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Editorial 82Jas Barley, Chair of the Editorial BoardImplementing a successful positive reinforcement training protocol in laboratory-housed dogs 83Laura E.M. Scullion Hall and Sally RobinsonThe RSPCA focus on severe suffering project 89Penny HawkinsPAPER SUMMARY TRANSLATIONS 93TECH-2-TECHThe evening of the sudden heart attack – a patient’s personal account of how the skills of doctors and medical 101research, saved his lifeGary MartinicPreparing a paper presentation for Congress 111Institute of Animal TechnologyInformation and instructions for preparing a poster for Congress 115Institute of Animal TechnologyLet’s talk about ‘Culture of Care’ 117Nikki OsborneAS-ET SPECIAL TRAVEL BURSARY ESSAY ENTRIESDo present United Kingdom regulations covering experiments using laboratory animals strike the right balance 119between the interests of human health and laboratory animal welfare?Jessica WoodDo present United Kingdom regulations covering experiments using laboratory animals strike the right balance 123between the interests of human health and laboratory animal welfare?Everest OnwubikoDo present United Kingdom regulations covering experiments using laboratory animals strike the right balance 126between the interests of human health and laboratory animal welfare?Jamie BarrattPOSTER PRESENTATIONSMore breeding for less surplus: the advantages of using B6CBA to supply F1 mice 129Leila Thomas, Colin Travis, Claudia Watson, Terry Brown, Brendan Doe and Francesca FlackNon-human primate housing facility at Newcastle University 131Stevie O’KeefeDo different strains prefer different enrichments? 133Rhiannon AtkinsonInnovative enrichment for the common marmoset (Callithrix jacchus) 136Matteo Battilocchi, Garr y Fulcher, Zoe Windsor, Aaron Smith, Ria Fisher and Claire PearceEvaluation of p-Chip (Pharmaseq) microchip as an identification method for Xenopus tropicalis Biological 138Research Facility (BRF) The CrickLucy FernRefinement of rodent identification by implementation of an ear tagging system 141Jacqui Swan, Matt Smith, Noelia Lopez-Salensansky, Richard Marais and Lisa DoarInstructions to Authors 145Index to Advertisers xvCONTENTSVol 15 No 2 August 20 16iATW PROFILEAnimal Technology and Welfare aims to be the medium for animal technologists and all those concerned with the care and welfare of animals used for research purposes tocommunicate ‘best practice’. ATW especially aims to promote and develop the 3Rs particularly in respect of Refinement. More importantly, ATW promotes the generally accepted‘4th R’, Responsibility. The responsibility that all animal technologists have in ensuring dissemination of ‘best practice’ to every institution using animals in research. ATWenjoys a unique position as the scientific publication for the leading organisations (IAT and EFAT) for the welfare of animals in research.Editor: Jas Barley atweditor@iat.org.ukATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page i

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iiIAT REPRESENTATIVESOFFICERSPresidentDr Robin Lovell-Badge FRSImmediate Past PresidentProfessor Sir Richard Gardner MA PhD FSB HonFIAT FRSVice-PresidentsDavid Anderson MRCVS, Stephen Barnett BA MSc FIAT CBiol FSBRAnTech, Brian Cass CBE, Miles Carroll PhD, Gerald Clough BScPhD EurBiol CBiol MSB SFZSL, Paul Flecknell MA Vet MB PhD DLASDipLECVA MRCVS, Sue Houlton BVSc MA DVR DVC MRCVS,Wendy Jarrett MA, Judy MacArthur-Clark CBE BVMS DLAS FSBDVMS (h.c.), DipECLAM FRAgS DipACLAM MRCVS,Fiona McEwen BSc BVM&S MSc MRCVS, Tim Morris BVetMed PhDDipACLAM DipECLAM CBiol FSB CertLAS MRCVS, Vicky Robinson,José Orellana BVSc MSc, Clive Page PhD BSc,Gail Thompson RLATG, Robert Weichbrod PhD RLATG,Lord Robert Winston FMedSci DSc FRCOG FRCP FRCS Ed FSBLife MembersCharlie Chambers MIAT RAnTech, Roger Francis MSC FIAT RAnTech,Pete Gerson MSc FIAT RAnTech, Cathy Godfrey FIAT RAnTech,John Gregory BSc (Hons) FIAT CBiol FSB RAnTech, Patrick HayesFIAT DipBA RAnTech, Robert Kemp FIAT (Hon) RAnTech,Keith Millican FIAT CBiol MSB, Phil Ruddock MIAT RAnTech,Ted Wills HonFIAT RAnTech, Dorothy Woodnott FIATHonorary MembersAndy Jackson MIAT, Brian Lowe MSc FIAT RAnTech, Sue McHughBSc FIAT, Terry Priest FIAT, RAnTech, Trevor Richards BEM MIAT,David Spillane FIAT, Pete Willan DMS FInstLM MIATMembers of CouncilKen Applebee OBE, Matthew Bilton, Charlie Chambers,Steven Cubitt, Simon Cumming, Andy Cunningham, Haley Daniels,Glyn Fisher, Nicky Gent, Cathy Godfrey, Alan Graham, Linda Horan,Sam Jameson, Elaine Kirkum, Adele Kitching, Sarah Lane,Theresa Langford, Norman Mortell, Steve Owen, Wendy Steel,Allan Thornhill, Lynda Westall, Carole Wilson, Adrian WoodhouseCouncil OfficersChair: Ken Applebee OBE FIAT CBiol FSB RAnTechVice Chair: Norman Mortell BA (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: Cathy Godfrey FIAT RAnTechChair Registration & Accreditation Board: Wendy Steel BSc (Hons)FIAT RAnTechChair ATW Editorial Board: Jas Barley MSc FIAT RAnTechBulletin Editor: Sarah Lane MSc FIAT RAnTechAssistant Bulletin Editor: Carole Wilson BSc MIATBranch Liaison Officer: Lynda Westall BSc (Hons) FIAT DMS RAnTechEFAT Representative: Charlie Chambers MIAT RAnTechCouncil Website Coordinator: Allan Thornhill FIAT RAnTechDiversity Officer: Haley DanielsIAT INFORMATIONAnimal Welfare Officers and LABA Representatives:Andy Cunningham, Matt Bilton, Simon Cumming, Nicky GentATW/Bulletin Editorial Board: Jas Barley, Patrick Hayes,Elaine Kirkum, Sarah Lane, Lynda Westall, Carole WilsonBoard of Educational Policy: Glyn Fisher (Chair), Steven Cubitt(Secretary) Adele KitchingBoard of Moderators: Cathy Godfrey (Chair), Haley Daniels(Secretary) Moderators: Gary Childs, Joanna Cruden, Nicky Gent,Linda Horan, Anthony Iglesias, Sue McHughCommunications Group: Norman Mortell (Chair), Elaine Kirkum,Theresa Langford, Allan Thornhill, Lynda Westall, Adrian WoodhouseRegistration and Accreditation Board: Wendy Steel (Chair),Sarah Lane (Secretary) Theresa Langford: Ken Applebee,Charlie Chambers, John Gregory, Cathy Godfrey, Stuart Stevenson,Carol WilliamsObservers: Charles Gentry (Certificate Holders Forum),Adrian Deeny (LASA), Ian Mason (Home Office), Ngaire Dennison(LAVA)Congress Committee: Alan Graham (Chair), Haley Daniels,Linda Horan, Adele Kitching, Allan ThornhillAdvertisement Managers: PRC Associates LtdEmail: mail@prcassoc.co.ukIAT OFFICERS MAY BE CONTACTED VIA:IAT Administrator: iat101@btconnect.comOR VIA THE IAT WEBSITE AT: www.iat.org.ukOR VIA THE REGISTERED OFFICE:5 South Parade, Summertown, Oxford OX2 7JLAlthough 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.© 2016 Institute of Animal TechnologyAll rights reserved. No part of this publication may be reproducedwithout permission from the publisher.BRANCH SECRETARIES 2016Aberdeen: Ms Donna Wallace aberdeenbranch@iat.org.ukCambridge: Ms Fran Flack cambridgebranch@iat.org.ukEdinburgh: Ms Janice Young edinburghbranch@iat.org.ukHertfordshire & Essex: Ms Helena Box hertsessexbranch@iat.org.ukHuntingdon, Suffolk & Norfolk: Ms Jo Martin hssbranch@iat.org.ukIreland: Mr Colin Travis irelandbranch@iat.org.ukLondon: Ms Amanda Dickson londonbranch@iat.org.ukMidlands: Mr Ian Fielding midlandsbranch@iat.org.ukNorth East England: Ms Rachael Handisides and Ms Joanne Bland northeastbranch@iat.org.ukNorth West: Ms Gail Morrisey cheshirebranch@iat.org.ukOxford: Mr Adrian Woodhouse oxfordbranch@iat.org.ukSurrey, Hampshire & Sussex: Ms Lesley Hughes shsbranch@iat.org.ukWest Middlesex: Ms Wendy Steel westmiddxbranch@iat.org.ukWest of Scotland: Ms Linda Horan westscotlandbranch@iat.org.ukATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page ii

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August 2016 Animal Technology and Welfare82EditorialJas BarleyChair of the Editorial BoardBefore I continue please accept my deepest apologies for the delay of this issue and the reduced content. Unfortunatelyduring our lifetimes all of us sooner or later will experience periods when our priorities will be dictated by events beyondour control and my life took such a turn which has meant that all things other than absolute necessities had to be puton hold. Now that life is returning to a more usual state (I hesitate to use the term normal), rather than delay furtherwhilst I prepare additional content I’ve decided to issue what I have in order to get back on schedule as soon as possible.I’d like to take this opportunity to thank the Institute’s Council for their forbearance and support during this difficult timeand also thank our advertisers together with Sue Ojakowa and Warwick Printing for their support and patience.This issue takes no particular theme but makes a break from our normal type of content by including a very moving andinformative article on how one of our regular contributors, Gary Martinic, survived a heart attack thanks to modernmedicine. His doctors made use of knowledge and technology, much of which was developed due to research involvinganimal use. If you have any doubts regarding the value of our work I’m sure Gary would be only too happy to reassure usof its worth. Glad to have you still on board Gary and look forward to further contributions from you in the future.Another new type of article is from Nikki Osborne’s blog. The included blog is on Culture of Care something that we allneed to consider and continually monitor in order to provide the best welfare for the animals in our charge and to preventthe phrase just becoming lip service. There are other blogs on the web that discuss topics of interest to our membersand subscribers, so if you have a favourite that is relevant to our readership please let me know and I’ll see if anythingis available that others may benefit from. Although we assume that everyone these days has access to the internet thisis not the case, especially for those working in developing countries. It has also been brought to my attention that somemembers are unable to use a computer due to medical conditions e.g. some forms of epilepsy.A more ‘regular’ type of paper is provided by Laura Scullion Hall and Sally Robinson on implementing a successful positivereinforcement training protocol in laboratory-housed dogs. Although, when compared to the numbers of rodents used,dogs are one of the less used species in the UK, they make an important contribution in many fields of research bothwithin the UK and globally. Training of animals to participate willingly in procedures is one method of refining procedures.Despite the acknowledged need for such training in the laboratory-housed dog and the availability of training protocols forthe pet or working dog, few published studies exist detailing the implementation of training protocols. This paper outlinesa positive reinforcement-based protocol and describes the implementation in a busy pharmaceutical industryenvironment.A further article from a regular contributor to Animal Technology and Welfare (ATW), namely Penny Hawkins of theResearch Animals Department at the RSPCA is also offered. This time the report concerns the focus by the RSPCA ontheir Severe Suffering project, of which the Institute is a major supporter. I would commend readers to take time to readthe report and see what steps can be implemented in their own establishments.Further posters from Congress 2016 are included in this issue and it is my intention to publish the remaining submissionsthat have been made available to ATW in the December issue. You will have recently seen the call for Platformpresentations and Posters for Congress 2017. If you have already registered your interest in taking your first stepstowards speaking in public there is some useful guidance on both presenting and preparing a poster on the IAT websitewithin the Congress section. However, hard copy of this guidance is included in the Tech-2-Tech section of this issue.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.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page ix

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August 2016 Animal Technology and Welfare83Implementing a successful positivereinforcement training protocol inlaboratory-housed dogsLAURA E.M. SCULLION HALL1and SALLY ROBINSON21Behaviour and Evolution Research Group, Psychology, School of Natural Sciences,University of Stirling, Stirling FK9 4LA2Drug Safety and Metabolism, AstraZeneca, Alderley Park, Macclesfield, Cheshire SK10 4TFCorrespondence: laura.hall@stir.ac.ukSummaryTraining is recognised as important for laboratoryanimals, both to improve welfare and increase theefficiency of conducting husbandry and regulatedprocedures. Despite the acknowledged need for suchtraining in the laboratory-housed dog and theavailability of training protocols for the pet or workingdog, few published studies exist detailing theimplementation of training protocols. Refinement ofrestraint for procedures and husbandry can improvewelfare. This paper outlines a positive reinforcement-based protocol and describes the implementation in abusy pharmaceutical industry environment. Sixty-sixnaive dogs held as stock were trained in brief, once-weekly training sessions for four weeks. Data wererecorded on behaviour and progress in training. Theresults support the implementation of brief, structuredtraining to increase welfare and compliance. Thescoring tools used also identified a small number ofdogs which failed to respond to training, providing ameans for staff to allocate dogs best suited to short-or long-term studies.KeywordsLaboratory-housed dog, training, Positive ReinforcementTraining (PRT), 3Rs, refinement, welfareWork carried out at Drug Safety and Metabolism,AstraZeneca, Alderley Park, Macclesfield, Cheshire.IntroductionThe use of dogs in laboratoriesDogs are a commonly-used second species in thesafety testing of new medicines with over 3,500 usedin the UK in 20131and over 100,000 used globallyeach year.2Dogs are given additional protection underthe Animals (Scientific Procedures) Act, 1986 (ASPA)and there is an ethical and scientific imperative tomaximise the benefits while minimising the harmsassociated with their use in scientific testing.Welfare, or the ability of an individual to cope with itsenvironment,3is acknowledged to influence the qualityof the data obtained from the use of animals inscientific procedures. Refinement, one of the guiding3Rs4principles of humane experimental practice, isdefined as‘Any approach which avoids or minimises the actualor potential pain, distress and other adverse effectsexperienced at any time during the life of theanimals involved and which enhances theirwellbeing.’5Refinement, therefore, must apply to all aspects of thelifetime of a laboratory-housed dog, not only the timeduring which it is on-study. Improving welfare mustbegin while dogs are held at stock, during which timeacclimatisation and basic habituation are normallyconducted, although this varies across industry.A multi-faceted Welfare Assessment Framework wasdeveloped for the laboratory-housed dog,6incorporatingmeasures of affect, behaviour, mechanical pressurethreshold and cardiac output. It was found that dogswith negative welfare had a negative affective state, alower tolerance for mechanical pressure, higher bloodpressure and more negative welfare indicatingbehaviours in the home pen and greater cardiac andbehavioural changes when exposed to positive andnegative behavioural challenges. In particular, theresponse of the dogs to restraint6suggested thatrefinements to the technique were necessary.Positive Reinforcement Training(PRT)Training of laboratory-housed dogs, in particular tableATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 83

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training, is common although techniques and frequencyof training vary considerably. Habituation (repeatedexposure to an aversive event) is common practice forregulated procedures in the laboratory setting and mayresult in a decreased behavioural response to theaversive stimulus or event.7However this may notrepresent actual habituation but rather a “freezing”response and cooperation, while internally, arousal hasnot decreased.8It is commonly recommended thatsome form of “habituation” take place before a study,7however the interpretation of its use varies and there iscurrently no standardisation in the use ofdesensitisation within the laboratory environment forthe dog.9This may be due to a lack of understanding ofthe distinction between habituation and desensitisationand lack of a structured programme to implementdesensitisation. As such, negative reinforcement (NRT)training is often more commonly used than positivereinforcement. NRT may involve the use of anunpleasant stimulus and as such encourages fear,resistance and avoidance, all of which are undesirablestates in an in vivo model of a healthy human.Positive Reinforcement Training (PRT) is a form ofoperant training, in which the subject (dog) has toperform an active behaviour in order to receivefeedback. The addition of an appetitive stimulus toincrease the occurrence of a behaviour.The use of PRT rather than classical conditioning (e.g.habituation or desensitisation) has the benefit ofincreasing the choice and control7,10of the subject, bothof which are known to be important to good welfare.Requiring the subject to make an active choice toexpress a behaviour allows it a sense of control, whilealso increasing the predictability of an event. PRT isfavoured over NRT for its effectiveness and the use ofonly appetitive stimuli, rather than aversive stimuli, isconsidered more ethically acceptable.PRT is well-described as an effective technique in thepet dog11or working dog,12,13as well as in otherlaboratory species,13yet there are few experimentalaccounts of the benefits in the case of the laboratory-housed dog. A study of refinements to the protocolused in dosing by oral gavage14found that amongstother refinements, PRT mitigated the negative changesin welfare associated with restraint and oral gavage.Habituation, which occurs by NRT, was found to resultin decreased welfare and increased time to dose.Increased positive staff contactWhile operant learning is the focus of this study, sociallearning in dogs must also be accounted for whendeveloping a training plan, given dogs’ affinity forunderstanding our intention,15emotions,16gaze,17gestures18and vocal communication.19Dogs also utilisesocial referencing to interpret unfamiliar situations.20Positive human contact is known to benefit dogs in anumber of behavioural,21physical22and physiological23ways. Specifically in the laboratory-housed dog, periodsof positive staff contact can increase overall welfare inthe home pen.6,24For these reasons, this trainingprotocol was designed to not only achieve a specifictraining goal but to use the contact between dogs andstaff to improve dog welfare.AimsThe aims of the present study were (1) to investigate theefficacy of a training protocol for laboratory-housed dogsand (2) to evaluate the benefits of the training protocolin relation to the feasibility of its implementation. It washypothesised that behaviour on the table, and the stageof training demonstrated by each dog would improvefrom Week 1 to Week 4. It was also hypothesised thatthe design of the training protocol and the tools used tomeasure progress would prove useful to staff andincrease overall ease of interaction with the dogs.MethodsAnimals and housingA total of 66 AZ strain beagle dogs (27 male, 39female) took part in the training protocol. At the startof training, dogs were aged between 12-24 months. Alldogs were naive to regulated procedures and receivedno training other than handling during weekly healthchecks.Dogs were housed in same-sex groups of up to 10 dogsin two interconnecting pens of approximately 15m2each. A variety of chew toys were provided asenrichment and there were raised platforms of varyingheights within the home pens. Dogs were allowedaccess to outdoor exercise areas containing climbingframes at least once per week.Training protocolNo structured training protocol had previously been inplace. Before the delivery of the training protocolbegan, staff received training in the welfareassessment and training of laboratory-housed dogsincluding both theory and practical learning.The overall goal of training was for dogs to exhibit thebehaviour ‘calm sitting while restrained’ when placedon the table. A combination of luring, shaping andstationing was used in the training protocol. Allbehaviour offered by the dogs was voluntary, with staffinstructed to avoid physical manipulation of the dogs.Stages of training were devised and are shown inTable 1. It was necessary for a dog to have met thecriteria of all previous stages of training beforeprogressing to the next. Staff were also provided withphotographic guides to the behaviours exhibited ateach stage of training and troubleshooting forunexpected behaviours.Implementing a successful positive reinforcement training protocol in laboratory-housed dogs84ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 84

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Implementing a successful positive reinforcement training protocol in laborator y-housed dogs85The training protocol was delivered to each dog onceweekly for four weeks. Each training session wasbetween 2-5 minutes with the time decreasing as staffproficiency in training increased. Four members of staffwere chosen as trainers and in addition to usualhusbandry duties they undertook training duringweekday afternoons. Staff worked in pairs with oneconducting the training and the other recordingbehaviour and stage of training data on the recordsheet. Typically, one room each of male and femaledogs would be trained per day.Data collectionTrainers recorded both the stage of training achievedeach week (A-G; Table 1) and behaviours exhibited(range -18 to +6; Table 2). All behaviours were scoredas occurring not at all, some of the time or all of thetime.EthicsThis design and implementation of this study wasoverseen by scientific and veterinary staff. Noregulated procedures were conducted and therefore noadditional approval was required under ASPA.ResultsChange in behaviour on the tableFigure 1 shows the increase in scores across the fourweeks of training. Training scores for all dogssignificantly improved from a mean score of -3.41 inWeek 1 to +3.64 in Week 4 (Wilcoxon signed ranks testZ=6.483, p<.001). Male and female dogs only differedsignificantly in behaviour score in Week 3 (MannWhitney U test, U=347.5, p=.015), see Figure 2.Stage Description––––––––––––––––––––––––––––––––––––––––––––––––––A Does not accept treats (nervous or excited).B Accepts treats from handler.C Calm and relaxed on table.D Attempts sitting behaviour.E Shows brief sits.F Can maintain a longer sit.G Sits well and tolerates gentle restraint.––––––––––––––––––––––––––––––––––––––––––––––––––Table 1. Stages of trainingFigure 1. Behaviour scores over four weeks of trainingfor all dogs.Figure 2. Behaviour scores over four weeks of trainingby sex.Figure 3. Training stage reached over four weeks oftraining for all dogs.Stage of training reachedStage of training also significantly improved from Week1 to Week 4 (Wilcoxon signed ranks test, Z=-6.722,p<.001). In Week 1, 46 dogs were at Stage A or B oftraining (mean = Stage B). Only four dogs were at StageATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 85

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Implementing a successful positive reinforcement training protocol in laboratory-housed dogs86E or F. After the final training session in Week 4, fourdogs were at Stage A and 10 were at Stage B. In Week4, the mean stage of training was E with 48 dogs (73%)at Stages E-G of training (Figure 3). Male and femaledogs did not differ in stage achieved during Weeks 1-3,however in Week 4, females had a higher mean stagethan males (Mann Whitney U test, U=367.5, p=.032),see Figure 4.Suitability of the table training toolThree male dogs and four female dogs had consistentlynegative behaviour scores across the four weeks oftraining with scores in Week 4 ranging from -15 to -3.These dogs also did not progress beyond Stage B oftraining. It was decided that training would continue foras long as feasible in order to determine if additionalsessions would be beneficial.These dogs were given additional training over thefollowing six weeks by the end of which all dogs had aminimum behaviour score of 0, however they remainedat training Stage A or B. From these results, it appearsthat the response of dogs to training in the first fourweeks predicts the success of training. Those dogswhich had a negative behaviour score in Week 4 did notrespond well to additional training.These results support two important findings. Firstly,that four weeks of brief training sessions is sufficientto improve the behaviour and training of most dogs with73% achieving Stages E–F of training. Secondly, thetable training tool is a sensitive measure ofresponsiveness to training, with dogs which hadconsistently negative behavioural scores failing torespond to an additional six weeks of training. Thismeans that using the table training tool, dogs which areunlikely to adapt to regulated protocols and otheraspects of being on-study, can be identified. It isaccepted that dogs which fail to adapt and thereforehave negative welfare should not be assigned to long-term studies for ethical and scientific reasons. Thistable training tool should therefore be useful totechnical and veterinary staff in assessing thesuitability of dogs for long-term studies.DiscussionResults of training protocolThe results of the training protocol showed a significantimprovement in both behaviour and stage of trainingover the four weeks of training. Other than in Week 3male and female dogs did not differ significantly inbehaviour score. However, female dogs were at ahigher mean stage of training in Week 4. This suggeststhat females dogs may respond better to training. Themean stage of training in Week 4 was E (shows briefsits) which means that most dogs were exhibiting calmbehaviour and volunteering the desired behaviour.Behavioural indicators of welfare also improvedsignificantly from Week 1 to Week 4, with a mean scoreof +3.64 of a maximum +6. This indicates that theprotocol has been successful in improving the welfareof the dogs while being handled on the table. Ourprevious research e.g.2,14suggests that brief restraintcan have a negative welfare impact on dogs but alsothat positive reinforcement training can mitigate thisimpact.Many of the dogs used in this study were latertransferred to a contract research organisation wherethey were used in long-term colonies. Staff noted thatthese dogs were easier to handle and better adapted toregulated procedures than dogs from other sources(personal communication April 2015), although thiswas not quantified using the tools in this study. Thislasting effect suggests that the investment in trainingresulted in long-term benefits to dogs and staff.Ease of implementationStaff reported the training protocol to be easy toimplement and the results to be readily observable.Assigning four dedicated staff to training wasdetermined to be a contributing factor to the overallsuccess of the protocol. This allowed staff to quicklybecome proficient in the techniques and to maintainconsistency between training sessions. The length oftraining sessions decreased as staff became moreproficient and as dogs’ training progressed, with manydogs needing no more than two minutes of training perweek. This was determined to be an acceptableincrease in duties for staff.Use of scoring toolsThe tools used to score behaviours and stages oftraining sessions were developed from those used in.14Figure 4. Training stage reached over four weeks oftraining by sex.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 86

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Implementing a successful positive reinforcement training protocol in laborator y-housed dogs87These tools proved to be easy-to-use for staff andsensitive to changing behaviour in the dogs. Further,the behavioural scoring proved to be useful inidentifying dogs which did not respond well to training(7/66). The majority of dogs responded well to fourweeks of training. However those that did not failed torespond to additional training.It is recognised that dogs which fail to adapt to theevents associated with being on-study should not beassigned to long-term studies or re-use colonies,however, there has been no evidence-based method ofdoing this. Anecdotally, technical staff who work withdogs have expressed a desire to identify such dogs inorder to assign them to appropriate studies of shorterduration, or terminal studies. The use of thebehavioural scoring tool described here should assiststaff in decision making.ConclusionsThe results of this study show a clear improvement inboth behaviour and training across four weeks. Themajority of dogs had reached a satisfactory level oftraining by the fourth week, while the use of positivereinforcement training also improved the welfare of thedogs while on the table. Dogs which failed to respondto four weeks of training also failed to respondsufficiently to additional training, meaning that thebehavioural scoring tool can be used to identify thesedogs and assign them to appropriate studies. Staffwere able to incorporate the protocol into regular dutiesand quickly became proficient in the trainingtechniques, which increased the success of the study.Given the benefits of implementing positivereinforcement training and the ease of implementation,it is recommended that a protocol such as this isimplemented in dog units.AcknowledgementsThe authors are indebted to LB for her assistance andenthusiasm in implementing the preceding study andover-seeing the collection of data and to CD, KH andEW for carrying out training.References21Home Office. (2013). Statistics of Scientific Procedureson Living Animals in Great Britain 2012. Crown Copyright.22Hall, L.E. (2014). A practical framework for harmonisingwelfare and quality of data output in the laboratory-housed dog. PhD Thesis, University of Stirling, Stirling,UK.23Broom, D. (1986). Indicators of poor welfare. BritishVeterinary Journal; Vol 142 No 6, 524-526.24Russell, W. and Burch, R. (1959). The Principles ofHumane Experimental Technique. Universities Federationfor Animal Welfare, Wheathampstead, UK.25Buchanan-Smith, H.M., Rennie, A. and Vitale, A. et al.(2005). Harmonising the definition of refinement. AnimalWelfare Vol 14 No 4, 379-384.26Hall, L.E., Robinson, S. and Buchanan-Smith, H.M. (inprep) A welfare assessment framework for the laboratory-housed dog: linking welfare and data output.27Laule, G. (2010). Positive Reinforcement Training forLaboratory Animals. In: The UFAW handbook on the careand management of laboratory and other researchanimals. Universities Federation for Animal Welfare,Wheathampstead UK.28Ruys, J., Mendoza, S., Capitanio, J. et al. (2004).Behavioral and physiological adaptation to repeated chairrestraint in rhesus macaques. Physiology and Behavior;Vol 82 No 2-3, 205-213.29Prescott, M.J., Morton, D., Anderson, D. et al. (2004).Refining dog husbandry and care: Eighth report of theBVAAWF/FRAME/RSPCA/UFAW Joint Working Group onrefinement. Laborator y Animals; Vol 38 (SUPPL. 1), S1:1-S1:94.10Overmier, J., Patterson, J. and Wielkiewicz, R. (1980).Environmental contingencies as sources of stress inanimals. New York: Plenum Press.11Hiby, E., Rooney. N. and Bradshaw, J. (2004). Dogtraining methods: Their use, effectiveness and interactionwith behaviour and welfare. Animal Welfare; 13 No 1, 63-69.12Fjellanger, R., Andersen, E. and McLean, I. (2002). Atraining program for filter-search mine detection dogs.International Journal of Comparative Psychology; Vol 15No 4, 278-287.13McKinley, J., Buchanan-Smith, H.M., Bassett, L. et al.(2003). Training common marmosets (Callithrix jacchus)to cooperate during routine laboratory procedures: Easeof training and time investment. Journal of Applied AnimalWelfare Science; Vol 6 No 3, 209-220.14Hall, L.E., Robinson, S. and Buchanan-Smith, H.M.(2015). Refining dosing by oral gavage in the dog: Aprotocol to harmonise welfare. Journal of Pharmacologicaland Toxicological Methods; Vol 72, 35-415Téglás, E., Gergely. A., Kupán, K.E., Gergely, A., Kupán,K. et al. (2012). Dogs’ gaze following is tuned to humancommunicative signals. Current Biology; Vol 22 No 3,209-212.16Müller, C.A., Schmitt, K., Barber, A.L. et al. (2015). Dogscan discriminate emotional expressions of human faces.Current Biology; Vol 25 No 5, 601-605.17Gácsi, M., Miklósi, Á., Varga, O. et al. (2004). Arereaders of our face readers of our minds? Dogs (Canisfamiliaris) show situation-dependent recognition ofhumans attention. Animal cognition; Vol 7 No 3, 144-153.18Riedel, J., Schumann, K., Kaminski J. et al. (2008). Theearly ontogeny of human-dog communication. AnimalBehaviour; Vol 75 No 3, 1003-1014.19Rossano, F., Nitzschner, M. and Tomasello, M. (2014).Domestic dogs and puppies can use human voicedirection referentially. Proceedings of the Royal Society B:Biological Sciences; Vol 281 No 1785, 20133201.20Merola, I., Prato-Previde, E. and Marshall-Pescini, S.(2012). Dogs’ social referencing towards owners andstrangers. PloS One; 7(10): e47653.21Hennessy, M., Voith, V., Hawke, J. et al. (2002). Effectsof a program of human interaction and alterations in dietcomposition on activity of the hypothalamic-pituitary-adrenal axis in dogs housed in a public animal shelter.Journal of the American Veterinary Medical Association221(1): 65-71.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 87

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89August 2016 Animal Technology and WelfareThe RSPCA focus on severe sufferingprojectPENNY HAWKINSResearch Animal Department, RSPCA, Wilberforce Way, Southwater, West Sussex RH13 9RSCorrespondence: penny.hawkins@rspca.org.ukSummaryThe RSPCA Research Animals Department has anongoing project that focuses on reducing and avoiding‘severe’ suffering which has been well supported bythe Institute of Animal Technology (IAT). This shortarticle describes two current initiatives; an onlineinformation resource and a ‘Road map’ pack designedto help establishments act on severe suffering at alocal level.The web-based resource, ‘Focus on severe suffering’,is endorsed by the IAT, Laboratory Animal ScienceAssociation (LASA) and the Laboratory AnimalVeterinary Association (LAVA). It sets out the factorsthat can contribute towards making a proceduresevere, explains how these can be addressed and linksto other websites and resources for further informationand practical guidance. The ‘Road map’ pack is linkedto Animal Welfare and Ethical Review Body (AWERB)tasks and comprises short presentations with notesand workshop materials that can be used to helpestablishments audit their procedures and focus onrefining any that can cause severe suffering.KeywordsSevere suffering, ethics, refinement, practicalguidanceIntroductionAll levels of severity in animal research and testing area concern to the RSPCA and the scientific communityalike but severe suffering is obviously the greatestconcern of all and is a top priority for us. Tacklingsevere suffering has been a theme throughout theRSPCA’s work for some years, particularly since ourproject on pain assessment1(which progressed into ourwork on welfare assessment)2began in the late 1990sand the severe suffering project has been ongoingsince 2012.Ensuring that we are working effectively on the issuehas required a great deal of research, liaison andcollaboration with individuals working in science, theHome Office and bodies such as the IAT, LASA andLAVA. The RSPCA is extremely grateful for the supportwe have received from all of these people andorganisations which has enabled us to produceresources that will help to reduce and avoid severesuffering, as well as getting the issue on the agenda atscientific meetings in the UK and overseas.Background to the severeresourcesIn addressing severe suffering, we are advocatingparallel approaches that are essentially a veryfocussed application of the 3Rs. Ideally, proceduresinvolving severe suffering would be replaced or avoidedaltogether – the RSPCA’s principal goal. Wherever thisis not currently possible, it is clearly essential toreduce the numbers of animals experiencing severesuffering and to refine procedures so that suffering isno longer severe or at least less severe. Findinginnovative and practical ways to refine severeprocedures is the main emphasis of the project, asrefinement is a key area of expertise for the RSPCA andis an immediately achievable ‘R’.To identify the best way to work on the issue, weneeded information on the actual severity ofprocedures – the level and nature of harms caused toanimals. Although this is now published in the annualHome Office Statistics3(Figure 1, Table 1) and thisinformation is very welcome with respect to88038 23202 11647 344 RegulatoryBasic researchTranslational/appliedresearchProtection of the naturalenvironmentFigure 1. Procedures reported as ‘severe’ actualseverity by purpose in Great Britain for 2015.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 89

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The RSPCA focus on severe suffering project90transparency about animal use, it is at quite a basiclevel and more detail is necessary on the types of‘models’, tests and procedures that have the potentialto cause severe suffering.Source: Home Office Statistics, Table 3.2.3Mice arecurrently the most likely species to experience severesuffering; a total of 101,494 experimental procedureson mice were reported as ‘severe’ actual severity for2015.Thanks to the high level of constructive liaison that wehave with the scientific community and the regulatorand a great deal of literature searching, we have beenable to produce a list of procedures that are especiallylikely to cause severe suffering, which has enabled usto identify procedures and models that require morespecific guidance and to formulate a more genericapproach to addressing the issue.Two work streamsThe RSPCA’s work to produce guidance on refiningspecific procedures or ‘models’ that may cause severesuffering involves setting up Expert Working Groups toconsider the necessity and justification for animal use,identify potential harms throughout the animals’ livesand recommend ways of alleviating these based on thescientific literature and current good practice. Groupmembers include animal technologists, vets, scientistsfrom industry and academia and a Home OfficeInspector.These Expert Working Groups are a natural follow-onfrom the BVAAWF/FRAME/RSPCA/UFAW* Joint WorkingGroup on Refinement (JWGR), which had a similarapproach and membership and produced its first reporton blood sampling back in 1993. Throughout the 23years of the JWGR and more recently with the ExpertWorking Groups, animal technologists and the IAT haveprovided strong support, input and advice, helping usto provide practical guidance that has had real impact.So far, we have produced four reports on refiningprocedures involving the Experimental AutoimmuneEncephalomyelitis (EAE) mouse model,4seizures,convulsions and epilepsy,5sepsis6and rheumatoidarthritis.7A fifth Expert Working Group is currentlyworking on spinal cord injury. Although the RSPCA andother bodies such as LASA and the NC3Rs are alsoaddressing these specific procedures, this is a longterm collective project so the RSPCA has created moregeneric guidance in the form of our ‘Focus on severesuffering’ web pages.The web pagesThe web-based resource, ‘Focus on Severe Suffering’(rspca.org.uk/severesuffering) was developed with andis endorsed by the IAT, LASA and LAVA (Figure 2). Theaim is to help relevant staff and committees or groupswithin establishments to address severe sufferinglocally. It became obvious to us at an early stage thatdifferent audiences required different information, sowe defined three target groups; animal technologistsand Named Veterinary Surgeons (NVSs), researchersand Animal Welfare and Ethical Review Body (AWERB)members. Each of these has a separate ‘pathway’ tofollow within the web pages which includes the mostPurpose Number of procedures––––––––––––––––––––––––––––––––––––––––––––––––––Batch potency testing 144,957Immune system 4,389LD50 and LC50 3,952Multisystemic research 2,918Human infectious disorders 2,869Oncology 2,060Cardiovascular blood and 1,928lymphatic systemNervous system 1,804Other type of toxicity or 1,230safety testHuman cancer 920––––––––––––––––––––––––––––––––––––––––––––––––––Table 1. Top ten purposes of severe experimentalprocedures involving mice in Great Britain, 2015.–––––––––––––––––––––––––––––––––––––––––––––––––––––* British Veterinary Association Animal Welfare Foundation, Fund forthe Replacement of Animals in Medical Experiments, UniversitiesFederation for Animal WelfareFigure 2. The RSPCA Focus on severe suffering webpages.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 90

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The RSPCA focus on severe suffering project91appropriate materials to help them fulfil their roles intackling severe suffering, although any visitor canaccess each of the pathways.For each one, the page begins by considering whatmakes a procedure ‘severe’, breaking this down intothree main causes:G some procedures or ‘models’ are more likely to besevereG a combination of factors can increase the level ofsuffering to severe; i.e. the cumulative severityeffect or mortality may involve severe sufferingWe have set out the factors that can contribute towardsmaking a procedure severe, explaining how these canbe addressed and linking to other websites andresources for further information and practicalguidance. Further content focuses strongly on welfareassessment, applying refinements and humaneendpoints, as these are all fundamental to avoidingand reducing severe suffering. Each of these key areasis addressed from a different angle for animaltechnologists/NVSs, scientists and AWERB members.For example, information relating to refinement ispresented from a different aspect within each of thethree pathways. The route for animal technologists andNVSs focuses on useful information sources, theimportance of evaluating refinement to ensure that it isimproving welfare and the value of the AWERB as aforum for identifying and discussing refinements andtheir evaluation throughout the project and at mid- orend-term reviews. For scientists, the pages begin witha definition of refinement and explain the legalrequirement to implement this, before setting out thesources of information. It also acknowledges theimportance of ensuring that refinement is compatiblewith the science and highlights how useful advice canbe from both the Named Information Officer (NIO) andthe AWERB when seeking to avoid or reduce severesuffering. The refinement pages for AWERB membersalso begin with a definition, moving on to list which ofthe AWERB’s tasks are relevant to addressing severesuffering and include the essential role of the NIO.The pathways for welfare assessment and humaneendpoints take a similar approach, in which we haveaimed to present both information and approaches insuch a way that they will be meaningful and relevant tothe different target audiences.The road mapOne of the resources that can be downloaded from theweb pages is a Road map resource pack, which isintended for use by internal bodies such as the AWERB,a 3Rs Group or other animal care and use or ethicscommittees, although it has also proved useful as atraining aid and for use by individual scientists.The ‘Road map’ is a series of practical steps that willenable establishments to identify ways to reduce, avoidand ultimately end severe suffering, which wedeveloped following consultation with a number ofresearch establishments. This approach is stronglylinked to relevant AWERB tasks and uses currentguidance from both the Home Office and the EuropeanCommission. We have taken this approach because wewanted to help ensure that current guidelines andrequirements were effectively implemented andapplied, without adding additional tasks.A key principle of the Road map is an ‘audit’ ofprocedures to establish how well current refinementpractices are working and to identify any areas wherefurther refinement can be applied. In order for this tobe achieved it is essential that researchers work withtheir animal technologists and named personsdeveloping a team approach to the alleviation ofsuffering. For further information, please see the‘resources’ box within the severe suffering web pages(rspca.org.uk/severesuffering).ConclusionWe hope that you find the severe suffering resourcesuseful and we would also like to encourage you to bringthe resources to the attention of your establishments,for example by making the AWERB or named personsaware of them. Although they address severe suffering,much of the content will help with refinement ingeneral.The RSPCA regards the Focus on severe sufferingpages and the Road map resource very much as worksin progress, which will need regular updates to ensurethat they are user-friendly, current and feasible. Weplan to update the pages quarterly and wouldespecially welcome any feedback that you may have, tohelp us develop and improve them – please emailresearch.animals@rspca.org.ukAcknowledgementsThanks to the IAT, LASA, LAVA, Home Office and thescientific community for their support for this projectand to the IAT for the opportunity to speak at Congress2016.References1Hawkins, P. (2002). Recognising and assessing pain,suffering and distress in laboratory animals: a survey ofcurrent practice in the UK with recommendations.Laboratory Animals, 36(4), 378-95.2Hawkins, P., Morton, D.B., Burman, O. et al. (2011). Aguide to defining and implementing protocols for thewelfare assessment of laboratory animals: eleventhreport of the BVAAWF/FRAME/RSPCA/UFAW Joint WorkingGroup on Refinement. Laboratory Animals, 45, 1-13.3Home Office. (2016). Annual Statistics of ScientificATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 91

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93PAPER SUMMARYTRANSLATIONSINHALTVERZEICHNISINHALTVERZEICHNISUmsetzung eines erfolgreichen, auf positiverVerstärkung beruhenden Trainingsprotokolls für imLabor untergebrachte HundeLAURA E.M. SCULLION HALL1und SALLY ROBINSON21Behaviour and Evolution Research Group, Psychology (Verhaltens- undEntwicklungsforschungs-Gruppe, Psychologie), School of Natural Sciences, University ofStirling, Stirling FK9 4LA2Drug Safety and Metabolism (Arzneimittelsicherheit und Stoffwechsel), AstraZeneca,Alderley Park, Macclesfield, Cheshire SK10 4TFKorrespondenz: laura.hall@stir.ac.ukAbstractTraining für Versuchstiere gilt als wichtiger Beitrag zur Verbesserung des Tierwohls und zur Effizienzsteigerung bei derDurchführung von Tierhaltungs- und vorgeschriebenen Versuchsverfahren. Trotz der anerkannten Notwendigkeitsolchen Trainings von im Labor untergebrachten Hunden und der Verfügbarkeit von Trainingsprotokollen für den Haus-oder Arbeitshund wurden bisher nur wenige Studien veröffentlicht, die sich mit der Umsetzung vonTrainingsprotokollen beschäftigen. Durch die Senkung der Belastung bei Versuchsverfahren und Tierhaltung kanneine Verbesserung des Tierwohls erreicht werden. Dieser Artikel umreißt ein auf positiver Verstärkung beruhendesProtokoll und beschreibt seine Umsetzung in einem arbeitsintensiven pharmazeutischen Produktionsumfeld.Sechsundsechzig untrainierte, als Bestand gehaltene Hunde erhielten über vier Wochen hinweg einmal wöchentlichkurzes Training. Daten zum Trainingsverhalten und -fortschritt wurden erfasst. Die Ergebnisse unterstützen dieDurchführung kurzen, strukturierten Trainings als Mittel zur Verbesserung von Tierwohl und Compliance. Mit denverwendeten Scoring-Tools konnte auch eine geringe Anzahl von Hunden identifiziert werden, die nicht auf Trainingansprachen, womit das Personal die Möglichkeit erhielt, die jeweils für Kurz- bzw. Langzeitstudien optimal geeignetenHunde auszuwählen.Schlagwörter: Im Labor gehaltener Hund, Training, positives Verstärkungstraining, 3R-Prinzip, Refinement, TierwohlAugust 2016 Animal Technology and WelfareATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 93

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94Paper Summar y TranslationsDas RSPCA-Projekt „Focus on Severe Suffering“(Beurteilung schwerer Belastung durchTierversuche)PENNY HAWKINSResearch Animals Department, RSPCA, Wilberforce Way, Southwater, West Sussex RH13 9RS,GroßbritannienKorrespondenz: penny.hawkins@rspca.org.ukAbstractEin laufendes Projekt der Abteilung „Versuchstiere“ der britischen Royal Society for the Prevention of Cruelty toAnimals (RSPCA) beschäftigt sich schwerpunktmäßig mit der Minimierung der Belastung und der Vermeidung von„schwerem“ Leiden und genießt große Unterstützung des Institute of Animal Technology (IAT). Dieser kurze Artikelbeschreibt zwei aktuelle diesbezügliche Initiativen. Dabei handelt es sich erstens um eine Online-Informationsquelleund zweitens um eine „Roadmap“, die Einrichtungen dabei unterstützen sollen, auf lokaler Ebene im Hinblick aufschwere Belastungen zu handeln.Die webbasierte Informationsquelle „Focus on Severe Suffering“ wird vom IAT, der Laboratory Animal ScienceAssociation (LASA) und der Laboratory Animal Veterinary Association (LAVA) unterstützt. Sie legt die Faktoren dar, dieVerfahren zu einer schweren Belastung für das Versuchstier machen können, und erläutert deren möglicheBewältigung. Sie ist verlinkt mit anderen Websites und Ressourcen mit weiterführenden Informationen undpraktischer Anleitung. Die „Roadmap“ nimmt Bezug auf die Aufgaben des Animal Welfare and Ethical Review Body(Gremium für Tierschutz und ethische Überprüfung, AWERB). Sie enthält Kurzpräsentationen mit Anmerkungen sowieWorkshop-Materialien, die Forschungseinrichtungen zur Auditierung und zur Verbesserung ihrer Verfahren nutzenkönnen, die potenziell eine schwere Belastung verursachen.Schlagwörter: Schwere Belastung, Leiden, Ethik, Refinement/Verminderung der Belastung, RoadmapATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 94

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August 2016 Animal Technology and Welfare95CONTENU DE LA REVUECONTENU DE LA REVUEMise en œuvre d’un protocole de dressage deschiens de laboratoire basé sur un renforcementpositif efficaceLAURA E.M. SCULLION HALL1et SALLY ROBINSON21Groupe de recherche sur le comportement et l’évolution, Psychologie, École des Sciencesnaturelles, Université de Stirling, Stirling FK9 4LA2Innocuité des médicaments et métabolisme, AstraZeneca, Alderley Park, Macclesfield,Cheshire SK10 4TFCorrespondance: laura.hall@stir.ac.ukRésuméLe dressage est reconnu comme étant important pour les animaux de laboratoire, tant pour améliorer leur bien-êtreque pour augmenter l’efficacité de l’élevage et le respect des procédures réglementaires. Bien qu’il soit reconnunécessaire de procéder au dressage des chiens de laboratoire et malgré les protocoles de dressage actuellementen vigueur pour les animaux domestiques et les chiens d’utilité, il n’existe que quelques études publiées décrivanten détail la mise en œuvre de ces protocoles de dressage. La réduction des contraintes liées aux procédures et àl’élevage peut améliorer le bien-être des animaux. Cet article présente un protocole basé sur le renforcement positifet décrit sa mise en œuvre dans un milieu pharmaceutique industriel très actif. Pendant quatre semaines, soixante-six chiens innocents faisant partie d’un élevage ont été dressés dans le cadre de brèves séances de dressageorganisées une fois par semaine. Des données ont été enregistrées afin d’évaluer leur comportement et les progrèsobservés lors du dressage. Les résultats de cette étude soutiennent la mise en œuvre de séances de dressagerapides structurées afin d’améliorer le bien-être et l’obéissance des animaux. Les outils d’évaluation utilisés ontégalement permis d’identifier un petit nombre de chiens qui n’ont pas répondu positivement au dressage, offrantainsi au personnel de laboratoire un moyen de sélectionner les chiens les mieux adaptés aux études à court ou àlong terme.Mots-clés: chien de laboratoire, dressage, dressage basé sur le renforcement positif, 3Rs (Trois R), amélioration,bien-êtreATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 95

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96Paper Summar y TranslationsÉtude de la RSPCA sur le projet lié aux gravessouffrancesPENNY HAWKINSService des animaux de laboratoire, RSPCA, Wilberforce Way, Southwater, West SussexRH13 9RS*Correspondance: penny.hawkins@rspca.org.ukRésuméLe service des animaux de laboratoire de la RSPCA développe actuellement un projet étudiant les moyens quipermettent de réduire et d’éviter les « graves » souffrances et bénéficiant du soutien de l’Institute of AnimalTechnology (IAT). Ce court article décrit deux initiatives actuellement mises en œuvre : une ressourceinformationnelle en ligne et un « programme détaillé » conçu pour aider les établissements à prendre des mesuresau niveau local dans le but de réduire les graves souffrances.La ressource Web intitulée Focus on severe suffering (Étude sur les graves souffrances) a été approuvée par l’IAT,l’Association des Sciences de l’Animal de Laboratoire (LASA ou Laboratory Animal Science Association) etl’Association de Médecins vétérinaires pour les Animaux de Laboratoire (LAVA ou Laboratory Animal VeterinaryAssociation). Elle présente les facteurs pouvant contribuer à ce qu’une procédure entraîne de graves souffrances,elle explique comment gérer ces facteurs et elle comprend des liens vers d’autres sites Internet et ressources offrantdes renseignements supplémentaires et des conseils pratiques. Le « programme détaillé » est lié aux missions del’Animal Welfare and Ethical Review Body (AWERB) et inclut de courtes présentations contenant des notes, ainsi quedes documents de séminaire pouvant aider les établissements à vérifier leurs procédures et à améliorer celles quipeuvent entraîner de graves souffrances.Mots-clés: Graves souffrances, Éthique, Amélioration, Programme détailléATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 96

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97August 2016 Animal Technology and WelfareINDICE DE LA REVISTAINDICE DE LA REVISTAImplementación de un protocolo fructífero para unentrenamiento con refuerzo positivo en perros delaboratorioLAURA E.M. SCULLION HALL1y SALLY ROBINSON21Grupo de investigación sobre evolución y comportamiento, Psicología, School of NaturalSciences, University of Stirling, Stirling FK9 4LA2Seguridad de medicamentos y metabolismo, AstraZeneca, Alderley Park, Macclesfield,Cheshire SK10 4TFCorrespondencia: laura.hall@stir.ac.ukResumenEl entrenamiento se considera una actividad importante para los animales de laboratorio, tanto para mejorar subienestar como para incrementar la eficiencia a la hora de realizar procedimientos de cría y pautados. A pesar dereconocer la necesidad de dicho entrenamiento para perros de laboratorio y de existir protocolos de entrenamientopara el animal de compañía o perro de trabajo, hay pocos estudios publicados al respecto que detallen laimplementación de protocolos de entrenamiento. Un replanteamiento de unos procedimientos y de unas actividadesde cría menos restrictivas puede mejorar el bienestar. Este estudio explica un protocolo basado en el refuerzopositivo y describe la implementación en un entorno activo del sector farmacéutico. Se entrenó a 66 perros sinpreparación, que se conservaban como reserva, mediante sesiones cortas semanales durante cuatro semanas. Sefueron registrando los datos sobre el comportamiento y los avances en el entrenamiento. Los resultados respaldanla implementación de un entrenamiento breve y estructurado para incrementar el bienestar y el cumplimiento. Lasherramientas de evaluación utilizadas también sirvieron para identificar a un pequeño número de perros que norespondió positivamente al entrenamiento, con lo que el personal pudo asignar los perros que mejor se adaptabana los estudios de corto y largo plazo.Palabras clave: perros de laboratorio, entrenamiento, entrenamiento de refuerzo positivo, 3R, replanteamiento,bienestarATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 97

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98Paper Summar y TranslationsEl RSPCA se centra en el proyecto de sufrimientogravePENNY HAWKINSResearch Animals Department, RSPCA, Wilberforce Way, Southwater, West Sussex RH13 9RSCorrespondencia: penny.hawkins@rspca.org.ukResumenEl Departamento de Investigación de Animales (Research Animals Department) del RSPCA lleva a cabo un proyecto,centrado en reducir y evitar cualquier sufrimiento "grave", que cuenta con el total apoyo del Institute of AnimalTechnology (IAT). Este breve artículo describe dos iniciativas actuales: una fuente de información en línea y unadocumentación de “hoja de ruta” diseñada para ayudar a las distintas organizaciones a actuar en casos desufrimiento grave a escala local.La fuente basada en la web, ‘Focus on severe suffering’, está respaldada por el IAT, Laboratory Animal ScienceAssociation (LASA) y la Laboratory Animal Veterinary Association (LAVA). Establece los factores que pueden contribuira hacer que un procedimiento pueda ser grave; explica cómo solucionar esta situación y proporciona enlaces a otraspáginas web y fuentes para más información y guías prácticas. La documentación de “hoja de ruta” está vinculadaa las tareas del Animal Welfare and Ethical Review Body (AWERB) y está compuesta de breves presentaciones, notasy material de talleres, que pueden utilizarse para ayudar a las distintas organizaciones a evaluar sus procedimientosy a centrarse en replantear cualquier procedimiento que pudiera causar un sufrimiento grave.Palabras clave: Sufrimiento grave, ética, replanteamiento, hoja de rutaATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 98

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99August 2016 Animal Technology and WelfareINDICE DELLA REVISTAINDICE DELLA REVISTAImplementazione di un effettivo protocollo diaddestramento basato sul rinforzo positivo nei canida laboratorioLAURA E.M. SCULLION HALL1e SALLY ROBINSON21Behaviour and Evolution Research Group, Psychology, School of Natural Sciences, Universityof Stirling, Stirling FK9 4LA2Drug Safety and Metabolism, AstraZeneca, Alderley Park, Macclesfield, Cheshire SK10 4TFCorrispondenza: laura.hall@stir.ac.ukAbstractL’addestramento ricopre un ruolo parimenti importante per gli animali da laboratorio, in quanto può risultare in unbenessere migliorato e accrescere l’efficienza delle procedure zootecniche e regolamentate. Nonostante si riconoscala necessità di tale addestramento per i cani da laboratorio e della disponibilità di protocolli specifici per i canidomestici o da lavoro, sono pochi gli studi in essere in cui viene approfondita l’applicazione di tali protocolli. Ilraffinamento dell’equilibrio tra procedure e allevamento può migliorare il benessere. Questo articolo delinea unprotocollo basato sul rinforzo positivo e ne descrive l’implementazione in un ambiente farmaceutico molto attivo. Inbreve, sessantasei cani naïve tenuti come scorta sono stati ammaestrati una volta a settimana per un periodo diquattro settimane. Quindi, sono stati rilevati i dati sul comportamento e i progressi conseguiti. I risultati confermanol’uso di un addestramento breve ma strutturato mirato ad accrescere il benessere e l’obbedienza. Gli strumenti divalutazione utilizzati hanno identificato, inoltre, un numero limitato di cani meno ricettivi all’addestramento,consentendo allo staff di scegliere quelli più consoni a studi a breve o lungo termine.Parole chiave: cane da laboratorio, addestramento, addestramento a rinforzo positivo, 3 R, Raffinamento, benessereATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 99

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100Paper Summar y TranslationsFocus della RSPCA sulla grave sofferenzaPENNY HAWKINSResearch Animals Department, RSPCA, Wilberforce Way, Southwater, West Sussex RH13 9RS,Regno UnitoCorrispondenza: penny.hawkins@rspca.org.ukAbstractIl Research Animals Department della RSPCA sta conducendo un progetto incentrato sulla riduzione e l’eliminazionedi sofferenze ‘gravi’, che ha ricevuto ampio sostegno dall’Institute of Animal Technology (IAT). Questo breve articolodescrive due iniziative attualmente in corso: una risorsa informativa online e una ‘roadmap’ elaborata per aiutare leistituzioni ad agire sul problema della sofferenza grave a livello locale.La risorsa presente sul web, ‘Focus on severe suffering’, è riconosciuta dall’IAT, dalla LASA (Laboratory AnimalScience Association) e dalla LAVA (Laboratory Animal Veterinary Association). Delinea i fattori che possono contribuireall’inasprimento della sofferenza in una procedura, spiegando come affrontarli e fornendo link ad altri siti web erisorse che mettono a disposizione ulteriori informazioni e guide pratiche. La tabella di marcia o ‘roadmap’ èassociata alle mansioni dell’Animal Welfare and Ethical Review Body (AWERB) e racchiude brevi presentazionicorredate di note, nonché materiale per workshop destinato alle istituzioni, che possono utilizzarlo per sottoporre adesame le loro procedure e concentrarsi sul miglioramento di quelle che potrebbero causare gravi sofferenze.Parole chiave: Grave sofferenza, Etica, Raffinamento, RoadmapATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 100

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101TECH-2-TECHHaven’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.The evening of the sudden heart attack –a patient’s personal account of how theskill of doctors and medical research savedhis life!‘I will never forget the evening of the 4thOctober 2010, when I clinically died twice’GARY MARTINICCentre for Transplant & Renal Research, Westmead Institute for Medical Research,Westmead, NSW 2145 AustraliaCorrespondence: gary.martinic@sydney.edu.auBackgroundIt was approximately 7pm on the 4th October 2010 andI was driving home with my two daughters from a karatetraining session that we had just completed. The dojowas only a short 15 minute drive from home. Although Ihad been regularly training in Karate for nearly 15 yearsprior, this particular training session was my first nightback into the sport after a 3-4 year break (though I hadattended a few intermittent training sessions inbetween). During the more than 1.5 hours of quiteintense training on that night, I have to admit that, as a‘black belt’, I was pushing myself to train at the samelevel of intensity as my colleagues, who had beentraining regularly. At various stages of this training, I feltthat it was getting beyond my capabilities but I did notfeel any discomfort.More than halfway into the training and afterwards, Ihad to pause as I felt tired and found myself out ofbreath. I did manage to complete the training sessionbut did not feel well afterwards. Within 5 minutes of thedrive home, I had to pull into a side street and I startedto vomit violently which was accompanied by profusesweating and the onset of strong chest pain. Once Ihad vomited I felt a little better and continued driving.After we arrived home, I quickly parked the car in ourdriveway and explained to my wife, who was waiting atthe door that I have been feeling very, very ill and thatI just needed to sit in our terrace area for a fewminutes, to see if whatever I was feeling was going tosettle down. I managed to sit for no more than a fewminutes when I started to feel crushing, central chestAugust 2016 Animal Technology and WelfareATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 101

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Tech-2-Tech102pain which felt ‘like an elephant was sitting on mychest’. Immediately recognising these symptoms, Igrabbed the phone and called an ambulance describingmy symptoms to the person on the other side of thephone.Once done, I walked calmly to the room where my wifewas and explained to her that I WAS having a heartattack and that I’ve called an ambulance already and itwas on its way. My wife, in disbelief, said that it wasunlikely to be a heart attack and that it was probablyonly ‘over-exertion’, as she knew that I had a habit oftraining at quite an intense level. I explained to her thatwhat I was feeling was ‘real’ and when she saw that Iwas serious she quickly asked what she could do andgot our car ready to follow the ambulance to our localhospital. I also told my wife to let my daughters knowthat everything would be alright and that I just neededa ‘check-up’. Luckily for me, it was the early eveningwith little traffic on the roads and the ambulanceshowed up in less than 10 minutes. Once they arrived,they asked me what I was feeling and I explained tothem that I felt that I was having a heart attack. Eventhe ambulance officers told me that it was probablyunlikely due to my age (45) and that I looked young andfit but they did still take me into the ambulance andadministered an injection of morphine just in case, aswe hurriedly made our way to Liverpool Hospitalemergency ward. It’s comical to note that in all of thistime I was still dressed in my karate outfit!What happened in emergency triageAs I arrived into the emergency room, I was lying on anexamination table with my wife standing beside me.The nurse and the attending doctor asked me what Iwas feeling and I started to explain my symptoms, justas I did, I blacked out at that point. As I found out later,this was my first heart attack (or in medical terms,Acute Myocardial Infarction (AMI) caused by insufficientblood flow to my heart due to an artery blockage.Apparently at this point my heart stopped, so themedical staff immediately jolted me with a 200 Joule(J) electric shock using a transducer and then rushedme into the Cardiac Catheter Laboratory (Cath Lab).Upon arriving in the Cath Lab, I found myself lying onthe examination table semi-conscious and I could hearand see one doctor working on my right femoral artery(and being supervised by another more senior doctoron the other side of the room) while a male nurse wasstanding next to the stretcher to my left. After beinghere for only a few short minutes, I heard the juniordoctor say to the senior one, ‘this one’s not working’(referring to the long balloon catheter he had insertedinto my femoral artery) to which the senior doctorreplied, ‘try the larger size’. Shortly after this, Iapparently blacked out again (yes, my heart stoppedworking again) but not completely, as I was stillconscious enough to know what was generally going onaround me. Before I knew it, I felt this sudden type ofelectrical shock to my chest, it was the 200J electrictransducer being used again! But this time I certainlyfelt it! Let no one tell you differently, as someone thathas experienced this, I can tell you that if you receivethis shock when you are semi-conscious, it is myopinion, ‘one of the scariest things on earth’ toexperience. The analogy is similar to a ‘shotgun blastto the chest at point-blank range’. However, the irony ofthis is that without the transducer being used, I wouldnot be alive today.Clinical presentation and follow-upThis section explains the course of the medicaltreatment that I received. While there is some medicalterminology used, I have tried to present much of thefollowing in plain English language as much as waspossible at the time.A 45 year old male patient presented to the rooms at20:00 hrs on the 4/10/10 with onset of centralcrushing chest pain with associated diaphoresis(sweating) and vomiting. He was alert and orientatedwith Blood Pressure (BP) of 110/60 and Heart Rate(HR) at 80 beats per minute (bpm). Heparin infusionwas commenced at 21.10 hrs. Patient experienced aVentricular Fibrillation arrest at approximately 21:20hrs, which was reverted with one 200J shock. He wastransferred to the Cardiac Cath Lab at 21:50 hrs. Theright posterolateral coronary artery was stented withBare Metal Stent (BMS) and an Intra-aortic BalloonPump was inserted during the procedure as BP waslow. Post-procedure the patient remained well with nofurther chest pain. He was commenced on severalmedications including; Aspirin 100mg daily, Clopidogrel75mg daily, Crestor 10mg daily and Ramipril 1.25mgdaily and was referred for outpatient cardiacrehabilitation. He was discharged well on 9/10/10 tofollow up with his cardiologist 2-4 weeks later. Patientalso had a Echocardiograph.Intra-aortic Balloon Pump is a mechanical circulatoryassist device that increases myocardial oxygenperfusion while at the same time increases cardiacoutput. It increases cardiac output and increasescoronary blood flow and therefore myocardial oxygendelivery (Source: Internet)Clinical intervention detailsReason for admission: Chest pain.Principal Diagnosis: Inferior STEMI complicated by VFarrest.Additional Diagnoses/Complications: PmHx; smoker,denies IHD, DM, HTN Indication: acute myocardialinfarction – primary.Procedure: coronary artery study going ontopercutaneous transluminal angioplasty (PTCA).Current medications: nil.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 102

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103Tech-2-TechAnaesthetic: Lignocaine 10ml.Procedural medications: Heparin sodium 4500 U.Coronary artery study going onto PTCA was performedpercutaneously, using the right femoral artery underlocal anaesthesia. The screening time was 10.4 minand 150 ml of lopromide was used. The procedure wascomplicated by ventricular tachycardia.Haemodynamics: BSA: sq.m. Rhythm: Sinus rhythmAorta (s/d/m) 90/59/74 (pressures in mm Hg).Angioplasty: 1 vessel attempted. Right posterolateral-prox. Third: 15mm long type Bl lesion.Equipment: Z2 JR 4.0 6F, Route wire, Export AspirationCather AP 6 Fr, NC Sprinter 3.0 mm/12 mm, Micro-Driver 2.75/18 mm.Dilatation: 4 inflations were used with a maximumpressure of 12 Atm for a maximum of 18 sec. Nodissection occurred. The lesion was stented. STsegment changes occurred in the inferior & posteriorleads.Outcome: Stenosis pre: 95%, post: 0%. The procedurewas successful.Coronary angiography: the coronary anatomy was rightdominant.Artery discreetly diseased left main: origin 40%Discreetly diseased LAD: proximal third 30%.Normal LCx diffusely diseased RCA: proximal third 30%Discreetly diseased R postero/ateral: proximal third95% feasible; graftable.Comments: bare metal stent. IABP inserted.Conclusions: Coronary artery disease; SuccessfulPTCA Recommendation: Angioplasty.Physical recovery and progressFollowing my AMI (Figure 1) my condition wassuccessfully treated and I was left to recover in hospitalfor 6 days. During this time, I met my cardiologist, DrCon Arronis, who followed up my condition a number oftimes while a barrage of electrocardiographs (Figure 2)medical reports (Figure 3) and pathology tests wereconducted, and would continue to be. The initial ECGshowed infero-lateral ST elevation (lead Il, Ill and aVFwith ventricular fibrillation with reciprocal changes aVLand VI-3 indicates transmural ischemia in the rightFigure 1. Coronary angiogram shows a narrowing(black arrow) in right posterolateral coronary artery.Figure 2. An example of one of my electrocardiographs.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 103

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Tech-2-Tech104coronary artery region (ST elevations have been circledon the ECG; see Figure 2). Consecutive pathologyshowed my results to be within normal ranges for mostparameters, however apart from my ‘troublesome’postolateral coronary artery, there were two otherarteries that were 40% and 30% occluded respectivelyhence my cardiologist commenced statin therapy and afew other drugs. I stayed on these drugs for the first 12months, with the exception of Ramipril (125mg), whichmade me very nauseous and so was discontinued.Most people after an MI usually take 2-3 months tocompletely recover before returning to work. I only took6 weeks. I also did not take up the offer of cardiacrehabilitation as I felt I did not need it. I wasdetermined to recover ASAP and the first positive thingfrom the whole experience (if ever an MI can be‘positive’) was that I stopped smoking.In the end, I had an inferior infarct on the 4th Oct. 2010.This relates to plaque rupture in the context ofvulnerable plaque and the above risk factors forischaemic heart disease (IHD). I was fully treated forsecondary prevention and have been well since thattime. According to my cardiologist report of the 10thMar 2011, my Ml was in fact undetectable ontransthoracic echocardiography and I had received astent to the culprit artery. The rest of my CAD diseasewas minimal and my lipid profile was excellent. Hence,if I adhered to the lifestyle changes suggested, i.e. quitsmoking, exercise regularly, eat sensibly, etc., and if IECHOCARDIOGRAPHY REPORT Name: MARTINIC, G.Date: 1/12/2010 Address:Lab. number: 31513 Sex: MRhythm: Sinus rhythm Age: 45Image quality: good DOB:Tape: DLP38-39/ Referrer: DocIndication: Post PTCAM mode dimensionsAortic root 29 mm (<38) RV diastole 23 mm (<30)AV excursion 22 mm (>10) LV septum 9 mm (<12)Left atrium 37 mm (<40) LV posterior wall 9 mm (<12)MV EF slope mm/sec (>70) LV diastole 56 mm (35-56)MV ESS mm (<7) LV systole 39 mm (20-40)MV excursion mm (>20) LV fract short 30% (28-38)M mode, 2D and DopplerNormal left ventricular size and functionNormal right ventricular size and functionNormal left atriumNormal right atriumNo pericardial effusionAortic valve: Morphologically normalDoppler mapping: No regurgitationMitral valve: Thickened with mobile leafletsDoppler mapping: Trivial regurgitationLV diastolic parameters:E/A velocity radio: 0.7Normal pulmonary valveTricuspid valve: Morphologically normalDoppler mapping: No regurgitationConclusions1. Normal left ventricular size and systolic function2. No significant valvular abnormalitiesCardiac sonographer: Final report Reporter:Figure 3.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 104

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Tech-2-Tech105maintained my lipid profile and blood pressure to withinthe target range prescribed by my cardiologist, then arecurrence of CAD was likely to remain low. I had alreadyreturned to my normal duties and to my previous levelof exercise gradually. I was also prescribed Fenofibrate40mg early on for a borderline/low HDL level (althoughmy LDL profile at 1.7 mmol/L was good), which mycardiologist felt would be good to take as a target forsecondary prevention but in the end I have to confessthat I never really ended up picking up this prescription,mostly because I did not want to be taking ‘too many’different medications.Results on ongoing lipid chemistryprofilesResults are presented in Table 1 covering the periodSeptember 2010 to August 2012, where lipidchemistry was being routinely monitored over a 6-month period and Table 2 covering the period April2013 to August 2015, where lipid chemistry was beingroutinely monitored over this period. As can be seenfrom Table 1, my initial total cholesterol dated Sept2010 was elevated at 5.5 mmoI/L (though notexcessively high). This was reduced quickly to 3.1mmol/L by November 2010 due to the commencementof statin therapy and it was generally maintained overthe period of the next nine months. My total cholesterolreading increased to 4.0 mmoI/L in March 2012though and generally stayed at that level over the next12-13 months to April 2013. From June 2014 my totalcholesterol level dropped to 2.8mmoI/L and at the lasttesting in Aug 2015 the results showed that it hadremained relatively stable at that level, albeitincreasing slightly to 2.9 mmoI/L. (Note: the acronym‘N/T’ in the tables denotes a sample that was ‘nottested)’.Both tables also show that Low Density Lipoprotein(LDL; or the ‘bad cholesterol’) over the period of fiveyears in which this parameter was tested, showed a‘rollercoaster’ effect where LDL was initially recordedwithin normal range with a reading or 1.7 mmol/Lbetween 2010 to 2011 but then increased to 2.6mmol/L in 2012, staying at this same level at thebeginning of 2013 before dropping significantly in June2014. It then increased again slightly in August 2015.By comparison, High Density Lipoprotein (HDL; or the‘good cholesterol’) generally remained quite stablevarying from 0.9 mmol/L (from 2010), peaking slightlyLipid 27 Sept 10 23 Nov 10 15 Feb 11 15 Aug 11 14 Mar 12 20 Aug 12––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Total cholestrol 5.1 3.1 3.1 3.3 4.0 4.1––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––LDL N/T 1.7 1.7 1.7 N/T 2.6––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HDL N/T 0.9 0.9 0.9 N/T 1.0––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Triglycerides 2.2 1.2 1.2 1.29 1.12 1.15––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Cholestrol/HDL N/T 3.4 3.4 3.3 N/T 4.1Ratio––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Table 1. A summary of half-yearly lipid chemistry results September 2010 – August 2012.Lipid 15 Apr 13 17 Jun 14 4 Aug 15––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Total cholestrol 4.1 2.8 2.9––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––LDL 2.6 1.2 1.4––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––HDL 1.0 0.93 0.93––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Triglycerides 1.02 1.36 1.27––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Cholestrol/HDL ratio 4.1 3.0 3.1––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––Table 2. A summary of annual lipid chemistry results April 2013 – August 2015.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 105

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Tech-2-Tech106higher at 1.0 mmol/L between 2012 and early 2013before returning to their almost previous level of 0.93mmoI/L as of August 2015. My triglyceride profileinitially showed a level of 2.2 mmoI/L in September2010 which was slightly above normal range (< 2.00mmoI/L) however this parameter dropped to 1.2mmoI/L, almost half of the previous level when testedin November 2010 (after commencement of statintherapy) and this level was essentially maintained overthe next three years rising to its highest reading of 1.36mmol/L in June 2014 before falling to 1.27 mmoI/L inAugust 2015, almost a year later. My cholesterol toHDL Ratio profile initially showed a level of 3.4 mmoI/Lin November 2010 which remained stable till August2012 when it rose to 4.1 mmol/L, remaining at thatlevel until June 2014 when it fell to 3.0 mmol/L, andessentially staying stable at 3.1mmol/L in August2015. All of these pathology results reflect that factthat medical management of my condition by mycardiologist was essentially achieving its target aims. Itwill be interesting to see how my lipids fare in the nextround of pathology testing which is scheduled forAugust 2016. Despite that fact that the abovepathology results tend to fall within normal ranges forall parameters tested, it needs to be pointed out that Iinitially commenced taking‘Crestor’ statin therapy on 10mg and continued at thisdose rate for the first 12 months. This was increasedto 20mg in the 2011 and continued until 2012, beforemy dose rate was again increased to 40mgapproximately at the beginning of 2013. I haveremained on this dose rate until the present day.The damage that smoking nicotinedoes to your heartEven though I was not a heavy smoker per se, I was aregular ‘social’ smoker and had been for over 25 yearsas I enjoyed it, particularly together with an alcoholicdrink and often in a social gathering of friends or anight ‘out on the town’. Knowing that smoking was badfor me, I had tried many times to give up withoutsustained success. However, after my MI, I quickly (andcompletely) gave up smoking ‘cold turkey’. Of course Iknew that smoking was damaging to one’s health but Idid not know just how damaging it was from a‘mechanism of action’ point of view. My cardiologistexplained to me that after years of smoking a plaquebecomes ‘brittle’ and small fragments of it (similarly aswith a blood clot) can break off from the main diseasedplaque. When this happens the fragments (or clots)travel in the blood circulation and where they lodgebecomes a potentially Ife-threatening problem. Forexample, if the plague fragment lodges in the bloodvessels of the heart, this often leads to an MI and if itlodges in the blood vessels inside the head, this oftenleads to a stroke (see Figure 4).Figure 4. A series of images which show how atherosclerosis essentially develops in three stages. The first imageshows a normal artery. The second image shows an artery with significant damage and a significant degree of bloodflow restriction. Blood products and connective tissue continue to build up and begin to bulge into the arterychannel. In the third image all three layers of the artery wall are affected, and some of the area has begun to calcify,forming a mass of dead tissue, which when either a blood clot, or a fragment of travelling ruptured plaque almostcompletely blocks off the artery channel leading to death of tissue. (Source: Heart Research Institute, Sydney,Australia).ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 106

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Tech-2-Tech107What do the statistics say aboutheart disease in Australia and inthe UK?According to figures provided by the Heart ResearchInstitute, based in Sydney, Australia (and closelyaffiliated with the Heart Foundation of Australia) around55,000 Australians suffer an Ml annually and 9,000Australians die of the same annually. It is estimatedthat 90% of Australians have one modifiable risk factorfor heart disease. Poor diet, low physical activity,smoking, drinking excess alcohol, carrying excessweight and high blood pressure all contribute tocardiovascular disease – the leading cause of death inAustralia. From a prevention point of view, one canimprove one’s heart health by quitting smoking,drinking less and excercising regularly. Smokers areabout 4 times more likely to die from heart diseasethan non-smokers and passive-smoking is linked to anincreased risk of dying from heart disease. Australiangovernment health guidelines recommend that healthymen and women drink no more than two standarddrinks of alcohol on any day, that they eat 375g ofvegetables and 300g of fruit per day, and that they do2.5 hrs of exercise each week. Comparatively, basedon National Health Service information2more than73,000 deaths occur from Ml annually and about 1 in6 men and 1 in 10 women die from Coronary HeartDisease (CHD). There are an estimated 2.3 millionpeople living with CHD and around 2 million peopleaffected by angina pectoris (the most commonsymptom of CHD).Advances in medical research andthe increased skill-sets of doctorsIn comparison to earlier times in our history, we havemuch to be thankful for regards the advanced state ofmedicine today. Although I was fortunate that goodtiming played a factor regarding how quickly emergencymedical help arrived for me, I am certain that had Isuffered this AMI say 50 years ago, I would more thanlikely have died. I was also fortunate in that the smallmedical team of doctors, nurses and para-medics whohad attended to my ‘medical emergency’ were highlycompetent, skilled and experienced in the medicalprocedures that they routinely performed, such as thePTCA that I had performed on me. The fact that this life-saving medical technique was available, is thanks tomedical research.Medical research over many decades and more thanany other factor, has been responsible for the high levelof medical care that we receive today. As someone whohas worked in the field of medical research ever sinceI left school, firstly as a technician, then later as ascientist and laboratory administrator, it is re-assuringand satisfying to know that the work we do in the labsis, as I found, really life-saving and sometimes evenlife-changing, as it was for me. It is interesting how lifeworks sometimes. The irony of it all was that I hadpreviously worked at the Heart Research Institute, oneof the leading medical research institutes in thecountry, which was solely dedicated to researchingheart disease. I even obtained my higher degree (MScHons) in the medical area of atherosclerosis researchwhich was the very subject of all my researchinvestigations in the lab, however little did I know at thetime that this insidious disease was to potentiallythreaten my life ‘down the track’.With regards to life saving cardiovascular medicaltechniques, it was Andreas Gruntzig3in 1977, who firstperformed the PTCA or put simply, dilatation of astenotic artery, which is in current times a commonlyused non-surgical method for the treatment of arteriesobstructed by atherosclerotic plaques. Perhapsironically again, I have carried out research work myselfon the testing of various surgical techniques, one ofthem describing the aortic balloon injury4approach(which was published in this same journal in 2004) forthe purposes of surgical refinement and potentialfuture application in treating dangerous atheroscleroticplaques.The views of other patients abouttheir heart attackIn doing this research, I quickly became interested inwhat other survivors of a life threatening heart attackfelt about their experiences. I began to searching outarticles on their experiences, firstly, to see if theirphysical and psychological experiences were similar tomine, and secondly, to understand how they now feltabout their lives post-event? I found that the physicalsymptoms which most described were similar to mine,for example, feeling suddenly very unwell, hot andsweaty and overwhelmed by incredible and agonisingpain which continued to get worse.56Despite a variationof causes which lead to their heart attacks, forexample, some patients suffered a tear in their arterywall, others suffered an atherosclerotic plaque ruptureand yet others had a spontaneous coronary arterydissection (a very rare medical condition), all howeverdescribed a generally similar symptom picture.But how were they now living day-to-day and what weretheir emotional and psychological views on their illnesspost-event? More specifically, what were their attitudesand beliefs now about their cardiovascular disease?Again, I found similarities in the views of most patientstowards their disease, in that immediately following theheart attack most felt scared about what they had beenthrough and, about the possibility of suffering anotherone which may have had the potential to kill them. Theywere also grieving the loss of their health saying that itwas a huge knock to their confidence.5However, withina few short months post-event, many patientsATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 107

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Tech-2-Tech108expressed surprisingly positive attitudes about theirday-to-day lives. Some examples include the inspiringstory of the 22 year old medical student from Austin,Texas who suffered a life threatening heart attack whilecycling, who survived and then in April 2015 hadparticipated in athletic research which attempted toanswer the question of how far young, athletic heartattack survivors could push their own physicalendurance limits on high-intensity stationary bikeexercises.6Another example was the 51 year old man,who being neither young at the time of his heart attacknow remotely athletic, did manage to lose 70 poundsand train well enough to complete a full marathon onthe 26th anniversary of his heart attack.7Anotherexample was that of a gentleman who at the age of 39years had triple bypass surgery due to severe coronaryartery blockages. At the time he weighed 195 pounds,and confessed to minimal physical activity, a bad dietand long working hours. After his major surgery hedecided to turn his life around. He began to eat betterand rest more and took up cycling four times per week.Within a short period he dropped in weight to 145pounds, as he spent his weekends cycling between 80-145 kilometres. As a result, his cholesterol levelsdropped significantly, his energy levels and general‘zest for life’ have massively improved. He concludedthat he felt better now at 46 years of age, than he didat 39.8There are many such similar examples where Mlsurvivors had positively ‘taken their lives back’.The above examples should hopefully inspire those thatmay have a fear of recurrent attacks that they cancompletely recover and prove that it is possible forheart attack survivors to return to a highly physical andactive lifestyle, if they were willing to take responsibilityfor their own health.5Previous work by behaviouralresearchers has shown that health and illness-relatedcognitions can be segmented into those that; concernthe condition itself, concern the treatments for thecondition and those that concern the patient’sattempts to manage or resolve their illness (and thecircumstances it provokes).9Their conclusions is thatthe behaviours of individuals are affected by theattitudes and beliefs they hold. If individuals thinksomething is appropriate for them they may do it; it not,they don’t.9Personal reflectionsWhile I have never been a regular church-goer, I havealways believed in ‘God’. This whole experience hastaught me that life is very ‘fragile’ and that you will neverreally know ‘when your number is up’ until you face thatvery moment and it is interesting how lots of emotionsand sad feelings go through your head immediately post-event. For example, following an Ml I found myselfoverwhelmed by the sad thoughts about those closestto me, my children, my wife and my other familymembers and how I came so close to never seeing anyof them again and what I should have said to them butdid not get time to. I also experienced thoughts like,“why have I suffered this life threatening disease soyoung?” And, “I must not have much time left, as it isonly a matter of time before my next Ml?”. Some cardiacpatients can actually develop depression after the event(according to nursing and cardiac rehabilitation staff).Other common thoughts that I had early on were, “whyam I here?, why am I sharing this cardiac ward with allthese ‘old people’?”. “At 45 years of age I am much tooyoung to be here, I should not be here yet.” Of course,most people that experience a severe Ml (and thatsurvive it) may express different thoughts, feelings andemotions, these are just mine.Where to from here?At the time of writing this article for ATW, I noted thatmore than five years had passed since my MI and I ammore than happy to report that I am still feeling ‘fightingfit’ as I continue to work at my full time medical researchjob, my secondary appointment as an Officer andinstructor in the Australian Air Force Cadets, to continueteaching young and underprivileged children the sport ofKarate, to write (something I have always enjoyed doing)and most importantly of course, to spend quality timewith my family. Despite the feelings I felt immediatelypost-event (and for quite a few months afterwards), Ihave learned that the fact that one has suffered a severeMI does not necessarily mean that one is ‘doomed’ tosuffer another one automatically. Over time, I havegradually come to the realisation that it is really all abouthow you look after yourself post-event, that can stronglyinfluence the chances of a recurrence. It is maintainingsensible lifestyle choices, keeping fit, keeping stress toa minimum, spending more time doing the things thatare important to you (ie. more time with family, andpursuing your interests ‘which you never made time forpreviously’), and of course keeping a positive attitudeand a ‘healthy outlook on life’. I have no idea how much‘time I have left’, not too many people generally do but Iremain grateful for every day that I get and believe thatif I continue on the sensible path of following mycardiologist’s lifestyle advice and continue to thinkpositively, I should still have a few years left to enjoy onthis earth! However, having strong faith, I also like tobelieve that ‘the Almighty’ was looking over me that verynight as well.AcknowledgementsI would sincerely like to thank the team of Doctors,Nurses and Paramedics from the EmergencyDepartment of Liverpool Hospital who provided me withexcellent acute care. I would particularly like to expressmy deepest gratitude to Dr K. Kadappu and Dr R.Rajaratnam of the Liverpool Hospital CardiacCatheterisation Laboratory, who performed thesuccessful cardiac intervention which saved my life onthe evening of the 4th October 2010.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 108

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Tech-2-Tech109I would also like to sincerely thank my cardiologist, Dr.Con Arronis, MBBS(Syd), FRACP, DDU, FCSANZ, FASE,for his excellent ongoing clinical care and advice thathe has provided over the last six years. I also expressmy appreciation to him for his interpretation of theseclinical findings, as well as his guidance and support inthe writing of this article, which will I hope, educateother sufferers of this insidious disease and give themrenewed hope that they can completely recover andonce again lead a fit and active lifestyle, provided theyalso take some measure of responsibility for their ownhealth. I wish all of the medical staff above continuedsuccess, health, happiness and prosperity, for thetremendous work they do on a day-to-day basis.References1Heart Research Institute website:see http://www.hri.org.au/about-heart-disease/preventionaccessed 09 Feb 20162National Health Service (UK) website:see http://www.nhs.uk/Conditions/Coronary-heart-disease/Pages/lntroduction.aspx accessed 09 Feb 20163Gruntzig, A. (1976). Percutaneous dilatation ofexperimental coronary artery stenosis – description of anew catheter system. Klin Wochenschr (German), June 1:54 (11): pp. 543-54Martinic, G. (2004). Surgical methodology for theprocedure of Aortic Balloon Injury in rabbits (Oryctolaguscuniculus) – a technical guide for untrained researchworkers. Animal Technology & Welfare, April issue, pp. 21-28.5Bee, S. (2013). How to get your life back after a heartattack at age 36: Sally Bee knows how tough it is to battlelong-term illness. Her insights can help you get the besttreatment. Daily Mail Australia, July 2nd edition.6American Heart Association News (2015). Young heartattack survivor pushes limits in new research. AmericanHeart Association, April 3rd edition.7Thompson, J. (2015). Uncategorized Blogs. AmericanHeart Association, April 3rd.8Papanikolaou, G. (2015). Uncategorized Blogs. AmericanHeart Association, April 7th.9Hirani, S.P. and Stanton P. Newman (2005). Patient’sbeliefs about their cardiovascular disease. Hear t Journal,Vol. 91 (9): pp. 1235-1239.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 109

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August 2016 Animal Technology and Welfare111Preparing a paper presentation forCongressINSTITUTE OF ANIMAL TECHNOLOGY5 South Parade, Summertown, Oxford OX2 7JLCorrespondence: congress@iat.org.ukIntroductionHaving completed your research work you may wish topresent it to colleagues within the industry. Presentinga paper at the IAT Congress may seem daunting but itis good experience and may help your career path.When presenting, you have a limited time in which toget your ideas across. Your aim is to ensure your workis understood, giving the audience a clear message totake away and think about. So deciding on the contentof your presentation, how you want to deliver it and itsvisual representation, all need careful consideration.Planning is the secret of success. This applies to mostthings and certainly to preparing a paper. It isimportant to start with clear objectives, know what youwant to achieve and what you are trying to convey. Thefollowing sections discuss the major areas you shouldconsider when preparing your presentation.Your audienceThe IAT Congress is open to everyone from TraineeAnimal Technicians to Senior Facility Directors, TradeRepresentatives and Research Workers. This meansthat your audience will have a broad range ofexperiences and knowledge. This will have an impacton how you structure your presentation and its content.Make your paper too technical and Trainee AnimalTechnicians will fall asleep, make it too basic and themore knowledgeable audience member will switch off.A good introduction and background, together with agood summary will ensure that you cater for everyone.The technical content should then only form perhaps30% of the paper. When preparing an introduction try tomake it imaginative and succinct, as this will help toengage your audience.Always allow time for a summary. This is an importantpart of your presentation because although theaudience’s attention may wander during the main bodyof the seminar – the 30% bit – when they hear thewords “Finally, in summary…” their attention will bedrawn back to you.ContentThe content of your presentation will be disciplinedependent. Some guidelines on the standard structureof a scientific based project are listed below. You do nothave to adhere to these headings but they are listedhere in a logical order:1. IntroductionState why you did the work.2. Materials and methodsState how you did the work and describeexperimental procedures used.3. ResultsOutline what you discovered.4. DiscussionDiscuss whether your results are what youexpected; do they agree or disagree with those ofothers?5. ConclusionsSummarise the key points of your presentation.6. Future workWhat next? Outline your plans for the follow up tothis work or any new projects you are considering.7. AcknowledgementsBriefly acknowledge your supervisor and anyonewho may have helped you with your work.8. QuestionsIt is common for the Chair of the session to invitequestions from the audience at the end of yourpresentation.Visual aidsThere are many factors that contribute to a goodpresentation and visual aids are definitely one.However, although good slides do not necessarily makea good presentation, bad slides can ruin one.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 111

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Tech-2-Tech112Microsoft PowerPoint, 35mm slides and video clips canbe used for presentations at Congress. It is up to youto decide which method is best for your purposes.Please note that Overhead Projectors (OHP) are not anoption at IAT Congress.LengthIt is good practice to keep to the time you are allocatedfor your presentation.Here are some tips to help you achieve this:– Any topic can be presented in any amount of time.– It’s better to narrow down rather than widen thescope of your talk.– As a general rule use a maximum of one slide perminute.– This also depends on the slide content and thelength of the talk – 30 slides in 30 minutes, forexample, is not recommended.– Allow more time for text slides than for images– If images are used you need to allow time to explainthe image if it is not clear.– When presenting graphs, allow time to explain whatthe x-axis and y-axis are, before describing thedetails of the result.– If your talk is too long, redesign it:GGdecide what can be removed and what impactthe remaining content will have GGcondense two slides into one, withoutcompromising their legibilityGGdon’t use too many slides as you will findyourself flashing through them during thepresentation. Practising your presentationGiving yourself the opportunity to run through yourpresentation beforehand is an essential part of yourpreparation. It can highlight omissions or mistakes andalso increase your confidence when you come topresent on the day. Rehearsing in front of colleagues or friends isparticularly useful for your first few presentations, untilyou start to gain more confidence. You can either runthrough your talk as you would on the day or ask themto interrupt with tips as you go along. Thinking through your talk is not sufficient. You need topractise so you can both duplicate the stress felt infront of an audience and judge how long yourpresentation will be. As you stand in front of anaudience, you will find you talk more quickly, so thepresentation might be shorter than you expect. Speakloudly and clearly – this will prevent you from speakingtoo fast if you are nervous. Learn the order of your slides as this will prevent youfrom looking surprised as the next slide is displayed. Tips for the preparation of your slides Regardless of the methods you use to produce andproject your slides, the key thing is to make themas legible as possible to everyone in theauditorium. Use the following guidelines whenproducing your presentation: G Text– Do not use full sentences… but Do not overabbreviate either.– DO NOT USE CAPITALS FOR THE WHOLESLIDE … but DO USE CAPITALS foremphasis.– Use a maximum of six words in the title,seven lines of text per slide and sevenwords across the slide. – Do not use too many colours or acomplicated background. G Tables– Use no more than 4 columns and no morethan 7 rows. – In general, most tables produced forjournals should be redesigned andsimplified for use in a presentation. G Graphs and Charts– Use horizontal labels on graphs, even forthe y-axis.– Minimise the number of tick marks andnumber labels.– All legends, captions and axis labels shouldhave an adequate font size. Use the 1/50thRule for the minimum font size i.e. if thelong axis of the graph is 20cm when printed,the smallest letter should be 4mm (or 20 pt).– If there are multiple lines on a graph, ensurethey are clearly labelled and easilydistinguishable. – Do not use 3D if 2D will do.– Histograms or pie charts are better thantables.– Use colour to highlight parts of a complexdiagram. G Animation and Illustrations in PowerPoint– Animation can be useful for emphasis. – Do not use it too much as this can bedistracting and confusing. G Duplicate material– If you need to refer to the same slide morethan once, in different contexts throughoutyour presentation, it is better to duplicatethe material. This will prevent you fromhaving to search through the slides youhave already shown. ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 112

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Tech-2-Tech113Practise answering questions from your colleagues; thinkin advance about the questions you may get asked. A rehearsal on your own in the room where you aregoing to give your presentation is also useful. It helpsyou to familiarise yourself with the acoustics andtechnical aspects ahead of time. During the rehearsalcheck your slides for timing, legibility and correct orderand orientation. Ask the Congress Committee memberresponsible for presentations if you would like torehearse before your big day. Audio Visual Technicianswill also be on hand to help you with your slides or yourPowerPoint presentation. Presenting your paper There is a lot to think about when presenting,particularly remembering what you are going to say ata time when you will be feeling nervous. Here are a fewthings to consider to ensure that the presentation runsas smoothly as possible. Controlling nerves – Even the most experienced presenters get nervousbefore talking to conference audiences, so you arenot alone. – Being nervous is a natural response and one thatactually works to improve performance. – It is simply a part of delivering a good presentationand can help to keep you on your toes. – Try not to worry too much about nerves you may feelbefore you start. – The benefits of being well prepared and wellpractised cannot be underestimated and will helpreduce nerves on the day. – Use positive self-talk. – Take a deep breath before you start andconcentrate on the key message that you want youraudience to take away. – Some people find that memorising their openingsentence is a useful way to get the talk started. – Think about your body language: if you have atendency to gesticulate, think about what you aregoing to do with your hands. – Try not to panic, speak slowly, keeping your head up.– Dress appropriately so you feel comfortable andconfident when you stand up to speak. Before your presentation Prior to giving your presentation it is useful to: – Familiarise yourself with the control buttons, lightswitches, slide controls, and microphone. – Find and familiarise yourself with the pointer. Avoidusing a laser pointer if your hands shake. – Think about where to stand. – Test the light levels for slides. An Audio Visual Technician will be on hand to help youwith all of this.Delivering your presentation Here are some tips you could follow when deliveringyour presentation: DO:  Take a few deep breaths before walking to thepodium to calm your nerves  Face the audience  Smile at the audience and the personintroducing you when you start yourpresentation  Speak slowly and clearly  Project your voice  Be calm and in control  Use slides as your notes  Talk through graphs and images  Establish eye contact with a few friendly faces  Plant a mentor in the front row for visualsupport  Make use of positive and negative feedbackfrom the audience  Keep the audience awake by: interleaving images or diagrams with texthanding round hard copies or brochures showing small pieces of equipment inviting audience participation Carry on talking if the computer or slideprojector failsDON’T: Panic Mumble Avoid eye contact  Scowl Appear nervous Read from written text Pace up and down Fidget with the microphone Jangle keys or change in your pocket Tell jokes that could fall flat Ignore your audience  Stare into space above your audience  Focus on: Your slides/the floor/your shoes or aparticular person  Give out copies of your slides before your talk:this will distract the audience from what youare saying  Go out drinking heavily the night before yourpresentation ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 113

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Tech-2-Tech114Answering questionsIt is common to allow time at the end of your talk toanswer questions from the audience. During yourpreparation it is always useful to anticipate thequestions that may arise and what your answers mightbe. Tips for using PowerPoint G Control the slideshow yourself:– You can set the programme to scroll throughthe slideshow itself using a timer but it isbest to switch this option off. If you take abit longer to do one slide or are interrupted,you will lose the thread of your talk. G Use a pointer where appropriate– Do not wave the pointer around the lectureroom. – The computer mouse could also be usedand may make it easier to maintain eyecontact with your audience. G Try to prevent a shaky pointer– Rest your arm on the podium.– Support your pointer hand with the otherhand.– Switch the pointer off when not in use. Tips for using 35mm slides G Make sure your slides are in the correct orderand orientation– This will need to be done when you load theprojector, before you start your talk. – A red dot is usually placed at the bottom lefthand corner of the slide mount. When slidesare placed in the carousel the slide isrotated through 180 degrees so that all thered dots can be seen and this confirms thatthe orientation of the projected slide iscorrect. – Make use of the facilities and the AudioVisual Technicians at the venue.G Use a pointer where appropriateTips for answering questions G Listen carefullyG Do not interrupt the questioner in midsentence G Be prepared to rephrase their question:– This gives you time to think of your answerand a chance for people at the back to hearthe question. G Keep your answers shortG Confess your ignorance, if you do not knowthe answer– Do not try to bluff your way through ananswer, remember you may be talking topeople who have more knowledge of thesubject than you.– Refer the question to your supervisor oranother colleague if they are present, butmake sure you agree this with them first. G Deflect hostile questions– These rarely occur, but if the questionerappears to be hostile, do not rise to the baitand have an argument in public. – Suggest politely that you continue thediscussion after the presentation.– The session Chair is there to control thequestioning. ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 114

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115August 2016 Animal Technology and WelfareInformation and instructions for preparinga poster for CongressINSTITUTE OF ANIMAL TECHNOLOGY5 South Parade, Summertown, Oxford OX2 7JLCorrespondence: congress@iat.org.ukIntroductionPreparing and presenting a Poster at IAT Congress canbe just as effective and as rewarding as an oralpresentation but without the nerves associated withtalking in front of hundreds of industry colleagues. Thefollowing guidelines will help you to prepare a goodposter ready for Congress – and it may even win thePoster Prize! Preparation1. Before you start you need to remember that yourposter is presenting highlights of your work. 2. Always read through the information that theCongress Committee provide, including thesubmission form. 3. Here you will find specific requirements of themeeting and the size and orientation of the posterboards. 4. It may be useful to mark out this area when you areplanning, to get an idea of the space available. 5. Think about how you want to present your poster.For example, it could be a series of A4 sheets(often mounted on card or laminated) or a printed,glossy poster. 6. Talk to people at work about the facilities availableto you and the time and costs involved. 7. You then need to work out the content. Read thestudy that you have done or the plan of the studythat you are about to undertake and ask yourself: G are the statements or plan of work accurate? G what data do you need to illustrate your findings? G what are the key points you want to communicate? Guidelines on technical content– Posters from within the UK that describeexperimental procedures must clearly state that thework was performed under the prevailing principlesand authority of the Animals (Scientific Procedures)Act, 1986.– Posters submitted from outside the UK mustexplicitly state what legislation and/or ethicalapproval the work has been carried out under.– Posters describing surgical techniques withrecovery should include details of post-operativecare and any analgesic therapy. – Posters that describe experimental proceduresshould be explicit in defining any benefits to animalwelfare.– Papers and posters must include any adverseeffects to the animals and the steps taken tomitigate and minimise such effects, including whenapplicable analgesic therapy or humane end points. Remember, your poster should be a stand-alone, self explanatory representation ofyour work that is relatively simple and easyto follow. Structure and designG posters are a visual communication of yourresearch so try to keep text to a minimum G use graphics, such as photos, figures and tables to‘tell your story’ G avoid over complicated images G your findings need to be clear and also visible froma short distance away G try to guide your audience through the research bypresenting information in a logical sequenceG use arrows or numbers to direct them A typical content and layout for a poster are shownbelow but you do not have to follow this exactly.Figure 1. Typical poster layoutATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 115

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Tech-2-Tech116TitleG the title should be short and attention grabbing ifpossible G it should be clear from a distance of three metres G use bold, black typeface (about 24 font size) G author names should be slightly smaller G include your facility name and logo if you wish Abstract and introduction G display a brief abstract exactly as it was submittedto the Congress Committee G include a brief introduction to your poster or work ifyou think it adds something MethodsG depending on your work, this could be called‘materials and methods’ or ‘study detail’ forexample G keep this brief and include photos and graphics ifnecessary ResultsG the results of your work should form the major partof your poster G ensure that graphs and charts are self-explanatoryand keep additional text to a minimum Discussion/conclusionG keep this brief G present as a numbered or bulleted list G suppliers of products can be acknowledged in thissection however the poster should contain noobvious company branding logos Remember, at least one author should beavailable during the display session to talkabout the work in more detail. Visual impact1. Visual impact of the poster is important.2. Avoid clutter – a clean, simple design is mostappealing. 3. Think carefully about the use of colour and how youare going to arrange the information before youcommit to the design. 4. Background – try a single colour or two to threerelated colours for different sections (muted shadesare often best).5. Areas of white or empty space can be used todifferentiate elements of the poster.6. Vary the size and spacing of sections to addinterest.7. Outline or alter the background to graphics foremphasis – dark images look good against palecolours and vice versa. SummaryG your poster should present the highlights of yourwork G make information clear and only as complex as itneeds to be G keep text to a minimum G make the design interesting and not too cluttered orover the top – remember that in the end yourresearch is the most important thing G give credit where it is due – contributors, co authorsand sponsorsG create an A4 handout – mini version of the poster,collection of PowerPoint slides or brief summary ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 116

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117August 2016 Animal Technology and WelfareLet’s talk about ‘Culture of care’NIKKI OSBORNEResponsible Research in PracticeCorrespondence: info@responsibleresearchinpractice.co.ukAccessed from: http://www.responsibleresearchinpractice.co.uk/blog.htmlSomething that I’m being asked to talk about a lotlately is ‘culture of care’ (CoC). So what is it – and howcan people set about ensuring that they have a goodone?The first thing to say is that, in my opinion, CoC is ameaningless phrase unless defined. There is also no‘one size fits all’ CoC to aspire to. Instead, eachresearch organisation needs to consider what CoCmeans to them, their students and staff. Havingdefined CoC at a local level it then becomes somethingmeaningful and its true value can be realised.So what might a CoC include? Anyone asked to talkabout CoC, particularly in the context of a researchsetting, will probably mention one or more of thefollowing points:– CoC can be used to express the duty of care,responsibilities, expectations or ethicalconsiderations placed upon researchers whoseresearch involves living subjects. This may behuman or animal subjects or more broadly, researchthat has an impact upon the world or theenvironment within which we live – and that we relyon to continuing living.– CoC can also be used to describe the work ethic orresearch practices within a given researchorganisation. In this context, CoC is about how theresearch framework and management practicessupport its staff and students to do great research(i.e. is ethical, useful and reproducible) but alsofulfil their potential to become the best researchersthat they can.– CoC is sometimes also viewed in a much broader,more holistic context, at which point it becomesmore about the stewardship and preservation ofscientific enterprise. This involves inspiring the nextgeneration, providing role models for researchers,being open and honest about what research entailsand maintaining a reputation of science as beingresponsible, trustworthy and for the greater good.Now that you have some idea of what CoC couldencompass, we can to start thinking about how it canbe translated into practice.For me, there are four key foundation stones uponwhich a good CoC can be built: commitment, educationand training, communication and management.1. CommitmentHaving defined what CoC means in an organisation andwritten it down, the process of taking collectiveresponsibility for achieving it can begin. There needs tobe widespread awareness of policies, procedures andexpectations that, together with a system of educationand training, support its fulfilment.2. Education and trainingHaving education, training or mentoring opportunitiesavailable to support lifelong learning is a must. Thishelps every student or staff member to understandwhat CoC means for them at each stage of their career,what they are expected to contribute, why and how theywill be supported to fulfil this expectation.3. CommunicationEffective communication that reaches and isunderstood by all, is essential to cut through the noisePicture credit: RSPCAACommitmentSupportiveManagementTraining &EducationEffectiveCommunicationATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 117

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Tech-2-Tech118of day-to-day work pressures and challenges.Researchers are often fighting to keep up to date withpublished findings and protocols, so time is of apremium. It is important therefore that organisationssupport (and even reward) students and staff to havean awareness of broader research integrity issues andresponsible conduct practices in order to inform theirown research conduct. This fits well with the ethos thatorganisations have a critical role in developingresponsible researchers and supporting them to be thebest that they can.4. ManagementStrong, supportive and positive leadership is essentialif everyone is to share the sense of collectiveresponsibility for achieving a good CoC. Managers musttake responsibility for ensuring that appropriatestandards of integrity, responsible conduct andbehaviour are implemented. Good monitoring will helpalert them to problems and identify potential issuesearly, so that assistance and support can be offered tothose who need it.–––––––––––Some of you at this point may just be looking for somemore reading around the concept of a ‘culture of care’so for you I can recommend:– The culture of scientific research in the UK (Dec2014). The Nuffield Council of Bioethics conducteda series of engagement activities during 2014 toinvestigate the culture of scientific research in theUK and this is the report of the findings.– The concordat to support research integrity (July2012). This concordat contains five commitmentsthat the signatory organisations believe everyoneengaged in research should be able to agree withregardless of their field of research. The thirdcommitment (pg15-16) is the most relevant to thisblog, talking about embedding a culture of researchintegrity.– Reproducibility and reliability of biomedicalresearch: improving research practice (Oct 2015).This symposium report discusses the relationshipbetween the culture and incentives embeddedwithin the scientific community and researchreproducibility (pg 52-54).– Promoting a culture of care – pg 55-58 in theLASA/RSPCA Guiding Principles on good practicefor Animal Welfare and Ethical ReviewBodies (Sept 2015). This chapter talks from anAWERB perspective about how to go about definingthe culture of care, the structural and behaviouralelements that can be a feature of a culture of careand suggests activities that can help in developingan establishments’ culture.– Identification and management of patterns of low-level concerns at licensed establishments (Dec2015). This Animals in Science Regulations Unitadvice note provides a definition of a good culture ofcare for establishments with licenced authority touse animals in research.– Creating a culture of care (Aug 2014). This NC3Rsblog post discusses culture of care from theperspective of a contract research organisation.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 118

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119AS-ET SPECIAL TRAVEL BURSARYESSAY ENTRIES Do present United Kingdom regulationscovering experiments using laboratoryanimals strike the right balance betweenthe interests of human health andlaboratory animal welfare?JESSICA WOODOxford University Biomedical Services, The Old Observatory, South Parks Road, Oxford OX1 3RQCorrespondence: jessica.wood@bms.ox.ac.ukWinning entry from the AS-ET Fourth Special Travel Bursary 2016IntroductionThe objective of this essay is to discuss whethercurrent UK regulations governing animal research hasan appropriate balance between the interests of humanhealth and animal welfare. To achieve this, the currentlegislation will be reviewed and case studies will beexamined, to explore the issue.LegislationThe Animals (Scientific Procedures) Act, 1986 (ASPA)has been regarded as one of the strictest legislationsin the world.1The legislation has been in effect in theUK since 1986 and amended in 2013, to meet thestandards of the European Directive.2The current ASPAregulations ensure that the welfare of animals isthoroughly considered with the use of an ethicalframework. The ethical framework which includes theharm/benefit analysis, must be implemented intoresearch justification. Severity bands/humane endpointsThe legislation also includes severity bands of nonrecovery, mild, moderate and severe. The severitylimits ensure that animals under a specified severityprotocol do not experience unnecessary sufferingwhilst allowing for research to be conducted. To ensurethese severity bands are not breached, humaneendpoints are established for procedures with thepotential to cause suffering. A number of non-invasiveobservational methods including changes in physiology(temperature and weight,3modified behaviour (reducedmovement,4increased thirst and drinking5andmeasuring blood saturation,6are currently beingutilised as criteria to identify humane endpoints. Humane endpoints establish clear criteria in whichscientific data is achievable at the earliest point of aprocedure, minimising unnecessary suffering causedby advanced pathology and death. Developments suchas these have allowed for death to no longer berecognised as an acceptable endpoint.2Includinghumane endpoints ensures animals experience theleast amount of suffering, whilst preventing data frombeing lost.Harm/benefit assessmentThe harm benefit analysis is required to be conductedby project licence applicants, which is reviewed by theHome Office prior to project licence approval, underASPA legislation. The analysis is a qualitative measureto determine whether a project licence has balanced itsscientific enquiry with animal welfare. The analysisdetermines the potential scientific gains to people,August 2016 Animal Technology and WelfareATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 119

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AS-ET Special Travel Bursary Essay Entries120animals or the environment, including the likely harmscaused to animals. The two main likely harmsexperienced by animals are; project related harms andcontingent harms, accumulated throughout theirlifetime in the laboratory environment.7All sources ofharm are considered to ensure the licence implementsthe 3Rs to aspects of the animals’ experienceincluding; transport, housing, husbandry, restraint,procedure and humane killing. A decision is then madeby individuals with varying perspectives, to ensure thedecision is on a neutral and moral basis as to whetherthe likely harms to the animals are justified by thebenefits. The assessment of harms and benefits arejudged on a case by case basis and must be anevolving process, to accommodate continualadvancements in scientific understanding of animalwelfare, refined techniques and social opinion, whichdevelop over time.8Whilst the method of evaluating ofharm-benefit analysis is continually debated, it is clearthat the current process aims to use current knowledgeto balance the needs of scientific discovery and that ofanimal welfare. If a licence is rejected, the proposedproject licence either cannot prove its value or justifyanimal use. The rejection of licences ensures that onlystudies able to justify animal use for research areapproved.The 3RsThe legislation’s key purpose is to regulate the use ofanimals used in science, by enforcing strictestablishment, project and personal licences. Theselicences ensure that the animals experience theminimal adverse effects possible. The 3R guidance setout by Russel and Burch9plays a pivotal role injustification of the project licence, setting thestandards for the humane use of animals in scientificresearch.The 3Rs comprise of:– Replacement – Developing alternative methods toanimal testing.– Reduction – Minimising the number of animalrequired with compromising the quality of research.– Refinement – refining methods to maximise animalwelfare.ReplacementThe 3R principles consequently pressure thedevelopment of alternative and refined scientificmethodologies. Stem cells,10 cell culturing11and refinedmethods12are just a few examples of techniques whichhave been progressed, benefitting medicaladvancement and reducing the requirement of animalmodels in research. While the 3Rs encourage thedevelopment of alternative methods, many arecurrently unable to fully replace the use of animals;therefore the implementation of the other two Rs ismore relevant to current laboratory animal welfare.Reduction of PrimatesPrimates play a fundamental role within research andas such their use requires strict regulation under theASPA legislation. In particular, great apes includingchimpanzees, gorillas and orang-utans are exempt frombeing used in science, as it was deemed unethical dueto their high cognitive ability and complex social lives.13However, according to the European parliamentdelegation, the vital role marmoset and macaqueshave in furthering research into human medicines anddiscoveries in neurological functions which cannotcurrently be replicated by other means. Consequently,the use of these primates is permitted if there isscientific evidence that the aims cannot be reachedwith other species.14ReductionGenetically altered (GA) animals play a fundamentalrole within research to directly determine the functionof genes within the human body, focusing on replicatingclinical symptoms similar to human diseases. GAmodels have enabled research to refine methods andreduce numbers of animals, to gain scientifically validresults.15A research project that developed a mouse model tostudy a protein disorder attributed as a feature ofAlzheimer’s and type II diabetes was previouslyinduced through chronic administration ofinflammatory agents under a moderate procedureseverity. The resulting models did not exhibit allsymptoms of the disease observed in humans andvariability between animals was high making theresearch difficult to reproduce. A GA model wasconsequently created to refine this method with onlyone injection required to stimulate the production ofthe protein of interest. As a result, the procedure hasa reduced severity rating of mild and study samplesizes decreased from 12 to 8 animals. The study alsogained new insights into the disease as side effects ofthe injections previously used to simulate the disorderwere diminished.15However, the benefits of using GA animals in researchmust outweigh the potential welfare problems includingunknown phenotypes which are still being discovered.17Nonetheless, there has not been a clear distinctionbetween the welfare issues associated with creatingGA models and those caused by other diseasemodels.18RefinementThe humane treatment of laboratory animals is alsovital for ensuring reproducible scientific results.Handling stress has been linked as a source ofvariation between animal studies, the variation iscaused by stress affecting the behaviour andphysiology of experimental animals.19,20TheATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 120

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AS-ET Special Travel Bursar y Essay Entries121consequences of handling stress induced variabilityhave the potential of masking/altering true results andattributed to the large number of animals required for avalid result.21,22 A study compared willingness of miceto approach their handler with three handling methods;pickup by tail, by home cage tunnel or external tunnel.Willingness to approach both tunnel methods wasgreater and consequent stress response of the mice ina test maze was lower compared to mice handled bythe tail.12 Refining husbandry and care techniques canlead to less variation in data and increase the validityof the results. This study highlights the importance ofbalancing scientific aim with animal welfare. ConclusionIt is clear that the overall aim of the ASPA legislation isto balance welfare and science. This is achieved byregulating the use of animals in scientific procedureswhich require licences and special justification with theintegrated consideration and implementation of the3Rs. Key aspects of the legislation include the licenceswhich permit animal work and the harm/benefitanalysis which must justify that the likely scientificbenefits outweigh the likely suffering to animals. Whilethe legislation may seem restrictive and heavilyfocused on animal welfare, it is reflective of our ethicalresponsibility to manage the well-being of experimentalanimals, ensuring they experience the least amount ofpain, suffering, distress and lasting harm. The 3Rs are at the core of the legislation, playing a vitalrole in developing refined methodologies which arerequired to justify that the most suitable methods areused in animal research. As a result, improvedtechniques have advanced research animal welfarethrough reduced numbers, refined husbandry andreplacement of harmful methods and higher species. Ithas also been observed that the 3Rs are integral forimproving the integrity of the data gained from suchstudies. The reduction and replacement of animal usehas also led to the development of novel techniques,resulting in reduced welfare related variation, thusimproving the overall validity of associated animalexperiments. Considering the points discussed above, the legislationdoes appear to reflect the need to balance animalwelfare and human health objectives. Given the need tobalance interests of human health and laboratoryanimal welfare, it is my opinion that theserequirements are interlinked and it is therefore in theinterest of scientific advancement to continue toimprove the welfare of animals in research.References21House of Lords. (2002). Select Committee on Animals In Scientific Procedures Report. Accessed [online] 26th May 2016. Available at: http://www.publications.parliament.uk/pa/ld200102/ldselect/ldanimal/150/15004.htm#a422Directive 2010/63/EU of the European Parliament andof the Council. (2010). On the protection of animals usedfor scientific purposes. Accessed (online) 12th June2016. Available at: http://eur-lex.europa.eu/legal-content/EN/TXT/?uri=celex%3A32010L006323Hankenson, F.C., Ruskoski, N., van Saun, M., Ying, G.-S., Oh, J., and Fraser, N.W. (2013). Weight Loss andReduced Body Temperature Determine Humane Endpointsin a Mouse Model of Ocular Herpesvirus Infection. Journalof the American Association for Laborator y AnimalScience, 52(3), p277-285.24Littin, K. et al. (2008). Towards humane end points:behavioural changes precede clinical signs of disease ina Huntington’s disease model. Proceedings of the RoyalSociety B, 275, p1856-1874.25Wood, N.I. et al. (2008). Increased thirst and drinking inHuntington’s disease and the R6/2 mouse. BrainsResearch Bulletin, 76, p70-79.26Verhoeven, D., Teijaro, J.R., and Farber, D.L. (2009).Pulse-oximetry accurately predicts lung pathology and theimmune response during influenza infection. Virology,390, p151-156.27Home Office. (2015). The harm-benefit analysis process.Accessed (online) 13th June 2016. Available at:https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/487914/Harm_Benefit_Analysis__2_.pdf28The Animal Procedures Committee. (2003). Review ofcost-benefit assessment in the use of animals inresearch. Accessed (online) 13th June 2016. Available at:https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/119027/cost-benefit-assessment.pdf29Russell, W.M.S., Burch, R.L. and Hume, C.W. (1959).The principles of humane experimental technique.10Biddle, A. et al. (2016). Phenotypic Plasticity DeterminesCancer Stem Cell Therapeutic Resistance in OralSquamous Cell Carcinoma. EBioMedicine, 4, p138–145.11Date, K. et al. (2013). Tumour and microparticle tissuefactor expression and cancer thrombosis. ThrombosisResearch, 131(2), p109-15. 12Gouveia, Kelly and Jane L. Hurst. (2013). Reducingmouse anxiety during handling: effect of experience withhandling tunnels. PloS one, 8(6).13Knight, A. (2008). The beginning of the end forchimpanzee experiments?. Philosophy, Ethics, andHumanities in Medicine, 3(1), p.1.14European Parliament. (2009). Limiting animal testingwhile not hindering scientific research. Accessed [online]5th June 2016, Available at: http://www.europarl.europa.eu/sides/getDoc.do?type=IM-PRESS&reference=20091207IPR66096&language=EN15Simons, P. (2011). A refined mouse model to studysystemic amyloidosis in NC3Rs: Research review. 2011.P45-47. Accessed (online) 9th June 2016. Available at:https://www.nc3rs.org.uk/sites/default/files/documents/Corporate_publications/Research_Reviews/Research%20Review%202011.pdf16Understanding Animal Research. (2015). Breeding andGM mice. Accessed (online) 6th June 2016. Available at:http://www.understandinganimalresearch.org.uk/how/areas-research/breeding-and-gm-mice/ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 121

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123Do present United Kingdom regulationscovering experiments using laboratoryanimals strike the right balance betweenthe interests of human health andlaboratory animal welfare?EVAREST ONWUBIKOCancer Research UK – Beatson Institute, Switchback Road, Bearsden, Glasgow G61 1BDCorrespondence: evarest@research-series.comOverviewThe importance of and ground breaking evidence fromthe utilisation of animal models in life science researchand development of new pharmaceuticals cannot beover emphasised. Animal research can only be carriedwhen there are compelling benefits to health andacceptable ethical and welfare standards are met. Overthe last decades the use of animals in pre-clinicalstudies has witnessed considerable changes whichhave further enhanced the standard of animal welfareand the way scientific research is carried out.The UK has one of the highest standards of laboratoryanimal welfare in the world and this has beenexemplified in the way the Home Office controls theauthorisation of establishment, project and personallicences. The role of ethical review committees isunderpinned by making sure that potential benefits ofany experiment outweighs any suffering to a laboratoryanimal.The use of animals in research is now underpinned by“3Rs” and this has enshrined the welfare of animals atthe heart of any experiment. The acceptance of the concordant of openness on theuse of animals in research will improve publicconfidence on the use of laboratory animals.The regulationsThe Animals (Scientific Procedures) Act, 1986 (ASPA) –is an Act of the Parliament of the United Kingdom whichregulates the use of animals in research.1The Cruelty to Animals Act, 1876 regulated animalexperimentation before the onset of ASPA in 1986.1Thecurrent Act (ASPA) has continued to be very robust sinceits inception in 1986. A very vital aspect of laboratoryanimal welfare is contained in the Act and it states that1‘if certain criteria are met’ then animals will be used.1The government select committee in 2002 describedASPA as the “tightest system of regulation in the world”in relation to use of animals for research.1,2The UKcontinues to play a key role in Europe and it issued astatement supporting the European Directive2010/63/EU (“Directive”) on the protection of animalsused for scientific purposes3– and the key of thisdirective is enshrined in the summary as “whereacceptable ethical and welfare standards can be met”then animals can be used. The Animals (ScientificProcedures) Act, 1986 (ASPA) as amended (2012) afterthe transposition of the European Directive4hasbecome more robust in relation to training andsupervision of the animal care staff. The UKgovernment in connection with the EU commission areinvesting heavily on the “3Rs” and alternativeapproaches and these efforts have led to variousmethods going through scientific validation through toregulatory acceptance.5The “3Rs” are now built intoevery project licence.2This is a constant reminder toevery individual working on a project licence to devise away to reduce the number of animals if no alternativereplacement is present and refine their techniques toensure any pain caused to these animals is minimal.The Act, as an all-encompassing document, madeprovisions for “additional controls”2which include; “re-use of protected animals”, “killing animals at conclusionof regulated procedures”, “prohibition of public displays”and “use of neuro-muscular blocking agents”. Theseadditional controls were specific to safeguard the welfareof laboratory animals by avoiding and/or minimising painto these animals where possible.August 2016 Animal Technology and WelfareATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 123

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AS-ET Special Travel Bursary Essay Entries124The regulations have aided a judicious use of animalsin research as much as they have standardised andimproved animal welfare. The welfare of laboratoryanimals as enshrined in the Act and all other workingdocuments are optimum and subject to routine reviewto ensure every aspect of the Act is fulfilled. Atpresent in the UK, all ongoing animal experimentationsare only possible because there has not been adependable and validated non-sentient alternativehence, we continue to utilise laboratory animals in thelife science sector.Laboratory animal welfareThe welfare requirements of laboratory animals areenshrined at the heart of ASPA and are evident in allthe working documents from establishment to projectsand to personal licences.The regulation ASPA clearly sets out the importance ofanimal welfare and the need to incorporate 3Rs inevery procedure where animals are used. The AnimalWelfare Act of 20066and also the correspondingAnimal Health and Welfare (Scotland) Act, 2006 haveaided the need to keep the care and welfare oflaboratory animals optimum.In recent years animal research has benefitedimmensely from improved understanding of thecardinal points of animal welfare and these have beentranslated into the outcome of animal experiments. Agood study design focusing on the welfare of animalshave been known to yield good research results andthis has helped to reveal the impact of optimumanimal welfare on the outcome of life scienceexperiments.Laboratory animals have also benefited hugely fromthe regulation through the legal requirements for thepresence of a Named Veterinary Surgeon (NVS) andNamed Animal Care and Welfare Officer (NACWO) andNamed Training and Competence officer (NTCO).2,4TheNVS and the NACWO provide 24 hour on-call care toensure the health of these animals is optimum andthat the care and husbandry requirement are of thehighest standard.There is also a Home Officer Inspector2attached toevery establishment who is independent from thescientific research to ensure that the welfare of theseanimals is monitored and that the ongoing scientificprocedures are in accordance with relevant projectlicence(s) and/or ASPA requirements.There has been an improved culture of care inestablishments licensed to conduct experiments onanimals and this has been aided through legislationand subsequently has lead to improved training,supervision, awareness and openness to animalresearch. The role of NTCOs in all licensedestablishments have also contributed to an improvedwelfare of laboratory animals and also to the quality ofcare provided to these animals through trained animalcare, breeding and the technical staff.The emergence of “the Grimace scale”7as a welfareassessment tool is a step forward in early detection ofpain in laboratory animals. This is now a governmentbacked initiative and will prove beneficial in ensuringstandard and optimum animal welfare if theapplication is holistic.As required by the Act and all the project licences, theholders of any project of work ensure that all licenseesperform their work in accordance with conditions oneach project licence. This ensures animal welfare andcares are optimum.ConclusionThe present UK regulations continue to strike the rightbalance between interest of human health and that oflaborator y animal welfare as regards to animalexperimentation. The UK government through itspowers responded to changes in moral climate byintroducing policies that have ended some types ofanimal experimentation such as;– Cosmetic testing is illegal in the UK; and there hasbeen no testing of household products carried outusing animals in the UK in 2012.8,9The use ofanimals in products and their ingredients, alcoholand tobacco has ceased in the UK. – Related policies were implemented regarding theuse of Great apes (orang-utans, bonobos, gorillasand chimpanzees) – and research utilising theseanimals are now banned in the UK.9The UK has continued to be vocal on alternatives tothe use of animals and they have gone further to funda number of research and/or projects in ‘alternativesto animal use’ through the National Centre forReplacement, Refinement and Reduction of Animals inResearch (NC3Rs).6,10The number of partners in biomedical research isincreasing year in and year out and this is prompting theproduction of compliance statements as can be seen onproducts from biotechnological companies which nowhave statements supporting good animal welfare andcare standard. The use of animals in life scienceexperiments have continued to deliver expectation in theUK by signing up to openness and constantlyundergoing strict check to ensure compliance.Although, it is difficult to rule out breaches completely,the Home Office is always prompt to deal effectivelywith non-compliance where it surfaces.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 124

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AS-ET Special Travel Bursar y Essay Entries125The stringent regulation of animal experimentation inthe UK and the degree of openness should particularlyenhance the public acceptance of the use of animalsand also improve the ethical frame work in animalresearch. The present UK regulations of the animalexperimentation industry strike the right balancebetween interests of human health and animal welfaresince animal use is only possible where there are noviable alternatives and when the benefits outweigh anypotential suffering to laboratory animals.References21Animals (Scientific Procedures) Act, 1986. Available at:http://www.legislation.gov.uk/ukpga/1986/14/contentsRetrieved 07 June 2016.22Select Committee on Animals In Scientific proceduresReport. Home Office (UK), The Stationery Office. (2002).Available at: http://www.publications.parliament.uk/pa/ld200102/ldselect/ldanimal/150/15004.htm#a4Retrieved 03 June 2016.23Statement supporting European Directive 2010/63/EU(“Directive”) on the protection of animals used for scientific purposes (2010). Available at:http://www.wellcome.ac.uk/stellent/groups/corporatesite/@msh_publishing_group/documents/resources/wtp058790.pdf Retrieved 13 may 2016.24The ethics of research involving animals (2005):Available at: http://nuffieldbioethics.org/wp-content/uploads/The-ethics-of-research-involving-animals-consensus-statement.pdf Retrieved 13 May 2016.25National Centre for the Replacement, Refinement andReduction of Animals in Research. Pioneering BetterScience. Available at: https://www.nc3rs.org.uk/Retrieved 07 June 2016.26Animal Welfare Act (2006). Available at:http://www.legislation.gov.uk/ukpga/2006/45/contentsRetrieved 13/may 2016. Retrieved 27 May 2016.27National Centre for the Replacement, Refinement andReduction of Animals in Research. Mouse Grimace Scale.Available at: https://www.nc3rs.org.uk/mouse-grimace-scale Retrieved 07 June 2016.28Understanding Animal Research. Forty Reasons why we need animals in research. Available at:http://www.understandinganimalresearch.org.uk/about-us/science-action-network/for ty-reasons-why-we-need-animals-in-research/ Retrieved 13 June 2016.29National Centre for the Replacement, Refinement andReduction of Animals in Research. The Science ofcosmetic testing without the use of animals 2014.Available at: https://www.nc3rs.org.uk/news/science-cosmetic-testing-without-use-animals Retrieved 07 June2016.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 125

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126Do present United Kingdom regulationscovering experiments using laboratoryanimals strike the right balance betweenthe interests of human health andlaboratory animal welfare?JAMIE BARRATTFrancis Crick Institute, The Mill Hill Laboratory, The Ridgeway, Mill Hill, London NW7 1AACorrespondence: jamie.barratt@crick.ac.ukCurrently in the UK animal welfare is governed by theAnimals (Scientific Procedures) Act, (1986) (ASPA) andThe Animal Welfare Act (2006). The legislation beingreviewed here is ASPA which specifically coversresearch on animals. It is widely regarded as thestrictest policy of its kind globally which can bringabout problems as well as solutions to the ethicalminefield surrounding research using animals. Thedrive for such a piece of legislation arose from therecognition of sentience in other non-human speciesand their ability to suffer. There are generally three viewpoints with regards toanimal welfare.1Animal rights activists believe thatanimals should be treated with the same respect givento humans and therefore should not be used byhumans. On the opposite end of the spectrum arethose that believe we have a right to use animals aswe wish because they are lesser beings. Finally thereare those which are in the middle of the two extremes.These people see that animals should be treated withrespect but that we need to use them in agricultureand research or we would suffer. This approach isUtilitarianism and is the basis of ASPA.Animal testing compromises animal welfare but isessential for promotion of human health. This is seenin almost all medical achievements such as theeradication of polio, which has saved millions of livesbut also used many laboratory animals. ASPA wasintroduced to strike a good balance between savinglives through research and ethical treatment ofanimals in research. A good balance would allowresearch to go ahead whilst maintaining animalwelfare at the highest levels possible. This isimportant as an imbalance in either direction wouldlead to either unethical testing using animals orrestrict further progresses in human health.In its simplest form ASPA outlines the responsibilities ofan animal research establishment and their governingbodies.2This allows employees to see a clear chain ofcommand. Each of the roles listed is specially trainedand given guidance on their expectations and limitationsin regards to ASPA. Whilst training is a requirement toachieving these roles the legislation that must becomplied to is complex. Efforts have been made by theHome Office to make these more user friendly but theline is not always as clear cut due to the overlaps inroles or lack of effective communication between thesepeople in establishments.The 3Rs were introduced as a measure to replaceanimals in research with non-animal alternatives suchas cell lines, reduce the number of animals used inresearch or refine the level of welfare the animalsreceive from their establishments.3The 3Rs can beimplemented by all staff involved with the animals. Inthe first instance the prospective project licence holdermust show in their application for a project licence thatthey have considered alternatives, that they havelooked for ways to decrease the amount of animalsthey need and also that they are using the besttechniques both in experimentation and husbandry toensure they are providing the best level of welfare tothe animals. Humane end points are a good refinementas they allow all staff directly involved with the animalsto review the animal condition which should minimiseunnecessary suffering. Animal care staff can alsorefine husbandry protocol and techniques. The additionof enrichment specific to the species used can reduceaggression and stereotypical behaviour which are bothindicators of substandard welfare conditions. The 3Rsalso have a good impact on the research as it ensuresthat the best relevant techniques are being applied toacquire a sound test of the hypothesis.4The use ofCRISPR/Cas9 technology in the creation of newAnimal Technology and Welfare August 2016ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 126

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127complex genotypes has allowed researchers to reducethe number of waste animals used under protocols asthe desired cohort is more readily available.5(Editor’snote: CRISPR is a genetic perturbation (disturb orderange) technique that allows for sequence-specificrepression or activation of gene expression).ASPA also requires each establishment to run anAnimal Welfare and Ethical Review Body (AWERB).These bodies are attended by the Named VeterinarySurgeon (NVS), Named Animal Care and WelfareOfficers (NACWO), researchers and lay persons. Thebody reviews prospective project licences and alsoreview work being undertaken at the establishment byanimal care staff and researchers.6The inclusion ofboth research staff and animal care staff along with alay person ensures that there is no bias towards thescience or welfare in prospective or current workcreating a good balance in output from the body. Byextension the AWERB is also expected to promote aculture of care within the establishment. This alongwith the involvement of the Named Training andCompetency Officer (NTCO), Named Information Officer(NIO) ensures that staff are well prepared for their roleand are aware of what to report and who to report to. For a project license to be granted the application isreviewed by the AWERB and passed on to the HomeOffice.6Both of these parties will analyse the harms inwhich the project will inflict on the animals and thebenefits it will bring to human health, this ensures thatthe project is ethically sound and the research is bothrelevant and has not already been performed to thesame standard. Writing a research application takes alot of time as does reviewing its ethical grounds. Ifsufficient evidence is not provided the application willneed further editing before it may be considered whichmay also involve a change in experimental protocol.Such an arrangement whilst good for animal welfarecould impose delays to research being carried out. Having specially trained employees to perform andsupervise experiments or care for animals involves alot of money. Added to this the cost of providing at leastthe minimum housing outlined in the codes of practiceby ASPA and ensuring environment conditions aresuitable for the species being housed makes animalresearch very expensive. This may limit research inhuman health to larger organisations or increase theprice of the finished treatment. Both of these wouldlimit the success of animal research as fewer liveswould be saved and therefore the initial harm benefitanalysis potentially biased.The policing of ASPA is carried out by inspectors. Thereare currently 17 full time equivalent inspectors workingfor the Home Office each of which cover on average10.5 establishments, 158.4 projects and around 1000personal license holders.7This would appear to be a lotof work as they not only inspect the work undertaken ateach establishment but also review new applicationsfor licenses and advise establishments on ASPA. Asidefrom inspections, non-compliance to ASPA regulationscan also be self reported and in 2014 this constitutedthe majority of non-compliance cases. Inspectors visitestablishments according to their relative risk of a non-compliance. The risk value is ascertained by looking atthe severity level of procedures undertaken, size of theestablishment and history of non-compliance.Therefore, if there were to be a breakdown incommunication at a previously well compliant, smallestablishment with mostly mild severity protocol thepoor levels of animal welfare could potentially gounnoticed for a long period of time or rely on theresponsible person’s honesty resulting in animbalance. ASPA provides protection for any living vertebrate otherthan man but including cephalopods. This definitiondoes not therefore provide protection for all animalsused in research.2The fruit fly Drosophiliamelanogaster and the multiple species of mosquito areboth frequently used in research however theirnociception capabilities are either not understood ordebated.8It is commonly accepted that vertebratesbeing higher mammals are afforded this protectioneven if their ability to feel pain is a matter of debate aswith fish.9,10The difference in cases seen above may bedue to speciesism which ranks animals according tothe social status or level of sentience. However,speciesism has afforded even greater protection forvulnerable species in the wild and the great apes forwhich the use of is either highly regulated or banned forthe latter.The use of animals in research is a necessary step toachieving both a higher understanding of human healthand in creating new means of controlling or curinghuman disease. It is also well understood that animalsshould be treated ethically. Striking a balance betweenresearch and welfare can be a daunting task. TheHome Office via the use of ASPA has created a systemwhich although complex, allows researchers andanimal care staff to become involved in the process ofbalancing human health and animal welfare. This hashad a negative impact on the science through theincrease in costs and delays to research. However, dueto the constant internal and external reviewing ofprojects involving animals, the law is able to evolveallowing for improved techniques and husbandry tobetter address the research into human health whilstnot implicating welfare.References21Morrison, A. (2009). An Odyssey with Animals. New York:Oxford University Press22Animals (Scientific Procedures) Act, 1986. London.Animals in Science Committee.AS-ET Special Travel Bursar y Essay EntriesATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 127

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Spot the difference129IntroductionThe Sanger Institute Research Support Facility (RSF)provides 50 to 100 pseudo-pregnant F1 hybrid mice aweek to surgery for use as embryo recipients. Thisrequires maintaining a pool of female F1’s from whichindividuals in oestrus are selected for mating withvasectomised stud males. During a standard refresh ofthe background colonies mating trios (F0) were crossedto provide offspring from C57Bl6/J (B6) and CBA/Ca(CBA) parental colonies. Reciprocal crosses: B6 x CBAand CBA x B6 were set to produce F1 offspring. Thisallowed use of male and females from the two parentalstrains during the expansion phase for supply. F1 miceproduced and their body weights are important supplycriteria, only females reaching 19g by 8 weeks of ageentered into the pool. To improve F1 recipient litteringperformance this weight criteria was increased to 20gresulting in fewer females reaching and maintaining therequired weight. Observations and data collection bytechnicians managing the pool supported the originalpre-refresh breeding strategy that the B6CBAF1 gainedand maintained weight better in comparison to theCBAB6F1. As part of the continual colony review anddata analysis it was decided to return to breeding F1’sexclusively to the B6CBA.POSTER PRESENTATIONSOriginally presented at:IAT Congress 2016More breeding for less surplus: theadvantages of using B6CBA to supply F1 mice*LEILA THOMAS, COLIN TRAVIS, CLAUDIA WATSON, TERRY BROWN,BRENDAN DOE and FRANCESCA FLACKThe Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, Cambridgeshire CB10 1SA*Correspondence: lt6@sanger.ac.ukAugust 2016 Animal Technology and WelfareFindingsImproved weight gain Compared to the CBAB6F1 a significantly higherproportion of B6CBAF1 mice (p<0.001) reach 20g at 6to 8 weeks old for use in the recipient pool. As a resultthere is less wastage of mice not achieving weight. A significantly larger percent of B6CBA F1’s achievedweight at 6 weeks old (p<0.05). Therefore there is areduction in “growing on” stock cages and lessATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:50 Page 129

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Poster Presentations130time/labour spent reweighing at 7 and 8 weeks old.(Figure 1).A reduction was also noted in the numbers of miceculled because they dropped back under 20g while inthe pool.As the average weight of recipients supplied to surgeryincreased, an improvement was observed in thenumbers born from transferred blastocysts after ES cellinjection. This improvement may result in a reduction ofrepeat surgeries being required to obtain viablefounders (Table 1). Contrary to this, when heavierrecipients are used for transfers of embryo afterCRISPR cytoplasmic injection the litter size is reduced.It is believed this is due to the fact that these injectionsare made into E0.5 zygotes, which are transferredhigher up the oviduct than ES cell blastocysts. Inheavier mice this is an area which can have additionalfatty deposits that make the transfer surgerytechnically more difficult. It is expected technicalproficiency will improve with experience.Figure 1. Comparison of weight in CBAB 6F1 andB6CBAF1.Table 1. Average litter size after embryo transfer into F1recipients.Reduced breeding and breeding surplus Reduced numbers of F1 Whilst using both the reciprocal crosses, for every 100female F1 recipients making weight per week, 161females needed to be weaned. By using only theB6CBA this drops to 137 F1 females weaned per week.A reduction of 24 females and therefore 24 lesssurplus males per week. Reduced numbers ofreciprocal crosses consequently the numbers ofbreeding trios producing F1 has been reduced by 24 forevery 100 F1 supplied to the pool.Increase in parental C57Bl6/J (B6) and CBA/Ca(CBA) breedingOriginally the B6 and CBA colonies provided equalnumbers of mice for use in the reciprocal crosses.Mating only B6 females with CBA males means theparental B6 colony has to supply all the females usedin these mates with no mating of its males. And viceversa for the supply of CBA males. However the increase in parental breeding andsurplus sex animals did not outweigh the reductionsin F1 production. Annually there are an additional 780mice produced by the parental colonies which are notused in the trio breeding. Setting this against areduction of 2,496 F1 mice, we have a net saving of1,716. Table 2. There is also more likelihood of finding an alternate usefor surplus B6 and CBA than for surplus F1.Table 2: Net changes in parental colony breeding.SummarySystematic observation made by technicians noticing asmall difference in weight gain has supported the 3R’sby maintaining a: – Reduction in numbers of F1 mice culled not makingpool weight.– Reduction in stock cages and time/labour spentreweighing. – Reduction in pooled mice culled underweight/sentto surgery underweight. – Improved litter sizes from transferred blastocysts. – Reduction in number of F0 breeders and reductionin F1 surplus. – Though there is an increase in surplus from theparental colonies, there is an overall reduction inthe numbers of mice required. Acknowledgements Team 83 RSF Technicians and Team 121.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 130

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133August 2016 Animal Technology and WelfareAbstractThere is a wide variety of enrichment available forlaboratory mice but do different strains of mice prefercertain enrichments? Enrichment should stimulate and encourage naturalbehaviour. If the correct enrichment is used it willminimise the occurrence of stereotypical behaviours,such as; fighting, bar biting, barbering andsomersaulting. This study was aimed at discovering if C57BL/6J andBALB/c mice have a preferred type of enrichment. Thefindings will help identify the right type of enrichmentfor each strain and ultimately aid in keeping laboratorymice happy and health. IntroductionProviding high levels of animal welfare is very importantin the animal science industry. The Animals (ScientificProcedures) Act, 1986 includes Codes of Practice inwhich the environmental conditions and cage sizes forthe animals are stated. It also stated that speciesspecific enrichment should be given but can we go onebetter and go strain specific and provide the bestenrichment for our animals?The purpose of this project was to investigate if certainenrichments were preferred by different strains ofmice. This was done so that laboratory animals can begiven the highest level of welfare. The aim is to providethe mice with the best source of stimulant for them.This will minimise stereotypical behaviours whichmeans happy and healthy animals and which in turn willhelp give more accurate results in scientific studies. Study detail– To identify which type of enrichment certain strainsof mice prefer, we tested two stains of mice withfour types of enrichment. – Types of enrichment chosen for testing werecardboard igloos, cardboard tunnels, red Perspexhouses and red Perspex tunnels on C57BL/6J andBALB/c mice.– Each cage held 25 animals with each enrichmentbeing used in a cage of males and females per strain.– The mice used will be five weeks of age starting thetrial and 7 weeks at the end.– In each cage 65 g of woodchip lined the bottom ofan MB1 (NKP) clear cage. – The cage contained the chosen enrichment and twochew blocks, 7 g of paper wool was also provided inthe stainless steel lid as well as diet.– The enrichment was trialled for a period of threeweeks. Every Monday during the study the cageswere cleaned and replaced with the same amount ofclean woodchip.– The chosen Perspex enrichment was cleaned andreturned to the same cage of animals and newreplacements for the cardboard enrichmentprovided. – Fresh paper wool and chew blocks were also given. – All cages were kept in the same area of the sameroom, so that the conditions they were exposed to,temperature, humidity, sound, lighting and smell,were as similar as possible.Photos were taken of each cage of animals at the sametime every day, including weekends. An ethogram willbe compiled at the same time 3x a day and once at theweekend.The parameters measured will be:G are the mice using the enrichment yes/no?G inside or on top of it?G how many are using the enrichment?G is there evidence of damage (soiled/ chewed)?G has the enrichment moved?G has the paper wool been used?G have they made a nest in the enrichment orelsewhere in cage?G has the chew blocks been used?G does the enrichment need to be changed?Do different strains prefer differentenrichments?RHIANNON ATKINSONEnvigo, Shaw’s Farm, Station Road, Blackthorn, Bicester OX25 1TP*Correspondence: adrian.woodhouse@envigo.comExhibited at IAT Congress 2016ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 133

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134Poster PresentationsResultsThe graphs show the amount of animals per strainsusing the chosen enrichments. (The results are theaverage from the three weeks that they were trialled).– Both strains used the paper wool but the BALB/c’sshowed more of an interest than the C57BL/6JConclusion– The results show that both strains used the Perspexhouse the most with the igloo being used by theC57BL/6J male.– Both tube enrichments had similar results and wereused the least.– When recording my results I noticed that theC57BL/6J’s liked to rest/sleep in a large group andwere not very active. They tried to fit as many ofthem as possible into the house and igloo and therest that didn’t fit would stay close outside. – The BALB/c’s were more active and liked to playand were also the ones that did the most damageto their enrichments.– The cardboard enrichment was more damaged thanthe Perspex enrichment by the end of each week.Even though the cardboard enrichment got damagedthe most, it did not affect how enriching it was tothe mice. It may not be aesthetically pleasing to usbut it shows that the enrichment is doing its job andenriching the animals. – The above table shows that the BALB/c’s movedtheir enrichment the most. This might be due to thefact that they are a more active strain of mouse. – The Perspex house was moved the least, thecardboard igloo was moved the most and the tubeenrichment had similar results.– All cages were given paper wool and two chewblocks. Every cage used the chew blocks and everycage pulled the paper wool into the cage. TheBalb/c’s seemed more interested in the paper wooland were the first ones to pull it into the cage. TheC57BL/6J’s did not seem that interested and pulledit through over the course the week. However, theywere the only ones to use it with their enrichment(Perspex house and cardboard igloo). The results to this trial prove that there is strainpreference when it comes to enrichment but this is notthe case when looking at gender preference.– The C57BL/6J strain prefers an enrichment thatthey can hide in so the Perspex house and igloosare ideal. – The BALB/c strain prefers a cardboard basedenrichment, so that they can chew and make it theirown. – The cardboard enrichments had the most damageseen compared to the Perspex items but all chewblocks from all cages had been used.Cardboard Cardboard Perspex PerspexIgloo Tube Tube HouseBalb/c Male 83% 40% 46% 8%Balb/c Female 75% 31% 29% 13%C57/BL/6J Male 38% 13% 21% 13%C57BL/6J Female 25% 23% 8% 19%ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 134

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136Animal Technology and Welfare August 2016Environmental enrichment is essential for captivehoused Marmosets (C. jacchus). As part of theenrichment programme at King’s College London (KCL)and in keeping with the guidelines of Animals (ScientificProcedures) Act, 1986 (ASPA) and the 3R’s (Russelland Burch), technologists have taken an active role indesigning innovative and beneficial items to encouragethe expression of natural behaviours and maximise theusable cage space. Making use of readily available items found within ourfacility, staff designed and constructed a simple, cost-effective, durable and safe plastic hammock which waspiloted with both naive and Parkinsonian affectedmarmosets over a 6 month period. Following preliminary observations we identified thatthe hammocks had a positive effect on the generalwell-being of the marmosets, showing higher activitylevels and cage usage on a daily basis. This design hasnow been implemented as a standardised enrichmentthroughout our marmoset colony.Materials and methodsThe established enrichment used within our marmosethome cages at KCL includes wooden ladders, nestboxes, turrets and perches, all helping to fulfil theirnatural primate behaviours.All the materials used in the project were readily availablein our facility. Materials used for the new enrichmentincluded plastic boxes, wooden poles, carabiners (dogclips) and cable ties. We were able to re-use plastictransport boxes that are able to accommodate twomarmosets. The plastic hammock is easily produced by drilling holesinto both sides of the wooden poles and into the bottomof the box, then using cable ties to hold the woodenpoles in place. Carabiners can then be used to attachthe hammock to the top of the cage. Four holes drilledinto the base of the hammock also act as a drain duringthe cage cleaning process. (Figure 1)Innovative enrichment for the commonmarmoset (Callithrix jacchus)*MATTEO BATTILOCCHI, GARRY FULCHER, ZOE WINDSOR, AARON SMITH, RIA FISHER and CLAIRE PEARCEDepartment of Biological Services, School of Biomedical Sciences, Kings College London, Guys Campus, London SE1 1UL*Corresponding author: matteo.battilocchi@kcl.ac.ukExhibited at IAT Congress 2016Figure 1. Custom made hammocks built by KCL staff.ResultsThe use of this innovative enrichment as a type ofRefinement has shown several benefits:G increase of social activities; playing, huddling andresting (Figure 2)G positive locomotory behaviours; leaping, climbingand swinging (Figure 3)G improved safety; no sharp edgesG promotion of social interaction betweenconspecificsG ease of cleaning; easy to wash without removingfrom the cageATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 136

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137Poster PresentationsConclusionOver the trial period in the Biological Services Unit, wewere able to demonstrate the functionality and socialimportance of these plastic hammocks. Marmoset well-being has been considerably enhancedby this refinement and combined with the simplicity ofthe design we have now implemented the use of theplastic hammock as a standard piece of enrichment toencourage natural behaviour and social interaction.Additionally, compared to the standard enrichmentpreviously in use, we found that when the marmosetsare provided with additional useable cage space, itallowed for an increase in visibility for technologistsduring routine health checks.Due to high level of safety, the plastic hammocks aresuitable for use with marmoset neurodegenerativemodels e.g. Parkinsonism.AcknowledgementsWe would like to thank Sarah Rose, Louise Lincoln andthe Biological Services staff at King’s College Londonfor participating in this work.We would also like to thank Kenneth Applebee andStephen Woodley for their comments on the draft ofthis poster.References1Jackson, M.J. (2001). Environmental enrichment andhusbandry of the MPTP treated common marmoset.Animal Technology 52(1): 21-28. ISSN: 0264-4754.2Lakshminarasimha, R. and Singh, V.R. (2009).Environmental enrichment for common marmosets(Callithrix jacchus jacchus) at Mysore zoo. Zoos’ Print24(10): 12-14. ISSN: 0971-6378.3The Animals (Scientific Procedures) Act 1986.4http://www.marmosetcare.com/care-in-captivity/what-is-enrichment.html5Russell, W.M.S. and Burch, R.L. (1959). The Principles ofHumane Experimental TechniqueFigure 2. Increased social activity, playing.Figure 3. Locomotory behaviours including leaping,climbing and swinging.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 137

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138Animal Technology and Welfare August 2016AbstractAt the Mill Hill Laboratory (The Francis Crick Institute)there is no standard form of identification for individualXenopus tropicalis, unlike the Xenopus Laevis sp whichhave individual patterns and can therefore bephotographed.If an individual X. tropicalis needs to be identified (e.g.a transgenic individual with a desired gene), it may benecessary to keep animals in individual tanks. Howeversingle housing is not an ideal identification method asthis has welfare and space implications.The objective of this study was to test the p-Chip(Pharmaseq) as a possible method for individualidentification of Xenopus tropicalis in live animals.IntroductionThe p-Chip was selected for the trial based on: the sizeof needle, ease of insertion, size of chip (smallest),simplicity of reading the chip, and durability.A preliminary trial was performed in December 2012with X. tropicalis cadavers to test the most appropriatesite for the insertion of the p-Chip. Two possibleinsertion sites were tested:– Intramuscular in rear legs (dorsal surface). – Subcutaneous (interdigital membrane in rear legs).The subcutaneous insertion into the dorsal lymph sacwas later considered and chips were also inserted intothis site in two cadavers and two live X. tropicalis. The intramuscular insertion was technically easier thanthe subcutaneous insertion. We thought that theintramuscular location could prevent migration of the p-Chip from the insertion site.When the initial readability was later tested in thesetwo insertion sites on dead X. tropicalis, none of the p-Chips inserted intramuscularly were readable,whereas 8 out of 10 of those inserted subcutaneouslywere readable after insertion. The intramuscular routeof insertion was discarded as a method to be used inlive animals. p-Chips were inserted subcutaneously in live X. tropicalis and their readability checked regularly fortwelve months. Animals were regularly checked for anyadverse effects related with the p-Chip and/or theinsertion process.Materials and methodsp-Chips and reader needed for the trial were providedby Pharmaseq. (www.pharmaseq.com). BRF providedthe cadavers, the live animals and the staff to performthe trial. The animals used were from the Mill HillLaboratory Wild type colony of X. tropicalis. p-Chip microchip characteristics G p-Chip is the smallest available chip on the market(0.5mm x 0.5mm x 0.1mm)G needle size is 0.6 mm diameterG weight 0.085mgG supplied in pre-loaded disposable applicatorG 9 digit readingG chips are compatible with all common imagingtechniques (CT scan, MRI, cone beam etc.) and canwithstand irradiation doses of 40 Gyp-Chip subcutaneous insertionmethod 21. Readability of the p-Chip was checked beforeinserting into the animal.Evaluation of p-Chip (Pharmaseq) Microchipas an identification method for Xenopustropicalis biological research facility (BRF)The CrickLUCY FERNThe Francis Crick Institute, Mill Hill Laboratory, The Ridgeway, Mill Hill, London NW7 1AACorrespondence: lucy.fern@crick.ac.ukExhibited at IAT Congress 2016ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 138

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139Poster Presentations22. The animals were anaesthetised by immersion in asolution containing 1g/l of MS-222 and 2g/l ofsodium bicarbonate. 23. After immersion in the anaesthetic solution, theresponse of the animals to stimuli was testedregularly (every 20 seconds) and the animals wereremoved from the anaesthetic solution onceunresponsive (loss of ‘righting reflex’) to preventoverdose of anaesthetic. 24. The frogs were then rinsed by immersion in freshwater from the system and wrapped in wet tissuepreviously soaked in fresh system water tomaintain moisture of the skin and prevent the skinfrom drying out during the insertion procedure. 25. The anaesthetised frog was held with one handkeeping one of the rear legs in a stretchedposition. 26. The p-chip applicator was inserted subcutaneously(from proximal to distal) on the dorsal surface ofthe metatarsal area and moved towards the 5th or4th toe. 27. The plunger was then pushed to deliver the chipsubcutaneously in the 4th or 5th toe and theapplicator slowly removed.28. The insertion site was observed to detect anybleeding or damage.Figure 1. p-Chip before use.Figure 2. Insertion of p-Chip subcutaneously into thedorsal surface of the metatarsal area.29. The readability of the p-Chip was checkedimmediately after insertion and if we could notread the chip, another p-Chip was inserted in theanimal’s other leg.10. After insertion (and confirmation of readability) theanimals were allowed to recover inside an emptytank while wrapped in wet tissue.11. Animals were returned to their original tanks oncethey had fully recovered from anaesthesia, whichtypically lasted 20-30 minutes.12. Animals were checked daily for any adverse effectsfor two weeks.p-Chip readingsReadings of the p-Chips were performed withoutanaesthesia, just by restraining the animal. Most of thetime this procedure was performed by two people butit can be performed by just one person who is highlyskilled at restraining the frogs.Figure 3. Position for reading p-Chip. ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 139

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140Poster PresentationsInitially readings of the p-Chips were performed everytwo weeks after insertion for a two month period. Thenon a monthly basis for up to twelve months.Results During the study two frogs were found dead butfollowing NVS examination this was not attributed tothe p-Chip or the insertion procedure.The p-Chip appears to be a reliable method over timeas only 5 out of 33 chips were not readable at 4months after insertion. This could be due to migrationof the chip more than failure of the chip itself.In spite of being a fairly reliable method for individualidentification of X. tropicalis, the drawback of the p-Chipis that it is not a visual method. This means that eachindividual animal in a tank must be restrained and itschip read when looking for a specific animal.Considering that tanks can be big enough to hold up to50 frogs, this can become a time consuming task andmeans additional handling of the frogs.ConclusionThe p-Chip seems to be a reliable method for individualidentification of X. tropicalis in some laboratorysettings/experimental conditions but does notdecrease handling and any associated distress.AcknowledgementsYolanda Saavedra-Torres, Clare Brazill-Adams, KimDemetriou and Jean-Philippe Mocho.DiscussionWe believe that the insertion of the p-Chipsubcutaneously at the toes on the rear legs is a reliablemethod for individual identification of X. tropicalis. It isimportant though that the insertion is performedalways in the same site to know where to locate thechips in subsequent readings. The animals toleratedthe p-Chips well, without any overt signs of pain,suffering, distress or lasting harm. The insertionprocedure is quick when staff are fully trained andcompetent. We attribute the non-readability of some ofthe p-Chips to different causes:1. Insertion too deep due to inexperience of theperson performing the insertion at the beginning ofthe study.2. Damage of the p-Chip during the insertionprocedure. The subcutaneous insertion into the dorsal lymph sacwas later considered and chips were also inserted intothis site in two cadavers and two live X. tropicalis. Thechips in the live X. tropicalis were not readable about amonth after insertion, most probably due to migration.The subcutaneous site at the toe level offers a verynarrow space between the skin and the underlyingtissues for the chip to migrate but there remains somedegree of migration.Number of cadavers used10(Bothintramuscular andsubcutaneoussites were tested)Number of livefrogs31 (subcutaneousroute only)33/40 chips 3 live frogsshowedsubcutaneoushaemorrhageafter insertionprocedure,without any signsof pain ordiscomfort. Theyrecovered in 4days. No otheradverse effectswere observed.28 out of the 33chips that werereadable afterinsertionremainedreadable 12months afterinsertion.The other 5 werenot readable 4months afterinsertion.4 out of the 28 p-Chips that arestill readable 12months afterinsertion havemoved from theinsertion site onone toe to adifferent toe.Readability afterinsertionAdverse effectsdue to insertionprocessReadability overtimeMigration of p-ChipNumber of animals used p-ChipsATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 140

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141August 2016 Animal Technology and WelfareAbstractLimitations to the well-established technique of earnotching restrict its usability for some of our in vivostudies.We aimed to refine our identification methods to allowgroup housing of patient xenograft experiments whichwere previously singly housed to guarantee tracking ofclinical samples. Additionally we wished to implementa new identification system for experiments requiringrolling recruitment.Ear tagging is a robust system that allows greatflexibility and reduces the stress of re-identificationpost-randomisation. We also anticipated fewer errorsduring routine procedures.We initiated a study in which we compared ear notchingand tagging in two different strains of mice (NSG andC57Bl/6J). We observed a significant strain differencewith the C57Bl/6J mice losing nearly all of their tagsand obtaining significant damage scores over 2-3weeks. In contrast, the NSG mice did not present anyproblems and we have since successfully rolled thetags out into larger experiments.Introduction Our Institute currently uses ear notches which run from1-7 (Figure 1a), these can be introduced by using asimple tool that punches a semi-circular ‘notch’ intoeither ear (Figure 1a and b). However, notched earshave a known propensity to develop tears, leading touncertainty in identification (Figure 1c). The ear taggingsystem we chose uses small, flat, unobtrusive tagsthat come in 5 different colours, ranging from 1-999(Figure 2a and b). Refinement of rodent identification byimplementation of an ear tagging systemJACQUI SWAN,1MATT SMITH,2NOELIA LOPEZ-SALENSANSKY,3RICHARD MARAIS2and LISA DOAR11Biological Resources Unit, Cancer Research UK Manchester Institute, The University ofManchester, Wilmslow Road, Manchester M20 4BX2Molecular Oncology, Cancer Research UK Manchester Institute, The University of Manchester,Wilmslow Road, Manchester M20 4BX3The Royal Veterinary College, Royal College Street, London NW1 0TU Correspondence: jacqueline.swan@cruk.manchester.ac.ukExhibited at IAT Congress 2016Figure 1a.Code Mouse’s left Mouse’s rightNumber Ear punch Ear punch––––––––––––––––––––––––––––––––––––––––––––––––––1 LEP –––––––––––––––––––––––––––––––––––––––––––––––––––2– REP––––––––––––––––––––––––––––––––––––––––––––––––––3 2 x LEP –––––––––––––––––––––––––––––––––––––––––––––––––––4– 2 x REP––––––––––––––––––––––––––––––––––––––––––––––––––5 LEP REP––––––––––––––––––––––––––––––––––––––––––––––––––6 2 x LEP REP––––––––––––––––––––––––––––––––––––––––––––––––––7 LEP 2 x REP––––––––––––––––––––––––––––––––––––––––––––––––––ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 141

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Poster Presentations143Behaviour Definition––––––––––––––––––––––––––––––––––––––––––––––––––Cage If any of the mouse’s cage mates mates show interest in the ear tag/notch either sniffing or nibbling it––––––––––––––––––––––––––––––––––––––––––––––––––Head Where the head moves from one sideshake to the other in rapid movement––––––––––––––––––––––––––––––––––––––––––––––––––Scratching Using any limb to scratch at theear or tags––––––––––––––––––––––––––––––––––––––––––––––––––Other Any other behaviour that could be associated with the impact of the procedure or having an ear tag e.g. vocalisation or grooming––––––––––––––––––––––––––––––––––––––––––––––––––Figure 3a. An ethogram to show the behaviours anddefinitions used on the tally charts.Score Definition––––––––––––––––––––––––––––––––––––––––––––––––––0 No damage––––––––––––––––––––––––––––––––––––––––––––––––––1 Small hole/very slight tear but difficultto see––––––––––––––––––––––––––––––––––––––––––––––––––2 Slight tear––––––––––––––––––––––––––––––––––––––––––––––––––3 Obvious tear damage––––––––––––––––––––––––––––––––––––––––––––––––––4 Significant tear damage––––––––––––––––––––––––––––––––––––––––––––––––––5 Severe tear damage––––––––––––––––––––––––––––––––––––––––––––––––––Figure 3d. Scoring of the impact of ear tag.Score Definition––––––––––––––––––––––––––––––––––––––––––––––––––0 Normal, no sign of redness or scabbing––––––––––––––––––––––––––––––––––––––––––––––––––1 Small hole/very slight tear but difficultto see––––––––––––––––––––––––––––––––––––––––––––––––––2 Very slight redness around the tag––––––––––––––––––––––––––––––––––––––––––––––––––3 Slight redness and/or scabbing around tag––––––––––––––––––––––––––––––––––––––––––––––––––4 Very red, large scabbed area, some swelling apparent and/or signs of bleeding––––––––––––––––––––––––––––––––––––––––––––––––––5 Very red and scabby, area swollen and/or bleeding––––––––––––––––––––––––––––––––––––––––––––––––––Figure 3e. Scoring of damage to ear. Table showing the scoring system to assess thedamage caused to ears after tagging/notching.Figure 3b. Daily observations – cage mates. Graph showing tally of cage mate interactions made inthe first 5 post notching/tagging.Figure 3c. Daily observations – head shaking. Graph shows a tally of head shakes in the first 5 dayspost notching/tagging.MethodsWe set up an experiment using 20 femaleC57BL/6JOlaHsd (C57BL6) and 20 female NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice, all 6-8 weeks of age.Each cohort was divided into two groups. The firstgroup (10 C57BL6 and 10 NSG) were tagged using theear tag system and the second group (10 C57BL6 and10 NSG) were notched as per our standard protocol.Each animal was observed immediately followingnotching/tagging for a total of 3 minutes, based on apre-determined ethogram (Figure 3a). Observationswere made every 15 seconds on a tally chart. The 3minute behaviour observations were continued out today 5 (Figure 3b and c). Observations of damage/impact scoring were also carried out up to day 5, withadditional observations recorded on day 7 and 14(Figure 3d–g). Two technicians were used to recordobservations, cages were chosen at random each timeto avoid bias.Number of incidentsATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 143

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Poster Presentations144Figure 3g. Graph showing the damage caused to earsby tagging/notching of mice.Figure 3h. Graph showing the amount of tags missingin C57BL6 mice during study.Figure 3f. Graph showing the impact caused bytagging/notching of mice.Statistical analysis was carried out using a 2-wayANOVA with matched observations, followed by Šidák’smultiple comparison test. Results– There is a significantly higher impact score in thenotched NSG group than either the tagged NSGgroup or the notched C57Bl6 group, when lookingover the length of the study (p= <0.001).– There is a higher impact score in the tagged C57Bl6group than the notched group, but only from day 7onwards and this is not significant over the full time-course.– There is a significantly higher damage score seen inthe tagged C57Bl6 mice than any other groups,when looking over the length of the study(p=<0.001).Conclusions– More head shaking and investigation by cage mateswas seen in all tagged mice but this was only for thefirst observation after which there was nodifference.– C57Bl6 mice did not tolerate the ear tags at all well.They chewed the tags out, causing damage to theears (see Figure 3g and h). Although impact scoreswere generally low, this jumped on day 7 whichcoincided with the day most of the tags were pulledout.– The NSG mice on the other hand all kept their tagsin and had low damage and impact scores. In fact,impact scores were significantly lower in the taggedmice – the tags seemed to cause less reddeningand soreness than traditional ear notching. NSGmice are therefore a good candidate for the use ofear tags.ATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 144

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145Subjects considered for publication may include originalarticles, technical notes and reviews pertaining to allaspects of animal science and technology, managementand education. The Editorial Board wishes to offerparticular encouragement to papers leading toimprovements in environmental enrichment, the generalcare and welfare of the animals used, in particular thosespecies and strains exhibiting harmful genetic defects,and papers describing refinements in techniques, areduction in the number of animals that need to be usedor alternatives to animal use.Papers describing experimental procedures will only beaccepted for publication if authors clearly state that theprocedures conform to the prevailing principles andCodes of Practice of the Animals (Scientific Procedures)Act, 1986. Papers submitted from outside the U.K.,should state what legislation and/or ethical approvalthe work has been carried out under. In addition, authorswho describe surgical techniques with recovery shouldinclude details of post-operative care and any analgesictherapy provided. All submissions should follow theARRIVE (Animal Research: Reporting of In VivoExperiments) guidelines (Kilkenny C, Browne WJ, CuthillIC, Emerson M, Altman DG (2010) Improving BioscienceResearch Reporting: The ARRIVE Guidelines forReporting Animal Research. PLOS Biol 8(6): e1000412.doi:10.1371/journal.pbio.1000412) The Editorial Board 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 Institute ofAnimal Technology.Articles may be submitted either electronically or by hardcopy as follows:ElectronicArticles should be submitted in Word format with doublespacing to the lines and all pages serially numbered.Any photographs or graphs must be submitted asseparate files and conform to the format in point 4)below.The relevant article must clearly indicate wherephotographs and/or graphs are to be inserted.Address for submission: atw@iat.org.ukHard copyThe original manuscript plus two copies should be sentto the address below together with a copy on disk (CD orDVD).All sheets should be typewritten on one side in doublespacing and serially numbered.Any photographs or graphs should be supplied asoriginals and conform to the format in 4) below.Address for submission: Journal Editorial BoardChairman, 5 South Parade, Summertown, Oxford OX27JL.No responsibility will be accepted for loss or damage tosuch articles. Electronic files of submissions are required together withseparate files of photographs and any graphics thatappear in the manuscript. Electronic submissions shouldbe sent via email via atw.iat.org.ukalternatively,manuscript plus two copies may be sent as hard copy tothe address below. All sheets should be typewritten onone side in double spacing with 4 cm margins andserially numbered. Additionally, a copy on disk should beprovided or sent by email via atw@iat.org.ukArticles for submission should be sent to: JournalEditorial Board Chairman, 5 South Parade, Summertown,Oxford, OX2 7JL.No responsibility will be accepted for loss or damage tosuch articles.Format1). The first sheet of the article should contain thefollowing:i. the full title of the paperii. the initials and last name of the author(s)iii. the full address of the department(s) andinstitution(s) where the work was carried out.iv. the address for correspondence if different to above.2). For the remainder of the paper, the text should beclear and concise and, where appropriate, sub-dividedunder the following headings:i. Summaryii. Introductioniii. Methodsiv. Resultsv. Discussionvi. Acknowledgementsvii. References3). Measurements should be given in metric units – seeThe use of S.I. Units (1969) British Standards Institutionpublication and spelling should follow that of the OxfordInstructions to AuthorsAugust 2016 Animal Technology and WelfareATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 145

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146English Dictionary. Abbreviations must be defined in fullat their first appearance in the text. The 24 hour clockshould be used for times. Words to appear in italic typeshould be underlined. Designation of inbred strainsshould be in accordance with the International Index ofLaboratory Animals, 6th edition, compiled, edited andpublished by M.W. Festing, 1993.4). Photographs should have clear and well contrastedtone values and be in colour. All illustrations, charts (e.g.histograms and graphs) and photographs should besubmitted separately and bear on the reverse side theauthor’s name, a number corresponding to the order inwhich it appears in the text e.g., Figure 1, and an arrowpointing to the top.Illustrations, charts and photographs supplied on diskshould be in JPEG, TIFF or EPS formats and have aresolution of no less than 300dpi.The captions for illustrations, charts and photographsshould be typed in double spacing in numerical order ona separate sheet of paper.5). References: Only essential references should beincluded. Authors are responsible for verifying themagainst the original source material. ATW uses theVancouver referencing system: references should beidentified in the text by superscript Arabic numbers e.g. 12after any punctuation and numbered and listed atthe end of the paper in the order of when they are firstcited in the text. Automatic numbering should be avoided.References should include the names and initials of upto six authors. If there are more than six authors, only thefirst three should be named, followed by et al.Publications for which no author is apparent may beattributed to the organisation from which they originate.Simply omit the name of the author for anonymousjournal articles – avoid using ‘Anonymous’. References should be set out as follows:Journals:- Surname and initials of author(s) (date), title ofarticle. Name of journal in full, volume number, first andlast page numbers.e.g. Saigeman, S. (1998). Environmental enhancementof cats – what? why? how? Animal Technology, Vol 49,No.3, 145-154.Books:- Surname and initials of author(s) (date), title ofbook. Name of publisher, Town of publisher.e.g. Flecknell, P.A. (1987). Laboratory AnimalAnaesthesia. Academic Press, London.Chapter from a multi-author book:- Surname and initialsof chapter author(s) (date), title of chapter. In: title ofbook (surname and initials of book editors). Name ofpublisher, Town of publisher, first and last page numbersof chapter.e.g. Gregory, J.A. (1985). Principles of AnimalHusbandry. In: Laboratory Animals – An Introduction forExperimenters. Second Edition. (Tuffrey, A.A.). John Wiley& Sons Ltd., Chichester, 87-105.Papers accepted for publication but not yet publishedshould be included in the list of references followed by‘(in press)’. Papers in preparation, personalcommunications and unpublished observations shouldbe referred to as such in the text only. ContentPapers describing procedures involving the use ofanimals should always include full details of the animalsand husbandry conditions used. These would be asfollows:AnimalsSpeciesBreed or strainSexAge and weight at start of procedureGenetic status: inbred; outbred; hybrid; mutantSourceMicrobiological status: conventional; specifiedpathogen free (define which pathogens animals arefree from); gnotobiotic (define which micro-organismsare present)Quarantine or acclimatisation periodHusbandry during procedureType of housing: material; size; cage type if relevantNumber of animals per cage or unitBedding: type; quality; any pretreatmentType of system: conventional; barrier; ventilated rack;isolatorEnvironmental temperature (°C ± range)Relative Humidity (% ± range)Lighting: natural; artificial (state hours of light anddark)Ventilation: number of air changes per hourPeriod of acclimatisation before start of procedureFeed: type; composition; any pretreatment; amount;frequencyWater: type; quality; any pretreatment; amount;frequencyScientific procedureNumber of animals and any pretreatmentTime of day of procedure(s)Quantity and frequency of any samplesStatisticsTests used should be namedReprintsFree reprints are no longer provided but the ATW EditorialBoard are happy to provide PDF files of articles afterpublication. Use of these files is subject to Copyrightrestrictions. Instructions to AuthorsATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 146

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INDEX TO ADVERTISERSINDEX TO ADVERTISERS August 2016Allentown Inc ..................................................................................................................OBCAnimals in Science – Education Trust ...................................................................................ivBell Isolation Systems .......................................................................................................viiiContec ................................................................................................................................vCongress 2017 .............................................................................................................xiv, xvEnvigo ..............................................................................................................................IFCInstitute of Animal Technology ..............................................................................vi, vii, xi, xiiiIPS Product Supplies Ltd ..................................................................................................IBCLBS .....................................................................................................................................xPFI Systems ........................................................................................................................iiiTecniplast UK ......................................................................................................................ixVet-Tech Solutions ...............................................................................................................xiATW Aug 28Oct.e$S:Animal Technology and Welfare 7/11/16 12:51 Page 152