologist Exchange Programme: A Level 6 Higher Education project28Stakeholder feedbackquestionnaire:IAT CouncilFigure 9. Feedback questionnaire IAT WebsiteCoordinator.Questions Comments/answers–––––––––––––––––––––––––––––––––––––––––––––––––––––––––How do you think theTechnologist programmehas changed AnimalTechnology?–––––––––––––––––––––––––––––––––––––––––––––––––––––––––What do you think arethe strengths/weaknesses of theAnimal TechnologistProgramme?–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Do you feel sufficientinformation about theproject was providedto you in a timelymanner?–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Do you feel you weresufficiently consultedduring the life of theproject?Figure 10. Website Coordinator questionnaire.Questions Comments/answers–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Was the timeframegiven to design thewebpage of theTechnologist programmesufficient?–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Do you feel sufficientinformation about theproject was providedto you in a timelymanner?–––––––––––––––––––––––––––––––––––––––––––––––––––––––––Do you feel your viewson the design and layoutof the webpage weretaken into accountand was the projectlead open to yoursuggestions?–––––––––––––––––––––––––––––––––––––––––––––––––––––––––To what extent wereyour needs or concernsin relation to the projectmet?Conclusions/lessons learnedWorking on this project has been a challenge; It hasinvolved a closer working relationship with somemembers of Council than before which has given meconfidence in my ability to work with people on aApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 28

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Animal Technologist Exchange Programme: A Level 6 Higher Education project293Betts, P.W. (1999). Supervisory Management. 7thedition, pp 285-347 Pub. Longman ISBN-10:0582418771, ISBN-13: 978-05824187764 Last accessed: 24/01/20185 last accessed 20/01/20187Chartered Management Institute (2015). Managingbusiness communication, Profile Books, LondonApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 29

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31PAPER SUMMARYTRANSLATIONSINHALTVERZEICHNISINHALTVERZEICHNISTrainingsprojekt: Intravenöse Injektion, Teil 1 –Planung und Umfang. Ein Hochschulprojekt derStufe 6 [Abschlussjahr]CARMEN ABELABiological Services Facility, Faculty of Infectious and Tropical Diseases, London School ofHygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, GroßbritannienKorrespondenz: intravenöse (i.v.) Injektion über die laterale Schwanzvene ist ein Verabreichungsweg beim Einsatz von Tieren inder medizinischen Forschung. Es ist die bevorzugte Art zur Abgabe von Substanzen, insbesondere wenn eineVermeidung des First-Pass-Effekts durch den Leberstoffwechsel erwünscht ist. Sie wird auch verwendet, um denvorgeschlagenen Weg der endgültigen Verabreichung zu simulieren. Sie ist jedoch technisch schwierig und erforderteine spezielle Ausbildung. Ungenauigkeiten während der Injektion sind häufig, weshalb Forscher davon absehen,diese Art zu nutzen, und stattdessen lieber auf üblichere Verabreichungswege wie die intraperitoneale Injektion oderdie Schlundsonde zurückgreifen.Die Verwendung von lebenden Tieren zum Erwerb technischer Fähigkeiten verbietet sich ethisch und gesetzlich, undin der Regel wird technische Kompetenz durch die Praxis mit Kadavern erworben. Ziel dieses Projekts ist es,festzustellen, ob es notwendig ist, Tierkadaver – tote Mäuse – für das Training zu verwenden oder ob ein alternativesTrainingsinstrument ebenso effektiv ist. Das Konzept eines Trainingsprojektes ist integraler Bestandteil desErgebnisses und muss, damit es erfolgreich sein kann, fristgerecht, innerhalb des Budgets und nach Spezifikationerfolgen.Schlagwörter: Training, intravenöse Injektionen, Maus, Kadaver, technische FähigkeitenApril 2019 Animal Technology and WelfareApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 31

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Paper Summary Translations32Tiertechnologen-Austauschprogramm:Tiertechnologen-AustauschprogrammMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road, London W12 ONN,GroßbritannienKorrespondenz: auf einem Hochschul-Projektmanagementauftrag der Stufe 6 [Abschlussjahr]AbstractDem Institute of Animal Technology wurde zur Kenntnis gebracht, dass es der biomedizinischen Industrie an einerformalen Möglichkeit zum Austausch zwischen Tiertechnologen verschiedener wissenschaftlicher Einrichtungenmangelt, um weitere Kenntnisse und Fähigkeiten erwerben zu können. Das Animal Technologist Exchange Programme(ATEP) wurde entwickelt, um Tiertechnologen eine strukturierte Möglichkeit zum Austausch zu bieten.Das Programm steht allen Ebenen von Tiertechnologen offen, einschließlich Auszubildenden, und die Austauschekönnen von unterschiedlicher Dauer sein.Schlagwörter: Tiertechnologen, Austauschprogramm, Fähigkeiten, fachliche WeiterbildungApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 32

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April 2019 Animal Technology and Welfare33CONTENU DE LA REVUECONTENU DE LA REVUEProjet de formation sur les injectionsintraveineuses: Partie 1 – Planification et portée duprojet. Un projet d’enseignement supérieur deniveau 6CARMEN ABELABiological Services Facility, Faculty of Infectious and Tropical Diseases, London School ofHygiene and Tropical Medicine, Keppel Street, Londres WC1E 7HTCorrespondance:ésuméLes injections intraveineuses (i.v.) effectuées via la veine caudale latérale représentent l’une des méthodesd’administration sur les animaux utilisés dans la recherche médicale. Il s’agit de la technique privilégiée pourl’administration de substances, en particulier lorsque l’on tente d’éviter un effet de premier passage parmétabolisme hépatique. Elles sont également utilisées pour reproduire la méthode d’administration finale proposée.Elles sont néanmoins difficiles à réaliser sur le plan technique et nécessitent une formation spécialisée. Les erreurspendant les injections sont fréquentes, ce qui dissuade les chercheurs d’employer cette méthode et les encourageà utiliser plutôt les techniques de dosage plus courantes telles que l’injection intrapéritonéale ou le gavage oral.L’utilisation d’animaux vivants en vue d’acquérir des compétences techniques est interdite d’un point de vue éthiqueet légal, et les compétences techniques sont habituellement obtenues en utilisant des cadavres d’animaux. Ce projeta pour objectif de déterminer s’il est nécessaire d’utiliser des cadavres d’animaux (souris mortes) à des fins deformation, ou bien si un outil de formation alternatif est aussi efficace. La conception d’un projet de formationdéterminera le succès de ce programme, et afin que ce dernier soit réussi, elle doit respecter des délais temporels,des contraintes budgétaires et les spécifications requises.Mots-clés: Formation, injections intraveineuses, souris, cadavres, compétences techniquesApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 33

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Paper Summary Translations34Programme d’échange des technologues animaliers:Programme d’échange des technologues animaliersMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road, Londres W12 ONNCorrespondance:é sur une mission de gestion de projet « HE de niveau 6 »RésuméIl a été porté à l’attention de l’Institut des technologies animales que le secteur biomédical ne possédait aucuneméthode formelle permettant aux technologues animaliers de communiquer avec leurs collègues basés au seind’autres établissements scientifiques afin d’acquérir des connaissances et des compétences supplémentaires. Leprogramme d’échange des technologues animaliers (ATEP ou Animal Technologist Exchange Programme) a été conçudans le but de fournir un cadre structuré favorisant la communication entre les technologues animaliers.Ce programme est offert aux technologues animaliers de tous niveaux (y compris les stagiaires), et les échangespeuvent être d’une durée variable.Mots-clés: Technologues animaliers, programme d’échange, compétences, formation professionnelle continueApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 34

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April 2019 Animal Technology and Welfare35INDICE DE LA REVISTAINDICE DE LA REVISTAProyecto de formación sobre inyeccionesintravenosas: Parte 1: Planificación y alcance.Proyecto de Educación Superior de Nivel 6CARMEN ABELABiological Services Facility, Faculty of Infectious and Tropical Diseases, London School ofHygiene and Tropical Medicine, Keppel Street, Londres WC1E 7HTCorrespondencia: inyección intravenosa (i.v.) a través de la vena lateral de la cola es una vía de administración que se usa al utilizaranimales para la investigación médica. Es la vía preferente para el suministro de sustancias, especialmente cuandose quiere evitar el efecto de primer paso a través del metabolismo hepático. También se utiliza para imitar la víapropuesta de administración final. Sin embargo, resulta técnicamente complicada y requiere formaciónespecializada. Suelen darse imprecisiones durante la inyección, de forma que los investigadores prefieren evitarutilizar esta vía en favor de vías de dosificación más comunes como la inyección intraperitoneal o la sonda gástrica.El uso de animales vivos para obtener conocimientos técnicos está ética y legalmente prohibido, de modo que seobtienen aptitudes técnicas utilizando cadáveres. Con este proyecto, se pretende aclarar si existe la necesidad deutilizar cadáveres de animales para la formación, como ratones muertos, o si existen otras herramientas deformación alternativas igual de eficaces. El diseño de un proyecto de formación es fundamental para los resultadosy para que tenga éxito este debe ser puntual, ajustarse al presupuesto y cumplir las especificaciones.Palabras clave: formación, inyecciones intravenosas, ratón, cadáveres, aptitudes técnicas.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 35

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Paper Summary Translations36Programa de intercambio de tecnólogos deanimales:Programa de intercambio de tecnólogos deanimalesMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road, Londres W12 ONNCorrespondencia: en una asignación de gestión de proyectos HE de nivel 6ResumenSegún se ha hecho saber al Institute of Animal Technology, el sector biomédico carece de un modo formal medianteel cual se puedan producir intercambios entre tecnólogos de animales en otros establecimientos científicos a fin deampliar conocimientos y aptitudes. El Animal Technologist Exchange Programme (ATEP) se ha diseñado para ofreceruna forma estructural de intercambio de tecnólogos.El programa está abierto a tecnólogos de animales de todos los niveles, incluidos aprendices, y los intercambiospueden tener varias duraciones.Palabras clave: tecnólogos de animales, programa de intercambio, aptitud, formación, formación continua.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 36

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April 2019 Animal Technology and Welfare37INDICE DELLA REVISTAINDICE DELLA REVISTAProgetto di formazione sulle iniezioni intravenose:parte 1 – pianificazione e scoping. Progetto diLivello 6 di formazione superioreCARMEN ABELABiological Services Facility, Faculty of Infectious and Tropical Diseases, London School ofHygiene and Tropical Medicine, Keppel Street, Londra WC1E 7HTCorrispondenza:’iniezione intravenosa (i).v. tramite la vena caudale laterale è una modalità di somministrazione utilizzata in ricerchemediche che fanno uso di animali. È il metodo preferito per la somministrazione di sostanze, soprattutto se si vuoleevitare l’effetto del primo passaggio attraverso il metabolismo epatico. Vi si fa ricorso anche per simulare la via disomministrazione finale proposta. Tuttavia, risulta difficoltoso da un punto di vista tecnico e richiede una formazionespecialistica. Durante l’iniezione, si verificano comunemente errori vari che scoraggiano i ricercatori dall’usare talemetodo a favore di forme di dosaggio più comuni, quali l’iniezione intraperitoneale o la sonda orogastrica.Per motivi etici e legali, l’uso di animali vivi per l’acquisizione di competenze tecniche è severamente proibito e, disolito, la competenza tecnica si sviluppa facendo pratica sui cadaveri. Questo progetto si prefigge di determinarel’effettiva necessità di utilizzo di cadaveri di animali per la formazione (ovvero topi morti) o se uno strumentoformativo alternativo possa rivelarsi ugualmente efficace. L’ideazione di un progetto di formazione fa parte integrantedell’esito e, per favorirne la riuscita, deve avvenire entro i limiti temporali stabiliti, rientrare nel budget e rispettarele specifiche impostate.Parole chiave: Formazione, iniezioni intravenose, topo, cadaveri, competenze tecnicheApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 37

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Paper Summary Translations38Animal Technologist Exchange Programme:programma di scambio per stabularistiMATTHEW BILTONCBS Imperial College London, Hammersmith Campus, Du Cane Road, Londra W12 ONNCorrispondenza: su una ricerca di project management del Livello 6 di formazione superioreAbstractAll’Institute of Animal Technology è stato segnalato che il settore biomedicale non disponeva di un canale di scambioformale tra stabularisti di istituti scientifici diversi che permettesse loro di acquisire ulteriori conoscenze ecompetenze. L’Animal Technologist Exchange Programme (ATEP) è stato progettato per fornire un quadro strutturatoentro cui gli stabularisti possono effettuare gli scambi.Il programma è aperto a stabularisti di ogni livello, apprendisti inclusi, e offre scambi di durata diversa.Parole chiave: Stabularisti, programma di scambio, competenze, formazione, Sviluppo professionale continuoApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 38

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39Haven’t the time to write a paper but want to get something published? Then read on!This section offers readers the opportunity to submit informal contributions about anyaspects of animal technology. Comments, observations, descriptions of new or refinedtechniques, new products or equipment, old products or equipment adapted to new use,any subject that may be useful to technicians in other institutions. Submissions can bepresented as technical notes and do not need to be structured and can be as short or aslong as is necessary. Accompanying illustrations and/or photos should be high resolution.NB. Descriptions of new products or equipment submitted by manufacturers are welcomebut should be a factual account of the product. However, the Editorial Board gives nowarranty as to the accuracy or fitness for purpose of the product.Using software to tackle the 3Rs –A scientist’s perspectiveCHRIS W.D. ARMSTRONG1,21Overwatch Research, Ormeau Baths, 18 Ormeau Avenue, Belfast BT2 8HS2Centre for Cancer Research and Cell Biology, Queen’s University Belfast, Belfast BT9 7AECorresponding author: chris@overwatchresearch.comApril 2019 Animal Technology and WelfareTECH-2-TECHIntroductionThe most recent figures have shown that 50% of pre-clinical research findings cannot be replicated by anindependent laboratory. In the USA alone, this meansthat of the $56.4 billion spent every year, half isessentially wasted on ‘bad science’.1These figures arethought to be representative throughout the world withsimilar rates of irreproducibility evident in Europe andAsia. Consider this in the context of in vivo experiments– half of the animals subjected to experimentationproduce results that are totally unreliable.Of course, this is not the fault of the animal andresponsibility falls entirely on the humans involved incontrolling the study. Mistakes are made duringprocesses such as study design, data collection anddata analysis. However, regardless of where themistake is made, the justification for using the animalafter this point is non-valid. The knock-on effect of thisis even worse. Additional animals are recruited to thestudy or entire experiments are repeated to correctprior mistakes.This is an all too common occurrence that could beeasily tackled using modern resources. Experimentalprocesses should be automated to eliminate human-input and ensure that results are reproducible.Moreover, we can take this further and directly addressother aspects of in vivo studies that also relate to the3Rs. This requires the combined effort of all interestedparties: technologists, scientists, facility managers andveterinary surgeons. To facilitate this, we need totaltransparency so that identified issues can be actedupon immediately. Put simply – the informationassociated with every animal (measurements,observations etc) should be visible to everyone toensure that animal welfare remains at the forefront ofour thoughts.Dozens of innovative products enter the market everyyear with a claim to have a direct impact on the 3Rs. ItApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 39

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40Tech-2-Techdoes not take long to see that the majority of these arereliant on specific hardware. However, the previouslymentioned problems that are experienced in researchlaboratories worldwide could benefit from a much moreaccessible solution. This is where the importance ofsoftware should not be overlooked.The Animal Technologist andscientist experience comparisonWhen considering how improvements can be made tothe 3Rs, it is crucial to understand how basic problemsarise in the first instance. Animal Technologists workwithin the vivarium on a daily basis and quickly developlasting expertise in the area. This can be as simple asbeing able to identify animal welfare issues earlier; ordeveloping competent injection techniques thatminimise stress for the animal.However, for scientists such as PhD students andpostdoctoral researchers, it is not uncommon for themto go long periods of time without conducting any invivo-related work. For example, a student couldundertake a two-day course followed by a 10-minutemultiple-choice exam and be awarded with a personallicence to allow experimentation on animals. Theycould then not perform any studies for months (or evenyears) before stepping back in to an environment inwhich they lack competency. The result:G Poor experimental technique leading to the studybeing unnecessarily repeated.G Lack of experienced judgement to identify animalhealth issues as early as possible.G Missing basic warning signs such as weight loss oranimals progressing beyond the experimental end-point.G Forgetting to perform routine measurements ormaking mistakes in the measurement process,causing gaps in data collection and leading to moreanimals being recruited to the study to compensate.It is important to say that there are many experiencedscientists who have developed more than adequatecompetency in this area. However, for inexperiencedresearchers, there are obviously basic mistakes thatcould be eliminated by improving the visibility ofwarning signs throughout the experiment. Moreover,these warning signs could easily be made apparent totechnologists and facility managers as a second line ofdefence for catching welfare issues early.Spreadsheets, spreadsheets,spreadsheetsTo address this problem, we sought to understand themechanisms used by researchers to manage theirexperiments. Perhaps unsurprisingly, 148 out of 150research facilities confirmed that their scientists reliedon spreadsheets.Let us consider the impact that this has on a standardsubcutaneous tumour experiment. An averageexperiment might involve 100 mice measured threetimes per week over a six-week period. If the ellipsoidcalculation method is used then three dimensions(width, length and depth) are recorded for everymeasurement alongside a regular weight assessment.At the end of the six-week study, scientists are left witha spreadsheet encompassing 7,200 numerical values.Why is this method not fit for purpose?Measurements are often recorded in to a notebook asthey are per formed before transferring to aspreadsheet. This produces two opportunities for error.Firstly, if you have ever performed callipermeasurements of a tumour, you will know that it isphysically impossible to restrain the animal, performthe measurement and record the result simultaneously.Often researchers will attempt to quickly measure eachdimension (to two decimal places) before placing theanimal back in to the cage and recording themeasurements from memory. This is an obvious sourceof error across a range of in vivo experiments.Secondly, as the measurements are transferred to aspreadsheet later, transcription errors are inevitable.Furthermore, it is not uncommon for scientists toperform daily measurements with no accessible copyof results from previous days. This means that theyhave no basis for comparison when assessing changesin body weight. Even if they are pro-active and ensurethat previous measurements are visible – many willresort to performing calculations in their head to detectthresholds such as 20% weight loss.Finally, there are obvious limitations of spreadsheetssuch as the inability to randomise animals in to studygroups based on a scientifically-accepted format. If thisis performed incorrectly, it will eventually be detectedwhen attempting to publish the results of the study andmay be sufficient to cause an entire repeat of theexperiment – subjecting more animals to unnecessaryprocedures.All of these problems can be solved with softwareinnovation.Communication is keyIf we are to truly revolutionise the in vivo experimentprocess then there must be constant communicationbetween technologists, researchers, facility managersand Named Veterinary Surgeons (NVS). Unfortunately,this is not the case in the majority of laboratories. Infact, I am often amazed by the number of scientistswho do not even know the name of the NVS in theirfacility.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 40

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Tech-2-Tech41When communication does occur, it is almost entirelyreliant on email. There is no feedback to tell if amessage has been received, read or acted upon. Often,the original email does not include enough informationsuch as: animal location; previous measurements orobservations associated with the animal; or action thathas already been taken. This can lead to emails goingback and forward – all whilst the animal requiringattention is made to wait.In an ideal scenario, all involved parties should haveimmediate access to measurements or observationsrecorded against any animal in the facility. This allowsfor quick identification of trends such as decreasingbody weight or the rapid progression of disease. Infacilities that have tried to make this possible, it oftenmeans constantly providing an updated version of thestudy spreadsheet. Unfortunately, if 20 active studiesare being performed then this process is reliant on 20unorganised spreadsheets containing tens ofthousands of data points. Inevitably, importantproblems get overlooked.This is an obvious area where software can make ahuge impact. The ability to organise large amounts ofdata into an understandable format and instantly flaganimals which pass pre-defined thresholds should notbe undervalued. Importantly, access to this informationcan be shared with all relevant parties prior tocommencing the study so that communication isautomated and instantaneous.The modern era of softwareWe live in an era where software plays a huge role ineveryday life. Nearly 3 billion people worldwide own asmartphone and rely on a multitude of applications tocontrol their finances, social interactions, access tonews and much more. However, not only have webecome reliant on these software solutions; we alsoexpect a high standard of design, navigation, features,data security and reliability. This can be collectivelyreferred to as the ‘user experience’ and oftendetermines our decision to use one application overanother.Unfortunately, industries such as life sciences oftensuffer from this rapidly evolving model. Technologistsand scientists alike have become accustomed to usingout of date software that would not have looked out ofplace 15-20 years ago. With such high expectationsdictated by other modern software experiences, theresult is often frustration and a lack of willingness touse the application.The solution requires a software developmentapproach that is user-focused. Nothing can competewith physically going in to a vivarium to directlyexperience the frustrations faced by the staff daily.Speaking with technologists and scientists can allowfor rapid identification of problems on both sides thatcan then be directly addressed during software design.This modern approach has resulted in softwaresolutions being implemented across the life scienceindustry in recent years. Examples include: Labstep, asoftware product that is transforming scientificresearch via the implementation of a digital laboratorynotebook; Biorender, an application that has allowedscientists to produce professional cell biology diagramsin minutes; and Benchling, a molecular biology suiteand bioregistry that has already been adopted by topinstitutions such as Harvard University and theUniversity of Oxford.We would be foolish to think that in vivo studies couldnot benefit from a similar solution. Animal welfare andexperimental integrity could be dramatically improvedwith the right product. Therefore, the 3Rs could bedirectly addressed by software innovation.Over-focused software design foranimal researchOverwatch is an in vivo experimental tracking platformthat has been built specifically for researchers,technologists, facility managers and veterinarians.There are several distinct problems directly relating tothe 3Rs that Overwatch can play a role in solving.ProblemInadequate study design is a common feature of in vivoresearch meaning that experiments have to be repeated.SolutionAt Overwatch, we take care of study design and providescientifically-accepted templates to the researcher.Figure 1. Users have total control when pre-assigningthresholds that will be notified throughout the study.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 41

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Tech-2-Tech42These templates form the basis of the study andensure that the relevant information is collected andanalysed correctly. Importantly, templates can becustomised to suit the needs of different facilities. Theuser has total control to pre-define animal identificationmethods, weight loss thresholds and experimentalendpoints (Figure 1).ProblemData is recorded incorrectly when taking basicmeasurements which results in unreliable outcomes.More animals will be recruited in to the study tocompensate; or in a worst-case scenario, the entireexperiment will be repeated.SolutionWith in vivo experimentation, accurate data collection isthe single most important consideration alongsideanimal welfare. Currently, the process should never bereliant on manually recording into laboratory notebooksor spreadsheets. Automation of this process isrequired to ensure experimental integrity and preventthe usage of more animals to gain reliable results.Overwatch is fully compatible with Bluetooth or USB-enabled devices such as scales and callipers. Thismeans that data is instantly sent to the software withno risk of error. Not only is this important for error-reduction; users have reported time-savings of up to30% when performing routine measurements (Figure2). Moreover, custom data uploads allow researchersand technicians to upload data directly from imagingsystems including x-ray, BLI and mCT. This ensuresconsistency across all studies and removes anyconcern regarding the reliability of the results.ProblemData is often transferred between laboratorynotebooks, spreadsheets and analysis software togenerate graphs. Mistakes are common whentransferring large amounts of data meaning that theresult can be inaccurate. These inaccurate results areoften used when deciding to perform future in vivoexperiments meaning that more animals areunnecessarily used.SolutionThe optimal solution to this problem is removing anyhuman input. With Overwatch, data is automaticallygraphed as it is collected ensuring that no mistakesare possible. These graphs can be exported inseconds, taking away the stress of data analysis(Figure 3).ProblemIt is not uncommon for an animal to lose weight or passan experimental end-point and go unnoticed.Measurements associated with animals that areenrolled in studies are often not visible to anyone otherthan the researcher. This has serious implicationstowards animal welfare and can even result in ethicalboundaries being passed.SolutionWith animal welfare-related issues, there should notbe any possibility of missing an important threshold.With Overwatch, if a measurement is performed thatpasses pre-defined weight loss or disease limits, anotification will be displayed instantly. Thisnotification allows for immediate identification ofproblems and will remain present until action hasbeen taken (Figure 4).Figure 2. Measurements can be automaticallytransmitted to the software using Bluetooth-enabledequipment. Every measurement is date and timestamped.Figure 3. Graphs are updated automatically as data isentered.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 42

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Tech-2-Tech43ProblemAside from metrics that can be directly measured(weight, disease burden etc) it is also important tohave a full medical or observation record associatedwith each animal. Often a medical issue will beobserved but there will be no available information totell if this is new or has presented previously. Having afull record of observations allows for action to be takenmuch sooner.SolutionOverwatch allows technologists and vets alike to enterobservations for any given animal at any time. Thesewill be permanently available within the animal profileto build a reliable medical history that can be accessedinstantly (Figure 5).ConclusionIt is time to bring in vivo facility processes in to thedigital age. Experiments involving animals should be asautomated as possible to increase confidence inresearch findings and prevent unnecessary futurestudies. Data (measurements, scans andobservations) from these experiments should be visibleto not only the researcher, but also the technologistsand NVS. Overwatch can address many of these issuesand directly help facilities to implement the 3Rs.References1Freedman, L.P., Cockburn, I.M. and Simcoe, T.S. (2015).The Economics of Reproducibility in Preclinical Research.PLoS Biol 13(6): e1002165. 4. If a measurement passes pre-definedthresholds, a notification will be displayed to alert youimmediately.Figure 5. Observations can be recorded against allanimals and are easily accessed.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 43

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45AbstractLiterature is replete with studies on stressors tolaboratory rodents; knowing the impact research andhusbandry interventions, such as routine cage-changing, have on rodents’ stress levels, physiologyand behaviour, we hypothesised that animalsrecovering in a familiar environment would have animproved recovery compared to those recovering in aclean cage. Using weight as an indicator, our studiesproved a home cage provides a better recoveryenvironment than a clean one. Habituated animalswere above their pre-surgery weight a week laterwhereas animals returned to a barren cage lost mostweight overall and were still below their pre-surgeryweight seven days post-operatively.IntroductionAn increase in heart rate is acknowledged as ameasure of stress;1when moved to a clean cage, arodent’s heart rate increases twofold for over an hourand for this reason it can be argued as one of the moststressful procedure: for example blood sampling, whichinvolves nicking the tail with a sharp blade, causes arodents heart rate to raise for 35-40 minutes,2incomparison a cage base change causes a longer rise inheart rate (45-60 minutes) and also an increase inphysical activity.3A healthy animal is better equipped to cope with thestress a change in husbandry can cause, for thisreason, we suggest any alteration from the animal’sstandard routine should be pre-emptive to the surgicalprocedure.Cage change before surgeryTypically in the past animals have recovered post-surgery, in a ‘vet bed’ lined cage inside a heatedchamber to provide a comfortable area to regainconsciousness and vital functions, before beingreturned to a clean cage.4Although this practicepromotes good hygiene, because rodents are reliant onolfactory cues for recognising and communicating withcage mates5it can be highly disruptive and stressfulrecovering in an unfamiliar environment.Bearing this in mind, we suggest it is best practice todo a cage change two to twenty-four hours before theprocedure because after two hours the animal’scardiovascular output will have returned to itsbaseline.3This acclimatisation period ensures theoccupants are habituated to their new cage while alsobeing in clean accommodation for recovery.MethodHousing and husbandryThe animals were housed in a barrier facility, whichoperates on being clear of pathogens according to TheFederation of European Laboratory Animal ScienceAssociations (FELASA) Guidelines andRecommendations.6The specified pathogen freeanimals were housed in individually ventilated cages(IVCs). The standard cage contents and environmentalenrichment was aspen bedding, cotton nestingmaterial, a permanent shelter, a raised cardboardtunnel and a chew stick. For social enrichment, themice were housed in groups of 2-6 individuals per cage.The animals were on a 12h light and dark period with aANDREW BLAKE TRIBUTE AWARD2019 ENTRYGive us a break!EMMA FILBYMira Building, University of Cambridge, University Biomedical Services,Charles Babbage Rd, Cambridge CB3 0FSCorrespondence: for the Andrew Blake Tribute Award 2019April 2019 Animal Technology and WelfareSponsored by The Association of the BritishPharmaceutical IndustryApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 45

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Andrew Blake Technician Award46dimming phase at dawn and dusk; the environmentparameters were in accordance with the Home OfficeCode of Practice.736 mixed sex C57 BL/6 mice, aged 13-16 weeks old,held under Animal [Scientific Procedures] Act 1986having an identical peripheral nerve lesion were splitequally into two groups; all surgical procedures werecarried out according to the LASA Guiding Principles forPreparing for and Undertaking Aseptic Surgery (2010).81. Group 1: Returned to home cage (‘habituated’) withthe aim to test our hypothesis.2. Group 2: Returned to a clean cage (‘non-habituated’) to act as controls, based on normalpractice.Experimental procedurePre-Operative PreparationsWe checked that the induction box has been flushedthrough with oxygen so that no isoflurane was present.To oxygenate the animal, we administered pureoxygen, 0% isoflurane. Next, 5% isoflurane and 100%oxygen was administered in order to achieve asmooth, rapid induction. Once unconscious, theanimal was maintained at 1.75-2.5% isoflurane and100% oxygen.The first line of pain relief would usually be Non-Steroidal Anti-Inflammatory Drugs (NSAIDs), however,this would interfere with the studies aim. Inflammationis important because remyelination is being studiedand the inflammatory response initiated bydemyelination is an important trigger forremyelination.9,10Because we need to see inflammationto trigger remyelination we cannot use anti-inflammatory pain relief so an opioid is used.We administer analgesic appropriate to weight:SUBSTANCE ROUTE DOSE DILUTION––––––––––––––––––––––––––––––––––––––––––––––––––Buprenorphine Subcutaneous 0.1ml per 100g 1:6For any surgery longer than 20 minutes we alsoadminister saline, 1ml per mouse subcutaneously inacknowledgement that their internal tissues will dry outwhen exposed to the air.SurgeryThe lower back was incised, the sciatic nerve identifiedand crushed. Post operatively animals could experiencesubtle defects such as the ability to use and move thehind limbs resulting in dragging of the lower operatedlimb and slower movements around the cage, anyadverse effect would resolve within 1 week. Generallythe recovery was expected to be uneventful. The animalswere humanely killed 4 weeks after the procedure.Recovery EnvironmentsGroup 1 had a cage base change the day before thesurgical procedure. On the day of surgery the animalswere taken to the pre-operative area and moved into aclean cage base with a small amount of soiled materialtransferred. The original home cage was placed insidethe heated chamber with diet in gel form and theanimals were returned to it post-operatively.Group 2 were brought into the pre-operative area intheir home cage. A clean base was placed inside theheated chamber with diet in gel form and the animalswere returned to it post-operatively.We took a bodyweight measurement pre-surgery andthen recorded their bodyweight daily for three days thenone final measurement a week later. Using bodyweightdata as an indicator, we wanted to compare the weightloss in animals habituated to their diet and cageenvironment before surgery to those returned to a newcage with no prior exposure to food types with the aimof finding which recovery environment was better.ResultsNormalising the data to show the increase/decrease inweight post operatively.Figure 1. Plot showing all the data, data for eachindividual mouse in the background showing variation.Figure 2. Data ‘split’ by gender.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 46

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Andrew Blake Technician Award47Looking at the mean values:DAY GROUP MEAN––––––––––––––––––––––––––––––––––––––––––––––––––0 Habituated 21.067 Non-habituated 21.100 Habituated 24.986 Non-habituated 24.32DAY GROUP MEAN––––––––––––––––––––––––––––––––––––––––––––––––––0 Habituated 21.061 Non-habituated 20.200 Habituated 24.981 Non-habituated 23.71– The habituated animals are marginally above theirpre surgery weight where the non-habituated onesare still below.– The lowest weight was recorded on day 1 for bothgroups. The non-habituated animals lost most weight.– Gender has no impact on the negative effects of nohabituation.DiscussionSurgeons and Animal Technologists should be aware ofthe impact of research and husbandry interventionsand that the rat’s stress response can affect theirphysiology, behaviour and consequentially dataquality.11We propose a balance where this proven beneficialhabituation can be achieved without compromisinghygiene.1. Change the cage base two to twenty-four hoursbefore the procedure as after two hours theanimal’s cardio vascular output will have returned toits baseline.32. Animals due for surgery are moved into a cleanbase, a small amount of soiled material istransferred. The transfer of some existing substrateis important for social hierarchy.5The occupants’heart rate is still elevated as even when soiledbedding is transferred, the familiar smells do notcounter the novelty of the new cage.33. When no longer occupied, move original homecage into heated chamber to be a warm, familiarrecovery environment.The stress of the healthy animals pre-surgery isoutweighed by benefits and improved recoveryachieved by returning the recovering animals to theirhome cage.ConclusionIf we have animals in a stressed state, they experiencephysiological changes and their opioid receptors will beblocked12meaning that they experience pain at a higherintensity as they are less responsive to pain reliefnegatively impacting their welfare and subsequentrecovery.Our review indicates deviating from the currentprocedures would reduce stress and benefit animals inresearch. Having animals with lower stress levelswould mean that welfare is promoted, recoveryimproves and their clinical relevance as an animalmodel is increased. Surgeons and AnimalTechnologists should be aware of the impact ofresearch and husbandry interventions and that therats’ stress response can affect their physiology,behaviour and consequentially data quality.11Additionally, in accordance with the 3Rs concept(reduction, replacement and refinement) the decreasein variables would also reduce the number of animalsrequired in an experimental protocol.AcknowledgementsMira Animal Care Staff Mira Building, University ofCambridge, University Biomedical Services.Dr David Price Research Associate (Statistics),University of Cambridge, Department of VeterinaryMedicine, Disease Dynamics Unit, Madingley Road,Cambridge.Hélène Gautier Department of Physiology,Development and Neuroscience, University ofCambridge, United Kingdom.References1Meijer, M.K., Spruijt, B.M., van Zutphen, L.F.M. andBaumans, V. (2006). Effect of Restraint and InjectionMethods on Heart Rate and Body Temperature in Mice.Laboratory Animals 40(4): 382-91.2Gerdin, Anna Karin et al. (2012). Experimental andHusbandry Procedures as Potential Modifiers of theResults of Phenotyping Tests. Physiology and Behaviour106(5): 602-11.3Duke, J.L., Zammit, T.G. and Lawson, D.M. (2001). TheEffects of Routine Cage-Changing on Cardiovascular andBehavioural Parameters in Male Sprague-Dawley Rats.Contemporary topics in laboratory animal science/American Association for Laborator y Animal Science40(1):17-20., P. (2015). Laboratory Animal Anaesthesia. 4thEdition. Pub. Academic Press. Hardcover ISBN:9780128000366, eBook ISBN: 9780128005781Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 47

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Andrew Blake Technician Award485Brennan, Peter A. and Kendrick, K.M. (2006).Mammalian Social Odours: Attraction and IndividualRecognition. Philosophical transactions of the RoyalSociety of London. Series B, Biological sciences361(1476): 2061-78. & rendertype=abstract.6FELASA. (2014). FELASA recommendations for the healthmonitoring of mouse, rat, hamster, guinea pig and rabbitcolonies in breeding and experimental units. Laborator yAnimals, 178-192.7Featherstone, H.O. (2014, Dec 17). Code of practice forthe housing and care of animals bred, supplied or usedfor scientific purposes. Retrieved from Guiding Principles for Preparing for andUndertaking Aseptic Surgery (2017). LASA 2017 GuidingPrinciples for Preparing for and Undertaking AsepticSurgery. A report by the LASA Education, Training andEthics section. Eds. Lilley and M. Berdoy., A., Lathia, J.D., Zhao, C., Wells, K., Rao, M.S.,Ffrench-Constant, C. and Franklin, R.J. (2006).Inflammation stimulates myelination by transplantedoligodendrocyte precursor cells. Glia. 54. 297-303.10.1002/glia.2010Zhao, C., Li, W.W. and Franklin, R.J. (2006). Differencesin the early inflammatory responses to toxin-induceddemyelination are associated with the age-related declinein CNS remyelination. Neurobiol Ageing. 2006 Sep;27(9):1298-307.11Castelhano-Carlos, M.J. and Baumans, V. (2009).“The Impact of Light, Noise, Cage Cleaning and in-House Transport on Welfare and Stress of LaboratoryRats.” Laborator y animals 43(4): 311-27.é Le Roy, E. L.-B.-P. (2011). Endogenous OpioidsReleased During Non-Nociceptive Environmental StressInduce Latent Pain Sensitisation via a NMDA-DependentProcess.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 48

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49AimTo investigate the effect of altered water hardness onZebrafish embryonic development and survival ratesfrom point of fertilisation until day three postfertilisation.IntroductionAs ectothermic poikilotherms, Zebrafish (genus Danio)are significantly more reliant on the environment forbodily function than endothermic homeotherms suchas man and mice.As the environment in which Zebrafish live, waterquality parameters are measured regularly. Waterquality is the number one factor affecting the healthand welfare of Zebrafish. (Table 1)All water has a hardness level – those low in mineralsare referred to as soft water and are alkaline in nature.Those high in minerals are referred to as hard waterand are more acidic in nature. Zebrafish are considereda hard water species.Water chemistry is a fairly complex thing as all waterparameters are interconnected (Figure 1). All waterquality measurements are interdependent on oneanother.POSTER PRESENTATIONSOriginally presented at:IAT Congress 2018Impact of water hardness on Zebrafishearly stage developmentCAROLE WILSON, PAUL BARWOOD, VISILIA MOICHE,JENNA HAKKESTEEG and KAREN DUNFORDUniversity College London, Zebrafish Facility, Division of Biosciences, Gower Street, LondonWC1E 6BTCorrespondence: 2019 Animal Technology and WelfareTable 1. Water parameters commonly used forZebrafish.Figure 1. Water quality is highly dependent on itscomponents; each can influence and change another.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 49

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Poster Presentations50MethodsWe undertook five separate trials to determine theeffect of various components of water hardness on thesurvival rate, growth and development of early stageZebrafish embryos (Table 2). We then placed fertilisedone-cell embryos into each mix and kept them thereuntil four days post fertilisation (dpf) to assess survivaland development rates (Figure 2).Table 2. The different mixes used for five afterhardness trials.THE FIVE DIFFERENT WATER HARDNESS TRIALS––––––––––––––––––––––––––––––––––––––––––––––––––Trial 1Altering ratios of TH cations – Ca2+, Mg2+, maintaining TH at 200mg/L; maintaining KH at 50 mg/L in a ratio of 1:1––––––––––––––––––––––––––––––––––––––––––––––––––Trial 2Altering the levels of TH but always maintaining a ratio ofcations at 1:1; maintaining KH at 50 mg/l in a ratio of 1:1––––––––––––––––––––––––––––––––––––––––––––––––––Trial 3Altering KH levels but maintaining a ratio of 1:1; maintaining THat a 0:0 ratio of Ca2+: Mg2+––––––––––––––––––––––––––––––––––––––––––––––––––Trial 4No KH; Altering ratios of TH cations – Ca2+, Mg2+maintaining at200 mg/L––––––––––––––––––––––––––––––––––––––––––––––––––Trial 5Increased the TH to 900 mg/L whilst also altering ratios ofcations of Ca2+: Mg2+; maintaining KH at 50 mg/LResultsTrial 1. Percentage survival at 0 and 4 dpf. High levelsof mild heart oedema Ca2free trial (data not shown).Trial 2. Percentage survival at 0 and 4 dpf. High levelsof debris on chorions at 1200mg/L TH. No survival at2 dpf at 0mg/L TH. (Data not shown).Trial 3. Percentage survival at 1, 2 and 4 dpf. Highlevels of debris in water and chorions at 400mg/L KH.(Table 4).Figure 2. Left: water of five different water hardnesslevels were made. Right: embryos that were placed intothe different mixes.Total hardness is the sum of multivalent cations(commonly magnesium and calcium). We usedPotassium bicarbonate and Sodium bicarbonate tocreate carbonate hardness (carbonates, bicarbonatesand carbonic acids in solution). In each trial we useddifferent levels of total and carbonate hardness and insome, varied the ratios of calcium and magnesium inthe total hardness. We used Magnesium sulphateheptahydrate and Calcium sulphate dihydrate.Table 3. Water hardness has three component parts:total hardness, carbonate hardness and non-carbonatehardness. In this trial we excluded non-carbonatehardness as it is usually included as a component oftotal hardness.Table 4. Survival as significantly different between50mg/L and 200 and 400 mg/L. **Indicates p<0.01(one way ANOVA with Tukeys HSD post-test). Data areshown as mean ± SD.Trial 4. Percentage survival at 1, 2 and 3 dpf. Lowsurvival at 2 dpf in both Ca2and Mg Free samples.Varying levels and types of embryo abnormalities overall other samples. (Table 5)Trial 5. Percentage survival at 1, 2 and 3 dpf. Low orno survival at 2 dpf in both Ca+and Mg free samples.Varying levels and types of embryo abnormalities overall other samples. High levels of debris in water and onchorions in all samples. (Table 6)Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 50

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Poster Presentations51DiscussionCalcium and magnesium are essential fordevelopment, growth and survival. Both calcium andmagnesium are required for bone growth, blood clottingand other metabolic reactors. These trials appear toconfirm that both are imperative for development – thetrials in which one or the other was missing resulted instatistically significant low survival rates. Somewhatstrangely when no TH or KH was present (thereforewater with no minerals in) the embryos still developedto 3 dpf: perhaps this is because the developingembryo is enclosed and protected from the water bythe chorion and reliant on the yolk for nutrition. Olderfish would not be able to survive in this very pure water.When calcium and magnesium become too high theyprecipitate out of the water and formed crystals (figure3) which was seen especialy in Trial 5. As well asforming crystals in the water, it also clung to thechorions and gave high levels of embryo defomites,suggesting it was penetrating the chorion anddamaging the embryo. (Figure 8).When the results were compiled (Figure 7), bettersurvival rates with no abnormalities or precipitationwere observed in Trial 2, suggesting that slightly higherTH levels may increase embryo viability.The orange square indicates normal water hardnessconditions run at UCL. The red squares indicate waterhardness conditions that potentially should be rejectedon the grounds of debris in the water, pH of the waterand abnormalities in the embryos. The green squaresneed further investigation and were chosen because noembryonic abnormalities were found, nor debris in thewater and had an overall high final survival rate.Figure 3. Crystals precipitating out in the trials water.Table 7. First Row indicates trial number. Second rowindicates the ratio of magnesium calcium or in Trials 2and 3 amount of carbonate hardness used. Third rowindicates final survival rates. Fourth row indicates thefollowing: debris = water debris, unhat = unhatchedembryos; ab = embryoni abnormalities; pigles =pigmentless embryos. Fifth row indicates pH of thewater in the trial 3, 4, 5 (not tested in trial 1, 2).Table 5. There was no significant difference in survivalbetween seven different Mg/Ca ratios. (One-wayANOVA). Data are shown as mean ±SD.Table 6. Survival was significantly different betweenalmost all the ratios. NS Indicates no significantdifference. (One-way ANOVA with Tukey’s HSD post-test). Data are shown as mean ± SD.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 51

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Poster Presentations52Further workFrom the overlaid table we were left with two potentialcandidates for further investigation. The resultssuggest that raising the levels of TH may improveembryo survival.An increase in total hardness would have to be testedon adult fish to establish if there were anyrepercussions for them. This seems unlikely asanecdotal evidence suggests that water hardnesslevels are generally held at higher levels in the USA.AcknowledgementsH. Callaway, E. Hitchcock, S. Eshmade.References1Lawrence, C. (2007). The husbandry of Zebrafish (Daniorerio): A review. Aquaculture 269 (1-4) pp 1-20.2Alestrom, P. (2013). Zebrafish: housing, husbandry andhealth monitoring recommendations (unpublished).Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 52

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April 2019 Animal Technology and Welfare53IntroductionThe increased scientific need for large numbers of goodquality embryos (Figure 1) has seen the rise of variousbreeding strategies. In this poster, a comparison wasmade between four different spawning methods. Fromboth a scientific and welfare perspective, a goodbreeding strategy is key; it should reduce stress in fish,allow the fish to express mate choice and preference,as well as to choose whether or not to spawn.Comparing Zebrafish embryo productionmethodsHEATHER CALLOWAY and CAROLE WILSONUniversity College London, Zebrafish Facility, Division of Biosciences, Gower Street, LondonWC1E 6BTCorrespondence: 1. Good quality embryos typically produced fromhealthy adults. No visible deformities nor debris in oraround the embryos.Figure 2. From top left to bottom right: MEM1, MEM2and TM1 and TM2. The embryos collected weremeasured in 2ml units in a Falcon™ tube.MethodsThree rounds of testing were carried out to comparedifferent methods: mass embryo method 1 (MEM1)and 2 (MEM2) and traditional method 1 (TM1) and 2(TM2). (Figure 2).In the first round comparisons of all four collectionmethod were per formed. Overall, 80 adult fishconsisting of 60 females and 20 males were used forMEM1, MEM2 and TM1. 20 pairs of adult fish wereused for TM2. Embryos were collected and counted foreach method using a sieve and a 15ml Falcon™ tubewhich was filled to the 2ml marker with embryos; thiswas done several times to obtain an average numberper ml.In the second round a comparison was made betweenMEM1, MEM2 and TM1. The embryos were collectedand counted using the same method as in the firstround.The third round was carried out comparing MEM1 andMEM2, specifically looking at the quality of embryosproduced.ResultsIn the first round we found no significant differencebetween number of embryos produced from eachmethod (Figure 3). In the second round, however, wefound a significant difference between the number ofembryos produced from MEM1 and those producedfrom TM1 (Figure 4).Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 53

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Poster Presentations54Percentage of total number of embryos bycollection methodIn the third round we found no significant differencebetween the number of embryos produced by MEM1and MEM2 (data not shown) but there was a significantdifference in the quality of embryos produced by thetwo different methods. (Figure 5a and 5b)Quality was measured by eggs that were discarded dueto debris or abnormalities. (Figure 6)Number of embryos produced and discardedfrom Mass embryo methodsFigure 3. The total number of embryos collected acrossall methods with no significant difference between anymethod.Figure 4. Above left: TM1 yielded the highest number ofembryos over the mass production methods in thesecond round. Right: Egg production as significantlydifferent between MEM1 and TM1. * indicates p<0.05(One-way ANOVA with Tukey’s HSD post-test) Datashown as mean ±SD.Figure 6. Above right: the quality of the embryos wasvisible under a microscope. MEM1 had debris with thechorion; MEM2 had various stages of embryos; TM1has more uniformity.Figure 5a. The total amount of embryos collected fromboth MEMs were not entirely viable and some werediscarded (light blue).Figure 5b. The total number of eggs discarded wassignificantly between MEM1 and MEM2. **indicatesp<0.01 (Student’s t-test) Data shown as mean ±SD.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 54

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Poster Presentations55DiscussionFrom this investigation, there is little difference in thenumbers of embryos produced in each method:however, there appears to be an embryo quality andwelfare issue between MEM1 and MEM2. Not only weremore embryos discarded from MEM1 but the fish alsohave no choice to spawn. In fact, it could be said thatthey were forced to breed which could cause stress inthe group. Compared to MEM1, MEM2 allows the fishto exhibit more natural breeding behaviour and therebycould lessen potential stress, as the structure of theunit allows the fish to choose between breeding or notby removing themselves from the breeding area.In adhering to the 3Rs, all factors should be consideredto determine the best method for breeding ofZebrafish, including potential stress. Whilst all currentmethods appear to provide a high number of embryosrequired by research, the structure of equipment mayhave a negative impact on both parents and embryos.Further workFurther work to be carried out will include modificationof MEM1 to allow a refuge away from the breedingarea. The aim of this will be to determine if it willproduce embryos of a higher quality than the currentmethod.AcknowledgementsMany thanks to the UCL Zebrafish Facility includingCarole Wilson, Karen Dunford, Jenna Hakkesteeg,Visilia Moiche, Paul Barwood, Stephen Eshmade andElise Hitchcock.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 55

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56Animal Technology and Welfare April 2019AimTo assess the impact of different feeding regimes onZebrafish development, refine husbandry and reduceanimal numbers.IntroductionThe reliabity and reproducibility of research utilisinganimal models is paramount. Ideally, a model organismis maintained in standardised conditons, thusminimising variablity. Being a poikilothermic ectotherm,Zebrafish are highly adaptable, they expressenvironmentally induced plasticity at all levels:genetic,individual and population.Zebrafish husbandry, particularly the diets and feedingpractices in use vary greatly, presenting variousrefinement opportunities. Nutritional requirementsdiffer at various life stages but Zebrafish are typicallyfed one type of dry diet that only varies in size notcomposition. Live foods are commonly used tosupplement dry diets, increasing survival rates andreproductive performance1and may constituteenvironmental enrichment. However, live food can actas a vector for pathogens and is another source ofvariability. Removing this aspect of their husbandry hasgreater implications than the effects on growth andsurvival.This poster highlights some of the variation that occursnot only because of different husbandry practices butalso from the technologists caring for the animals. Thelack of standardisation between facilities means thatthere are many possible husbandry refinements whichwill lead in a reduction in the number of animals beingused and improving welfare.MethodsIn Trial 1 Zebrafish were fed four different dietscontaining dry food, live brine shrimps (Artemia spp.),frozen copepods and live rotifer (Bracghionus plicatilis).Standard length (SL) was measured approximatelyevery 8 days with survival recorded at 28 days postfertilisation (dpf) and 56 dpf.Trial 2, Zebrafish were fed 5 different commerciallyavailable dry diets, all supplemented with live rotifer.SL and width measurements were taken approximatelyevery 6 days, weights were recorded at 112 dpf.Survival was recorded at 28 dpf and 56 dpf.Comparison of feeding regimes forZebrafish (Danio rerio)PAUL BARWOOD and CAROLE WILSONUniversity College London, Zebrafish Facility, Division of Biosciences, Gower Street, LondonWC1E 6BTCorrespondence: 1. A trial by Maley et al 2006 highlighted howZebrafish growth is affected by diet.Figure 2. Live food. From top left. Artemia, copepodsand rotifer.Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 56

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Poster Presentations57Results 1Human error affected this trial. The dry diet feedingwas found to be inconsistent, negatively impacting thereliability of the results, the diets supplemented withlive artemia or live rotifer had the best growth, smallestsize variation and survival, indicating that their use canhelp to mitigate against suboptimal use of dry food.Figure 7. Graph showing the average weights at 112dpf of the female and male Zebrafish reared on fourdifferent diets.Figure 8. The fish fed dry diets A, B and E had significantdifferences in the ratio of females and males.Figure 3. Chart showing the survival rates of Zebrafishat 28 dpf and 56 dpf that were reared using fourdifferent diets.Results 2At 56 dpf, the fish fed diet D were significantly shorterthan those fed diets A, B, C and E. There was nosignificant difference between diets A, B, C and E. Thesurvival was similar for all the diets used.The similar survival rates of all five dry diets andsimilar SLs at 5 dpf of fish fed dry diets A, B, C and Esuggest similar performance between the diets.However, differing sex ratios, SL at sexual maturationand highly varied female weights at 112 dpf, SL atsexual maturation and the highly varied females’weights at 112 dpf indicate greater phenotypic variationdue to nutrition. Dry diet D was found to be nutritionallydeficient, resulting in poor growth rates and causedskeletal deformities, its use as stopped at 56 dpf.Figure 4. Growth curve showing the average lengths ofZebrafish reared on eight different diets from bothtrials 1 and 2.Figure 5. Scatter plots of length against widthsuggesting that Zebrafish reared on different dietsreach sexual maturity at different lengths.Figure 6. The body width of a well-conditioned maturemale (left) is less than that of a female (right).Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 57

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Poster Presentations58The SL of Zebrafish reared on different diets wasmeasured and the averages plotted over time. Thiscreates a growth curve showing growth rates. Trials 1and 2 are shown together allowing for comparison. Trial1 and Diet D of Trial 2 were stopped at 56 dpf. (Figure4).The scatter plots of width against SL suggest thatZebrafish reared on different diets reach sexualmaturity at different sizes. The region where the datasplits indicates when the body proportions of femalefish change due to ovary development. (Figure 5).Figure 6 shows that differences in width of the malesand females.The weights of female and male Zebrafish wererecorded at 112 dpf. The females fed dry diet A weresignificantly heavier than those fed dry diets B, C andE. The females fed dry diet C also weighed significantlyless than those fed dry diets B and E. No significantdifferences were found between the weights of themales (Figure 7).At 56 dpf the zebrafish fed dry diets A, B and E hadsignificant differences in the proportion of females andmale fish (Figure 8).DiscussionAs we know, Zebrafish are very plastic and this studysuggests that nutrition can have a high impact not onlyon growth, as might be expected but also on sex ratios– which may be more unexpected. The impacts of poorZebrafish nutriton are poorly understood2but it isknown that dietary nutrients can interact with molecularmechanisms and modulate physiological functions. Theeffect food has on gene expression (nutrigenomics)and how genetic variation affects responses tonutrients (nutrigenetics) needs more study – usually theonly factors considered are survival, growth rate andfecundity. The adaptability of Zebrafish likely meansthat dietary and feeding practices cannot beconsidered in isolation. It is likely that smallenvironmental changes such as tank proportions andcolour, stocking density, water flow rate and flowdynamics, water temperature and quality, light intensityand spectrum, food quality and quantity and breedingregimes will produce subtle differences. Whencumulatively added they may not be so subtle and willsurely impact the outcomes and reproducibility of thescience.Further workFurther work will include repeating Trial 1 to obtainreliable results, along with testing differences betweenvarious strains to identify any potential genetic effects.Additionally, body composition testing between saidstrains can identify any morphological variables withinthe study. On top of the testing, this can also beapplied to the diet variation. Body condition scoringusing different parameters can also be measured, suchas body depth compared to length.AcknowledgementsH. Callay, J. Hekkesteeg, V. Moiche, K. Dunford, S.Eshmade and E. Hitchcock.References1Goolish, E.M., Okutake, K. and Lesure, S. (1999). Growthand survivorship of larval zebrafish Danio rerio onprocessed diets. NA J Aquaculture 61:189-1982Watts, S.A., Powell, M. and D’Abramo, L.R. (2012).Fundamental approaches to the study of Zebrafishnutrition. ILAR Journal 53(2) 144-160Apr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 58

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59April 2019 Animal Technology and WelfareBackground10% of us are infected by a seasonal influenza viruseach winter resulting in 500,000 deaths worldwide.The study of influenza transmission is therefore vitallyimportant and aids the prediction of possiblepandemics.The ferret model has long been the “gold standard”animal model for studying human influenza infectionand is used all around the world for transmissionstudies.Professor Wendy Barclay and her team have usedferrets at Imperial College for 10 years for studyinginfluenza. In this poster we describe current practiceand introduce how novel equipment can aid answeringkey questions in transmission, reducing and replacingsentinel ferrets in the process.Current practicesTo test transmission, the ferrets are housed as shownabove (Figure 1). 1 ferret (Donor) is infected with thevirus and then a contact sentinel introduced to thesame cage and a Respiratory Droplet (RD) sentinel intoUsing novel equipment to aid reductionand replacement of the ferret model ininfluenza transmissionTERESA BOREHAMCBS, Imperial College London, St Mary’s Campus, Paddington, London W2 1PGCorrespondence: for husbandry for experimental design and resultsFigure 1. Housing of ferrets to test transmission.Graphs 1. The idea – graphs A-E.Clinical signs (graph A and B) do not appear until day 2after infection of the donor ferrets, reaching a peak atday 6. Graph C shows virus detected in the nasalwashing of the donor animals for 6 days after infection,peaking at D1-D2 and again at D5-6. Sentinel animalswere RD exposed to infected donors at either the earlyor late time points. Only animals exposed early, beforeclinical signs became infected through transmission.This shows that although “coughs and sneezes spreaddiseases”, there can also be transmission without anyclinical adjacent cage. The RD sentinel cannot touch thedonor or contact but can breathe air that passesthrough the perforated grid. These studies are carriedout with 4 groups (total of 12 ferrets per study).The ideaApr LATEST:Animal Technology and Welfare 22/3/19 12:28 Page 59

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Poster Presentations60To develop this study further the group needed to lookat how small that transmission window was. The idealscenario would be to introduce new sentinels ferrets at12hr intervals up to day 6. This would result in the useof 104 ferrets (13 time points with 4 groups of donors)in which the sentinel ferrets are just like canaries in themine, scoring yes or no for infection. This was not anethical option.Using funding from the NC3Rs the team designed “TheInfluenza Virus Transmission Tunnel (IVTT)”. (Figure 2)Figure 2. Transmission tunnel.Figure 3. Ferret in position within ferret chamber.Figure 4. Ferret concentrating on the outlet pipe.With the IVTT system a donor ferret is infected and isplaced in the ferret chamber at set intervals (at leastonce a day) depending on the study requirements.Ferre Tone (skin supplement/treat) is placed aroundthe outlet pipe. The ferret will then willingly sit andfocus on this area. The ferrets are in the chamber for10 minutes. Any virus exhaled in breath is drawn downthe tube and deposits on to the plates of cells where itcan be counted and compared with the nasal washsamples.Air enters the ferret chamber from the top and thebottom and is directed into the IVT exposure tunnel, asillustrated by the block arrows. The ferret chamber canbe replaced by a nebulisation unit for in vitro studies.Cell culture plates at 30, 60 and 90 cm from the inletport can be accessed via drawers on the side of theapparatus. Temperature and humidity can be monitoredwithin the ferret chamber and cell tunnel. This givesadditional data on environmental parameters for virustransmission during the study.ResultsWith the IVTT the group could ask why transmission didnot occur on the D5-6 exposure despite there being apeak of virus shed in the nasal wash.Six ferrets were infected and assessed for exhaledvirus each day as well as nasal washed to monitor virusshed into the nose. The data below shows ferrets onlyexhaled infectious virus measured by the IVTT on D2 oroccasionally low levels D3 or D4 (solid grey bars) eventhough sometimes the amount of virus in the nose washighest on later days. This explains why animalsexposed during the D5-6 did not become infected viaRD transmission and confirms that the window for RDtransmission is very small.As well as answering questions as described above thissystem can also be used to evaluate why notransmission occurs with certain strains of virus. To dothis the virus is nebulised (rather than using a ferret).The group change the cell plates every 10 minutes tosee how long the virus stays infectious within the air. Iffor example the virus died after 10min in the air, RDtransmission between animals (and humans) would beunlikely to occur.Through using the IVTT system the group have beenable to cross validate against previous transmissionstudies in ferrets and reduce the number of sentinelferrets needed to check transmission levels.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 60

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Poster Presentations61Going forward – challengesThough the ferret is the gold standard in flutransmission there is no standardisation around theworld for how to house ferrets or for environmentalparameters and experiment design.This means that although the IVTT provides a uniqueopportunity to the group here at Imperial to answer keyquestions when testing RD transmission of differentstrains of influenza virus, cross validation will berequired before groups around the world start trustingand utilising similar systems themselves. In the shortterm this will not immediately reduce ferret numbers asthe IVTT studies will have to be run alongside ferretstudies, nor will it replace chains of transmission. Inthe long term, a reduction in ferrets will be possible asthe data from the IVTT is seen to be dependableespecially when testing new strains of virus.AcknowledgementsBarclay Laboratory – Wendy Barclay, Rebecca Frise,Anika Singanayagam, Lauren Parker, Jenny Shelly, RuthElderfield, Kim Roberts, Holly Shelton.CBS Staff – Kasia Smigielska, Christopher Mack, MarkPritchard, Colin Wells, Catherine Underwood.Contact Tess ( for husbandryqueries and Wendy ( forexperimental design and results.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 61

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62AbstractHealth tracking of animals is required legally under theAnimals (Scientific Procedures) Act 1986 (ASPA) toprevent pain, avoidable suffering, distress or lastingharm. This is important with Genetically Modified (GM)animals as they are more likely to have unexpectedphenotypes or health issues.The Sanger Mouse Database (MCMS) enables the userto add and track a wealth of information for everymouse in the facility. For example, mating details,genetic backgrounds and procedures can be accessedthrough reports that allow retrieval of specific data toanalyse.The dedicated health observation module is used fortracking welfare both at the mouse and colony level toassess the prevalence of an observation.This poster details what steps can be taken if a mouseis unable to complete our primary pipeline, howdatabase reports can be used to gain an overview ofhealth and welfare issues in GM colonies, and how wecan increase the scientific value of that information.MGP primary pipeline40 mice enter the MGP, Primary Phenotyping Pipelineeach week and go through a standardised set of testsencompassing dysmorphology, metabolism, bodycomposition, histology and haematology. All proceduresand mouse production are performed under theAnimals (Scientific Procedures) Act 1986.This enables colony health reports to be run on thedatabase – an overview of all health observationscaptured in the colony are broken down into thegenotypes and number of mice affected.Optimising health & welfare trackingusing the Sanger Mouse DatabaseJOSHUA DENCH, ROBBIE MCLAREN-JONES, VALERIE VANCOLLIE,EMMA SIRAGHER, DAVID TINO LAFONT, LAUREN ANTHONY,SIMON MAGUIRE, MARK GRIFFITHS, HANNAH WARDLE-JONES,CARL SHANNON, JAMES BUSSELL and CHRIS LELLIOTTThe Wellcome Sanger Institute, Wellcome Genome Campus, Cambridgeshire CB10 1SACorrespondence: Technology and Welfare April 2019Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 62

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63Poster PresentationsObservations withover 10% of micebeing affected areconsidered and shouldbe investigated furtherBy using the database reports and lookinginto the colony information, we can seethat this colony is sub-viable forhomozygous mice.Not many homs are being produced and thosethat are surviving to P14, appear prone tohydrocephalus. This is a welfare issue thatneeds to be addressed, so what can we do?Category ‘Other’ are micewithout a confirmed genotype.This means they are likely tohave been less than 9 days old(age of genotyping) when thehealth observation was recordedApr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 63

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Poster Presentations64ConclusionThe case study above is a great example of how byusing the mouse database we can easily assess thehealth of a colony, identify a potential welfare issue andact quickly by putting measures in place to preventmore animals from being affected.As the homozogotes (homs) from this colony are sub-viable, we made the decision to switch to phenotypingadult heterozygotes (hets). This does however presentanother opportunity to investigate homs by using adifferent route such as the Recessive Lethality Embryoscreen or the Neonate pipeline to gain more insight intothis gene at an earlier time point.This enables colony health reports to be run on thedatabase – an overview of all health observationscaptured in the colony are broken down into thegenotypes and number of mice affected.Categor y ‘Other’, are mice without a confirmedgenotype, this means they are likely to have been lessthan 9 days old (age of genotyping) when the healthobservation was recorded.It is important to consider the total number of mice inthe colony. In a new colony there might not be manymice of the particular genotype yet and the smallsample size may mean the percentage affected ismisleading. Another consideration is the distribution ofthe observation throughout the colony.Individual mouse details should be checked to see ifthe health observation is limited to a specific breedingpair, in which case it may be a hereditary issue.Often a health observation will be an isolated case,however by following the same protocol with any sickmice found in the pipeline, we can be confident inaccurately detecting any colony-wide health and welfareissues.GlossaryMGP Mouse Genetics ProjectMCMS Sanger Database Mouse Colony ManagementSystem.Sick Mouse A mouse deemed unfit to complete thepipeline due to health observation or discoveredphenotype.NaMPA Neurological and Morphological PhenotypicAssessment.IP-GTT Intraperitoneal Glucose Tolerance TestDEXA Dual Energy X-Ray Absorptiometry.AcknowledgementsMouse Informatics Group, Research Support FacilityGroup, MGP Mouse Transgenic Technologies Group.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 64

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April 2019 Animal Technology and Welfare65IntroductionNaomi Nakagata from the Kumamoto University inJapan has developed a new method of Ultra-superovulation to produce a greater number of oocytesfor in-vitro fertilisation (IVF) than the traditionalmethods using inhibin antiserum and equine chorionicgonadotropin to promote follicle growth. This newreagent was named Hyperova®.This method has been proven to enable the increasedproduction of fertilised oocytes via in vitro fertilisationcompared with the conventional superovulationmethod.1Traditional methods:Traditional superovulation of C57 Bl6 (J or N, CRl orOla) requires mice of optimal age to produce a goodyield of oocytes for IVF. 3-4-week-old mice give theoptimal yield but it is not always possible to use miceat this age, especially transgenic mice as it takes timeto ship frozen sperm, process ear clips for genotyping,obtain results and report these results.Application of Hyperova®(New method)Early trials by the Central Transgenic Core (CTC) in BVS,Edinburgh University showed the new reagent workedwell, exactly as described by Nakagata1but addedapproximately 30% on to the cost of a rederivation. Wetherefore decided to only use it as a refinement and toreduce the numbers of mice bred and speed up therederivation timeline in the case of:1. Only a small cohort of mice were available toprevent further breeding and speed up therederivation process.2. If we had aged Wild Type (WT) mice on the shelfwhich would normally be unsuitable forsuperovulation and would otherwise be wasted/culled.MethodIn 2016 CTC carried out 45 IVF’s to rederive lines intothe rederivation unit in BVS or to cryopreserveembryos. During this time Hyperova®was trialledalongside the traditional superovulation reagents(PMS/HCG). The protocol and timings of administrationof Hyperova®were identical to PMSG/HCG with the onlydifference being in the process of thawing Hyperova®which must be done 30 minutes in advance of injectionin a water bath.The use of Hyperova®to produce oocytesfrom aged WT mice and small groups oftransgenic mice for IVF rederivationJULIE THOMSON, AILSA TRAVERS, JACEK MENDRYCHOWSKIand EMMA ALLANUniversity of Edinburgh, Bioresearch and Veterinary Services (BVS), Hugh Robson Building,George Square, Edinburgh EH8 9XDCorrespondence: at University of Edinburgh 3Rs Symposium, May 2018Figure 1. 2-cell embryos produced via IVF usingHyperova.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 65

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Analysis of the IVF projects performed by CTC over thepast year (see Table 1) shows an overall reduction inthe number of donor animals by 10-20 mice per projectand a cost saving in breeding time, cage charges andgenotyping costs. In addition, we were also able to useolder donor females that would otherwise have to beculled.Although it is difficult to directly compare each IVFsession due to several inherent variables includingquality of sperm (which may be frozen or fresh) anddifferences in transgenic sperm fertility, backgroundand genetics of donors used we have identified a newsuperovulation regime that will help us promote the3Rs for rederivation projects.TimelineShowing the process from donor Ultra-superovulation tolive pupsConclusionHyperova®has given us the opportunity to Reduce thenumbers of donors we use: a minimum of 3 females ofany age up to one year can be used to rederive a strainby IVF.We have also been able to Refine the procedure aswe can use mice that would otherwise not be any usefor traditional superovulation due to being too old.Smaller groups of transgenic mice (minimum of 3) cannow be used. This saves researchers time and moneyand prevents more mice being bred as usually wewould need 8-10 donor females when usingPMSG/HCG.As Edinburgh is undertaking a range of new buildingprojects in the next 5 years this approach will refinethe rederivation strategy. We estimate that we canachieve a 50% Reduction in the donors we use andbreed for the IVF rederivation using Hyperova®. We canalso Refine this process and speed it up by performingthe IVF in advance of the rederivation andcryopreserving the embryos so that they can betransferred more efficiently.3Rs impact1. We can now utilise Aged donors: This represents asaving of approximately 10-20 mice per IVF sessionand 6-8 weeks breeding time as well as cagecharges and genotyping costs. The mice that cannow be utilised as donors would otherwise beculled. This is a Refinement as aged mice are notwasted and a Reduction in the numbers of miceused for breeding and superovulation.66Poster PresentationsTable 1. Comparison of Superovulation regimes.PMSG V’s #Lines Success Age Minimum Total TotalHyperova IVF’d rate range # of ##donors donors donorsused) used used––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––PMSG/HCG 26 100% 3-8 9 253 294weeks––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HYPEROVA/HCG 19 100% 8-26+ 3 141 137weeksTable 2. Comparison of different strains.Reagent Strain # lines Total #Oocytes #fertilised #live % pupsIVF’d # donors produced pups bornused born––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––PMSG/HCG C57 BL6 J 10 95 1895 858 112 13%––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HYPEROVA/HCG C57 BL6 J 1 10 312 91 20 22%OLA––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––PMSG/HCG C57 BL6 J 10 94 1639 928 162 17%––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HYPEROVA/HCG C57 BL6 J 6 66 1558 783 112 13%––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––PMSG/HCG C57 BL6 N 3 36 536 219 20 9%––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HYPEROVA/HCG C57 BL6 N 18 111 52 5 10%––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––PMSH/HCG Various 3 28 476 22 48 21%Transgeniclines––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HYPEROVA/HCG Various 9 57 1201 759 130 17%Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 66

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Poster Presentations672. We can now use much smaller groups oftransgenics (min 3): This also represents a savingof approx. 10-20 mice per IVF session and 6-8weeks breeding time as well as cage charges andgenotyping costs. This is also Refinement as agedtransgenic mice do not need to be wasted and aReduction in the numbers of mice used and bredspecifically for rederivation which would normallytake approx. 8-12 weeks.Next experimentFurther trials were carried out using Hyperova toproduce fertilised embryos for pronuclear andcytoplasmic injection.ResultsFertilisation rate with Hyper ova/HCG (10 mice) = 27%Fertilisation rate with PMCG/HCG (60 Mice) = 60%ConclusionFurther work must be done to optimise conditions toproduce fertilised embryos.DiscussionIt is difficult to compare the success rates ofHyperova/PMSG as the IVF success is also dictated byseveral variables:– The quality of the sperm: Different freezingprotocols are used in each institution with differentQuality check and assurances.– Transgenic (Tg) line differences: Some lines mayshow a phenotype which affects reproduction.– Age of donor: We are using donors as they becomeavailable so there is a range of ages in the data set.– Different media: For IVF we have a few differentprotocols and media at our disposal which we tailorto the quality of the sperm and whether it is fresh orfrozen.– Superovulation carried out in different units withdifferent protocols: Our data set presents oocyteyield from several different units within BVS.– Bought in/in house mice: We try to use ‘in house’mice as much as possible but in some occasionsmice were bought in from various vendors.AcknowledgementsIn 1998, Prof. Naomi Nakagata started the project ofMouse Bank (CARD R-Base), which collect, preserveand supply genetically engineered mice to efficientlyconduct the research using them, at Centre for AnimalResources and Development (CARD) in Kuma-motoUniversity. Now, CARD Mouse Bank archives more than3100 strains of genetically engineered mice andsupplies them to various scientists of domestic andinternational institutes. Our team developed variousreproductive techniques to improve mouse bankssystem and educate our knowledge and techniques ofmouse reproductive technology via domestic andinternational mouse sperm and embryocryopreservation workshop. Superovulation Using the CombinedAdministration of Inhibin Antiserum and EquineChorionic Gonadotropin Increases the Number ofOvulated Oocytes in C57BL/6 Female Mice huge thank you to all the technical staff in all BVSunits who helped us by doing superovulation injectionsout of hours and at weekends to facilitate the IVF. Theirdedication makes it possible for new developments inall aspects of research including the 3Rs to happen.References1Takeo, T. and Nakagata, N. (2015). Superovulation Usingthe Combined Administration of Inhibin Antiserum andEquine Chorionic Gonadotropin Increases the Numberof Ovulated Oocytes in C57BL/6 Female Mice. PLoSONE 10(5): e0128330. LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 67

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Animal Technology and Welfare April 201968IntroductionThe application of recombinase-mediated alleleconversion is widespread in model organisms. In orderto isolate the change caused by a mutation in a specificcell type comparison is often made between animalswhere loss of the gene in question is controlled byspatially or temporally restricted Cre expression andthe complete null mutation. Null mutation animals aretraditionally created by interbreeding the LoxPline withanimals transgenic for a ubiquitously expressed Crerecombinase in order to create the null allele in the 1-cell stage zygote.There are two consequences to this approach:(1) These breeding steps are time-consuming and(2) Interbreeding leads to mixed genetic backgroundsderived from each parental line.Breeding uses more animals simply to achieve theexperimental genetic status and takes extra time andmoney. Contamination of genetic backgrounds mayoccur because Cre driver lines are not available in manystrains. Months of back-cross breeding and alteredexperimental data can be the result.Allele conversion using cell-permeableCre recombinaseAILSA TRAVERS, JULIE THOMSON and MATT SHARPUniversity of Edinburgh, Bioresearch and Veterinary Services (BVS), Hugh Robson Building,George Square, Edinburgh EH8 9XDCorrespondence: at University of Edinburgh 3Rs Symposium, May 2018Figure 1. Comparison of traditional breeding versuspermeable recombinase to generate null alleles.((A) Importation of a conditional allele mouse line anda ubiquitous Cre driver line by in vitro fertilisation (IVF),followed by interbreeding and sib-mating to generatehomozygous null animals. (B) Fewer steps and animalsare required to import a conditional allele mouse linewhen Tat-Cre is used during the IVF stage to generatethe null allele. Sib-mating is required to generatehomozygous null animals. (C) If homozygousconditional (f) animals are available, then application ofTat-Cre to a small batch of embryos is all that isrequired to generate a cohort of null mice.)Figure 2. Diagrammatic representation of Crerecombinase. (Left panel – A dimer of Cre recombinasebound to double-stranded DNA. Right panel – A complexof two Cre dimers bound to a pair of LoxPrecombination targets prior to strand cleavage.)Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 68

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69Poster PresentationsWe are applying protocols to generate null alleles inthe homozygous state in all tissues of mice and ratsby delivery of a cell-permeable recombinase protein tothe zygote stage.1Tat-Cre (NLS-Cre; Excellgen,Rockville, MD) is a fusion protein between the basicpeptide of HIV-TAT and the bacteriophage Crerecombinase gene can be used to permeate livingcells to catalyse recombination in vivo. This methodresults in rapid allele conversion and minimises theuse of experimental animals when compared toconventional Cre transgenic mouse breeding, resultingin a significant reduction in costs and time withincreased welfare benefits.MethodsGeneration of oocytes and IVFNine 3-4 week old C57BL/6JOlaHsd or C57BL/6JCrlfemales per frozen sperm sample were super-ovulatedby intraperitoneal (IP) injection of 5IU of pregnantmare’s serum (PMSG) at 17:30 h (on a 12 h light/darkcycle, at 07:00on/19:00 off) followed 48 h later by anIP injection of 5 IU human chorionic gonadotrophin(hCG). Oviducts were dissected at approximately07:50 am on the day of the IVF and cumulus-oocytecomplexes were transferred into the IVF fertilisationdish containing HTF±GSH. An aliquot of 20 µL ofsperm from the pre-incubation dish was then added tothe fertilisation dish. After allowing 5 h for fertilisationto occur the embryos were washed and culturedovernight in HTF at 37°C, in 5% CO2in air.Tat-Cre treatment of 2-cell embryosTat-Cre was sourced from Excellgen (product code RP-7) and stored at –80°C. One-cell embryos werecollected and cultured overnight to 2-cell embryos. Adish for Tat-Cre treatment was prepared as a series offive drops on the morning of the treatment. Embryoswere washed twice in drops of HTF to remove bovineserum albumin and then incubated for 30-40 minutesin DMEM+ Tat-Cre (0.3 µM final concentration). Aftersufficient time for Cre to be taken into the embryosthe Cre drop was flooded with HTF containing BSA toallow easier manipulation of the embryos.Embryos were then washed twice in HTF drops beforebeing returned to the incubator prior to embryotransfer into a 0.5 days post coitum pseudo pregnantfemale recipient produced by mating to avasectomised male.SummaryThe published efficiency of Tat-Cre in recombiningconditional alleles in 2-cell embryos is greater than theefficiency of breeding with commonly-used ubiquitousCre expressing lines. Demonstration of Tat-Cre actionin this protocol will be available when offspring aregenotyped.Figure 3. Cre recombinase gene can be used topermeate through living cells to catalyse recombinationin vivo.This simple refinement to an established method willreduce animal usage by up to 84%, compared withnormal breeding. The need for ubiquitous expressingrecombinase lines will be replaced. It will cut the timetaken to achieve a null mutant cohort to 1/3 comparedwith breeding. As an alternative to the undesirablemixing of genetic backgrounds when the only Cre driverline available is not in the preferred genetic backgroundthis method will replace months of wasteful back-crossbreeding.References1Ryder, E., Doe, B., Gleeson, D., Houghton, R., Dalvi, P.,Grau, E. et al. (2014). Rapid conversion of EUCOMM/KOMP-CSD alleles in mouse embryos using a cell-permeable Cre recombinase. Transgenic Res. 23: 177-185).We have seen reduced viability of embryos whenconcentration or duration of incubation is increased.Rat embryos are very sensitive to Tat-Cre protein andimproved delivery conditions that are less toxic to therat embryos are required. These conditions are beinginvestigated.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 69

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Animal Technology and Welfare April 201970IntroductionThis is a collaborative project using a device, the MK-11 RAT Vaginal Impedance Checker (VIC),1(MuromachiKikai Co, Japan) to measure electrical impedance toprovide embryo donor and embryo recipient rats for usein making genetically modified rat models.Traditional methods of producing rat embryos haveproven to be less efficient.SmearingCytological examination of the vaginal epithelium[Papanicolaou stain] can be used to determine whetherrats are in oestrus. This is time consuming andrequires training and experience, making consistencyof interpretation during weekend cover difficult. Slidesmust be prepared/dried/stained, then examinedmicroscopically. Regular smearing can induce pseudo-pregnancy. We calculate that it would take 5-6 hours tosmear 75 rats to successfully identify 10 embryodonors and 5 recipients each day. This is not costeffective.Superovulation and mini-pumpimplantationOestrus can be induced and embryo productionincreased by superovulation. This is a licencedprocedure that involves administering immature ratswith large doses of exogenous sex hormones to hyper-stimulate the ovary. Surgical implantation of minipumps primed with hormone induce large numbers ofhigh-quality embryos, however, this is expensive andtime consuming. This protocol is an invasive procedure,the mini pumps must be surgically implanted undergeneral anaesthetic and complications such asinfection, inflammation, blood loss or anaestheticdeaths are risks which must be considered. There is noguarantee that embryos will be produced using thistechnique. Rats that do not copulate cannot be re-used. Simple intraperitoneal injection (ip) is analternative method that requires juvenile females but issomewhat unreliable and can fail to produce anyembryos, or yield low quality embryos with limitedcapacity to develop. However, the rate of copulation ishigh (60-75%) following induction of oestrus. Ratembryos do not develop well in culture, so weinvestigated embryo harvesting at blastocyst stage(E4.5) for ES cell injection to produce gene targetedrats. It has been shown by Akira et al. (1993)2that highratios of oestradiol/progesterone caused bysuperovulation, lead to acceleration of embryotransport. By E4.5 embryos cannot be found in thereproductive tract after superovulation (data not shown)therefore we had to explore other options to prepareblastocysts.Natural matingNatural mating produces high quality but lowernumbers of embryos. Selecting rats in oestrus by eyeis unreliable (unlike the mouse) so large number ofpairings are required to generate embryos by randommating. Around 1 female in 5 will plug. Therefore, toproduce 10 embryo donors and 5 recipients each daywe would require 75 male and females to be paired foreach injection session. We can expect 23% ofrandomly-paired animals to mate naturally.We identified a need to r efine the embryo productionprotocol but still yield good quality embryos from as fewrat mating’s as possible with high copulation successrates to reduce the number of animals used but alsoreduce the technician time and the number of cagesrequired.Approximately 18 months ago we were introduced tothe MK-11 Rat Vaginal Impedance Checker (VIC),1(Muromachi Kikai Co, Japan) and we were convincedthat this was the best technological initiative we hadseen in years that would:The use of a par-vaginal impedancechecker to improve rat plugging efficiencyJULIE THOMSON and WILLIAM MUNGALLUniversity of Edinburgh, Bioresearch and Veterinary Services (BVS), Hugh Robson Building,George Square, Edinburgh EH8 9XDCorrespondence: at University of Edinburgh 3Rs Symposium, May 2018Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 70

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71Poster Presentations1. Have a positive impact on animal welfare, byreplacing the need to perform surgery or administerhormone injections.2. Provide an efficient way to produce good qualityembryo donors from naturally mated rats with a highmating probability.3. Reduce the number of animals used as any non-mated rats can be returned to stock.4. Be an efficient way of producing pseudo-pregnantrecipients.How it worksThe VIC has been designed help to determine theoptimum day for mating during the oestrus cycle in rats.The electrical impedance of the epithelial cell layer ofvaginal mucosa is measured at a frequency of 1 kHz byinsertion of a special probe into vagina. In the pro-oestrus stage significantly, high impedance is producedcompared to that in the other stages of the oestruscycle. An impedance level of 3 kOhm indicates the pro-oestrus stage. The measuring range is 0 – 19.9 kOhm.The use of the probe is not a regulated procedure anddoes not require Home Office licencing. It is as simpleas plug checking and can be carried out by trained un-licenced technicians. Competent handling of rats is theimportant factor. Each rat is gently restrained and theprobe is inserted gently into the vagina so that bothmetal rings are inside (approximately 2 cm), it takesaround 2 seconds for a reading to stabilise on themonitor. The reading is recorded and the probe iscleaned with sterile water and a sterile tissue beforemoving on to the next rat. [See Picture 1]Picture 1. Probe in use.Injection Number Number Number Numberday of of of pups ofplugs recipients born chimeras–––––––––––––––––––––––––––––––––––––––––––––––––––––––––16291–––––––––––––––––––––––––––––––––––––––––––––––––––––––––24216 1–––––––––––––––––––––––––––––––––––––––––––––––––––––––––38215 0–––––––––––––––––––––––––––––––––––––––––––––––––––––––––43214 2–––––––––––––––––––––––––––––––––––––––––––––––––––––––––5 10 5 27 2–––––––––––––––––––––––––––––––––––––––––––––––––––––––––6 11 3 20 1–––––––––––––––––––––––––––––––––––––––––––––––––––––––––74318 2–––––––––––––––––––––––––––––––––––––––––––––––––––––––––89261–––––––––––––––––––––––––––––––––––––––––––––––––––––––––97221–––––––––––––––––––––––––––––––––––––––––––––––––––––––––10 10 5 21 4–––––––––––––––––––––––––––––––––––––––––––––––––––––––––11 5211 0–––––––––––––––––––––––––––––––––––––––––––––––––––––––––12 6419 5–––––––––––––––––––––––––––––––––––––––––––––––––––––––––13 5215 4–––––––––––––––––––––––––––––––––––––––––––––––––––––––––14 6328 7–––––––––––––––––––––––––––––––––––––––––––––––––––––––––15 6313 2–––––––––––––––––––––––––––––––––––––––––––––––––––––––––16 4213 2–––––––––––––––––––––––––––––––––––––––––––––––––––––––––17 8423 6–––––––––––––––––––––––––––––––––––––––––––––––––––––––––18 9423 3–––––––––––––––––––––––––––––––––––––––––––––––––––––––––19 11 4 21 6Table 1. Number of chimeras.ExperimentWithin the Unit, rats are set up daily to provideembryos for various researchers, in order to assessthe efficiency of the VIC we ran a small-scale trial.Instead of pairing random females with males, five ratswere probed using the VIC daily for 14 days. Thefemales showed no signs of stress and very soon gotused to the procedure which is as gentle as checkingfor the presence of a copulation plug. The resultsshowed that the rats had an oestrus cycle of 4-5 days.An impedance reading above 3 kOhm indicated to uswhen the rats were going into oestrus. Femalesidentified as being “in oestrus” according to animpedance reading of >3 kOhm using the VIC wereplaced with males and left overnight to mate.Copulation plugs were found in 82% of these cagesindicating that mating had taken place. The femalesthat failed to mate (and were possibly coming out ofoestrus) were returned to stock. In contrast, super-ovulated rats which fail to copulate cannot be re-used.ResultsWe have produced over 150 chimeras in the last year.[See Table 1]Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 71

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– Approximately 60 blastocysts per day were producedfrom 10 donors and transferred into 5 recipients.– On average, 4 chimeras were produced each day.– In this scheme, only 10 stud males are required and5 vasectomised males for one microinjection dayeach week.– On average 5 females are probed to produce 1embryo donor or recipient (i.e. 75 females for eachinjection day). [See Figure 1]– If we were using natural mating for this project, wewould require 50 studs.AcknowledgementsDr Matt Sharp, our thanks for his continued tirelessenergy and support.Dr Graham Thomas Director of BRR, with thanks for hissupport and encouragement.LF2 technologists, without whose commitment andexpertise this work would not have been possible.References1 This product has been developed in cooperationwith Chugai Pharmaceutical Co. Ltd.2Akira, S., Sanbuissho, A., Lin, Y.C. and Araki, T. (1993).Acceleration of embryo transport in super-ovulated adultrats. Life Sci. 53: 1243-51.72Poster PresentationsPicture 2. Example of chimeras.Picture 3. Happy mum with litter.Figure 1. Impedance reading changes with time toshow oestrus in 5 SD rats.Conclusion1. The VIC has made it possible for us to produceembryo donors and recipients much more efficientlythan when using traditional methods.2. The embryo quality is improved when compared totraditional methods.3. As technicians we are delighted that fewer animalsare being used and the method is far less stressfulto the rats also the researchers are delighted withthe numbers of chimeras being produced at a fasterrate. [See Pictures 2 and 3]4. All 3Rs were successfully achieved using thisapproach and we believe the collaborative nature ofthis project made for the best possible results. Theexpertise and knowledge of the animal technicianshad a massive impact on the outcome.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 72

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April 2019 Animal Technology and Welfare73IntroductionTo attempt to reduce, refine and replace, we haveemployed a several new approaches to rederive mousestrains:By using in vitro fertilisation (IVF) with fresh or frozensperm we can reduce the timeframe and number ofanimals required to re-derive a strain. This method ofimportation also reduces the stress on animals byreducing the need to transport live animals:a. Frozen sperm can be safely shipped in a dry shipperand IVF carried out at a time that suits the user.b. Theoretically fresh sperm in the form of caudalepididymis can be shipped from anywhere in theworld. The sperm is viable for up to 72 hours at 4°Cbefore the fresh sperm is used to carry out IVF andcryopreservation. However we have found that thetime taken for the shipments to arrive from Asia andAmerica in oil means that the sperm viability isunreliable. This approach works excellently withshipments from the UK.c. Animals from a unit anywhere in the world where theshipment time is too long or the unit does not haveaccess to sperm or embryo cryopreservationtechniques that have an unacceptable pathogenload and are not allowed into any unit within theBRR can now be imported (2 males only) straight toa quarantine room. On arrival these animals areculled and the caudal epididymis is dissected andstored under oil or in the preservation media, Lifor(an organ preservation solution) until IVF can beperformed the next morning.Refining mouse re-derivation by using invitro fertilisation (IVF) with fresh or frozensperm as opposed to embryosAILSA TRAVERS, JULIE THOMSON and MATT SHARPUniversity of Edinburgh, Bioresearch and Veterinary Services (BVS), Hugh Robson Building,George Square, Edinburgh EH8 9XDCorrespondence: at University of Edinburgh 3Rs Symposium, May 2018Picture 1. Sperm and egg pre-fertilisation.( 1. Time frame reduction.Table 2. Animal reduction required prior to rederivation.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 73

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Traditional methods:– Usually requires importation of live mice (2 breedingpairs).– Held in quarantine conditions.– Breeding to provide a cohort of 20-30 mice.– Takes approximately 4 months.– Wasteful as generally half of the mice generated arenot of a suitable genotype.Embryo transfersEmbryo recipients are still required, in general moreembryos are produced by IVF than by natural matingand more Generation Zero (G0’s) transgenic mice areproduced.Each transgenic strain responds differently to IVFconditions therefore we can use several variations ofmedia, antioxidants and protocols.Using sperm and IVF to re-derive mouse strains insteadof embryos is a great technological initiative because:1. Large scale breeding is not required to produce 1 or2 sperm donors. Reduction.2. It has a positive impact on the welfare of theanimals, by replacing the need to import liveanimals (in some cases). Refinement andReplacement.3. The timeframe for re-derivation is Reduced by 50%which saves money, time and resources.4. We now have the opportunity to rederive animalswith an unacceptable pathogen load for import intoour units. This helps the research community vastly.The only alternative before would have been to re-make the animals by pronuclear microinjection orES cell injection. Reduction.AcknowledgementsI wish to thank all BVS staff for their efforts andtechnical assistance which has made this project sucha success.74Poster PresentationsTable 3. Increased efficiency by using IVF.Table 4. Outlining differences.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 74

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April 2019 Animal Technology and Welfare75BackgroundSkin closure clips are widely used within the Universityof Edinburgh to close incision sites for variousprocedures in both rats and mice.In mice, skin closure clips work well with only a verysmall percentage of clips coming out after surgeryleading to the requirement to re-suture or cull. In thelast 7 years out of approximately 1500 mice only 2mice have had to be re-sutured or re-clipped underveterinary advice.However, in rats the skin closure clips very often comeout as a result of cage mates biting each other’sincision sites or for other unknown reasons. Onesolution is to singly house rats that have undergonesurgery. This is not ideal as rats are companion animalsand this, therefore, must be approved on a projectlicence.This is a widespread problem throughout and the HomeOffice require that improvements are made.ObjectiveThe Central Transgenic Core (CTC) for EdinburghUniversity have performed 500 embryo transfers over 6years in rats without encountering the problems of clipscoming out. Prior to 2016, only 1 rat was identifiedwhere cage mates had chewed the wound site post-surgery. The problem of clips coming out in rats has onlyoccurred in the last 12 months. Review of the mostrecent 20 embryo transfers performed by CTC showsthat 2 rats had to be culled and another 2 rats had toUsing subcuticular stitching in rats toreplace skin closure clips as a refinementJULIE THOMSON and WILLIAM MUNGALLUniversity of Edinburgh, Bioresearch and Veterinary Services (BVS), Hugh Robson Building,George Square, Edinburgh EH8 9XDCorrespondence: at University of Edinburgh 3Rs Symposium, May 2018Two systems widely used in unitsAbove: Reflex skin closure systemBelow: Michel clips with applicatorFigure 1a and 1b. Images showing subcuticular stitchwith Aberdeen knot.Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 75

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be removed from the isolator, anaesthetised and re-sutured (under veterinary guidance). This interventionwas unacceptable and had to be addressed.MethodTo overcome the recent problem of rat surgical clipscoming out post-surgery we tried using different typesand sizes of clips and applicators and re-trained staffin applying the clips, however this did not seem to help.An alternative method of suturing was researched anda short course on subcuticular stitching with Aberdeenknot was provided by one of us (William Mungall) usingcadavers to practice. YouTube videos were also vieweduntil competency was confirmed.ResultsThirty rat embryo transfers have been completed withsubcuticular stitching finished with the Aberdeen knot.All the incision sites were perfectly closed andinfection-free for the duration of the experiment. Noadverse effects were noted and rat recipientssuccessfully got pregnant and littered down. Althoughthe time taken to perform the stitching during embryotransfer surgery takes approximately 10-15 minuteslonger than to clip, we believe that the benefitoutweighs the increased time under anaesthetic.We tried using subcuticular stitching instead of clips inmice as a refinement to embryo transfer surgery butthe stitching takes approximately 45 minutes as theskin is so thin (almost like tissue paper). Although thetechnique was successful, the benefit for switchingsurgical closure techniques in mice did not outweighthe time taken. There is no problem with the skinclosure clips coming out in mice but we are constantlymonitoring and will revisit this as suturing is morerefined.Vicryl Rapide™The type of suture recommended for use in embryotransfer by the Named Veterinary Surgeon and usedthroughout the University for the subcuticular stitchesin rats is Vicryl Rapide™.76Poster PresentationsFigure 2. Stages of wound closure using subcuticularsuturing.Conclusion and 3Rs impactThe use of subcuticular stitching and Aberdeen knotsfor surgical procedures in rats (i.e. embryo transfer) isa refinement which Edinburgh is rolling out to all usersthat currently use skin closure clips via the SpecialistTraining Officer (William Mungall). It will reduce theneed for intervention and the number of failedexperiments due to clips coming out. For moreinformation regarding this technique please get intouch with Mel Leech (NTCO) and William Mungall(Designated Trainer).An excellent training video can be found at LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 76

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78Subjects considered for publication may includeoriginal articles, technical notes and reviews pertainingto all aspects of animal science and technology,management and education. The Editorial Boardwishes to offer particular encouragement to papersleading to improvements in environmental enrichment,the general care and welfare of the animals used, inparticular those species and strains exhibiting harmfulgenetic defects, and papers describing refinements intechniques, a reduction in the number of animals thatneed to be used or alternatives to animal use.Papers describing experimental procedures will only beaccepted for publication if authors clearly state that theprocedures conform to the prevailing principles andCodes of Practice of the Animals (Scientific Procedures)Act, 1986. Papers submitted from outside the U.K.,should state what legislation and/or ethical approvalthe work has been carried out under. In addition,authors who describe surgical techniques with recoveryshould include details of post-operative care and anyanalgesic therapy provided. All submissions shouldfollow the ARRIVE (Animal Research: Reporting of InVivo Experiments) guidelines (Kilkenny C, Browne WJ,Cuthill IC, Emerson M, Altman DG (2010) ImprovingBioscience Research Reporting: The ARRIVE Guidelinesfor Reporting Animal Research. PLOS Biol 8(6):e1000412. doi:10.1371/journal.pbio.1000412)The Editorial Board reserves the right to seekindependent advice on any aspect of the content of anarticle but the final decision on acceptance or rejectionremains with the Board.SubmissionMaterial submitted for publication will be consideredprovided that it is contributed exclusively to AnimalTechnology and becomes the property of the Instituteof Animal Technology.Articles may be submitted either electronically or byhard copy as follows:ElectronicArticles should be submitted in Word format with doublespacing to the lines and all pages serially numbered.Any photographs or graphs must be submitted asseparate files and conform to the format in point 4)below.The relevant article must clearly indicate wherephotographs and/or graphs are to be inserted.Address for submission: copyThe original manuscript plus two copies should be sentto the address below together with a copy on disk (CDor DVD).All sheets should be typewritten on one side in doublespacing and serially numbered.Any photographs or graphs should be supplied asoriginals and conform to the format in 4) below.Address for submission: Journal Editorial BoardChairman, 5 South Parade, Summertown, Oxford OX27JL.No responsibility will be accepted for loss or damage tosuch articles.Electronic files of submissions are required togetherwith separate files of photographs and any graphicsthat appear in the manuscript. Electronic submissionsshould be sent via email via,manuscript plus two copies may be sent as hard copyto the address below. All sheets should be typewrittenon one side in double spacing with 4 cm margins andserially numbered. Additionally, a copy on disk shouldbe provided or sent by email via for submission should be sent to: JournalEditorial Board Chairman, 5 South Parade,Summertown, Oxford, OX2 7JL.No responsibility will be accepted for loss or damage tosuch articles.Format1). The first sheet of the article should contain thefollowing:i. the full title of the paperii. the initials and last name of the author(s)iii. the full address of the department(s) andinstitution(s) where the work was carried out.iv. the address for correspondence if different toabove.2). For the remainder of the paper, the text should beclear and concise and, where appropriate, sub-dividedunder the following headings:i. Summaryii. Introductioniii. Methodsiv. ResultsInstructions to AuthorsAnimal Technology and Welfare April 2019Apr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 78

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79v. Discussionvi. Acknowledgementsvii. References3). Measurements should be given in metric units –see The use of S.I. Units (1969) British StandardsInstitution publication and spelling should follow that ofthe Oxford English Dictionary. Abbreviations must bedefined in full at their first appearance in the text. The24 hour clock should be used for times. Words toappear in italic type should be underlined. Designationof inbred strains should be in accordance with theInternational Index of Laboratory Animals, 6th edition,compiled, edited and published by M.W. Festing, 1993.4). Photographs should have clear and well contrastedtone values and be in colour. All illustrations, charts(e.g. histograms and graphs) and photographs shouldbe submitted separately and bear on the reverse sidethe author’s name, a number corresponding to theorder in which it appears in the text e.g., Figure 1, andan arrow pointing to the top.Illustrations, charts and photographs supplied on diskshould be in JPEG, TIFF or EPS formats and have aresolution of no less than 300dpi.The captions for illustrations, charts and photographsshould be typed in double spacing in numerical orderon a separate sheet of paper.5). References: Only essential references should beincluded. Authors are responsible for verifying themagainst the original source material. ATW uses theVancouver referencing system: references should beidentified in the text by superscript Arabic numberse.g.12after any punctuation and numbered and listedat the end of the paper in the order of when they arefirst cited in the text. Automatic numbering should beavoided. References should include the names andinitials of up to six authors. If there are more than sixauthors, only the first three should be named, followedby et al. Publications for which no author is apparentmay be attributed to the organisation from which theyoriginate. Simply omit the name of the author foranonymous journal articles – avoid using‘Anonymous’.References should be set out as follows:Journals:- Surname and initials of author(s) (date), titleof article. Name of journal in full, volume number, firstand last page numbers.e.g. Saigeman, S. (1998). Environmental enhancementof cats – what? why? how? Animal Technology, Vol 49,No.3, 145-154.Books:- Surname and initials of author(s) (date), title ofbook. Name of publisher, Town of publisher.e.g. Flecknell, P.A. (1987). Laboratory AnimalAnaesthesia. Academic Press, London.Chapter from a multi-author book:- Surname and initialsof chapter author(s) (date), title of chapter. In: title ofbook (surname and initials of book editors). Name ofpublisher, Town of publisher, first and last pagenumbers of chapter.e.g. Gregory, J.A. (1985). Principles of AnimalHusbandry. In: Laboratory Animals – An Introduction forExperimenters. Second Edition. (Tuffrey, A.A.). JohnWiley & Sons Ltd., Chichester, 87-105.Papers accepted for publication but not yet publishedshould be included in the list of references followed by‘(in press)’. Papers in preparation, personalcommunications and unpublished observations shouldbe referred to as such in the text only.ContentPapers describing procedures involving the use ofanimals should always include full details of theanimals and husbandry conditions used. These wouldbe as follows:AnimalsSpeciesBreed or strainSexAge and weight at start of procedureGenetic status: inbred; outbred; hybrid; mutantSourceMicrobiological status: conventional; specifiedpathogen free (define which pathogens animals arefree from); gnotobiotic (define which micro-organisms are present)Quarantine or acclimatisation periodHusbandry during procedureType of housing: material; size; cage type if relevantNumber of animals per cage or unitBedding: type; quality; any pretreatmentType of system: conventional; barrier; ventilatedrack; isolatorEnvironmental temperature (°C ± range)Relative Humidity (% ± range)Lighting: natural; artificial (state hours of light anddark)Ventilation: number of air changes per hourPeriod of acclimatisation before start of procedureFeed: type; composition; any pretreatment; amount;frequencyWater: type; quality; any pretreatment; amount;frequencyScientific procedureNumber of animals and any pretreatmentInstructions to AuthorsApr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 79

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80Time of day of procedure(s)Quantity and frequency of any samplesStatisticsTests used should be namedReprintsFree reprints are no longer provided but the ATWEditorial Board are happy to provide PDF files ofarticles after publication. Use of these files is subjectto Copyright restrictions.Instructions to AuthorsApr LATEST:Animal Technology and Welfare 22/3/19 12:29 Page 80

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INDEX TO ADVERTISERSINDEX TO ADVERTISERS April 2019AAALAC/IAT ..............................................................................................................ivAllentown Inc .......................................................................................................OBCAS-ET ...................................................................................................................xviiiAVID plc ...................................................................................................................viBell Isolation Systems Ltd ......................................................................................xviiDatesand Ltd .........................................................................................................IFCInstitute of Animal Technology ............................................................................xiv, xviIPS Product Supplies Ltd ........................................................................................IBCLBS ........................................................................................................................xvMarshall BioResources .............................................................................................iiiPFI Systems Ltd ......................................................................................................viiSpecial Diets Services ............................................................................................viiiSurrey Diagnostics Ltd ............................................................................................xiiiTecniplast UK Ltd ......................................................................................................xVet-Tech Solutions Ltd ................................................................................................v7KLV 6XPPHU   VWRI -XO\  RXU 6HQLRU $FFRXQW0DQDJHU IRU 6RXWKHUQ (XURSH 'DYLG 0D\R LV SODQQLQJ WR UXQWKH GLVWDQFH IURP 0LOQJDYLH WR )RUW :LOOLDP  PLOHV NPV LQ XQGHU  GD\V FDUU\LQJ DOO KLV JHDU DQG IRRG RQ KLVEDFN 2Q WKH ODVW GD\ KH ZLOO DGG D FOLPE XS WR %HQ 1HYLVZHDWKHU SHUPLWWLQJ%HFDXVH ,'(;; %LR$QDO\WLFV YDOXHV WKH LPSRUWDQFH RIDGYDQFLQJ HGXFDWLRQ DQG SURPRWLQJ H[FHOOHQFH LQ WKH FDUH DQGZHOIDUH RI DQLPDOV HVSHFLDOO\ WKRVH XVHG LQ VFLHQFH 'DYLG LVFRPPLWWLQJ WR WKLV FKDOOHQJH DQG KDV GHFLGHG WR IXQGUDLVH IRU$6-(7 $QLPDOV LQ 6FLHQFH ± (GXFDWLRQ 7UXVW $6-(7 KDVGLVWULEXWHG QHDUO\  LQ JUDQWV WR VXSSRUW HGXFDWLRQ DQGZHOIDUH DFWLYLWLHV 5HDG PRUH DERXW WKHLU ZRUN KHUHZZZDQLPDOVLQVFLHQFHHGXFDWLRQWUXVWRUJXN'RQDWLQJ WKURXJK -XVW*LYLQJ LV VLPSOH IDVW DQG WRWDOO\ VHFXUH7R GRQDWH SOHDVH YLVLWKWWSVZZZMXVWJLYLQJFRPIXQGUDLVLQJGDYLG-PD\R7+$1. <28 )25 <285 6833257 Apr LATEST:Animal Technology and Welfare 22/3/19 12:30 Page 86

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