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IAT Journal Animal Technology and Welfare G Themed Issue Farm species G Congress 2014 posters part two G RSPCA UFAW Rodent Welfare meeting report G AS ET Travel Bursary 2014 essays Official Journal of the Institute of Animal Technology and European Federation of Animal Technologists ISSN 1742 0385 Vol 13 No 3 December 2014

CONTENTS Vol 13 No 3 December 2014 Editorial Jas Barley Chair of the Editorial Board ix Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research cattle pigs sheep and poultry Penny Hawkins Sharon Brookes Susannah Parkin R Eddie Clutton Peter Gade Julie Lane Helen Proctor Joanne Edgar Isobel Vincent and Ute Weyer 155 Report of the 2013 RSPCA UFAW rodent welfare group meeting Penny Hawkins Secretary Paul Littlefair Huw Golledge Claire Richardson Sarah Allden Tania Boden Colin Hendrie Dominic Wells Nikki Osborne Marie Hutchison Stephen Ryder Maggy Jennings and Robert Hubrecht 165 PAPER SUMMARY TRANSLATIONS 181 TECH 2 TECH Welfare First How good is your induction programme Mathew Sanderson 189 AS ET SPECIAL TRAVEL BURSARY 2014 ESSAYS Advantages and disadvantages of animal technicians performing regulated procedures in research Jan Bilton 193 Advantages and disadvantages of animal technicians performing regulated procedures in research Karyn Heath POSTER PRESENTATIONS Advances in sheep anaesthesia within our facility Katie Blackwell 196 199 How best to train a sheep in research Kathryn Chambers 202 Pain management in sheep how we have developed our techniques to improve animal welfare Emma Tozer 204 The use of computed tomography CT and digital X ray in sheep Lisa Griffin 207 Do sheep need environmental enrichment Katie Amess 210 Alternative restraint of Gottingen minipigs for blood sampling Adrian Zeltner 213 Determination of disinfectant efficacy against the eggs of Syphacia species Alison Livsey 216 Increasing useable surface area in standard mouse cages Andrew Newman and Sabrina Brando 219 Application of molecular technologies on non invasive samples to enhance health monitoring programs of rodents in IVCs a comparison with a traditional approach Angela Kerton Savvas Piperelis Mandy Thorpe C Clarke Francisco Diaz K Henderson C Clifford Colin Dunn S Taft M Foa and R Norrington 223 You can t keep anything clean without getting something else dirty Care of the nude mouse as a sentinel strain in a non sterile IVC environment Mandy Thorpe Angela Kerton and Savvas Piperelis 226 Refined techniques for acclimatising female macaques to restraint Lee Reed Terri Jackson Lauren Dean and Stuart Baker 229 Instructions to Authors 230 Index to Advertisers xvii ATW PROFILE Animal 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 to communicate 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 ATW enjoys 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 uk i

IAT REPRESENTATIVES OFFICERS President Dr Robin Lovell Badge FRS Immediate Past President Professor Sir Richard Gardner MA PhD FSB HonFIAT FRS Vice Presidents David Anderson MRCVS Stephen Barnett BA MSc CBiol FSB RAnTech John Bleby TD JP DVetMed DLAS CBiol FSB MRCVS Brian Cass CBE Miles Carroll PhD Gerald Clough BSc PhD EurBiol CBiol MSB SFZSL Paul Flecknell MA Vet MB PhD DLAS DipLECVA MRCVS Barbara Mortimer BVetMed DLAS MRCVS Judy MacArthur Clark CBE BVMS DLAS CBiol FSB MRCVS Fiona McEwen BSc BVM S MSc MRCVS Tim Morris BVetMed PhD DipACLAM DipECLAM CBiol FSB CertLAS MRCVS Jos Orellana BVSc MSc Clive Page PhD BSc Sophie Petit Zeman PhD Gail Thompson RLATG Robert Weichbrod PhD RLATG Sheila Whitehead BVMS MSc CertLAS MRCVS Lord Robert Winston FMedSci DSc FRCOG FRCP FRCS Ed FSB Life Members Roger Francis MSC FIAT RAnTech Pete Gerson MSc FIAT RAnTech John Gregory BSc Hons FIAT CBiol FSB RAnTech Patrick Hayes FIAT DipBA RAnTech Robert Kemp FIAT Hon RAnTech Keith Millican FIAT CBiol MSB Phil Ruddock MIAT RAnTech Ted Wills HonFIAT RAnTech Dorothy Woodnott FIAT Honorary Members Andy Jackson MIAT John Lesley FIAT RAnTech Brian Lowe MSc FIAT RAnTech Ronald Raymond FIAT RAnTech Peter Russell FIAT RAnTech David Spillane FIAT Ray Thatcher FIAT RAnTech Pete Willan DMS FInstLM MIAT RAnTech Members of Council Ken Applebee OBE Jas Barley Kate Burton Charlie Chambers Steven Cubitt Andy Cunningham Glyn Fisher Cathy Godfrey Alan Graham John Gregory Linda Horan Elaine Kirkum Adele Kitching Sarah Lane Norman Mortell Steve Owen Wendy Steel Allan Thornhill Lynda Westall Debbi Young Council Officers Chair Steve Owen FIAT RAnTech Vice Chair Wendy Steel BSc Hons FIAT RAnTech Honorary Secretary Linda Horan Honorary Treasurer Glyn Fisher FIAT RAnTech Assistant Treasurer Charlie Chambers MIAT RAnTech Chair Board of Educational Policy Ken Applebee OBE FIAT CBiol FSB RAnTech Chair Board of Moderators Cathy Godfrey FIAT RAnTech Chair Registration Accreditation Board Charlie Chambers MIAT RAnTech Chair ATW Editorial Board Jas Barley MSc FIAT RAnTech Bulletin Editor Sarah Lane MSc FIAT RAnTech Assistant Bulletin Editor Elaine Kirkum MIAT RAnTech MIScT Branch Liaison Officer Lynda Westall BSc Hons FIAT DMS RAnTech EFAT Representatives Charlie Chambers MIAT RAnTech Council Website Coordinator Allan Thornhill FIAT RAnTech IAT INFORMATION Animal Welfare Officers LABA Representatives Sarah Lane Debbi Young ATW Bulletin Editorial Board Jas Barley Chair Patrick Hayes Editorial Assistant Elaine Kirkum Sarah Lane Lynda Westall Board of Educational Policy Ken Applebee Chair Steven Cubitt Secretary Sarah Lane Board of Moderators Cathy Godfrey Chair Glyn Fisher Secretary Moderators Gary Childs Joanna Cruden Nicky Gent Linda Horan Sue McHugh Communications Group Norman Mortell Chair Kate Burton Linda Horan Elaine Kirkum Allan Thornhill Lynda Westall Registration and Accreditation Board Charlie Chambers Chair Ken Applebee Gerald Clough John Gregory Cathy Godfrey Sarah Lane Ron Raymond Wendy Steel Secretary Steve Owen Stuart Stevenson Carol Williams Observers Charles Gentry Certificate Holders Forum Adrian Deeny LASA Kathy Ryder Home Office Lucy Whitfield LAVA Advertisement Managers PRC Associates Ltd Email mail prcassoc co uk IAT OFFICERS MAY BE CONTACTED VIA IAT Administrator iat101 btconnect com OR VIA THE IAT WEBSITE AT www iat org uk OR VIA THE REGISTERED OFFICE 5 South Parade Summertown Oxford OX2 7JL Although every effort is made to ensure that no inaccurate or misleading data opinion or statement appear in the journal the Institute of Animal Technology wish to expound that the data and opinions appearing in the articles poster presentations and advertisements in ATW are the responsibility of the contributor and advertiser concerned Accordingly the IAT Editor and their agents accept no liability whatsoever for the consequences of any such inaccurate or misleading data opinion statement or advertisement being published Furthermore the opinions expressed in the journal do not necessarily reflect those of the Editor or the Institute of Animal Technology 2014 Institute of Animal Technology All rights reserved No part of this publication may be reproduced without permission from the publisher BRANCH SECRETARIES 2014 Aberdeen Cambridge Edinburgh Hertfordshire Essex Huntingdon Suffolk Norfolk Ireland London Midlands North East England North West Oxford Surrey Hampshire Sussex West Middlesex West of Scotland ii Ms Donna Wallace Ms Fran Flack Ms Janice Young Ms Helena Box Ms Jo Martin Mr Colin Travis Ms Amanda Dickson Mr Ian Fielding Ms Rachael Handisides and Joanne Bland Ms Nicky Windows Mr Adrian Woodhouse Ms Lesley Hughes Mrs Wendy Steel Ms Linda Horan aberdeenbranch iat org uk cambridgebranch iat org uk edinburghbranch iat org uk hertsessexbranch iat org uk hssbranch iat org uk irelandbranch iat org uk londonbranch iat org uk midlandsbranch iat org uk northeastbranch iat org uk cheshirebranch iat org uk oxfordbranch iat org uk shsbranch iat org 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December 2014 Animal Technology and Welfare THE INSTITUTE OF ANIMAL TECHNOLOGY ETHICAL STATEMENT IN THE CONDUCT OF THEIR PROFESSIONAL DUTIES ANIMAL TECHNOLOGISTS HAVE A MORAL AND LEGAL OBLIGATION AT ALL TIMES TO PROMOTE AND SAFEGUARD THE WELFARE OF ANIMALS IN THEIR CARE AND TO RECOGNISE THAT GOOD LABORATORY ANIMAL WELFARE IS AN ESSENTIAL COMPONENT OF GOOD LABORATORY ANIMAL TECHNOLOGY AND SCIENCE Editorial Jas Barley Chair of the Editorial Board Welcome to the 2014 themed issue of Animal Technology and Welfare ATW which this year focusses on Farm Animals used in Research I am pleased to have been able to compile this issue because it is in direct response to a request for more information on farm species welfare made to the IAT at the 2013 RSPCA AHVLA Farm Animal Welfare meeting see ATW April 2014 The institute is grateful to have been able to respond to this request and our thanks as always go to contributing authors On a personal front I am delighted to see farm animals used in research situations awarded such consideration as is shown by the report of the second RSPCA AHVLA Farm Animals used in Research meeting and in the farm animal related posters reprinted from Congress 2014 Much of my working life has been involved farm species and for a large portion of my career the attitude to farm species used in the main differed little from that of commercial agriculture despite the fact that the environment in most cases was extremely different from that on a farm I hasten to add that animal technologists were not unique in this attitude as few veterinary surgeons in the late 1960s whether in research or private practice would have experience of analgesics for farm animal species unless perhaps with the exception of horses and the terms environmental enrichment had yet to come into common parlance Thankfully over the intervening years we have come to appreciate that sheep pigs goats cattle and poultry when in a research situation warrant welfare that is appropriate to this special environment and purpose Many establishments have chosen to contract out research necessitating farm species to specialists who can provide a more appropriate environment in addition to the specialist equipment required for working with larger animals Of course animal technologists who are well trained in the procedures required and appropriate handling techniques for larger animals are also essential The benefits and disadvantages to animals in having procedures carried out by animal technologists was the topic for Animal in Science Education Trust s AS ET Special Travel bursary and the winning essay by Jan Bilton and one of the runners up is included in the issue Further essays will be included in future issues of ATW Kathryn Chambers Congress 2014 poster How Best to Train a Sheep in Research shows how handling stress can be dramatically reduced in sheep used for research Posters from Katie Blackwell and Emma Tozer both illustrate how anaesthesia and analgesia regimes can be refined for use in sheep and Katie Amess poster discusses how environmental enrichment can be provided for sheep when it is not possible for them to be at pasture By no means less important is the use of highly specialised techniques such as Computed Tomography as demonstrated in Lisa Griffin s poster Pigs are not forgotten and an improved method of blood sampling in G ttingen minipigs is explained in Adrian Zeltner s poster which again not only reduces the potential for stress in both pigs but also on the technologist s back Other species have not been ignored as the winner of the LBS Biotech Best Poster award from Congress 2014 shows Here Andrew Newman and Sabrina Brando show how simple measures can increase the opportunity for mice to follow natural behaviours Finally the welfare of Rhesus macaques can be improved by refining techniques for acclimatisation is covered by Lee Reed and colleagues As always the information included in this issue demonstrates how the quest for a better understanding of the animals we care for is paramount to the work of both animal technologists and the researcher ix

December 2014 Animal Technology and Welfare Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research cattle pigs sheep and poultry PENNY HAWKINS1 SHARON BROOKES2 SUSANNAH PARKIN3 R EDDIE CLUTTON4 PETER GADE5 JULIE LANE6 HELEN PROCTOR7 JOANNE EDGAR8 ISOBEL VINCENT9 and UTE WEYER2 1 2 3 4 5 6 7 8 9 Research Animals Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Animal Health and Veterinary Laboratories Agency AHVLA Animal Sciences Unit Weybridge Addlestone Surrey KT15 3NB Canterbury College New Dover Road Canterbury Kent CT1 3AJ Royal Dick School of Veterinary Studies University of Edinburgh Easter Bush Campus Midlothian EH25 9RG Novo Nordisk A S Novo All 2880 Bagsvaerd Denmark National Wildlife Management Centre AHVLA Sand Hutton York YO41 1LZ World Animal Protection 222 Grays Inn Road London WC1X 8HB University of Bristol School of Veterinary Sciences Langford House Langford Bristol BS40 5DU Royal Veterinary College RVC Clinical Skills Centre Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Corresponding author penny hawkins rspca org uk Introduction This meeting was jointly convened by the RSPCA and AHVLA to bring together animal technologists researchers veterinarians and students with an interest in the welfare of cattle pigs sheep and poultry used in research and testing for a programme of talks and discussion sessions The meeting which was held in September 2014 addressed a range of topics including refining endpoints in avian influenza studies reducing farm animal numbers in research pain management in pigs housing refinements for singly housed pigs the use of cortisol levels to predict farm animal welfare promoting positive welfare for chickens and replacing ewes in education and training A discussion session on positive welfare in farm animals concluded the programme Refining endpoints in avian influenza studies Sharon Brookes AHVLA Avian influenza AI is caused by viruses of the family Orthomyxoviridae in the genus influenza A virus Many species of bird are susceptible to infection with influenza A viruses including aquatic birds a major reservoir chickens and turkeys Most isolates in chickens and turkeys have been of low pathogenicity LP low virulence but some influenza A viruses can be highly pathogenic HP causing morbidity and devastating mortality Outbreaks of avian influenza present significant animal health and welfare economic and human health concerns so research that aims to improve the understanding diagnosis treatment or prevention of the disease is essential However some of this research has the potential to cause severe suffering which is an ethical and animal welfare concern for us 155

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research The outcome of infection with the HP virus depends upon the bird s species and age characteristics of the viral strain involved and environmental factors There may be sudden death preceded by few or no overt clinical signs or birds may contract a more characteristic infection with variable clinical presentations including respiratory signs swelling of the sinuses and or head apathy reduced vocalisation markedly reduced feed and water intake nervous signs and diarrhoea Egg production can be markedly reduced in laying birds These clinical signs are associated with AI but they are not pathognomonic that is they are not definitive indicators of the disease The level of variability in responses also presents some real challenges for veterinary inspectors in the field and for scientists in experimental situations All naturally occurring virulent strains isolated to date have been either of the low virulence H5 or H7 subtypes but these may mutate and become virulent This means that a risk assessment has to be carried out to determine the level of biosecurity needed for laboratory diagnosis and poultry inoculation with characterisation of the HPAI and other Notifiable Avian Influence NAI viruses at biocontainment level BCL 3 and other LPNAI at BCL level 2 at least The AHVLA avian virology and mammalian influenza team carries out a small number of diagnostic pathogenicity studies such as Intravenous Pathogenicity Index IVPI studies but most of our in vivo studies involve infecting poultry species with avian influenza HP and LP viruses to determine susceptibility pathogenicity transmission and or disease intervention A specialist area of interest has been cross species transmission in avian hosts These data collectively provide evidence for assessing risk of disease introduction and dissemination within the UK poultry industry plus informing control strategies and business continuity AHVLA uses around 600 birds in AI studies per year i e 200 chickens turkeys and ducks and over the last five years we have worked to refine humane endpoints and establish indicators of mortality with the objective of reducing the numbers of sudden deaths to a minimum One obvious approach was to increase the frequency of clinical obser vations with three assessments per day for virus host combinations with the potential to cause severe disease especially in the case of pathogens such as HPAIV where birds often transition rapidly from inapparent clinical disease to sudden death We also introduced a binary score sheet for these studies Figure 1 which has helped to reduce the number of birds found dead and increase the number of sick or moribund birds humanely killed during the course of experiments 156 Figure 1 Poultry record sheet used during Avian Influenza This is a step in the right direction but we want to achieve further refinements with respect to identifying indicators of mortality and implementing humane endpoints Body temperature has been successfully used to reduce mortality in mammals such as rodents pigs and ferrets in disease and vaccine development research and we are exploring the potential to use this indicator in poultry In the case of AI we have used thermal imaging to establish that increasing pyrexia is followed by a rapid decrease in body temperature as the clinical score escalates This area is being further investigated via both thermal imaging and biothermal microchip profiling to gather baseline data and trial adaptations for BCL3 usage We are also making progress with alternative techniques such as in ovo and ex vivo organ culture models whilst we continue with our efforts to refine study protocols and reduce severity Could reproductive technologies reduce farm animal numbers in medical research Susannah Parkin and Jennie Litten Brown Canterbury College Sheep are currently used in many areas of biomedical research including cardiovascular studies kidney dialysis research and the development of artificial

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research lungs Transgenic sheep are also used in pharming in which animals are genetically modified to produce pharmaceutically active compounds in their milk such as factors IX and VIII to treat haemophilia human protein C to treat thrombosis and cystic fibrosis transmembrane conductance regulator CFTR A total of 45 790 sheep were used in regulated procedures in the United Kingdom in 2013 a 7 increase on the previous year 1 Given this increase it may be useful to consider whether the number of sheep used in medical research might be reduced using reproductive technologies RT such as 1 Sexing semen SS 2 Artificial insemination AI 3 Intracytoplasmic sperm injection ICSI an in vitro fertilisation procedure in which a single sperm is injected directly into an egg This procedure may be used for the production of transgenic animals 4 Embryo transfer ET the process by which fertilised embryos are flushed removed from a source ewe and transplanted into recipient ewes 5 Stem cells SC undifferentiated cells that can produce other cells that eventually make up specialised tissues and organs There are two major types of stem cells embryonic and adult 6 Nuclear transfer NT which was used to produce the first cloned mammals The nucleus of a somatic cell is transferred into an egg cell whose own nucleus has been removed which is then stimulated by an electric shock to divide and form an embryo 7 Transgenic TG animals who have had one or more genes inserted into their genome from the genome of another species These techniques could be used to either accelerate the process of breeding farmed species or reduce animal numbers This might include reducing wastage if a particular sex is required or reducing numbers by facilitating more predictable growth and development Other applications could comprise improving disease control producing animals with a known response to anaesthesia or compacting lambing time which could shorten trial length and reduce numbers due to reduced impact of environmental variants Cloning could be used to eliminate genetic variation The use of sheep in orthopaedic research may be taken as a case study Sheep bone is anatomically close to human bone so sheep are used to evaluate orthopaedic techniques such as developments in intramedullary nailing which is the standard method of internally stabilising closed diaphyseal long bone fractures 2 Current protocols for treating humans with such fractures involve immobilisation and surgery to facilitate bone healing but continuous monitoring of changes in the load distribution between the implant and surrounding bone could inform interventions and improve patient outcome Sheep have been used to evaluate a telemetric strain gauge positioned within the intramedullary nail 3 If crossbred sheep are used in such studies there are likely to be significant variations in relevant traits Using purebred sheep and RT AI and ET may give rise to animals with more consistent bone structures enabling experimental group numbers to be reduced Although there is potential to reduce animal numbers using RT many techniques involve procedures that can cause pain and distress For example obtaining gametes preparing females for pregnancy insemination procedures and removing embryos can involve restraint anaesthesia and surgery Health and welfare concerns have also been repor ted for transgenic and cloned animals These potential harms need to be weighed against the benefits of reducing animal numbers for each project In conclusion once the technologies have been developed to a stage where they can be used repeatedly and consistently RT could be used to reduce the number of farm animals in some studies However it is wor th noting that whilst these technologies may be used to reduce numbers that is not always the preferred option if harms to individual animals are increased Pain management in pigs undergoing experimental surgery R Eddie Clutton University of Edinburgh Estimated numbers of pigs used recently 2011 2013 in biomedical research are 53 260 Canada 61 384 USA and 77 280 EU 1 A proportion of these animals are involved in experimental surgical studies conducted under general anaesthesia some of which are terminal or unclassified while others involve recovery Recovery procedures carried out on pigs are associated with postoperative pain in human patients so the animal model should be treated in a similar manner to the modelled i e humans and provided with appropriate perioperative analgesia This reflects legal requirements to reduce or eliminate suffering4 and helps to improve the translational value of the research since human patients receive pain relief From an ethical standpoint it is also an injustice to use animals in scientific procedures without ensuring that all available refinements have been fully implemented We aimed to evaluate whether the model and modelled were receiving a consistent standard of care by conducting a structured literature review of perioperative pain management in the pig Bradbury et al submitted 5 The results are soon to be published in 157

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research the journal Laboratory Animals 5 so a brief overview will be set out in this report A total of 233 papers described recovery surgery procedures using pigs which would have been painful for humans Postoperative analgesia was explicitly described in just 87 studies 37 Postoperative pain assessment was described in 10 of papers and only one article described the use of a pain scoring system 6 Six articles which described the use of analgesics gave no information on the drugs involved including their identity and only 20 included all of the information on analgesic agents recommended by the ARRIVE guidelines 7 which set out good practice for reporting animal use the name of the drug dose frequency route and duration of treatment Our study showed that the reporting of postoperative pain management for pigs involved in biomedical research is poor and falls short of the level of detail necessary for study replication This of course raises the question of whether analgesia is actually not provided or its use is simply not repor ted Unfortunately it may be that the situation as reported does reflect current practice In a similar study of analgesia reporting in rodents the authors contacted researchers who had not included information on pain relief in their publications to ask whether they had administered analgesia but had not described the regimen in the materials and methods section 8 They found that in 71 of cases analgesia had not in fact been given If this is also the case for pigs this would indicate a serious animal welfare concern in many of the procedures described in the literature clear Perspex dividers with holes of 10 cm diameter drilled into them so that animals can have visual olfactory and tactile contact with their neighbours Figure 2 Chains for pulling are also set up so that an animal pulling the chain in one pen causes the other end to move within the neighbouring pen Figure 3 Figure 2 Snout to snout contact between pigs in neighbouring pens Photo credit Novo Nordisk Housing refinements and enrichment for single housed pigs Peter Gade Novo Nordisk Many pigs used in biomedical research undergo instrumentation with a device to facilitate dosing or sampling such as ear vein catheters or access ports This makes group housing quite a challenge as the inquisitive nature of the pig increases the risk of another pig manipulating damaging or even removing the device Once the device is inoperable the animal can no longer be used so single housing is often necessary to prevent pigs from being wasted and to avoid any pain or distress caused by other animals interfering with devices For naturally gregarious animals like the pig single housing is undoubtedly a welfare problem At our facility we have successfully introduced refinement initiatives to improve the welfare of singly housed pigs while achieving the scientific objectives We designed the pens so that the pigs do not feel isolated using 158 Figure 3 Chain enrichment for singly housed pigs Photo credit Novo Nordisk We also make good use of the areas adjacent to the home pens Pigs are allowed supervised exercise times with other animals both in corridors between the pens and outdoors in a designated exercise area as shown at http tinyurl com pfc75z8 This clip also shows outdoor enrichment in the form of brushes and a paddling pool which has since been filled with bark chips which the pigs use for rooting the pigs shown in the video are not individually housed as they have no exteriorised instrumentation Pigs regularly spend time outdoors for as long as the weather permits this although we cannot always let them out in cold weather and we have to make sure that they do not get sunburn in the summer

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research There are some increases in both staff workload and cost associated with these refinements but in our view these are by far outweighed by the increased welfare for the animals Can cortisol levels really predict the welfare of farm animals Julie Lane and Fiona Bellamy National Wildlife Management Centre AHVLA Stress is an important consideration with respect to farm animal welfare and disease control On farm outbreaks of diseases such as campylobacter in chickens are suspected to be more common in situations where there are higher levels of stress and laboratory studies have demonstrated that chronic stress reduces the body s ability to fight a variety of virus and bacterial infections So an effective and objective indicator of stress for livestock in a commercial setting is vital for economic as well as animal welfare and ethical reasons Robust indicators of stress are also essential for farm animals used in scientific procedures with respect to designing and evaluating refinements defining and implementing humane endpoints and assessing the actual severity of procedures Behaviours can be important and useful indicators of stress but can also be difficult to interpret and to measure objectively There are many physiological indicators that a body is under stress which lend themselves to more objective measurement but these often require instrumentation of the animal or restraint and blood sampling both of which can cause stress to the animal affecting the integrity of the data collected There are also usually financial and temporal constraints that limit the number of indicators that can be assessed It is important therefore to develop reliable appropriate and accurate indicators of animal welfare It has been established for nearly half a century that stressful experiences cause the synthesis and release of glucocorticoids such as cortisol or corticosterone from the adrenal gland It used to be necessary to obtain blood samples to measure glucocorticoid levels but non invasive techniques have been developed including the analysis of saliva and faeces We have used these methods to assess levels of cortisol in a wide variety of farm animals under many conditions and shown that cortisol can be an effective and accurate tool for assessing stress For example a study of sheep welfare during transport involved two groups of sheep transported by drivers using either a forward aggressive or a defensive driving style There were no significant differences in behavioural responses or heart rate between the two groups of sheep but salivary cortisol levels were significantly increased following transpor t in the sheep driven by the aggressive haulier In the above example it was the cortisol levels that showed animals were stressed when other indicators were not significantly increased This makes measurement of cortisol an attractive tool for helping to assess welfare In addition cortisol levels are not affected by an animal s social standing or normal levels of exercise or by diet However the use of cortisol is not without its issues and caveats which need to be identified and explored before use of these techniques is considered For example levels can be affected by blood sampling anaesthesia an animal s age or sex pregnancy infertility and the time of day as cortisol rises and falls according to circadian or ultradian cycles depending on species It is essential to understand how all of these factors interact and affect cor tisol level data especially now that the technology is becoming increasingly more sophisticated enabling very small concentrations to be measured in animal by products such as hair and milk The answer to the question Can cortisol levels really predict the welfare of farm animals is therefore yes provided that the context for the data is clearly understood and results are properly interpreted Measuring positive emotions in dairy cattle Helen Proctor and Gemma Carder World Animal Protection A sentient animal can consciously experience both positive and negative emotions As a result their feelings matter to both the animal and to us The importance of promoting positive emotions in animals as well as avoiding or minimising the negative emotions is increasingly recognised 9 10 Despite this we still know very little about the subjective minds of animals and much of what we do know is focussed on indicators of negative experiences and emotions such as pain and suffering 11 In 2013 we published a systematic review of the scientific literature where we searched for evidence of animal sentience 10 We found that not only is animal sentience more accepted than is often thought to be the case but most of the sentience traits utilised in research were negative ones such as pain fear and anxiety Knowledge of negative states in animals is important for improving animal welfare but this is only part of the issue We still need to develop our understanding of positive emotions and how animals express these so that we may promote and assess positive emotional states in the animals under our care 12 159

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research Emotions are subjective and personal states and are therefore difficult to interpret and measure especially in animals as we do not have a shared language However animals do feel experience and communicate emotions in fact emotions are essential in enabling animals to communicate with one another interpret situations correctly and facilitate appropriate responses 11 In this study we sought reliable measures of positive emotions in dairy cows testing the suitability of ear postures as a reliable measure of a positive low arousal emotional state in cattle To elicit this state we emulated allogrooming in 13 habituated dairy cows by stroking them on regions of their head neck and withers that have been shown to be preferred areas during both allogrooming and stroking 13 14 at the rate allogrooming typically occurs 13 Stroking calms cattle and has been shown to reduce cortisol levels15 and heart rate 16 The stroking stimulus was performed only to habituated cows and on a voluntary basis as the cows were able to move away at any point and were not pursued or followed These results suggest that ear posture could be a useful indicator for assessing low arousal positive emotional state in dairy cows although further work needs to be done to validate these results before ear postures can be used in routine welfare assessments The next steps will involve testing this indicator on other stimuli including on high arousal positive stimuli in order to further explore the effects of arousal Once validated ear posture could provide a non invasive easy and objective measure of emotional state in dairy cows These results also provide a helpful insight into positive emotions an area that is often neglected yet is essential to good animal welfare Further research into this important field needs to continue and our study demonstrates that such research can be carried out on existing commercial farms By conducting the research in this way we not only assured that the measure is valid in the industry setting but it allows us to utilise existing populations of animals and enables us to work with farmers to provide solution focussed animal welfare research A good life for chickens This study is in press elsewhere17 so a brief overview of the conclusions will be presented here We analysed video footage from the focal observations and found four distinct ear postures Figure 4 a d The duration of time spent in each of the postures was significantly affected by the stroking stimulus The alert ear postures 1 and 2 EP 1 EP 2 were performed for significantly less time during the stroking segment and the relaxed ear postures 3 and 4 EP3 EP4 were performed for significantly longer during the stroking segment The positive low arousal stimulus therefore caused significant differences in the time spent in each of the four ear postures a Ear posture 1 EP1 b Ear posture 2 EP2 c Ear posture 3 EP3 d Ear posture 4 EP4 Figure 4 Ear postures associated with alert and relaxed states in cattle Photo credit Helen Proctor World Animal Protection 160 Jo Edgar University of Bristol In 2013 129 448 domestic fowl were used in 129 538 scientific procedures in the UK 1 The majority 90 of procedures were for the purpose of applied veterinary research with most birds used in the production of infectious agents and parasitology Domestic fowl are also used in fundamental research 8 of procedures psychology 3 and pharmaceutical efficacy testing 9 The care and use of domestic fowl kept for scientific research is regulated by legislation and Codes of Practice that largely focus on the alleviation of negative aspects of welfare However it is becoming increasingly accepted that good welfare is not simply the absence of negative subjective states but also includes the presence of positive experiences such as pleasure e g references 11 and 18 This concept has been promoted by the Farm Animal Welfare Committee FAWC formerly the Farm Animal Welfare Council an advisory body to the government on farmed animal welfare In 2009 FAWC proposed that a good life could be considered in terms of additional opportunities for example access to a resource that an animal does not need for biological fitness but which is valued by the animal 19 FAWC identified four states Comfort Pleasure Interest and Confidence which are necessary for an animal to be considered to have a good life Figure 5 When considering whether animals experience feelings like these we can think about whether each might have a function for example in motivating behaviours that are important for survival such as seeking valuable resources or avoiding harms Some have argued that

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research Life See Figure 6 for a diagram summarising the elements of the resource tiers and reference 20 for a full explanation Figure 5 Quality of a life Photo credit Novo Nordisk feelings or emotions do not necessarily need to be consciously experienced but concern for animal welfare rests on the assumption that animals do experience negative emotional states There is a large body of evidence that has been used to infer negative emotional states in chickens such as those associated with pain and distress Of course emotions cannot be directly assessed in animals but there are physiological and behavioural indicators that can be used to infer how animals may be feeling For example hens show a strong motivation for mealworms over other feed rewards displaying arousal type behaviour and changes in their surface body temperature when they are anticipating a mealworm reward These behavioural and physiological changes during anticipation and consumption of rewards may provide indirect information about pleasure in chickens Comfor t behaviours like dustbathing may also be associated with a positive emotional state in domestic fowl On this basis we proposed that animals can be said to have a good life if their quality of life is substantially higher than that afforded by the current legal minimum standards of housing and care and includes positive experiences such as comfort and pleasure 20 We needed to determine which resources laying hens require to make them happy and how important each of these resources are to the birds As well as using published evidence the resources required for a good life were identified using the opinion of twelve experts from five academic institutions in the UK and New Zealand These were researchers with extensive experience and knowledge of farm animal behaviour and welfare including a sound publication record The experts also provided guidance on the relative ranking of resources i e which resources would be required to attain three tiers of higher welfare and leading towards a good life The resources identified by the experts were organised according to FAWC s four opportunities with a fifth opportunity added Healthy Figure 6 Elements of good life resource tiers for laying hens Following construction of the draft laying hen resource tiers a pilot study was undertaken to establish the validity reliability and feasibility of the framework for assessing whether birds had a good life within a given production system Twelve farms from different systems were visited and assessed according to the criteria within the tiers and an interview with the producer The average assessment time was 23 minutes with a range of 15 to 45 minutes and results were generally positive with the good life framework distinguishing between systems reliably The good life framework could also be applied to laying hens in a research setting Some elements make it more applicable to research animals than farmed animals given the generally higher standards of housing and care for the former Other criteria are less applicable for example in a farm setting access to the outdoors and natural light naturally lit verandas and well covered ranges score highly One obvious difference between farmed and laboratory hens is that the Healthy Life opportunity is often compromised for example within disease studies It may be possible to use the good life framework to see how the effects of procedures might be ameliorated by focusing on other resources that can still be provided Is ewe OK Improving sheep welfare by use of simulation Isobel Vincent Royal Veterinary College In their first year Royal Veterinary College students spend two weeks at an allocated sheep farm to gain 161

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research experience of the activities involved around lambing time The farms vary greatly in size housing facilities and husbandry methods so the resultant learning outcomes are likely to reflect these inevitable variations 21 Some students will already come from farming backgrounds but ever burgeoning numbers of undergraduates mean that providing sufficient opportunities to learn and develop practical skills such as lambing is increasingly challenging One way of addressing increasing numbers is to create simulated learning situations 22 These provide safe non critical environments where students can practise certain skills repeatedly until they feel confident Ewe simulators are used as teaching aids prior to students observing and assisting at lambing The simulators are made from plastic water tanks with a birth canal and uterus made of polythene tubing The tubing is fed through a life sized fused pelvis fixed by stainless steel bars in the centre of the tank Figure 7a The uterus can hold at least two cadaver lambs The tank is filled with warm water providing the realistic sensation of pressure around the lambs a One of the most impor tant components of this simulator system is the cadaver lamb However it is a constant logistical challenge to source enough on site cadavers for the large numbers of students taught lambs must be small enough to fit through the simulator birth canal and death cannot be due to disease Although they provide the best alternative to the live lamb they are of limited reusability due to post mortem changes and repeated births To address this a fully ar ticulating manikin lamb was commissioned in an attempt to supplement or replace cadavers 23 However students found the manikin less useful than cadavers and it was not as robust as had been hoped The practical class involves an initial demonstration of how to lamb a ewe using the simulator highlighting welfare aspects for example do not intervene too early have clean hands use lubrication be gentle and patient Figure 7b The class also covers abnormal presentations use of lambing aids and vaginal prolapse Students are divided into groups of four or five to practise on one of the four available simulators They are instructed to arrange the lambs in various positions in utero in order to gain confidence for real scenarios Those with prior lambing experience are encouraged to share their lambing tips with peers However for effective learning to occur staff circulate amongst the subgroups to give feedback24 and to ensure that any prior knowledge is in fact describing good practice formation of bad habits at this point could lead to real welfare issues For example delivering a lamb too quickly and roughly can cause internal trauma to the ewe leading to postpartum malaise and even death Students n 42 were sur veyed after this year s practical classes Over 83 said practising on the simulator helped their confidence with lambing and of those nearly 89 said it helped familiarise them with the process without worrying about injury to ewe or lambs b Figure 7 The ewe simulator a top view showing uterus b first year students practising on the simulators Photo credits Isobel Vincent RVC 162 This year in a continuing attempt to improve welfare of lambs a group of students n 75 were given the opportunity to practise tail docking and castration using elastrators on bespoke manikins just prior to carrying out the procedure on live lambs A survey showed that 95 felt more confident about elastrating real lambs after practice with the manikins Over 75 indicated that this was because they could not hurt the manikin could take more time and did not feel pressured to get it right first time These findings back up research showing that simulators are particularly useful if they include some sort of palpation25 and where they replace the live animal there are important welfare benefits

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research Discussion In the final discussion session at the end of the meeting delegates exchanged ideas about indicators of positive welfare in the animals they care for Discussions at the previous year s meeting had also touched on this topic indentifying playfulness and positive social interactions with other animals and humans as indicators of positive welfare in sheep cattle and pigs 26 The discussion at this year s meeting reinforced the importance of understanding animal behaviour especially with regard to observing and interpreting animals responses to humans This included understanding that animals do not always display behaviours that indicate pain or distress e g the sheep in the transport study where stress was only apparent when cor tisol levels were examined Delegates acknowledged the importance of using more than one indicator to reduce the risk of missing signs of suffering or wellbeing Delegates also discussed the application of the Three Ss good Science good Sense and good Sensibility as set out by the late Dr Carol M Newton 27 Many felt that taking play behaviour and positive approaches to staff as indicators of positive welfare was an example of applying good sense to human animal interactions as well as providing a useful early indicator that animals may be experiencing adverse effects for example if play behaviour ceases or an individual is reluctant to approach a familiar carer Overall action points The following action points are suggested on the basis of the talks and discussions on the day If you are responsible for using caring for or obser ving animals on studies where mor tality occurs set a goal to reduce this to a minimum This could include reviewing monitoring frequency implementing tailored assessment sheets and regularly reviewing these and actively seeking new indicators that will enable humane endpoints to be defined Consider whether the use of reproductive technologies could help to reduce animal numbers while still effectively addressing the scientific question In doing this carefully weigh the potential to reduce numbers against the harms caused to animals and or their offspring by the use of reproductive technologies If analgesia is withheld in a procedure that would be painful for a human question the justification for this either as an individual via a Named Person or through a committee such as the Animal Welfare and Ethical Review Body AWERB If you are a researcher always include information on refinements such as pain relief in your publications justifying this if you encounter any resistance from editors or peer reviewers If studies necessitate singly housing social animals research whether they would benefit from some visual tactile and or olfactory contact and if they would refine their housing to accommodate this Be aware that some species including many farm animals do not always display signs of pain or distress that are obvious to human observers Ask your Named Information Officer to periodically review the literature on welfare indicators including the use of non invasive cortisol samples If you work with or care for animals with ears pay attention to their ear postures and see whether these might be useful welfare indicators in your particular setting Look up the good life paper for domestic fowl to see how your facility compares to the different levels and whether there any improvements could be made If you are involved in training staff to assist with lambing or birth in other large animals consider using simulators as described above Initiate discussion at your facility about indicators of positive welfare including which behaviours people can identify and whether these could be included in formal welfare assessment protocols Acknowledgements Thank you to all of the delegates and speakers for their support and enthusiasm for the meeting especially with respect to the discussions Special thanks to Adrian Smith of Norecopa for summing up the day and chairing the final discussion session We are also grateful to AHVLA staff for their assistance which helped to make the day such a success References 21 22 23 24 25 26 Home Office 2014 Statistics of Scientific Procedures on Living Animals Great Britain 2013 http tinyurl com p447f3w last viewed 3 October 2014 Caeiro Potes J da Costa Reis J Capela e Silva F Relvas C Silv rio Cabrita A and Sim es J A 2008 The sheep as an animal model in orthopaedic research Experimental Pathology and Health Sciences 2 29 32 http tinyurl com lj3cfdn last viewed 3 October 2014 Wilson D J Morgan R L and Janna S W 2008 A single channel telemetric IM nail for in vivo measurement of fracture healing Poster No 1040 54th Annual Meeting of the Or thopaedic Research Society http www ors org Transactions 54 1040 pdf last viewed 3 October 2014 European Commission 2010 Directive 2010 63 EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes Official Journal of the European Union L276 233 Bradbury A G Eddleston M and Clutton R E submitted Pain management in pigs undergoing experimental surgery a literature review 2012 2014 Laboratory Animals Murison P J Jones A Mitchard L Burt R and 163

Report of the second RSPCA AHVLA meeting on the welfare of agricultural animals in research 27 28 29 10 11 12 13 14 15 16 17 18 19 20 21 Birchall M A 2009 Development of perioperative care for pigs undergoing laryngeal transplantation a case series Laboratory Animals 43 338 43 Kilkenny C Browne W J Cuthill I C Emerson M and Altman D G 2010 Improving bioscience research reporting The ARRIVE guidelines for reporting animal research PLoSBiol 8 e1000412 see https www nc3rs org uk arrive guidelines last viewed 3 October 2014 Richardson C A and Flecknell P A 2005 Anaesthesia and post operative analgesia following experimental surgery in laboratory rodents are we making progress Altern Lab Anim 33 119 27 Green T C and Mellor D 2011 Extending ideas about animal welfare assessment to include quality of life and related concepts N Z Vet J 59 263 71 Proctor H Carder G and Cornish A 2013 Searching for animal sentience A systematic review of the scientific literature Animals 3 882 906 download at http www mdpi com 2076 2615 3 3 882 last viewed 3 October 2014 Boissy A Manteuffel G and Jensen M 2007 Assessment of positive emotions in animals to improve their welfare Physiol Behav 92 375 397 Mellor D 2012 Animal emotions behaviour and the promotion of positive welfare states N Z Vet J 60 1 8 Schmied C Boivin X and Waiblinger S 2008a Stroking different body regions of dairy cows effects on avoidance and approach behavior toward humans J Dairy Sci 91 596 605 Schmied C Waiblinger S Scharl T Leisch F and Boivin X 2008b Stroking of different body regions by a human Effects on behaviour and heart rate of dairy cows Appl Anim Behav Sci 109 25 38 Hemsworth P and Barnett J 1989 The effects of handling by humans at calving and during milking on the behaviour and milk cor tisol concentrations of primiparous dairy cows Appl Anim Behav Sci 22 313 326 Waiblinger S Menke C Korff J and Bucher A 2004 Previous handling and gentle interactions affect behaviour and hear t rate of dair y cows during a veterinar y procedure Appl Anim Behav Sci 85 31 42 Proctor H S and Carder G 2014 Can ear postures reliably measure the positive emotional state of cows Appl Anim Behav Sci http dx doi org 10 1016 j applanim 2014 09 015 Yeates J W and Main D C J 2008 Assessment of positive welfare A review Vet J 175 293 300 Farm Animal Welfare Council 2009 Farm Animal Welfare in Great Britain Past Present and Future FAWC London download at http www fawc org uk pdf ppfreport091012 pdf last viewed 3 October 2014 Edgar J L Mullan S M Pritchard J C McFarlane U J C and Main D C J 2013 Towards a Good Life for farm animals Development of a resource tier framework to achieve positive welfare for laying hens Animals 3 584 605 download at http www mdpi com 20762615 3 3 584 last viewed 3 October 2014 Wenger E 1998 Communities of Practice Learning Meaning and Identity Cambridge New York Cambridge University Press 164 22 23 24 25 26 27 Meller G 1997 A typology of simulators for medical education Journal of Digital Imaging 10 3 Suppl 1 194 196 Vincent I and Thompson J 2011 Use of an articulating manikin lamb and ewe simulator to enhance and supplement teaching of ovine obstetrics 2nd Veterinar y Education Symposium University of Nottingham UK 13 14 July 2011 Issenberg S B McGaghie W C Petrusa E R Lee Gordon D and Scalese R J 2005 Features and uses of high fidelity medical simulations that lead to effective learning a BEME systematic review Medical Teacher 27 10 28 Parkes R Forrest N and Baillie S 2009 A mixed reality simulator for feline abdominal palpation training in veterinary medicine Studies in Health Technology and Informatics 142 244 246 Hawkins P Clutton R E Dennison N et al 2014 Report on the RSPAC AHVLA meeting on the welfare of agricultural animals in research cattle goats pigs and sheep Animal Technology and Welfare 13 1 43 56 Newton C M Quoted in Rowsell H C 1977 The ethics of biomedical experimentations pp 267 281 in The Future of Animals Cells Models and Systems in Research Development Education and Testing Washington D C National Academy of Sciences

December 2014 Animal Technology and Welfare Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting PENNY HAWKINS SECRETARY 1 PAUL LITTLEFAIR2 HUW GOLLEDGE3 CLAIRE RICHARDSON3 SARAH ALLDEN4 TANIA BODEN4 COLIN HENDRIE5 DOMINIC WELLS6 NIKKI OSBORNE1 MARIE HUTCHISON7 STEPHEN RYDER8 MAGGY JENNINGS1 and ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS International Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Newcastle University Newcastle Upon Tyne NE1 7RU UCB Pharma Ltd 208 Bath Road Slough Buckinghamshire SL1 3WE University of Leeds Institute of Psychological Sciences Leeds Yorkshire LS2 9JT Comparative Biomedical Sciences The Royal Veterinary College Royal College Street London NW1 0TU Mary Lyon Centre MRC Harwell Harwell Science and Innovation Campus Oxfordshire OX11 0RD Home Office Animals in Science Regulation Unit 2 Marsham Street London SW1P 4DF UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Corresponding author penny hawkins rspca org uk Introduction The RSPCA UFAW Rodent Welfare Group holds a oneday meeting every autumn so that its members can discuss current welfare research exchange views on rodent welfare issues and share experiences of the implementation of the 3Rs of replacement reduction and refinement with respect to rodent use A key aim of the Group is to encourage people to think about the whole lifetime experience of laborator y rodents ensuring that every potential negative impact on their wellbeing is reviewed and minimised To mark the 20th anniversary of the Rodent Meetings the 2013 meeting began with a presentation on animal welfare issues in China including the work of the RSPCA with respect to laboratory animals as the 20th wedding anniversary is the China anniversary Other speakers gave presentations on aversion to euthanasia agents assessing welfare and reducing suffering in rodent disease models and the impor tance of understanding rodent behaviour when designing experiments and husbandry and when interpreting data All the presentations end with some action points The meeting also included a special session on welfare assessment in genetically altered GA rodents with talks on current practice and an interactive discussion session Animal welfare in China Paul Littlefair International Department RSPCA Animal welfare is a rapidly evolving field and nowhere has this been more the case than in China over the past decade There are a number of identifiable factors which have raised the profile of animal protection issues in China in recent years Internal factors include a catalogue of widely publicised animal abuse incidents a sharp rise in companion animal keeping an increase in meat and dairy consumption and animal health crises as well as food safety and security issues These have all added to the momentum for improvements in the treatment of animals The debate has also been driven by external influences particularly as China s political and economic standing in the world has been boosted and its participation in 165

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting multilateral fora has had growing implications for animal welfare The space for this debate has been shaped by the global social media boom which has played a major role in exposing China s animal issues to both the domestic population and the international community Of all areas of animal use in China the field that has shown the most potential for integrating animal welfare improvements has been laboratory animal science The sector has grown rapidly since the impact of economic reforms began to be felt in the early 1980s and the field of laboratory animal sciences is also globalised to an extent generally not seen in other areas of animal use Since 1999 China has dramatically increased its share of scientific papers published worldwide Chinese scientists increasingly acknowledge that poor laboratory animal welfare may undermine the validity of scientific results and therefore the quality of the research China breeds around 19m laboratory animals a year compared to an estimated 20m in the US and 12m in the European Union EU If the country is to continue to excel in science and particularly in the bioscience fields then it needs to secure and maintain both scientific quality and competitiveness These economic and technological imperatives have enabled the RSPCA and others to promote good laboratory animal welfare as conducive to good science As early as 1988 the Ministr y of Science and Technology MOST issued a Statute on the Administration of Laboratory Animal Use which covered the basic needs of laborator y animals and the requirement for trained personnel This was followed in 1997 by the first explicit appearance of animal welfare and the 3Rs in MOST s Laboratory Animal Development Programme for the Period of the Ninth Five Year Plan From 2004 first Beijing and then several provincial laboratory animal administrations also included modest animal welfare references in their regulations While not laws in the strict sense these ministerial Statutes and municipal or provincial regulations operate as de facto legislation and as such represent the first appearance of animal welfare in China s regulatory measures This environment has provided opportunities for the RSPCA and others with an interest in laboratory animal welfare to partner Chinese institutions aiming to improve standards Since 2007 the RSPCA s International and Research Animal Depar tments have worked closely with government and academic institutions and laboratory animal science associations in both Europe and Asia to promote laboratory animal welfare particularly ethical review and the 3Rs This collaboration has included the sharing of materials the provision of conference speakers and the delivery of training Figure 1 166 Figure 1 RSPCA workshop on laborator y animal welfare and the 3Rs China Photo credit RSPCA In 2007 RSPCA materials on the housing and care of 14 laboratory animal species were translated into Chinese by the Chinese Association of Laboratory Animal Sciences CALAS and were launched at the 3rd Congress of the Asian Federation for Laboratory Animal Sciences AFLAS in September 2008 in Beijing At the same meeting staff gave presentations on ethics committees and the implementation of the 3Rs A two day workshop held in Xi an in 2009 focussing on good practice in laboratory animal care and the importance of sound experimental design in promoting the 3Rs attracted around 90 CALAS members scientists academics animal technologists and breeding establishment representatives In 2010 a two day course was given in Suzhou on Laboratory Animal Welfare and Alternatives Collaboration with CALAS continued in Yangzhou in 2012 with a presentation at the plenary of the association s annual meeting and in separate sessions on Refinement of Procedures and an overview of laboratory animal welfare in the UK The RSPCA is currently funding a project with support from NC3Rs to develop with CALAS a Chineselanguage version of the Procedures With Care website www procedureswithcare org uk produced by Newcastle University with the Institute of Animal Technology and NC3Rs The Chinese site was launched early in 2014 Action points If you work for a company that has facilities overseas including in China find out whether these operate according to UK or other good standards with respect to animal housing husbandry and care and ethical review If they do not consider how you could raise this issue and encourage levelling up for example by seeing whether staff could be encouraged to join organisations like CALAS and to use resources such as the Chinese Procedures With Care site

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting Aversion to rodent euthanasia agents Huw Golledge Newcastle University Millions of laboratory rodents are killed for scientific purposes each year This means that euthanasia is effectively the most commonly performed technique in the laboratory so it is critically important from both ethical and animal welfare aspects to ensure that it really does provide a good death for the animals concerned However controversy continues to surround the question of which methods for killing rodents are the most humane Directive 2010 63 EU 1 which regulates animal experimentation within the EU requires that animals are killed with minimum pain suffering and distress It lists permitted methods for humanely killing animals which in the case of rodents excluding foetal and immature forms are physical methods cer vical dislocation concussion and decapitation the last of these only if no other method is possible anaesthetic overdose a gradual chamber fill of carbon dioxide CO2 or inert gases argon and nitrogen Schedule 1 of the revised Animals Scientific Procedures Act ASPA governs the humane killing of laboratory animals in the UK but the UK chose not to include decapitation or inert gases in Schedule 1 due to welfare concerns about these methods 2 Concerns about CO2 Although the techniques listed in Schedule 1 are generally believed to be humane there are persistent concerns about some of them For example it is often suggested that the use of carbon dioxide may be inhumane since it can cause animals distress 3 Nonetheless it is still widely used especially when there are large numbers of animals to kill There are considerable incentives to use CO2 as it is cheap safe for the operator easy to use effective and does not contaminate tissues Despite these benefits for humans however there are some animal welfare questions that deserve serious consideration G G G does CO2 cause pain does CO2 cause distress is anything else better Carbon dioxide is painful to inhale when the concentration reaches 50 for humans4 and probably for rodents too This is because CO2 forms carbonic acid when it comes into contact with water which can occur on moist tissues such as the nasal passages trachea and eyes Being placed into a chamber prefilled with CO2 could therefore be extremely painful which is why Schedule 1 stipulates that a gradual chamber fill should be used Studies have shown that with a flow rate of around 20 chamber volume per minute rats and mice become anaesthetised before the concentration of CO2 reaches painful levels Unfortunately this does not solve the problem of distress associated with CO2 exposure Several studies have examined whether rats and mice find carbon dioxide aversive i e whether they find it unpleasant to the extent that they actively avoid it and will become distressed if they cannot get away This has been evaluated using approach avoidance tests in which rats and mice are first trained to enter a test chamber for a highly desired food reward e g Cheerios cereal and once they have learned to enter this chamber for a reward it is gradually filled with either the test agent or a flow of air as a control 5 6 When the incoming flow is just air the animals will generally finish the food reward However when the flow contains CO2 rats will leave at concentrations above 15 even if there are still some rewards left 6 7 Most studies like these have found aversion to relatively low levels of CO2 in rats and mice so although exposure to a gradual fill may not be painful it is likely to be distressing There is currently much debate about the level of distress caused by CO2 and whether it is such that CO2 killing is not humane Is anything better Anaesthesia with isoflurane prior to killing with CO2 or argon or another non inhalation method has been suggested to be a more humane alternative yet it is unclear whether isoflurane causes less aversion than CO2 Approach avoidance tests based on a single exposure to the gases as above have shown that isoflurane is less aversive than carbon dioxide8 and argon is considerably more aversive than either of the other two agents 6 It looked as though there was a simple scale of degrees of aversion but more recent research has complicated this On the basis of the mouse or rat s first exposure to isoflurane it is less aversive than CO2 but on second and subsequent exposures it is equally aversive to rats9 and possibly to mice although this has yet to be tested Thus aversion to isoflurane appears to be at least partially learned This has implications for studies that involve anaesthetising animals multiple times such as repeated imaging studies and also calls into question whether anaesthesia before switching to CO2 is truly a refinement for animals who have already been exposed to isoflurane NB other studies have found that the learned aversion persists for some time and is transferred to other volatile agents such as sevoflurane Ongoing research and new techniques There have been some criticisms of the conclusions of studies involving exposing animals to agents and measuring aversion For example an animal might leave the chamber when the concentration of the test agent reaches a certain level but does that mean it would cause significant distress if the animal was 167

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting unable to leave Is the gas really aversive or does it simply alter motivation or make the reward less palatable A technique that can be used to address these questions is conditioned place aversion which measures animals abilities to associate the environment where they have been exposed to an agent with their response to that agent If the exposure is aversive e g if a gas is unpleasant to inhale then the animal should learn to avoid the environment they associate with that exposure This is an especially convincing technique because it uses the animals memory of what has happened to them rather than the cues they experience at the time as the animal avoids the environment even when the agent is not present This is regarded as an indication of the affective or emotional state of the animal induced at the time of exposure A typical conditioned place aversion protocol presents animals with a choice between two chambers one with plain walls and a smooth floor and the other with distinguishing features such as striped walls and a textured floor Figure 2 Each animal is trained to expect a flow of air in one chamber and a flow of the test agent in the other by confining them in each chamber during training sessions Once training is complete the animal is placed into the apparatus with just air in both sides and allowed to move freely between the two chambers and the amount of time they choose to spend in each chamber is recorded If the test agent is aversive the animal should decide to spend significantly less time in the chamber that they have learned to associate with the agent aversive than CO2 confirming that the UK made the right decision when excluding iner t gases from Schedule 1 However care is needed when interpreting the results for isoflurane and carbon dioxide These are both apparently equally aversive but repeated exposures were necessary to train the animals so aversion to isoflurane could have been learned as outlined above This means that the aversiveness of isoflurane for na ve animals relative to CO2 could be overestimated using this technique What causes aversion to CO2 All of the above research raises the question of what actually causes the aversion that is displayed towards all three agents by mice and rats This is easiest to explain for CO2 Rats and mice are both burrowing species so their sensitivity to carbon dioxide is likely an adaptation to ensure that they can sense and be motivated to escape from pockets of accumulated CO2 In fact mice can smell and will avoid CO2 at concentrations of just 0 2 which is little above atmospheric levels 10 Breathing low levels of CO2 causes feelings of anxiety in humans and brief exposure to 20 CO2 also activates panic defence brain circuitry in rats11 via direct detection in the amygdala of the acidosis caused by inhaling carbon dioxide 12 The reason for the very marked aversion to argon is unknown but may relate to a heightened response to hypoxia or asphyxia In the case of the learned aversion to isoflurane it has been hypothesised that animals remember feeling nauseous when recovering which motivates them to escape when they smell the agent again The implications for repeatedly anaesthetising animals with gaseous agents also need to be further explored To conclude research to date suggests that no inhalation agent so far tested is likely to offer an entirely humane method for killing laboratory rodents Carbon dioxide is not ideal but it is unclear whether isoflurane is significantly better and inert gases are almost certainly worse A continued search for a humane method is required Figure 2 A typical conditioned place aversion experimental set up The rat is trained to expect a flow of test agent in the left hand plain chamber and a flow of air in the right hand chamber which has patterned walls and a textured floor Following training the animal is placed in the chamber with air in both sides and their position is monitored see text We have been using this protocol in research funded by the NC3Rs and UFAW to evaluate the comparative aversiveness of carbon dioxide isoflurane and argon Studies so far indicate that all three cause significant place aversion Argon is also significantly more 168 Note The second Newcastle Consensus Meeting on euthanasia was held in August 2013 and the forthcoming report in preparation at the time of writing will address the above issues in more depth along with physical methods and euthanasia of fish Action points If CO2 is used to humanely kill rodents at your establishment suggest that practice is reviewed taking the current literature into account This could be initiated by the Named Information Officer NIO or Named Animal Care and Welfare Officer NACWO reviewed by the AWERB or discussed more widely Ensure that the AWERB regularly reviews the literature on humane killing in general so that good practice is maintained in line with current thinking

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting Watch out for the publication of the second Newcastle report and bring it to the attention of your NIO Assessing welfare in mouse disease models Claire Richardson Newcastle University A key component in reducing suffering within disease studies is the implementation of appropriate humane endpoints Humane endpoints can be defined as criteria that allow early termination of experiments before animals experience significant harm whilst still meeting the experimental objectives see http www humane endpoints info The effective implementation of humane endpoints therefore depends upon recognising when the experimental objectives have been met unless severity limits are approached or exceeded Today it is thankfully widely accepted that death is not an acceptable endpoint Ideally humane endpoints should not be considered as predictors of imminent death but rather as ways of identifying animals in more moderate clinical states so that appropriate action can be taken to ameliorate suffering However the use of animals as disease models presents a problem with respect to refining and implementing humane endpoints It may be necessary to allow animals to become sick in order to answer the experimental question and this may cause pain and distress as the disease progresses But it is possible to set limits on disease progression and this is not only essential from an animal welfare aspect but will also minimise experimental variability leading to reductions in numbers Achieving these limits obviously requires an understanding of how far disease has progressed in each animal However different disease models have different effects such as pain malaise nausea or fatigue some of which are difficult to detect and assess It is essential that observations are made by experienced compassionate staff using carefully considered indicators and record keeping systems Work is also in progress to identify surrogate markers of disease progression as a way of further refining both scientific and humane endpoints Imaging is frequently advocated as a method of tracking disease progression but repeated general anaesthesia is typically required which may affect experimental outcomes and have welfare implications9 13 see also Golledge this paper Additionally imaging is expensive requires specialist skills and equipment and may not be carried out at a sufficient frequency to detect critical points in disease progression We therefore need to identify non invasive biomarkers e g subtle behaviours or clinical signs that can be used as specific indicators of disease progression 14 15 Beyond their importance in humane endpoints biomarkers that reliably predict the onset of clinical signs can also be used to introduce refinements at times when they are most needed by the animals We are developing an enhanced clinical monitoring approach funded by UFAW and the NC3Rs using minimally invasive radiofrequency identification RFID technology to try to define useful biomarkers to help track disease progression RFID chips which transmit body temperature are implanted into mice already involved in disease studies so that data can be obtained without creating disease models just for that purpose and without causing any additional suffering apart from implanting the chip So far this technology has been used to monitor temperature in mice used in lymphoma studies In this procedure tumour cells infiltrate the lymphoid organs and the mice develop enlarged lymph nodes spleens and thymus glands but the tumour burden cannot be measured directly Food and water consumption decrease as tumours grow and mice develop clinical signs of sickness including hunching huddling together staring coats dull demeanour and ears pulled back from baseline position A decrease in mean body temperature was then found to predict tumour burden effectively in these mice 16 As another example Radio Frequency Identification RFID chips were implanted into mice used in a study to develop therapies for liver fibrosis which involved bile duct ligation This is a severe procedure and several measures are in place to refine it to reduce suffering such as providing heated areas in the cage and special attention to perioperative care and pain relief In addition to the body temperature data transponder technology was also used to monitor individual water consumption In this case body temperature did not correlate with the level of liver fibrosis but drinking bout duration did as this was significantly reduced in animals with fibrosis The use of enhanced clinical monitoring to identify biomarkers to help track disease progression therefore shows considerable promise provided that the most relevant biomarkers are identified for each model and experimental question Action points Question and challenge the use of endpoints that involve animals suffering from advanced disease states Ensure that indicators of pain suffering or distress are tailored to each disease study and can detect subtle signs of suffering 169

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting Keep up with current information on new techniques and approaches to monitoring animals using these to detect suffering and refine humane endpoints Refining rheumatoid arthritis a joint approach Sarah Allden and Tania Boden UCB Collagen Induced Arthritis CIA Glucose 6 Phosphate Isomerase G6PI ar thritis and Collagen Antibody Induced Arthritis CAIA are all polyarthritis models that are commonly induced in mice However these models differ with respect to the way they are induced their speed of onset and the severity of the arthritis suggesting that generic approaches to welfare assessment and refinement would not be appropriate The introduction of the G6PI and CAIA models therefore prompted a re evaluation of existing welfare scoring sheets alongside husbandry refinements such as improvements to environmental enrichment in order to improve the overall welfare of animals used in arthritis studies at our facility The original record sheet we used for assessing CIA mice was based on Wolfensohn and Lloyd 17 with scores reflecting progression in paw inflammation This was useful for the CIA mice but it soon became apparent that it would have to be tailored for the G6PI and CAIA models This triggered discussions involving scientists animal technologists and the NACWO to determine how best to go about modifying the way in which these animals should be assessed and monitored Good communication between all and consensus with respect to indicators and the terminology used to describe them helped us to make significant refinements to both our monitoring systems and procedures To begin with the mice used in G6PI and CAIA studies were very closely observed by both researchers and care staff particularly with respect to weight loss and disease progression We collated the data to assess how many individuals lost weight below a certain criterion and how many had a maximum disease score in order to see how we might reduce the number of animals approaching these levels Proactive interventions were established whenever the need arose for example weight loss was reduced by diet supplementation and humane endpoints were defined to prevent severe arthritis The observations were used to create a scoring system using a standardised terminology for describing and logging obser vations which was specifically tailored to each model Some indicators were common to all models such as weight loss appearance coat condition and behaviour isolation from cage mates The humane endpoint for weight 170 loss was refined by reducing the maximum from 25 to 20 and including another endpoint of a 15 weight loss that had not begun to reverse after 5 days Other indicators varied between models For CAIA some indicators that occurred in the short term following the administration of Lipopolysaccharides LPS were added soft stools ocular and nasal discharge Problems with mobility and dehydration were added for G6PI The use of tailored indicators for each model also enabled us to reduce the length of time that animals spent on procedures thereby reducing suffering for example the CIA studies were successfully reduced from 30 days to 20 The benefits of the tailored scoring systems are summarised in Table 1 below Old sheets New sheets Model specificity No Yes Cumulative suffering Not addressed Maximum scores over time taken into consideration Weight loss Endpoint of 25 or 20 over 72 hours Endpoint of 20 or 15 over 5 days Adverse events No specific details Model specific observations on sheets Individual disease score Total disease score not taken into account Distress scoring based on number of swollen paws not severity Total mouse disease score captures severity more effectively Table 1 Summary of distress sheet refinements in polyarthritis studies Alongside the work to refine welfare assessment we have worked to develop the environmental enrichment provided for the animals DBA 1 male mice are used in the CIA studies and territorial behaviour was observed when a refuge was provided in the cage We have found that simply adding another house has solved the problem as the dominant mouse takes possession of one house and the rest of the group uses the other one One of the houses has an integral running wheel to encourage activity before and sometimes after disease induction Different kinds of nesting material have been trialled to see which one stimulates nest

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting making and is also best for inflamed paws paper shavings are now provided because they are less likely to tangle than wool shavings When mobility becomes restricted during the acute phase of the disease longer nozzles are added to the water bottles so that animals do not have to rear up and place additional pressure on their hind limbs Food is also made accessible around the cage including supplementary diets to reduce weight loss especially following administration of LPS Refinement is a continuous process and we are always seeking further modifications For example we have begun to use the Mouse Grimace Scale18 for welfare assessment and refining endpoints and we are introducing non disease modifying pain management in some of our studies All of these factors combine to result in a proactive approach to better animal welfare and improved experimental results Note as part of our work on reducing severe suffering the RSPCA has set up an Expert Working Group on applying the 3Rs in rheumatoid arthritis research using mice and rats Tania is a member of the group and a report is currently being produced if you would like updates on progress with this contact research animals rspca org uk Action points Consider whether there is a similar joint approach to welfare assessment and refinement at your establishment If not how could you encourage this Review how effectively welfare assessment protocols are tailored to species strains and procedures at your establishment If you are involved with using or caring for animals in arthritis research consider some of the husbandry refinements in this section and contact the RSPCA to express an interest in the forthcoming report Are behavioural scientists aware of the natural history of the animals they work with and does it matter if they re not Colin Hendrie University of Leeds Much attention is given to the physical conditions in which laborator y animals are housed including environmental enrichment but very little attention is given to meeting their social needs There are compelling animal welfare and scientific reasons for understanding and catering for the behaviour of laboratory animals including rodents In the case of commonly used rodents rats are a colonial species and both sexes generally do well when housed in groups Mice on the other hand are highly territorial and this is reflected in the fighting that is commonly seen when males are group housed In a typical cage of group housed male mice one individual will become dominant with the rest being subordinates If group housed male mice are exposed to inescapable aggression this will cause severe suffering Besides these welfare problems animals of each social rank display different behavioural physiological and immunological profiles which will inevitably increase the variance in any experiment and can even render the results meaningless 19 Understanding the social behaviour of rodents should therefore be a top priority for any scientist using these animals in research and testing but it is not A metaanalysis of 100 research papers using mice and rats published in 2010 11 found that 99 of these failed to take into account the social organisation of these animals and the one that did got it wrong 20 A further online survey was conducted to explore the level of knowledge that behavioural researchers possess about their study species This asked questions about the biology and behaviour of a range of species including the fox rat mouse elephant and tree kangaroo the tree kangaroo was included to detect respondents who looked up the answers on the assumption that few people are even aware these animals exist Answers from behavioural scientists working with laboratory animals were analysed and compared with those from scientists working in other areas and members of the public The study found that the behavioural scientists knew very little about the animals they worked with and had no specialist knowledge of these animals beyond being familiar with their Latin names and some trivial physical characteristics such as adult weight and longevity These findings have implications for scientific quality and translatability In particular lines of GA mice are created using the behaviour of standard laboratory mice as the wild type with little or no knowledge of the behaviour social organisation or natural history of the wild mice from which these laboratory strains were derived This leads to misinterpretations of the behaviour of GA lines there are many examples in the literature where normal mouse behaviour has been mistaken for the effects of genetic alteration Examples of this are male mice fighting when one animal is introduced into the home cage of another and speciesand strain specific variations in responses in common models of anxiety such as the elevated plus maze Although it is commonly assumed that inbred or GA laboratory strains are behaviourally far removed from their wild types natural behaviours are in fact strongly conserved and innate This is effectively demonstrated by the Ratlife documentary in which laboratory rats 171

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting were released into a semi wild enclosure see www ratlife org and the ability of laboratory mice to recognise the calls of murine predators 21 The above indicate that there is a major educational task ahead This needs to encompass initial training and Continuing Professional Development for researchers Available training and resources should be complemented with input from animal technologists who have exper tise in animal biology and behaviour that could help with the interpretation of behavioural tests and also with experimental design However achieving this depends on effective communication and collaboration and appropriate status afforded to animal technologists and care staff Action points Suggest discussion of this issue at your establishment including general awareness of the importance of understanding behaviour and review of whether there is sufficient communication between people with different expertise including researchers animal technologists veterinarians and the NIO If you are a researcher consider how familiar you are with the behaviour and biology of your study species Would you like more training and if so would this be readily accessible If you are an animal technologist consider whether you are satisfied with the training and CPD that you have received do receive with respect to animal behaviour and natural history Would you like more training and if so would this be readily accessible Welfare assessment of Genetically Altered GA rodent lines what is out there Dominic Wells Royal Veterinary College There are a number of reasons for assessing the welfare of GA rodent lines In common with conventional strains effective welfare assessment allows the prompt identification and alleviation of any health or welfare problems which gives rise to both scientific and animal welfare benefits There is also a legal requirement under the revised ASPA for the actual severity experienced by each animal to be retrospectively assessed and repor ted A fur ther reason specific to GA animal use is the acquisition of information relevant to maximising welfare that can be passed on to others for example as part of the GA passport 22 Genetic alteration has the potential to cause suffering if the GA line is a model of a disease that causes pain 172 or distress if the gene disruption leads to a physical impairment or if there is an unexpected adverse effect of a random integration or a targeted gene disruption As a result welfare assessment of GA rodents can raise some specific issues over and above observing and monitoring conventional lines For example it may be difficult or impossible to predict adverse phenotypes and any welfare impact these may have on the animal and some adverse effects may not become apparent until certain developmental stages In the early days of GA rodent creation and use in the early to mid 1980s there was a wait and see approach to welfare assessment in which records of increased morbidity and mortality were simply analysed retrospectively Practice has moved on considerably since then and a number of studies have examined the welfare of GA mice 23 26 Score sheets for assessing and monitoring the welfare of GA mice were proposed by Mertens and R licke27 and van der Meer et al 28 and a 2003 review of the literature by Jegstrup and colleagues concluded that there was a clear need to develop a generally applicable and practical protocol 29 Similar conclusions had also been reached by the Animal Procedures Committee APC the independent body that advised the Secretary of State on the implementation of the ASPA until the end of 2012 when it was superseded by the Animals in Science Committee The APC produced a repor t on biotechnology in 2001 making a number of recommendations relating to the welfare assessment of GA animals particularly mice In response major UK research funders established an expert working group to define a flexible protocol for welfare assessment 30 This suggests appropriate welfare indicators and repor ting formats emphasising the need for comprehensive assessments of each new line from neonates to adults These assessments should be revised whenever breeding onto a different genetic background breeding to homozygosity or crossing with another GA line The 2011 Joint Working Group on Refinement guide to welfare assessment31 provides additional guidance intended for those responsible for assessing both conventional and GA animals It emphasises the importance of a team approach with input from researchers animal technologists and veterinarians plus input from or discussion with local ethical or animal care and use committees such as the UK AWERB if appropriate A longer version of the guidance includes recommendations on training and a list of fur ther resources see also www nc3rs org uk welfareassessment for some useful links The Biotechnology and Biological Sciences Research Council BBSRC Cancer Research UK Medical Research Council and Wellcome Trust

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting Most recently the European Commission has produced some guidance on welfare assessment including actual severity assessment 32 with some worked examples including GA animals 33 The Home Office has also produced advice notes on severity assessment of GA animals and on actual severity reporting 2 However there are currently still some areas where more clarity is needed with respect to classifying actual severity in GA mice These include G G G distinguishing between mild and subthreshold severity differentiating between knowledge of a GA line i e what is expected or what is known at a cellular level and clinical observations of the animals how to classify the severity of a sudden death e g in GA models of some cardiomyopathies Action points Ensure that the literature on welfare assessment especially with respect to GA animals is regularly reviewed and its approaches and recommendations implemented where appropriate at your establishment How and where to start with GA severity assessment Nikki Osborne RSPCA Research Animals Department Article 54 of Directive 2010 63 EU requires Member States to collect and report statistical information on the actual severity of the pain suffering distress or lasting harm experienced by each animal To help Member States comply with what was for most a new requirement the European Commission held two expert working group meetings on severity assessment in 2012 The first considered the specific severity issues related to the creation production and maintenance of GA animals while the second addressed severity assessment more generally These meetings resulted in the Working Document on a Severity Assessment Framework that was endorsed by the National Competent Authorities for the implementation of Directive 2010 63 EU in January 2013 32 On the 9th September 2013 the Wellcome Trust Sanger Institute hosted a meeting on Severity Assessment and Actual Severity in GA Animals This was attended by a range of staff including managers animal technologists and Named Persons from NB The Statistical Environment Reporting Framework for Directive 2010 63 EU states that animals from genetically altered lines include transgenic knock out and naturally occurring or induced mutant animals and other forms of genetic alteration regardless of phenotype research establishments using GA animals in the Cambridgeshire and Oxfordshire area as well as representatives of the Home Office Animals in Science Regulation Unit Some but not all of the represented establishments took part in the Home Office pilot study of new statistical repor ting standards and draft guidelines which ran from August to October 2013 A full report of the meeting is currently being prepared the summary points below represent an overview of some of the meeting outcomes The reasons for assessing actual suffering were understood inclusion of the actual suffering experienced by the animal provides greater transparency and understanding of the impact of scientific procedures on animal welfare This is not an additional burden but takes the observations that animal technologists already make on a daily basis and uses them to inform the severity assessment in doing so their expertise in knowing what is normal and what is not normal for the animals in their care will be acknowledged and contribute to improving animal welfare Severity assessment encompasses G identifying when an animal is experiencing pain suffering distress or lasting harm G recording whatever it is that is not quite right and G deciding how severely the identified animal is suffering Anyone who believes that an animal is experiencing pain suffering distress or lasting harm should make a record of it Signs of pain suffering distress or lasting harm are currently detected G during routine husbandry and care procedures e g cage change identification G during experimental procedures G during routine obser vations or welfare assessment checks or G during phenotyping procedures Generally all animals are observed for some reason at some point once a day see Hutchison this paper Observations are commonly noted by G discussing with a colleague NACWO NVS G entering them either physically or electronically into a structured database or spreadsheet G writing them down and filing them G writing them down and storing them somewhere with communal access or G noting and discussing them with the project licence holder or NVS The important point is that all observations should be recorded and kept in some traceable and reviewable way 173

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting Everything that is not right should be recorded as seen but this does not involve making a diagnosis For example observations may include reluctance to be handled closed eye wound on tail unsteady walking gait nasal discharge reduced food water intake not interacting with cage mates To enable consistency records should use a controlled vocabulary see www mousewelfareterms org This will also enable trends to be identified over time and data to be reviewed or analysed in an informative way Observations and clinical signs that could be used to assess severity are listed in Table 2 These have been sorted into the high level categories set out in the EC guidance 32 High level categories Commonly used indicators for GA rodents Appearance Weight body condition general appearance coat e g piloerection discharges e g ocular nasal mouth urogenital masses or growths pallor facial expressions Body functions Respiration food and fluid consumption cold to touch moribund breeding related indicators e g parturition neurological signs grip response Environment Presence location and consistency of faeces and urine Behaviours Mobility vocalisation audible socialisation responses to cage mates and humans convulsions mutilation of self or others provoked behaviours changes in response to environment writhing hunching lethargy Procedurespecific indicators Death pre weaning mortality Free observations There should always be a facility to note any other observations that may be unexpected or impact on animal welfare suffering Table 2 Examples of commonly used welfare indicators for GA rodents Clear GA relevant definitions were suggested by the delegates attending the Wellcome Trust meeting in September for use when assigning severity classifications Sub threshold looks and behaves like a wildtype mouse and has the same housing and husbandry requirements Mild housing and husbandry needs to be 174 adapted to maintain the mice in a normal condition e g they may require IVCs adapted feeding methods special diets or supplementary feeding or adjustments to environmental temperature Legal requirements and points of guidance were clearly understood by the group 1 The actual severity that should be reported annually to the Home Office as of 2014 is the peak severity that an individual animal has experienced e g if an animal experiences mild severity which then drops back below threshold the actual severity following the end of the procedure should be reported as mild 2 If an animal is identified as having exceeded the severity limit of the protocol this must be reported to your HOI at the earliest opportunity with the understanding that actual severity assessment will not occur until after the procedure has ended 3 If an animal is found dead actual severity should be classified as severe unless there is evidence to the contrary Other aspects were less clear with some outstanding questions 1 If there is a 2 spontaneous death rate in the background strain and a 4 death rate in the GA line all animals found dead must be counted it is not permissible to discount the first 2 but can and if so should background effects be taken into account 2 Would repeated mild procedures constitute moderate suffering if so when and how should cumulative effects be taken into account 3 Is it possible and is there a need to distinguish between husbandr y and procedure related effects Action points Discuss the summar y points with colleagues including the difficult questions under the last bullet point This can start or further inform the process at your establishment with respect to reporting actual severity Consider whether your establishment has all the necessary protocols in place to assess severity affectively in GA animals This includes both day today welfare assessments and the assessment of actual severity after the procedure has finished If your establishment does not already use www mousewelfareterms org suggest that it does

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting The importance of welfare assessment in an ageing rodent programme Marie Hutchison Mary Lyon Centre MRC Harwell A major aim of the ageing programme at MRC Harwell is to produce mouse mutants that can be used to study diseases that affect ageing human populations This is because age related diseases are becoming increasingly common which is putting pressure on society as well as compromising the health and wellbeing of those who are directly affected Large cohorts of ageing wild type mice are housed at Harwell with over 5 000 aged mice in the current screening programme These animals can live for 18 months so effective welfare assessment is essential to detect signs of suffering due to an animal s age or other adverse phenotype Basic yet thorough routine cage side observations are essential when looking for any abnormalities or welfare concerns in ageing animals Significant amounts of time are allocated for daily observations of animals to check their behaviour and appearance with open cage checks for closer inspection as needed Comprehensive in house training and familiarity with the normal appearance and behaviour of a healthy animal are especially important helping to ensure that even very subtle differences can be recognised and monitored from the time they appear A full nose to tail inspection is carried out of each animal including an examination of head shape facial features limbs behaviour and activity body shape digits tail length coat condition and colour This assessment is carried out in addition to standard phenotyping protocols and is done at different time points at 4 weeks of age then every 6 weeks for animals under 16 weeks or over 1 year old or every 12 weeks for animals aged between 16 and 52 weeks Standardised wording is used to describe observations as in www mousewelfareterms org to ensure consistency and continuity These observations and the accurate records that accompany them enable more subtle details to be picked up which has been crucial in the discovery of new phenotypes and the refinement of humane endpoints All mice exhibiting an adverse phenotype or any other welfare concern are recorded and examined throughout their lives to monitor any progression or degeneration in condition Weighing regimes are an informative tool in assessing the health of an individual animal and frequent weight data collection is an essential element in assessing if an animal s health is deteriorating However a degree of weight loss can be normal in some strains as the animals age or a line may be lean but healthy Weight should therefore be interpreted in conjunction with other indicators although rapid weight loss is always a cause for concern For most strains animals are weighed according to the same time points as the morphology check above Frequency increases to weekly if a loss of 8 or more is noted daily if a loss of over 15 is noted and animals are humanely killed if more weight is lost or if there are other complications In addition to increased weighing frequency mice with weight loss are palpated for growths and undergo a full welfare assessment Assessing the welfare of aged animals comes with some challenges we often have to consider carefully whether an animal is sick or just old but otherwise healthy For example hair loss and thinning is observed in ageing mice Figure 3 Elderly animals are also prone to benign lumps and swellings under the skin these are carefully monitored and lesions sometimes appear which are treated with a topical cream for three days after which animals are humanely killed if there is no improvement Some dominance behaviours such as barbering can also become more marked in the long term and offenders may have to be separated from the group and housed individually for short periods Figure 3 Normal hair loss in a healthy ageing mouse Photo credit MRC Harwell The data that we gather through careful open cage checks and daily observations have all contributed to establishing an effective routine of cage side welfare assessment and phenotyping Some phenotypes require very careful welfare assessment in order to identify when an animal has reached the humane endpoint for the par ticular line For example unexpected mortality was observed in one line with no obvious predictors which was a serious concern However with enhanced monitoring it was found that gait changes and continuous eye blinking occurred before death which has enabled humane endpoints to be defined and mortality to be reduced The phenotype is still under investigation but is thought to include a cardiac or neurological disorder As another example careful daily monitoring of the weight of another line with higher mortality than expected has enabled 175

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting humane endpoints to be defined so that the severity limit has been reduced from severe to moderate This line has a renal phenotype The above approach has reduced the effects of adverse phenotypes experienced by the mice and it is hoped that the aged lines of mice will be of use in studying human disease Animal technologists should not undervalue the role they play in assessing the welfare of animals as their key observational skills and the ability to pass this information on to other research and technical staff is essential in helping to ensure that the animals welfare requirements are met Action points If you use or care for ageing rodents set a target for reducing mor tality levels by identifying new predictors of death and refining humane endpoints Make sure that your group is in contact with others using aged animals to exchange ideas and information about monitoring animals reducing suffering and improving welfare Actual severity assessment and retrospective reporting Steve Ryder Home Office Inspectorate As mentioned earlier Directive 2010 63 EU requires Member States to collect and repor t statistical information on the actual severity of the pain suffering distress or lasting harm experienced by each animal This has been transcribed into the revised ASPA which came into effect in January 2013 and resulted in some changes to the way in which the annual Return of Procedures is collected and reported see ASPA Sec 21A 1 2 a 2 The major changes are 1 procedures are counted once completed as opposed to when started 2 the actual severity of all procedures must be assessed and recorded after the end of the procedure Assessing actual severity is a requirement for the annual Home Office Return of Procedures and is also a prerequisite if the project licence permits re use as there are restrictions relating to the severity of previous procedures As of 1 January 2013 the Project Licence Holder is responsible for ensuring that the actual severity of procedures ending during 2014 is assessed and collated for submission to the Home Office It is essential to be aware that actual severity must be judged from the day to day records of each animal s health and welfare not the prospective severity of the protocol It should reflect the worst experience of each 176 individual in terms of the impact on the animal and not the technique applied In practice the prospective and actual severity may be the same but this will not necessarily always be the case Actual severity will be reported according to the same categories as those used for prospective severity classification mild moderate severe and non recovery but there will also be a subthreshold category in which the actual severity experienced by the animal was below the threshold for regulation a level of pain suffering distress or lasting harm equivalent to or higher than that caused by the introduction of a needle in accordance with good veterinar y practice As a hypothetical example animals were given an altered diet and some were also given an intraperitoneal ip injection of a drug where neither the drug nor the diet was expected to be harmful The prospective limit was mild The altered diet was found not to cause any adverse effects so the actual severity for those animals only receiving the diet was subthreshold The animals receiving the intraperitoneal injection experienced transient discomfort but the drug had no adverse effects so their actual severity was mild The Guidance on the Operation of ASPA includes sections on defining severity categories and the classification of actual severity is addressed in more detail in a separate advice document 2 Some difficult issues were identified during the drafting process how to take account of procedural and non procedural effects how different species and stages of development experience suffering found deads cumulative suffering and GA animals Some special issues that arise with GA rodents include neonatal deaths and how these should be classified poor fertility late weaning and innate morbidity or mortality in background strains Genotyping methods and some phenotyping protocols e g SHIRPA also have the potential to cause discomfort or distress thus adding to cumulative suffering The UK currently regards breeding of GA animals as a regulated procedure with most currently licensed with a mild severity limit At present virtually all breeding of GA animals is within the regulatory process In future however only the breeding of lines with phenotypes that are above threshold should be regulated which will likely require some rethinking of the definition of mild for GA animals Article 30 of the Directive requires that records are kept of the number and species of animals bred but not used in regulated procedures These should be kept for at least 5 years and made available to the competent authority on request The Home Office proposes to collect these data either annually or relating to one year in every five commencing in 2017 GA animals will be distinguished from wild type providing an indication

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting of the number of animals humanely killed because they do not have the required genotype or are surplus to requirements who voted worked at establishments housing at least 1 000 GA animals Figure 4 The Home Office Guidance to the revised ASPA and the advice document on severity assessment are both now complete and published on the Home Office website 2 There will also be the ability to revise and update the Guidance and advice document as required see https www gov uk research and testing usinganimals for further information and to check the current status of these documents and other relevant guidance and advice notes Action points G Ensure that you are familiar with the guidance on assessing and reporting actual severity G Ask your Home Office Inspector if you are unsure about how to categorise actual severity in any difficult cases G Remember that actual severity assessment should provide information to help progress refinement and reduce suffering it is not just about the statistics G Consider what role the AWERB might play or what kind of oversight it will have regarding assessing actual severity Interactive session assessing actual severity in GA mice Figure 4 Number of GA animals housed at delegates facilities Routine welfare checks were generally carried out once a day 60 of the vote or twice a day 22 Observations were recorded whenever they were made 25 or whenever there was something to record 60 in most cases In some instances 7 5 records were only made when action was required Records were kept on paper 42 votes and or electronically 39 and or verbally 23 Two delegates responded that observations were not recorded Steve Ryder and Penny Hawkins Following on from the presentations on assessing GA animals an interactive TurningPoint session was used to find out more about current practice at delegates establishments and provide some hypothetical examples of adverse effects that could be observed in a breeding colony of GA mice An overview of the outcome is presented below note that the numbers of responses vary according to the number of people who voted or abstained or whether more than one option could be selected These are included in this report as an example of the delegates views on the day to stimulate thought and discussion for guidance on actual severity assessment please refer to the Home Office Most of the 90 delegates were animal technologists 38 votes NACWOs 11 or researchers students 7 The remainder included other Named Persons Named Training and Competency Officers Named Information Officers and Named Veterinar y Surgeons establishment licence holders with one Home Office Liaison Officer and three regulators it was possible to vote for more than one role The majority worked at an academic establishment 22 votes Government agency 13 or medical or veterinary research institute 14 with fewer delegates from industry commercial companies and welfare organisations Most delegates Not all GA animals were reported to undergo specific formal welfare assessment at the cage side although most did if there appeared to be a welfare problem or there was some scientific interest Figure 5 With respect to the actual severity assessment following the end of the procedure five delegates voted that this should be done by the researcher only In contrast 39 voted that the judgement on actual severity should be Figure 5 Responses to the question Do GA animals undergo specific formal welfare assessment in addition to phenotyping 177

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting made by the researcher with input from others Animal technologists 35 votes and vets 37 received the most votes as those who should help to make the decision Study Directors received 14 votes and the AWERB 12 while eight delegates agreed most with the statement it depends Situation Number of delegates voting that actual severity was severe 2 year old breeding stock found dead The offspring of a GA mouse who is weaned normally has no observable adverse phenotype is ear notched for genotyping and killed by a Schedule 1 method as surplus stock A nude mouse ear notched for identification and maintained within a barrier facility with no evidence of illness before being killed by a Schedule 1 method at 3 months old A GA mouse ear notched for identification from a line that develops spontaneous tumours but who is killed by a Schedule 1 method at 8 weeks old before tumours develop A GA mouse who dies as a result of a one off spontaneous seizure 0 20 8 3 Severe 0 Table 4 Delegates views on severe severity in relation to the breeding and husbandry of GA mice Delegates could vote for as many of the options as they wished Number of votes 24 21 8 0 5 7 1 hour ago 8 Found dead is always severe no matter when last observed 5 4 2 hours ago 17 8 This morning it is now 4pm 34 Yesterday On Friday today is Monday 37 0 Table 5 Delegates answers to the question would an animal found dead be classified as severe if they appeared normal when last seen 1 2 11 34 Table 3 Delegates views on actual severity in relation to the breeding and husbandry of GA mice 178 37 None of these 44 Moderate 25 A GA mouse with overgrown teeth that need regular clipping 15 Mild A GA mouse displaying almost continuous circling behaviour from which they cannot be distracted A female GA mouse developing internal tumours causing 10 weight loss and slight abdominal swelling while feeding a litter Below threshold 27 Death of a GA mouse due to facility infection with MHV that did not kill any non GA mice The rest of the interactive session was devoted to discussion of some hypothetical but realistic cases relating to the breeding of GA mice Tables 3 and 4 It appeared that most delegates regarded an animal found dead as experiencing severe severity on the basis of Tables 3 and 4 and also a question that aimed to explore whether the time that had elapsed between the last check and the animal being found made a difference Table 5 Action points Discuss the examples in Tables 3 to 5 at your establishment or make up some more to explore

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting views on severity and consider how effectively the assessment process is working Include staff with a range of roles discussing the reasons for different views Think about how an informed choice will be made as to the level of suffering that may have been experienced by animals found dead on the basis of the type of procedures conducted locally Encourage colleagues not to be defeatist about the potential to reduce mortality as progress can be made by refining monitoring and reviewing records to identify indicators that can be used as humane endpoints 12 13 Acknowledgements 14 Thank you to the speakers and delegates for the very useful and interesting talks and discussions Thanks also to Steve Ryder for his valuable help with the discussion session 15 References 21 22 23 24 25 26 27 28 29 10 11 European Commission 2010 Directive 2010 63 EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes Official Journal of the European Union L276 33 79 See https www gov uk research and testing usinganimals last viewed 13 February 2014 Hawkins P Playle L Golledge H Leach M Banzett R Coenen A Cooper J Danneman P Flecknell P Kirkden R Niel L and Raj M 2006 Newcastle Consensus Meeting on Carbon Dioxide Euthanasia of Laborator y Animals Download at http www nc3rs org uk news asp id 292 last viewed 6 November 2013 Danneman P J Stein S and Walshaw S O 1997 Humane and practical implications of using carbon dioxide mixed with oxygen for anesthesia or euthanasia of rats Laboratory Animal Science 47 376 385 Makowska I J Vickers L Mancell J and Weary D M 2009 Evaluating methods of gas euthanasia for laboratory mice Applied Animal Behaviour Science 121 230 235 Niel L and Weary D M 2007 Rats avoid exposure to carbon dioxide and argon Applied Animal Behaviour Science 107 100 109 Kirkden R D Niel L Lee G Pfaffinger M J Makowska I J and Weary D M 2008 The validity of using an approach avoidance test to measure the strength of aversion to carbon dioxide in rats Applied Animal Behaviour Science 114 216 234 Makowska I J and Weary D M 2009 Rat aversion to induction with inhalant anaesthetics Applied Animal Behaviour Science 119 229 235 Wong D Makowska I J and Weary D M 2013 Rat aversion to isoflurane versus carbon dioxide Biology Letters 9 doi 10 1098 rsbl 2012 1000 Hu J Zhong C Ding C Chi Q Walz A Mombaerts P Matsunami H and Luo M 2007 Detection of nearatmospheric concentrations of CO2 by an olfactor y subsystem in the mouse Science 317 953 957 Johnson P Fitz S D Hollis J H Moratalla R Lightman S L Shekhar A and Lowry C A 2011 16 17 18 19 20 21 22 Induction of C Fos in panic defence related brain circuits following brief hypercarbic gas exposure Journal of Psychopharmacology 25 26 36 Ziemann A E Allen J E Dahdaleh N S Drebot I I Coryell M W Wunsch A M Lynch C M Faraci F M Howard M A 3rd Welsh M J and Wemmie J A 2009 The amygdala is a chemosensor that detects carbon dioxide and acidosis to elicit fear behavior Cell 139 1012 1021 doi 10 1016 j cell 2009 10 029 Workman P Aboagye E O Balkwill F Balmain A Bruder G Chaplin D J Double J A Everitt J Farningham D A Glennie M J Kelland L R Robinson V Stratford I J Tozer G M Watson S Wedge S R and Eccles S A 2010 Guidelines for the welfare and use of animals in cancer research British Journal of Cancer 102 1555 1577 Franco N H Correla Neves M and Olsson I A S 2012 How humane is your endpoint refining the science driven approach for termination of animal studies of chronic infection PLoS Pathogens 8 e1002399 Littin K Acevedo A Browne W Edgar J Mendl M Owen D Sherwin C W rbel H and Nicol C 2008 Towards humane end points behavioural changes precede clinical signs of disease in a Huntington s disease model Proceedings of the Royal Society B 275 1856 1874 Hunter J E Butterworth J Perkins N D Bateson M and Richardson C A 2014 Using body temperature food and water consumption as biomarkers of disease progression in mice with E myc lymphoma British Journal of Cancer doi 10 1038 bjc 2013 818 download at http www nature com bjc journal vaop ncurrent full bjc2013818a html last viewed 13 January 2014 Wolfensohn S and Lloyd M 2013 Handbook of Laboratory Animal Management and Welfare 4th edition Wiley Blackwell Oxford Langford D J Bailey A L Chanda M L Clarke S E Drummond T E Echols S Glick S Ingrao J KlassenRoss T LaCroix Fralish M L Matsumiya L Sorge R E Sotocinal S B Tabaka J M Wong D van den Maagdenberg A M J M Ferrari M D Craig K D and Mogil J S 2010 Coding of facial expressions of pain in the laboratory mouse Nature Methods 7 447 449 Hendrie C A and Pickles A 2013 The failure of the antidepressant drug discover y process is systemic Journal of Psychopharmacology 27 5 407 416 download at http tinyurl com qfy6c6m last viewed 4 November 2013 Hendrie C A and Pickles A R 2011 Behavioural scientists appear not to have specialist knowledge of laborator y animal behaviour Journal of Psychopharmacology 25 Suppl 8 A35 download at http tinyurl com obt3gmf last viewed 4 November 2013 Hendrie C A Weiss S M and Eilam D 1996 Exploration and predation models of anxiety evidence from laborator y and wild species Pharmacology Biochemistry and Behaviour 54 13 20 Osborne N Bussell J Carver R Chadwick C Enser B Haskings M A Hubrecht R Latcham J Maconochie M Mallon A M Norrington R and Wilson S 2010 GA Passports The Key to Consistent Animal Care Wellcome Trust Sanger Institute RSPCA Southwater Download at http www rspca org uk 179

Report of the 2013 RSPCA UFAW Rodent Welfare Group meeting 23 24 25 26 27 28 29 30 31 32 33 sciencegroup researchanimals implementing3rs gapass port last viewed 5 November 2013 Costa P 1997 Neuro behavioural tests in welfare assessments of transgenic animals In Proceedings of the 6th FELASA Symposium on International Harmonization of Laborator y Animal Husbandr y Requirements Royal Society of Medicine Press London pp 51 3 Francis R J 1997 Being a Transgenic Animal can Seriously Damage your Health MSc thesis University of Bristol Langford UK Dennis M B 2000 Humane endpoints for genetically engineered animal models ILAR Journal 41 94 98 Dennis M B 2002 Welfare issues of genetically modified animals ILAR Journal 43 100 109 Mertens C and R licke T 1999 Score sheets for the monitoring of transgenic mice Animal Welfare 8 433438 van der Meer M Rolls A Baumans V Olivier B and van Zutphen L F M 2001 Use of score sheets for welfare assessment of transgenic mice Laborator y Animals 35 379 389 Jegstrup I Thon R Hansen A K and Ritskes Hoitinga M 2003 Characterization of transgenic mice a comparison of protocols for welfare evaluation and phenotype characterisation of mice with a suggestion on a future certificate of instruction Laboratory Animals 37 1 9 GA Mouse Welfare Assessment Working Group 2006 Assessing the welfare of Genetically Altered mice Summar y in Laborator y Animals 40 111 114 or download in full at www nc3rs org uk page asp id 231 last viewed 6 November 2013 Joint Working Group on Refinement 2011 A guide to defining and implementing protocols for the welfare assessment of laboratory animals Laboratory Animals 45 1 13 European Commission 2012 Working Document on a Severity Assessment Framework Download at http tinyurl com 99oo3hv last viewed 6 November 2013 European Commission 2013 Worked examples of actual severity assessment Download at http tinyurl com ozwvkah last viewed 6 November 2013 180

December 2014 Animal Technology and Welfare PAPER SUMMARY TRANSLATIONS INHALTVERZEICHNIS Bericht zur zweiten RSPCA AHVLA Tagung zum Thema Tierschutz von landwirtschaftlichen Nutztieren in der Forschung Rinder Schweine Schafe und Gefl gel PENNY HAWKINS1 SHARON BROOKES2 SUSANNAH PARKIN3 R EDDIE CLUTTON4 PETER GADE5 JULIE LANE6 HELEN PROCTOR7 JOANNE EDGAR8 ISOBEL VINCENT9 und UTE WEYER2 1 2 3 4 5 6 7 8 9 Research Animals Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Animal Health and Veterinary Laboratories Agency AHVLA Animal Sciences Unit Weybridge Addlestone Surrey KT15 3NB Canterbury College New Dover Road Canterbury Kent CT1 3AJ Royal Dick School of Veterinary Studies University of Edinburgh Easter Bush Campus Midlothian EH25 9RG Novo Nordisk A S Novo All 2880 Bagsvaerd D nemark National Wildlife Management Centre AHVLA Sand Hutton York YO41 1LZ World Animal Protection 222 Grays Inn Road London WC1X 8HB University of Bristol School of Veterinary Sciences Langford House Langford Bristol BS40 5DU Royal Veterinary College RVC Clinical Skills Centre Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Korrespondierende autorin penny hawkins rspca org uk Abstract RSPCA und AHVLA veranstalteten im September 2014 gemeinsam eine Tagung f r Tiertechniker Wissenschaftler Tier rzte und Studenten mit Interesse am Tierschutz von zu Forschungs und Testzwecken dienenden Rindern Schweinen Schafen und Gefl gel In den Gespr chen und Diskussionen der Tagung ging es um unterschiedlichste Themen wie Endpunkt Verfeinerung der Gefl gelpest Studien Reduzierung der Zahl von Nutztieren f r die Forschung Schmerzmanagement bei Schweinen Verbesserungen der Unterbringung von einzeln gehaltenen Schweinen Nutzung des Cortisolspiegels zur Prognose des Wohlbefindens von Nutztieren F rderung positiver Tierschutzma nahmen f r H hner und Ersatz von Mutterschafen bei der Ausbildung Eine Diskussionsrunde ber positive Tierschutzma nahmen f r Nutztiere bildete den Abschluss des Programms 181

Paper Summary Translations Bericht zur Tagung der RSPCA UFAW Tierschutz Gruppe f r Nager 2013 PENNY HAWKINS SECRETARY 1 PAUL LITTLEFAIR2 HUW GOLLEDGE3 CLAIRE RICHARDSON3 SARAH ALLDEN4 TANIA BODEN4 COLIN HENDRIE5 DOMINIC WELLS6 NIKKI OSBORNE1 MARIE HUTCHISON7 STEPHEN RYDER8 MAGGY JENNINGS1 und ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS International Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Newcastle University Newcastle Upon Tyne NE1 7RU UCB Pharma Ltd 208 Bath Road Slough Buckinghamshire SL1 3WE University of Leeds Institute of Psychological Sciences Leeds Yorkshire LS2 9JT Comparative Biomedical Sciences The Royal Veterinary College Royal College Street London NW1 0TU Mary Lyon Centre MRC Harwell Harwell Science and Innovation Campus Oxfordshire OX11 0RD Home Office Animals in Science Regulation Unit 2 Marsham Street London SW1P 4DF UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Korrespondierende autorin penny hawkins rspca org uk Einleitung Die RSPCA UFAW Tierschutz Gruppe f r Nager veranstaltet jeden Herbst ein eint giges Treffen bei dem die Mitglieder ber Themen aktueller Tierschutz Forschung diskutieren und Erfahrungen und Meinungen ber nagerrelevante Tierschutz Probleme sowie ber die drei Prinzipien Vermeidung Reduktion und Verfeinerung des Einsatzes von Nagern austauschen Ein Hauptziel der Gruppe ist es zum Nachdenken ber die Gesamtheit der Lebenserfahrungen von Nagern als Laborversuchstiere anzuregen und sicherzustellen dass jede potenzielle Beeintr chtigung ihres Wohlbefindens gepr ft und auf ein Minimum gesenkt wird Das Meeting 2013 war die 20 Jubil umsveranstaltung der Nager Tagungen und wurde mit einer Pr sentation ber Tierschutzbelange in China eingeleitet Dabei ging es auch um die Arbeit der RSPCA in Bezug auf Laborversuchstiere Weitere Redner hielten Vortr ge ber Abneigung gegen Einschl ferungsmittel Beurteilung von Wohlbefinden und Reduzierung von Leiden bei Nagerkrankheitsmodellen sowie ber die Bedeutung des Verstehens von Nagerverhalten bei der Konzipierung von Versuchen und Tierhaltung und bei der Interpretation von Daten Am Schluss all dieser Pr sentationen standen Aktionspunkte Auf der Tagung fand auch eine Sondersitzung ber die Beurteilung des Wohlbefindens von genetisch ver nderten GV Nagern mit Vortr gen zu entsprechenden aktuellen Praktiken sowie eine interaktive Diskussionsrunde statt 182

December 2014 Animal Technology and Welfare CONTENU DE LA REVUE Rapport de la deuxi me r union de la RSPCA AHVLA sur le bien tre des animaux d levage en recherche bovins porcins ovins et volaille PENNY HAWKINS1 SHARON BROOKES2 SUSANNAH PARKIN3 R EDDIE CLUTTON4 PETER GADE5 JULIE LANE6 HELEN PROCTOR7 JOANNE EDGAR8 ISOBEL VINCENT9 et UTE WEYER2 1 2 3 4 5 6 7 8 9 Research Animals Department Service de la recherche animale RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Animal Health and Veterinary Laboratories Agency AHVLA Agence de la sant animale et des laboratoires v t rinaires Animal Sciences Unit Weybridge Addlestone Surrey KT15 3NB Canterbury College New Dover Road Canterbury Kent CT1 3AJ Royal Dick School of Veterinary Studies cole royale d tudes v t rinaires Dick Universit d dimbourg Easter Bush Campus Midlothian EH25 9RG Novo Nordisk A S Novo All 2880 Bagsvaerd Danemark National Wildlife Management Centre Centre national de gestion de la vie sauvage AHVLA Sand Hutton York YO41 1LZ World Animal Protection Soci t mondiale de protection des animaux 222 Grays Inn Road Londres WC1X 8HB Universit de Bristol cole des sciences v t rinaires Langford House Langford Bristol BS40 5DU Royal Veterinary College RVC Centre de comp tences cliniques Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Auteur ressource penny hawkins rspca org uk R sum La RSPCA et l AHVLA ont conjointement convoqu une r union rassemblant les technologues de la sant animale les chercheurs les v t rinaires et les tudiants ayant un int r t dans le bien tre des bovins porcins ovins et de la volaille utilis s dans la recherche et les tests lors d un programme de conf rences et de s ances de discussion La r union qui a eu lieu en septembre 2014 a port sur toute une s rie de th mes dont le raffinement des crit res de jugement utilis s dans les tudes men es sur la grippe aviaire la r duction du nombre d animaux d levage utilis s dans la recherche la gestion de la douleur chez les porcs l am lioration des locaux h bergeant des porcs individuellement l utilisation de niveaux de cortisol pour pr dire le bien tre des animaux d levage la promotion du bien tre positif de la volaille et le remplacement des brebis dans l ducation et la formation Une s ance de discussion sur le bien tre positif des animaux d levage a conclu le programme 183

Paper Summary Translations Rapport de la r union RSPCA UFAW 2013 du groupe de protection du bien tre des rongeurs PENNY HAWKINS SECR TAIRE 1 PAUL LITTLEFAIR2 HUW GOLLEDGE3 CLAIRE RICHARDSON3 SARAH ALLDEN4 TANIA BODEN4 COLIN HENDRIE5 DOMINIC WELLS6 NIKKI OSBORNE1 MARIE HUTCHISON7 STEPHEN RYDER8 MAGGY JENNINGS1 et ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Service de la recherche animale Groupe scientifique RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Service international RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Universit de Newcastle Newcastle Upon Tyne NE1 7RU UCB Pharma Ltd 208 Bath Road Slough Buckinghamshire SL1 3WE Universit de Leeds Institut des sciences psychologiques Leeds Yorkshire LS2 9JT Sciences biom dicales comparatives Royal Veterinary College Coll ge royal de sciences v t rinaires Royal College Street Londres NW1 0TU Centre Mary Lyon MRC Harwell Campus Harwell de science et d innovation Oxfordshire OX11 0RD Bureau de r glementation sur l utilisation scientifique des animaux Minist re de l Int rieur 2 Marsham Street Londres SW1P 4DF UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Auteur ressource penny hawkins rspca org uk R sum Le groupe de protection du bien tre des rongeurs de la RSPCA UFAW tient chaque automne une r union d une journ e afin que ses membres puissent discuter de la recherche actuelle en mati re de bien tre changer des vues sur les questions relatives au bien tre des rongeurs et partager des exp riences de mise en uvre des 3R de Remplacement de R duction et de Raffinement l gard de l utilisation des rongeurs L un des objectifs cl s du Groupe consiste encourager la r flexion sur l ensemble de l exp rience de vie des rongeurs de laboratoire assurant que chaque impact n gatif potentiel sur leur bien tre soit revu et r duit au minimum Pour marquer le 20 me anniversaire des r unions sur les rongeurs la rencontre de 2013 a commenc par une pr sentation sur les questions de protection des animaux en Chine introduisant notamment le travail de la RSPCA sur les animaux de laboratoire le 20 me anniversaire de mariage tant l anniversaire chinois D autres conf renciers ont pr sent des expos s sur l aversion que suscitent les agents d euthanasie valuant le bien tre et r duisant les souffrances des mod les de rongeurs malades ainsi que sur l importance de comprendre le comportement des rongeurs lors de la conception d exp riences et d levage et lors de l interpr tation des donn es Toutes les pr sentations se terminent par quelques points d action La r union comprenait galement une session sp ciale sur l valuation du bien tre des rongeurs g n tiquement modifi s GA accompagn e de d bats sur la pratique actuelle et d une session de discussion interactive 184

December 2014 Animal Technology and Welfare INDICE DE LA REVISTA Acta de la segunda reuni n de RSPCA AHVLA acerca del bienestar de los animales de granja en la investigaci n ganado cerdos ovejas y aves de corral PENNY HAWKINS1 SHARON BROOKES2 SUSANNAH PARKIN3 R EDDIE CLUTTON4 PETER GADE5 JULIE LANE6 HELEN PROCTOR7 JOANNE EDGAR8 ISOBEL VINCENT9 y UTE WEYER2 1 2 3 4 5 6 7 8 9 Departamento de Investigaci n Animal RSPCA Wilberforce Way Southwater West Sussex RH13 9RS AHVLA Agencia de Laboratorios de Salud Animal y Veterinaria Unidad de Ciencias Animales Weybridge Addlestone Surrey KT15 3NB Colegio de Canterbury New Dover Road Canterbury Kent CT1 3AJ Colegio Real Dick de Estudios Veterinarios Universidad de Edinburgh campus Easter Bush Midlothian EH25 9RG Novo Nordisk A S Novo All 2880 Bagsvaerd Dinamarca Centro Nacional para la Gesti n de la Fauna AHVLA Sand Hutton York YO41 1LZ Protecci n Animal Mundial 222 Grays Inn Road Londres WC1X 8HB Universidad de Bristol Facultad de Ciencias Veterinarias Langford House Langford Bristol BS40 5DU RVC Colegio Real de Veterinaria Centro de Aptitudes Cl nicas Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Autora del documento penny hawkins rspca org uk Resumen RSPCA y AHVLA celebraron una reuni n de mutuo acuerdo con el fin de congregar a tecn logos de animales de laboratorio investigadores veterinarios y estudiantes interesados en el bienestar de ganado cerdos ovejas y aves para realizar un programa de charlas y sesiones de debate El evento se celebr en septiembre de 2014 y en l se abord una amplia gama de tem ticas como por ejemplo puntualizar los criterios aplicados en los estudios de la gripe aviar la reducci n del uso de animales de granja en la investigaci n la gesti n del dolor en pr cticas con cerdos una mejora de las condiciones de las granjas de cerdos aislados el uso de cortisol para predecir el bienestar de los animales de granja el fomento del bienestar de las aves de corral y la sustituci n de ovejas por otras especies en la educaci n y en la formaci n El acto concluy con una sesi n de debate entre los asistentes acerca del bienestar de los animales de granja 185

Paper Summary Translations Acta de la reuni n del Grupo de apoyo al bienestar de los roedores RSPCA UFAW celebrada en 2013 PENNY HAWKINS SECRETARIA 1 PAUL LITTLEFAIR2 HUW GOLLEDGE3 CLAIRE RICHARDSON3 SARAH ALLDEN4 TANIA BODEN4 COLIN HENDRIE5 DOMINIC WELLS6 NIKKI OSBORNE1 MARIE HUTCHISON7 STEPHEN RYDER8 MAGGY JENNINGS1 y ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Departamento de Investigaci n sobre Animales Grupo cient fico de la RSPCA Real Sociedad para la Prevenci n de la Crueldad contra los Animales Wilberforce Way Southwater West Sussex RH13 9RS Departamento Internacional RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Universidad de Newcastle Newcastle Upon Tyne NE1 7RU UCB Pharma Ltd 208 Bath Road Slough Buckinghamshire SL1 3WE Universidad de Leeds Instituto de Ciencias Psicol gicas Leeds Yorkshire LS2 9JT Ciencias Biom dicas Comparativas Colegio Real de Veterinaria Royal College Street Londres NW1 0TU Mary Lyon Centre MRC Harwell campus Harwell para la Ciencia y la Innovaci n Oxfordshire OX11 0RD Ministerio del Interior ASRU Unidad de regulaci n del estudio de los animales en la Ciencia Marsham Street n 2 Londres SW1P 4DF UFAW Federaci n de universidades en pos del bienestar animal The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Autora del documento penny hawkins rspca org uk Introducci n El grupo de apoyo al bienestar de los roedores RSPCA UFAW se re ne una vez al a o cada oto o para que sus miembros puedan comentar y cambiar impresiones sobre estudios recientes acerca del bienestar animal y compartir experiencias sobre la puesta en pr ctica de 3R de sustituci n reducci n y mejora del uso de roedores en las pr cticas El Grupo pretende principalmente concienciar sobre la experiencia que tienen los roedores en su ciclo de vida en un laboratorio con el fin de detectar y reducir cualquier impacto negativo en su bienestar En conmemoraci n del 20 aniversario de las reuniones de este Grupo la celebrada en 2013 comenz con la presentaci n de la problem tica del uso de animales en China en la que se cit el estudio de RSPCA realizado acerca de los animales de laboratorio ya que su 20 aniversario es el aniversario de China Otros asistentes a la reuni n expusieron la aversi n a los m todos de eutanasia mediante la evaluaci n del bienestar y la reducci n del sufrimiento en modelos de enfermedad de roedores y mencionaron la importancia que tiene entender el patr n de comportamiento de los roedores a la hora de dise ar experimentos durante su cr a y en la interpretaci n de datos Al final de cada presentaci n se expuso un plan de acci n La reuni n tambi n incluy una sesi n especial sobre la evaluaci n del bienestar de roedores transg nicos GA por sus siglas en ingl s que se centr en las pr cticas actuales y que concluy con una sesi n de debate entre todos los asistentes 186

December 2014 Animal Technology and Welfare INDICE DELLA REVISTA Relazione sul secondo incontro tra RSPCA e AHVLA sul benessere degli animali da allevamento impiegati nella ricerca bovini suini ovini e pollame PENNY HAWKINS1 SHARON BROOKES2 SUSANNAH PARKIN3 R EDDIE CLUTTON4 PETER GADE5 JULIE LANE6 HELEN PROCTOR7 JOANNE EDGAR8 ISOBEL VINCENT9 e UTE WEYER2 1 2 3 4 5 6 7 8 9 Research Animals Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Animal Health and Veterinary Laboratories Agency AHVLA Animal Sciences Unit Weybridge Addlestone Surrey KT15 3NB Canterbury College New Dover Road Canterbury Kent CT1 3AJ Royal Dick School of Veterinary Studies University of Edinburgh Easter Bush Campus Midlothian EH25 9RG Novo Nordisk A S Novo All 2880 Bagsvaerd Danimarca National Wildlife Management Centre AHVLA Sand Hutton York YO41 1LZ World Animal Protection 222 Grays Inn Road Londra WC1X 8HB University of Bristol School of Veterinary Sciences Langford House Langford Bristol BS40 5DU Royal Veterinary College RVC Clinical Skills Centre Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Autore corrispondente penny hawkins rspca org uk Introduzione RSPCA e AHVLA hanno congiuntamente indetto un incontro incentrato su una serie di relazioni e sessioni di discussione al fine di riunire tecnici stabularisti ricercatori veterinari e studenti interessati al benessere di bovini suini ovini e pollame utilizzati per fini scientifici e sperimentali L incontro tenutosi a settembre 2014 ha affrontato diverse tematiche tra cui la messa a punto degli obiettivi negli studi dell influenza aviaria la riduzione del numero di animali da allevamento impiegati nella ricerca la gestione del dolore nei suini il miglioramento dei sistemi di stabulazione per i suini in stabulari individuali l utilizzo dei livelli di cortisolo per valutare il benessere degli animali da allevamento la promozione degli effetti positivi del benessere per il pollame e la sostituzione di ovini femmine nell addestramento La riunione si conclusa con una sessione di discussione sul benessere positivo negli animali da allevamento 187

Paper Summary Translations Relazione sull incontro del RSPCA UFAW Rodent Welfare Group Gruppo sul benessere dei roditori del 2013 PENNY HAWKINS SEGRETARIA 1 PAUL LITTLEFAIR2 HUW GOLLEDGE3 CLAIRE RICHARDSON3 SARAH ALLDEN4 TANIA BODEN4 COLIN HENDRIE5 DOMINIC WELLS6 NIKKI OSBORNE1 MARIE HUTCHISON7 STEPHEN RYDER8 MAGGY JENNINGS1 e ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS International Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Newcastle University Newcastle Upon Tyne NE1 7RU UCB Pharma Ltd 208 Bath Road Slough Buckinghamshire SL1 3WE University of Leeds Institute of Psychological Sciences Leeds Yorkshire LS2 9JT Comparative Biomedical Sciences The Royal Veterinary College Royal College Street Londra NW1 0TU Mary Lyon Centre MRC Harwell Harwell Science and Innovation Campus Oxfordshire OX11 0RD Home Office Animals in Science Regulation Unit 2 Marsham Street Londra SW1P 4DF UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Autore corrispondente penny hawkins rspca org uk Introduzione Durante il periodo autunnale RSPCA UFAW Rodent Welfare Group Gruppo sul benessere dei roditori indice annualmente un incontro della durata di un giorno permettendo ai suoi membri di discutere degli attuali studi di ricerca sul benessere di scambiare opinioni sulle problematiche inerenti al benessere dei roditori e di condividere esperienze sull applicazione del principio delle 3R sostituzione replacement riduzione reduction e perfezionamento refinement nell utilizzo di roditori a scopi scientifici Uno degli obiettivi principali del Gruppo incoraggiare le persone a considerare l esperienza di vita dei roditori da laboratorio nel suo insieme assicurando il controllo e la riduzione di ogni possibile impatto negativo sul loro benessere Per celebrare il 20 anniversario dei Rodent Meetings Incontri sui roditori la riunione del 2013 si aperta con una presentazione sulle problematiche inerenti al benessere degli animali in Cina incluso il lavoro di RSPCA per gli animali da laboratorio il 20 anniversario costituisce l anniversario della Cina Inoltre sono state esposte relazioni sull avversione agli agenti eutanasici valutando il benessere e riducendo il grado di sofferenza nei roditori utilizzati come modelli e con un determinato quadro patologico e sull importanza di comprendere il comportamento dei roditori nella delineazione degli esperimenti e dell allevamento e nell interpretazione dei dati Tutte le relazioni si sono concluse con la messa a punto di alcuni provvedimenti Durante l incontro si inoltre tenuta una sessione speciale incentrata sulla valutazione del benessere nei roditori geneticamente modificati GM con discussioni sull attuale prassi e un forum interattivo 188

December 2014 Animal Technology and Welfare TECH 2 TECH Haven 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 any aspects of animal technology Comments observations descriptions of new or refined techniques 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 be presented as technical notes and do not need to be structured and can be as short or as long as is necessary Accompanying illustrations and or photos should be high resolution NB Descriptions of new products or equipment submitted by manufacturers are welcome but should be a factual account of the product However the Editorial Board gives no warranty as to the accuracy or fitness for purpose of the product Welfare first how good is your induction programme MATHEW SANDERSON Agenda Resource Management Ltd PO Box 24 Hull HU12 8YJ Corresponding author mathews agenda rm co uk Introduction The welfare of the people who care for animals is at the centre of the Welfare First programme developed by Agenda Resource Management ARM as it is the company s belief that excellent animal welfare depends upon well trained and dedicated individuals who care passionately about animals and their welfare One of the key areas in the Welfare First programme is training and development which starts from the very first day a new employee starts work Employing a large number of animal technicians who work in more than 25 different locations throughout the United Kingdom we operate shared responsibility for health and safety under the Health Safety at Work Act 1974 between our clients and ourselves It is therefore vital that we ensure all of our technicians receive a thorough induction at the start of a new placement What is the point of induction When a new team member starts working at a facility it is essential to assist them to become productive and efficient as quickly as possible and staff induction training forms a pivotal part in this process One of the difficulties many new starters face is that they quickly become overwhelmed with information It is therefore important to remember that induction is a process that should take place over a number of weeks However although not a one day event some of the information is vital on day one Who needs an induction All employees require an induction The character of the induction will vary but all staff need some form of programme whether they are in academic technical professional or suppor t roles senior or junior appointments full or part time permanent or on a short term contract new to the organisation or an internal appointment The quantity of information depending on the role and or situation It is an ARM company requirement that contract technicians complete the induction part of their training records within the first two weeks of starting a new placement We believe that this is vital to enable them to become an integral part of the team within a facility and helps ensure that the health and safety requirements of both our clients and ourselves are met 189

Tech 2 Tech Some parts of the induction programme are essential under the Health and Safety at Work Act 1974 and others are required for the new starter to familiarise themselves with the physical environment the establishment s culture and the on site working procedures and policies We separate the induction part of our training records into five sections First Aid Health and Safety and Environment Introduction to the site Induction to the role Additional information These are broken down further into specific topics that are considered essential to the induction programme although not all topics will be applicable to every facility Consequently the information is reviewed after the initial two weeks of a new placement Overview of the induction process To ensure that our technicians are safe and have all of the information that they require when starting at a new placement we have completed a sur vey of the induction processes at twenty facilities Facilities included academic pharmaceutical and Government establishments with the following results Data has been anonymised and shown in the following table TOTAL First Aid Health and Safety and Environment Introduction to the site 190 How to access First Aid First Aid Equipment obtained through First Aiders First Aid Procedure Reporting Accidents and Incidents 20 20 20 Health and Safety Overview Health and Safety Policies and Procedures Health and Safety at Work etc Act 1974 Health and Safety Induction Biohazards and Clinical Waste Management and Storage Location and Access to Risk Assessments and COSHH Assessments Induction to COSHH Disposal of Sharps Handling Spillages Display Equipment Usage and Assessment if applicable Lone Working Gas Cylinder Storage and Use Manual Handling PPE and LAA 19 19 19 16 20 15 16 16 16 10 16 9 18 18 Full site tour Tour of the Facility Facility Access Given Organisational Structure of the Facility General Security Procedures Site Car Parking if applicable 18 20 17 20 20 12 20 20 19 Sickness absence procedure notification of absence Working hours lunch and break time s arrangements overtime and rotas Introduction to colleagues 19 20 20 Additional information What should be included in the induction programme Fire Alarm Procedure including emergency numbers First Aid and Near Miss Procedure Issue of Safety Shoes Introduction to the role and our technicians deliver high standards of animal welfare Computer use and access if applicable Waste Disposal Read and Understood SOPs Animal Rights Activists Security Awareness Training Security Guidance demonstration etc Social Media and Internet Security for Staff 16 20 15 16 8 11 10 Having reviewed the data gaps in induction have been filled by providing additional training on security awareness and security guidance as well as safety information on the use of social media to all of our staff via our Welfare First newsletter and additional elearning and online training We know that not all of our technicians are given computer access so this could be why they do not complete a Display Screen Equipment DSE assessment but we believe that it is essential to provide all of our staff with information on the use of display screen equipment and we provide this along with advice on manual handling Laboratory Animal Allergens and details of Personal Protective Equipment and face mask fit testing as part of our Agenda induction programme We have concluded that as a lot of facilities are now moving away from using gas cylinders to piped gas this is the reason why no training on gas cylinder storage and use is being completed but we do ask if they are using cylinders when we review the induction information after the initial two weeks Evaluating the induction process Evaluating the effectiveness of the induction programme is vital to ensure that all the areas have been covered fully This can be done by speaking with the individual the manager and the trainer at the induction conclusion By receiving feedback the induction programme can be reviewed and changed to meet both the individuals and organisations needs At ARM we review the completed induction part of our technician training records at every placement after the first two weeks and supplement with any additional information that we think is required Key points to remember 1 Induction should take place over a number of weeks even months During this time employees need to assimilate a great deal of information

Tech 2 Tech 2 Effective induction helps new employees to settle into their new job faster and become productive sooner 3 Do not overload An overly intensive informationgiving session is likely to be ineffective 4 Decide on priorities what must be said on the new starter s first day What can wait for the second week 5 Explain the purpose of knowing each new piece of information 6 Great animal welfare depends upon well trained and dedicated individuals who care passionately about animals and welfare Conclusions The majority of the twenty research facilities surveyed had excellent induction processes in place Gaps in Display Screen Equipment checks and Gas Cylinder Usage related to specific locations where computers and gas cylinders were not routinely used Security training guidance and social media advice were lacking and these are issues that can enable staff to feel safer protect their security and can help staff to avoid making social media mistakes and reduce online security risks 191

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December 2014 Animal Technology and Welfare AS ET SPECIAL TRAVEL BURSARY 2014 ESSAYS Advantages and disadvantages of animal technologists performing regulated procedures in research JAN BILTON St James Biomedical Services Clinical Sciences Building St James s University Hospital Beckett Street Leeds LS9 7TF Corresponding author j l bilton leeds ac uk Winning entry of the AS ET Special Travel Bursary 2014 In some research facilities animal technologists can have a dual role As well as providing daily care and husbandry for the animals they are responsible for they are often also licenced to carry out regulated procedures on those animals Regulated procedures are those procedures carried out on a protected animal for a scientific or educational purpose and which may cause that animal a level of pain suffering distress or lasting harm Because of this there are advantages and disadvantages of animal care staff per forming these procedures themselves An animal technologist who has progressed to obtaining their personal licence will have spent many hours caring for and handling a variety of species becoming relaxed confident and competent in handling and restraining animals correctly A natural handling manner combined with a developed skill set is a significant advantage when performing procedures Importantly this refines the experience for the animal as well as ensuring scientifically sound and accurate procedures Having chosen animal technology as a profession animal technologists are likely to have a natural empathy with the animals in their care and with experience of working on a variety of studies they will have been exposed to a wide range of potential adverse effects and observations and be quick to spot an animal in distress and know how to act on it Animal technologists also have the advantage of taking an unbiased view on the procedure they are performing their primary concern remaining with the welfare of the animals rather than the research In large facilities the procedure can be just one of many that will be performed that day by the staff on a variety of experiments It is possible that if the procedure was performed instead by the researcher themselves who has extensive knowledge of the history of the research an idea of the expected or desired outcome and the responsibility of future funding that the work done may lose some impartiality Having less detailed knowledge of the research taking place although always remaining knowledgeable regarding how and why this procedure is happening can be a positive tool for an animal technologist who is performing procedures that requires post procedure observations for example As they are not directly involved with the planning and histor y of the experiment they are likely to record accurate observations of what is seen without being influenced by previous or expected results Similarly a researcher may have only been working on a small number of animals and is likely to have been focussing on one par ticular disease or therapy 193

AS ET Special Travel Bursary 2014 essays Therefore they may not have been exposed to as wide a range of observations after effects or symptoms as an animal technologist may have seen Training and competency records are of high importance in animals units In the UK the Home Office Guidelines on the operation of the Animals Scientific Procedures Act 19861 issued March 2014 state that all people fulfilling a function under the Act records need to be kept in association with the Named Training and Competency Officer NTCO and animal technologists are of course included in this requirement Larger units may employ training officers whose full time role is to organise plan and carry out regular training as well as ensuring all staff training records are up to date Because of this structure the level of competence of animal care staff at a range of procedures is likely to be consistently high Scientists and research groups may not always have this level of infrastructure or standardisation of training as they can often be small teams learning methods from the group leader and training new colleagues or PhD students as and when they join the team Research groups are highly skilled and of course in the UK need to obtain modules 1 3 in order to perform regulated procedures and module 4 for surgical techniques but they may lack the valuable support given by a training officer From a logistical angle another advantage of animal care staff performing regulated procedures is that it limits the amount of visitors within a facility compared to situations when scientists research groups may enter an animal unit to carr y out a procedure themselves It is preferable for all units to reduce footfall to limit the risk of external contamination particularly important in a Specified Pathogen Free SPF facility and when performing aseptic techniques In the UK the use of animals in experiments is regulated under the Animals Scientific Procedures Act 1986 ASPA ASPA states that an animal which has undergone a regulated procedure remains the responsibility of the licensee who carried out the procedure for the duration of the study This is a further positive when animal technologists per form procedures themselves as they are also the primary care givers they are around the animals for the majority of the day and easily available if any complications should arise They are familiar with the animals normal behaviour and being based in the animal unit are in a good position to quickly pick up on any abnormalities or animals in pain or distress As well as the advantages of animal care staff carrying out procedures on animals in their care there are also some disadvantages that should be discussed Firstly from an ethical point of view there could be a conflict of interest when technologists are performing 194 procedures on animals in their care As primary carers a close attachment can form between the staff and their animals Although technologists have a clear understanding of the impor tance of animals in research it can still be hard particularly when working on experiments with a severity limit of moderate or severe where some level of pain or distress may be seen Any potential conflict between the roles of animal care provider and personal licensee in this situation could be minimised by ensuring the technologist is well informed about what they are doing and why Although animal care staff spend a majority of their time ensuring the care and welfare of their animals is optimum they also understand the reasons why it is sometimes unavoidable to use animals in research The conflict can also be reduced by ensuring a clear reporting procedure is in place if a technologist has concerns over any procedures taking place and ensuring NACWOs are at hand to offer advice or support At our facility we have a monthly Mouse Club where researchers with ongoing experiments at the University give presentations to the animal care staff explaining their work and plans for the future Steps like this can minimise the conflict that technologists may feel at times about their dual role as they begin to understand more about the benefits gained from the ongoing research They also have the opportunity to address the researcher with any concerns or questions they have A further disadvantage could be animal care staff lacking the specific skills to perform new or intricate procedures Usual routes of administration blood sampling techniques and basic surgeries are within the daily work of many licenced technologists but more specialised procedures may not be In this case it may be preferable for the researcher involved with experience and knowledge of the procedure to perform the work himself herself In this case competent animal care staff could be encouraged to observe the procedure as much as possible with the aim of learning the procedure themselves over time Due to the nature of their job and their familiarity handling animals technologists are often quick to pick up new techniques At larger research organisations there may unfortunately be a high turnover of animal care staff which could be seen as a disadvantage when animal technologists also perform procedures It is preferable to have the same person people per forming a procedure for the length of the study as much as possible This consistency may be less easy to achieve when using animal technologists rather than the work

AS ET Special Travel Bursary 2014 essays being undertaken by the research group who are likely to be invested for the duration of the period of work However if comprehensive and standardised training is given and detailed standard operating procedures are in place there should not be significant variance between licensees Although working on a variety of species and studies can be an advantage when discussing a technician s skills it may also be seen as a disadvantage when considering cross contamination risk A researcher entering a unit for the sole purpose of performing procedures on their own mice may be considered a cleaner option rather than using an animal technologist who may work in all areas of the unit and may have handled other animals during the course of the day However by observing the facility protocols wearing the recommended Personal Protective Equipment PPE when needed and taking care in showering or changing clothes when moving between areas on the same day this concern could be significantly minimised When discussing the advantages and disadvantages of animal technologists per forming procedures on animals in their care I feel that utilising the unique skills that animal technologists bring outweighs the disadvantages The experienced and sympathetic handling of animals that technologists provide is invaluable Fur thermore by retaining impar tiality towards experimental results approaching experiments with a fresh outlook and being part of an established training and competency system animal technologists have the scope to add real value to experimental research References 1 Guidance on the Operation of the Animals Scientific Procedures Act 1986 This document is available from the UK Home Office website at http www gov uk research and testing using animals Web ISBN 9781474100298 195

AS ET Special Travel Bursary 2014 essays Advantages and disadvantages of animal technicians performing regulated procedures in research KARYN HEATH University of Aberdeen Medical Research Facility Foresterhill Aberdeen AB25 2TD Corresponding author k heath abdn ac uk Essay for the AS ET Special Travel Bursary 2014 Introduction The use of laboratory animals in scientific research began over a century ago when vaccines for polio and rabies became available for production Since then animals have played an important role in unfolding vital information about the human and animal life processes This has helped in the advancement of medicine development of drugs and diagnostics for alleviating suffering in both human and animals As animal research has expanded the role of the animal technologist has also evolved from someone who learned about animal husbandry on the job with minimal supervision and instruction to a professional who is now recognised as a backbone of the entire research animal programme Animal technologists are expected to be responsible for the care and welfare of the animals as well as being licensed to carry out regulated procedures which bring with it many pros and a few cons Technicians are trained skilled and have a vast wealth of knowledge and experience which allows them to carry out procedures effectively and efficiently but there is also the danger that they may care too much which could lead to problems for not only the technologist but also the animal and the science It is important to look at both the advantages and disadvantages to ensure any variables that could affect the research are assessed Training An animal technologist s licence may cover a wide variety of procedures so that they can work in almost any area of research which can be very useful to a scientist It means there is always someone on hand that can aid them in their experiment allowing them to go about other parts of their work The animal technologist s key role is to safeguard the welfare of the animals while supporting the researchers so they 196 have to be knowledgeable in any specific procedures that are relevant to their duties They are trained to a high degree in all the procedures they perform and unlike a lot of researchers who may only have to do a procedure once they are constantly performing these techniques making them experts in their field of work However even with training performing a procedure on an animal that they have cared for perhaps from birth can be extremely difficult It can cause a great deal of mental stress which can lead to hesitation and someone who is hesitant is more likely to make a mistake that could cause the animal harm and affect the experiment However with more experience and training comes more confidence which will reduce the chance of problems occurring Although an animal technologist may be slightly more wary to do something that will cause the animal distress they will naturally be more confident with restraint techniques This in itself is beneficial as a poorly held animal is likely to be able to move which could have serious consequences The animal is also more likely to become stressed which can have many adverse effects Skills When an animal is stressed its body will release various hormones which are part of the animal s natural self protecting mechanism evolved over time as a response to dangerous situations These hormones such as cortisol and adrenalin will cause the heart to beat faster muscles to tighten blood pressure to rise and breathing to quicken This can have significant effects on metabolism cardiovascular activity immune function and behaviour which in turn can alter experimental results A research animal is always going to be undergoing stressful situations so it is vitally important to minimise this as much as possible

AS ET Special Travel Bursary 2014 essays The way in which an animal is interacted with is very important when trying to reduce stress Care staff are skilled in restraint and handling techniques and when doing a procedure they will be confident and precise meaning that the animals will be calmer and easier to handle Socialisation with staff is also beneficial and a technologist achieves this easily through frequent exposure This cannot only affect the ease of handling but also things like body weight and antibody responses There are many other ways that technologists are able to reduce stress Good housing and care often through environmental enrichment means animals live better lives with less stress Food rewards can be used to train animals to sit still meaning less restraint and animals are routinely kept in social groups which is more natural for them and keeps them calmer Although reducing stress is extremely important it has to be remembered that most experiments have a time scale It would be detrimental to put off procedures for long periods in order to try and reduce stress further so a decision has to be made by the technologist as to when it is appropriate to do the procedure within the time limitations This can be difficult especially if the procedure is likely to cause adverse effects The technologist may be inclined to put it off If this became an issue management may have to become involved to give the staff member a deadline that must be adhered to unless there was a logical reason for not Good communication between all the staff and the use of study plans can help the technologist stay focused Knowledge and experience Many procedures will cause the animal some sort of pain or distress and the minimisation of this improves both animal welfare and research quality The parameters for measuring the animal s responses can be shown through physiological biochemical and behavioural alterations The most commonly used by researchers are physiological parameters such as changes in body temperature heart rate respiration or body weight and some may also measure changes in the levels of hormones or similar factors However behavioural parameters are often overlooked or underestimated even though they are frequently the earliest signs of abnormality These parameters can be effective for detecting or grading pain and are sometimes inherent in collecting the biochemical or physiological data The effective use of behavioural change to assess pain is reliant on the observer being familiar with normal behaviours An animal care worker can make an assessment of pain and discomfort based upon observations alone because they have knowledge of behaviour and biology of the animals Non technical staff may misinterpret or fail to recognise these common clinical signs especially in species that do not react to pain in an obvious way The selfpreservation reflex of prey animal species is much more subtle and difficult to interpret than the familiar reaction of a domesticated animal or predator species An experienced technologist will not only be able to detect pain but they are better qualified to differentiate between different levels in terms such as mild moderate or severe This is a great advantage not only for the animal itself but for the scientist especially if the procedure being performed has almost instant results as subtle changes can be picked up very quickly However a common problem that can occur is anthropomorphism A technologist must be very careful to avoid making assumptions that animals share any of the same mental social and emotional capacities of humans and to rely instead on strictly observable evidence If they fail to do this observations may be skewed Care staff may also have problems when an experiment is relying on an adverse effect taking place and they may try to avoid doing a procedure in order to postpone an animal s suffering Equally if an animal is in pain or distress their first instinct will be to put it out of its misery as quickly as possible so it is important that whenever they are doing a procedure they are aware of any effects that may occur and what to do in these circumstances This will reduce the chance of an animal being euthanised prematurely The technologist should be given a grading system for the signs of pain so that they know when they should intervene Care staff should also be informed of the specific route for communicating concerns i e the researcher a supervisor or a veterinarian so that they can be assured that the animal s welfare will not be compromised This will help reduce the technologist s apprehension and will also allow the scientist to be confident that the technologist will carry procedures out without issue Conclusion Although there are a few disadvantages to a technologist performing procedures on the animals that they care for daily they are by far outweighed by the advantages and through good planning these disadvantages can be greatly minimised The use of technical staff benefits the research the researcher and the animal It not only saves time and money it also ensures that procedures are done by the most qualified staff Technologists are experienced and knowledgeable but more importantly they care about the animals which means better welfare This is not only ethically correct but it is good science 197

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December 2014 Animal Technology and Welfare POSTER PRESENTATIONS Originally presented at IAT Congress 2014 Advances in sheep anaesthesia within our facility KATIE BLACKWELL Research Centre Smith Nephew York Science Park Heslington York YO10 5DF Corresponding author katie blackwell smith nephew com Introduction Improvement of anaesthetic techniques is essential for refinement of studies and for the welfare of the animals within our facility The length of anaesthesia carried out on our sheep can vary from 1 hour up to 5 hours dependent on the type of procedure carried out In the past the parameters we checked whilst the sheep were anaesthetised were minimal However during the past 2 years we have researched and examined different techniques for anaesthetising sheep and monitoring the parameters whilst they are anaesthetised Previous practice 1 Sheep fasted overnight 2 Premedication ACP 0 1mg kg intramuscular We increased the dose to 0 3mg kg but there appeared to be no difference See Figure 1 Buprenorphine 0 01mg kg was given with the pre med as well as prophylaxis This premedication did not appear to have much effect on the sheep the sheep were still lively and had to be encouraged to the prep room from the animal room 3 Induction Propoflo 10mg ml IV to effect Figure 1 Sheep after receiving the previous premedication of ACP Once anaesthetised the sheep are intubated with an appropriate sized endotracheal tube 1ml of local anaesthesia is used if necessary to aid intubation 4 Maintenance Approximately 2 5 of Isoflurane and O2 N2O at 4 litres 2litres per minute respectively 199

Poster Presentations Stomach tube placed IV catheter placed in cephalic vein to maintain Intravenous access and give Saline fluid NaCl replacement Monitoring not recorded with a hand held pulse oximeter See Figure 2 oxygen saturation SpO2 pulse rate Monitoring is carried out using a T5 Mindray Multi parameter monitor See Figure 4 Every 5 minutes the following are recorded on a record of anaesthesia G G G G G SpO2 pulse rate end Tidal Carbon Dioxide ETCO2 breaths per minute temperature Figure 2 Hand held pulse oximeter Current practice 1 Sheep fasted overnight 2 Premedication Xylazine 0 1 0 3mg kg along with Ketamine 10mk kg Intramuscular Buprenorphine 0 01mg kg is given with the premed as well as prophylaxis Atipamezole 0 025mg kg to reverse the xylazine if necessary This premedication works much better giving us a recumbent sheep see Figure 3 which can be lifted on to a trolley and wheeled in to the prep room Figure 3 Sheep after receiving the premedication of xylazine and ketamine current 3 Induction Propoflo 10mg ml IV to effect Once anaesthetised the sheep are intubated with an appropriate sized endotracheal tube 1ml of local anaesthesia is used if necessary to aid intubation 4 Maintenance Approximately 2 of Isoflurane and O2 N2O at 4 litres 2 litres per minute respectively Stomach tube placed Positive pressure ventilators used on all sheep for surgical procedures See Figure 5 Intravenous i v catheter placed and sutured in Jugular vein to maintain i v access and give NaCl or constant rate infusion see below Constant rate infusions are used for the more invasive procedures during the procedure as follows Ketamine 20mg kg hr and Lidocaine 10mg kg hr 200 Figure 4 Anaesthetic machine and T5 Mindray Multi parameter monitor

Poster Presentations We also have a laryngoscope endotracheal tubes and an ambu bag available in the sheep holding area team member receives training Each cardiopulmonary resuscitation CPR techniques in Figure 6 Crash cart and defibrillator Conclusion The changes made to the anaesthetic protocols within our facility have greatly improved the welfare of the sheep and safety of the staff induction and recovery periods are far smoother and faster During the actual surgical procedure the animals are more closely monitored and parameters are recorded which can be referred back to if issues with recovery arise Figure 5 Ventipac ventilator The improvements have resulted in a dramatic reduction in anaesthetic related deaths the results are as follows Emergency procedure Two crash carts see Figure 6 are also available one in each theatre which are stocked with all the following equipment 3 2011 0 2012 0 2013 adrenaline lidocaine atropine endotracheal tubes fluids catheters syringes urinary catheters laryngoscopes chart of drug dosages paper and pen count down timer set at 2 minutes tape to secure catheter lace to secure endotracheal tube defibrillator stethoscope ambu bag for manual ventilation of the lungs What next The T5 monitors we currently use have the capabilities to allow us to also monitor invasive and non invasive blood pressure and electrocardiography ECG in the sheep The theatre team recently attended a course on ECG interpretation and are organising a training session with a veterinary anaesthetist This should allow us to utilise the monitors more therefore improving animal welfare Acknowledgements Emma Tozer Lisa Griffin and Kathryn Chambers 201

Animal Technology and Welfare December 2014 How best to train a sheep in research KATHRYN CHAMBERS Research Centre Smith Nephew York Science Park Heslington York YO10 5DF Corresponding author kathryn reek smith nephew com Introduction Methods Female Blue faced Leicester x Suffolk sheep are used in our in vivo experiments these experiments are for orthopaedic research To get the maximum data from a study we collect clinical observations force plate data gross pathology histology and or biomechanical testing We have included the use of a kistler force plate in some of the studies to collect loading force data from the operated limb The force plate records the force put through the operated limb over a set time period To achieve this data collection the animal is required to be in a chase system separated from the flock and in close proximity to handlers which creates a stressful environment for the sheep By introducing acclimatisation this procedure has been refined Acclimatise animals to new surrounding and technologists offer feed incentive e g grass nuts offer grass nuts on the ground progress to grass nuts in a bucket or scoop feeding grass nuts from hand Figure 2 Grass nuts Figure 1 Sheep grazing Objectives Sheep have strong herding instincts and live closely together in fields as a flock When sheep arrive they are unable to be housed individually and are nervous around the technicians Does acclimatisation allow the sheep to accept the technicians new environment and allow being handled individually for force plate data collection 202 Figure 3 Sheep feeding from scoop

Poster Presentations Figure 6 Sheep standing with operated limb on force plate Figure 4 Sheep hand feeding Training for force plate data collection Complete the above stages and acclimatise the animal to the Kistler equipment and chase system this acclimatises the animal to being on their own in the chase and requires them to remain still for a set time period The technologists will then move the sheep back and forth to simulate data collection When the animal has adapted to the surrounding and behaves in a stress free manner data for the study can be collected Figure 7 Sheep on plate with friend in front Results 1 The sheep are easy to handle and accept the technicians improving animal welfare and technician safety 2 Less stressed during live phase which improves rehabilitation 3 Successful acclimatisation to the equipment results in accurate data collection Conclusions The use of sheep acclimatisation has proven to be beneficial to the sheep technicians and study It has proven to reduce stress and potential injury to the sheep making handling easier for the technicians and improve the accuracy of data collection ensuring study requirements are achieved Acknowledgements Figure 5 Force plate set up Katie Blackwell Rachael Cawthorne Lisa Griffin and Emma Tozer 203

Animal Technology and Welfare December 2014 Pain management in sheep how we have developed our techniques to improve animal welfare EMMA TOZER Research Centre Smith Nephew York Science Park Heslington York YO10 5DF Corresponding author emma tozer smith nephew com Introduction Smith Nephew use female Blue faced Leicester x Suffolk sheep in their in vivo research As these projects are primarily for orthopaedic research the sheep must be skeletally mature so are used in the age range 18 months to approximately 4 years Orthopaedic surgery can vary from a simple implant to measure bone ingrowth Figure 1 to plating surgery Figure 2 using a fracture model Pain is managed post operatively by two main methods 1 Analgesia 2 Observations with a scoring system The type of surgery will dictate the analgesic regime and the post operative observations that need to be followed Figure 2 Fracture model with plate Analgesia Analgesia regimes are decided on an individual study basis the relevant personnel usually including NVS NACWO and study director who will develop a regime dependant on surgery type Two general regimes are used depending on the severity of the surgery n a Less invasive contained defect model sub articular model b More invasive fracture Anterior cruciate ligament model surgery Dose dependant analgesia based on animal weight will be given for a every 4hrs for 48hrs b every 4hrs for 72hrs and then every 8hrs for 72hrs Figure 1 Implant to measure bone ingrowth 204 These regimes can be added to if necessary for the full

Poster Presentations group or individual animals taking into consideration the exact procedure and how animals have reacted previously to similar surgeries In the past we administered all of our analgesics manually via intramuscular injections as pain relief may be required for up to 14 days this was most importantly painful for the animals Manual injections had to be carried out through the night meaning unsociable hours for staff Due to these reasons automatic ambulatory pumps namely CADD legacy pump Smiths Medical see Figure 3 were introduced to the unit allowing for improved animal welfare and happier technicians The pumps are programmed to dispense a dose at regulated intervals according to a selected time cycle The pumps have a cassette attached in which the drug to be administered is stored The drug is administered directly into a vein via a catheter inserted during surgery see Figure 4 the pump is held in situ on the animals back via a strap around its girth A head scarf is used to keep the tube from the pump to the catheter in place it also prevents the sheep from chewing the tube Observations and scoring All post operative animals follow the same general observation schedule See Figure 5 unless extra observations are added prior to the start of the experiment or an animal shows cause for concern Day 1 to 6 Week 2 Week 3 Week 4 Once daily Once every working day 3 evenly spaced occasions Once per week or until fully recovered Figure 3 CADDTM legacy pump Figure 5 Large animal observation sheet The observations carried out are listed below and reflect vital signs that may be affected by surgery These observations are scored from 0 animal showing no signs of discomfort through to 10 animal is showing signs of concern and must be monitored the NACWO and NVS must be informed G Figure 4 Catheter in jugular vein G respiration rate temperature 205

Poster Presentations G G G G G G appetite demeanour palpation of wound site pain standing ambulation In conjunction with these observations gait standing data using a force plate is analysed which helps identify lameness that cannot be seen by the human eye Refinements 1 Cadd Pumps allow analgesia to be dispensed automatically without the need for manual intramuscular injections This refinement has improved animal welfare due to reduction of pain and disturbance 2 Acclimatisation of animals prior to surger y involves a series of training and handling techniques This refinement enables the animals to become accustomed to the presence of animal technologists and the weekly routines it also enables technologists to become familiar with each individual animals behaviour which improves accuracy of post surgery observations 3 Harness this has been developed to keep the pump securely in situ for longer periods of time see figure 6 If the animal needs analgesia cover for up to a fortnight a harness will be used as it prevents possible slippage and rubbing instead of the normal single strap 4 Constant Rate Infusion extra anaesthesia and analgesia are given throughout more invasive procedures via an intravenous drip 5 Premedication for more invasive procedures a preemptive dose of non steroidal anti inflammatory pain relief and antibiotics are given the night before surgery additional to the normal premedication 6 Analgesia regimes for more invasive studies analgesia is given on an individual basis rather study basis 7 Local Anaesthetics for more invasive studies local anaesthetics are administered at the wound site to ensure more effective pain relief immediately post surgery Conclusions We believe that the refinements put into place have considerably improved our animals welfare and promotes a more efficient recovery These refinements have also relieved technologists working conditions in terms of unsociable hours and less time consuming techniques Acknowledgements Katie Blackwell Rachael Chambers and Lisa Griffin 206 Cawthorne Kathr yn

December 2014 Animal Technology and Welfare The use of computed tomography CT and digital x ray in sheep LISA GRIFFIN Research Centre Smith Nephew York Science Park Heslington York YO10 5DF Corresponding author lisa griffin smith nephew com Introduction Computed Tomography within the research environment is still relatively novel Radiographic imaging has been previously obtained by the use of Fluoroscopy digital xray and standard radiography The combination of both CT and Digital X ray increases the amount of information available from an individual animal CT imaging The sheep is placed on the CT table in dorsal recumbency and using Velcro straps is secured on the table see Figure 1 A specific protocol is selected to concentrate on the area of interest in this case the tibia Using this protocol a scan is acquired which is then saved on the system to be reviewed at a later date see Figure 4 One of Smith Nephew s main areas of research is looking at bone healing fracture repair Using both imaging techniques has not only proved to be very beneficial for pre screen animals for surgical suitability but also provides vital quantitative data throughout the live phase of the animal Objectives Investigating the implementation of the GE Brightspeed 16 Slice and the DR4000 Digital X ray Device figure 2 into our establishment and the positive outcomes that are achieved Aims for the future fur ther development of Osteoporotic Models using specifically designed Bone Mineral Density BMD software which is used in conjunction with the CT scanner Figure 1 Sheep secured onto CT scanner using velcro straps Digital x ray Methods Pre screening live phase imaging Selected sheep are premedicated to sedation using the following sedation regime Xylazine0 1 0 3mg kg along with Ketamine 10mk kg both administered intramuscularly Once the animals are successfully sedated they are placed on the CT table digital table for imaging to commence During scanning a surgical drape is placed over its eyes to ensure the animal remains calm The sheep is placed first in dorsal recumbency to obtain an Anterior Posterior image then is placed in Lateral recumbency to obtain a Medial Lateral image Beanbags are used to help with positioning of the animal A specific protocol is selected and images are acquired in both planes these images can be manipulated and measurements taken see Figures 3 and 5 When undergoing pre screen measurements and live phase scanning the sheep are allowed to gently recover once the images have been taken and are returned to their holding area 207

Poster Presentations Figure 2 DR4000 x ray system Figure 4 3D reconstruction Figure 5 Measurements taken on digital image Results Figure 3 Digital acquisition 208 CT Scanning is quick efficient and provides a variety of reconstruction options i e 3D sagittal axial etc Digital x ray software enables accurate pre screen measurements to be taken It was found by using both imaging techniques it provides a extensive amount of information not only looking at an animals suitability for surgery

Poster Presentations shape and formation of the bone in question and also enables accurate measurements to be taken before surgery commences Reduced risk to animal during imaging as sedation is used not full anaesthesia Reduction of animal numbers used as scans can be taken throughout the live phase therefore reducing time points throughout the study Conclusion Combining both imaging techniques has proved to be a safe and effective method of obtaining measurable data also enabling us to increase the amount of valuable information acquired from one animal Acknowledgements Katie Blackwell Emma Tozer R Cornell and Kathryn Chambers 209

Animal Technology and Welfare December 2014 Do sheep need environmental enrichment KATIE AMESS Research Centre Smith Nephew York Science Park Heslington York YO10 5DF Corresponding author katie blackwell smith nephew com Introduction Methods Smith Nephew use female Blue faced Leicester x Suffolk sheep in their in vivo experiments As these experiments are primarily for orthopaedic research the sheep must be skeletally mature so are used in the age range 18 months to approximately 4 years 3 different areas are used to house the sheep these are Trying to find enrichment products for sheep proved very difficult as most farm animal enrichment is aimed at pigs It was therefore decided to try enrichment products which are available for other types of animals and to adapt these in ways which the sheep would use them The items tried were The farm this is situated a few miles from the main site and consists of 11 fields of various sizes and a barn for housing in the winter 1 2 3 4 The barns these are on the main site The stock barn holds up to 80 sheep in pens of up to 10 This is where the sheep are kept prior to surgery The study barn holds up to 54 sheep in pens up to 3 This is where the sheep are kept after surgery Footballs Kong balls filled with nuts Ewe and lamb feed Boomer balls filled with nuts Old rabbit water bottles suspended from the ceiling upside down filled with nuts 5 Plastic drums empty mineral lick tubs with lids on The surgical unit the sheep are held here overnight prior to surgery and for up to 48 hours after surgery for close monitoring Up to 26 sheep can be held in the unit at a time Figure 2 Standard sheep nuts ewe and lamb feed Figure 1 Sheep at pasture Objectives Sheep are very social animals and choose to spend most of their time outdoors grazing in flocks When they are outside they do not require environmental enrichment The sheep used in our studies can sometimes be housed indoors for periods of up to 12 weeks post operatively either before being returned to the farm of if they are on a short duration study It was therefore decided to study these sheep to see if they would benefit from environmental enrichment 210 Figure 3 Giant sheep nuts

Poster Presentations 6 Hay nets filled with straw and giant nuts hidden in between layers of straw for the smaller pens 7 Apples suspended from the sides of the pens 8 Bales of straw in the pen for the sheep to climb on 5 Plastic drums when the plastic drums were first introduced into the pens the sheep showed a mild level of interest After that they showed no interest at all 6 Hay nets filled with nuts the sheep showed very high interest in the nets when all the nuts had gone the sheep spent a long time picking through the straw 7 Apples some sheep showed a high level of interest in the apples others showed no interest in them at all 8 Bales of straw in the pens when the bales were first introduced into the pens the sheep showed a very high degree of interest in the bales They spent a lot of time picking through the straw and climbing on top of them Figure 4 Sheep with Kong filled with sheep nuts Figure 6 Sheep emptying sheep nuts from rabbit bottle Figure 5 Sheep with Boomer ball Results 1 Footballs when the footballs were first introduced into the pens the sheep showed a mild level of interest After that they showed no interest at all 2 Kong Balls filled with nuts when these were introduced the sheep showed a high degree of interest emptied out all the food and then completely lost interest in them 3 Boomer balls filled with nuts again the sheep showed a high level of interest when the balls were first introduced and emptied some of the food out but then lost interest in them 4 Old rabbit water bottles filled with nuts as before the sheep showed a very high level of interest in these initially but once emptied of sheep nuts they then lost interest in them Figure 7 Sheep with empty rabbit bottle 211

Poster Presentations Acknowledgements Lisa Joplin Susannah Parkin Mike Smith and James Dodd Figure 8 Sheep with hay net Conclusions Although the most important enrichment for sheep is the social companionship of other sheep the sheep did seem to respond to some of the enrichment products tried in this trial In particular the sheep responded to the products involving food whereas if a food reward was not obvious the response was minimal The enrichment products that the sheep seemed to react to the most were G G G G hay nets filled with nuts rabbit bottles filled with nuts bales of straw filled with nuts apples These are the enrichment products that will be used by Smith Nephew in the future and are all easy to put in place Figure 9 Sheep on bales of straw 212 Figure 10 Never mind the toys where is the food

December 2014 Animal Technology and Welfare Alternative restraint of G ttingen Minipigs for blood sampling ADRIAN ZELTNER Ellegaard G ttingen Minipigs A S Soroe Landevej 302 DK 4261 Dalmose Denmark Corresponding author az minipigs dk Abstract Traditionally Minipigs are restrained in dorsal recumbency to have access to the blood vessels in the neck Figure 1 Minipigs like any other animal do not particularly like to be restrained and being laid on the back with the belly exposed is not a position animals prefer to be in With increasing age and weight this method creates some physical challenges to the technologists The force needed to control a resisting Minipig can be considerable as well as lifting and turning a larger Minipig on its back could touch some occupational health regulations issues The sling has been proven to be a valuable restraint for various procedures with the Minipig This study showed that with slight modification the sling can be used to restrain Minipigs for blood sampling and thus reduce strain on animals and personnel All procedures were carried out in compliance with current laboratory guidelines for good conduct and the project was conducted with approval of the Danish Animal Experiments Inspectorate according to Danish law Animals were housed at approved facilities and taken care of by qualified persons Materials and methods Figure 1 Pig restrained in dorsal recumbency The aim of this study was to test whether the sling could be used as a restraint in various blood sampling situations A standard sling was modified and several options explored to find the most satisfying design As there was a height adjustable table in the facility a frame was custom made to fit this device so an ergonomically suitable working position can be obtained The height is adjustable with an electric motor even when the Minipig is in the sling The actual sling is quite stretched in the frame and a cut out was made at the head end to such a degree that the manubrium sterni of the Minipig is exposed when it hangs in the sling Once the Minipig is placed in the Figure 2 Minipig restrained in sling Figure 3 Height adjustable table and sling frame 213

Poster Presentations sling the head is supported by an assistant and slightly raised so the neck is nicely stretched At the same time the table can be raised to give easy access to the vessels Sitting in a low chair the technician can now obtain a blood sample Figure 4 Frame showing sling in position Figure 7 Pig being placed in sling Figure 5 Sling Figure 8 Position of pig ready for sampling Figure 9 Sampling Results conclusion and discussion Figure 6 Showing position of the pig 214 One of our goals was to find a method that is practical and works as well in demanding settings like

Poster Presentations toxicokinetics pharmacokinetics sampling We could come up with a technique and design that works flawless in most cases if the Minipigs are properly acclimatised The Minipigs are calm do not appear to be stressed and no vocalisation was observed Less manpower than usual was required and we were able to take a sample every 2 minutes with time to spare The system was tested on males as well as females ranging from 5 to 25 kg It proved to be particularly successful in the range 10kg however after a certain size it is advised that two persons lift and place the Minipig in the sling Minipigs in the low weight range might be restrained by a sitting technician Figure 10 Figure 10 Restraint in a sitting position It is imperative to take your time when placing the Minipig in the sling You need to give this procedure the utmost attention when you do it the first time with the Minipig because the outcome of this first attempt will define the character of all the subsequent sling placements The technique of the actual sampling needs to be adapted to the new position Practice has shown that technicians adapt quickly to the new angle of view and even not so experienced technicians have no problems obtaining a blood sample with this type of restrain 215

Animal Technology and Welfare December 2014 Determination of disinfectant efficacy against the eggs of Syphacia species ALISON LIVSEY CONTEC INC Zl du Prat RP 3707 56037 Vannes Cedex France Corresponding author alivsey contecinc com Abstract 10 minutes later The objective of this study was to determine the rate of kill of Syphacia species eggs when treated with an aqueous ready to use disinfectant 2000 ppm hypochlorous acid The Syphacia eggs were collected from the perianal area of rats Flynn 1973 The host animals were either Han Wistar or Sprague Dawley rats The collected eggs were immersed in the hypochlorous acid for a range of exposure times then placed into a hatching medium and incubated to allow determination of viability compared to untreated control eggs The results showed that following exposure of the pinworm eggs to the hypochlorous acid 95 kill was achieved in 10 minutes 95 pinworm egg reduction Experimental method Parasite eggs were collected onto clear cellulose tape by pressing the tape against the perianal region of the rat Flynn 1973 The tape was attached to a microscope slide sticky side up to allow contact between the parasite eggs and the test substance hatch solution Prior to the application of the disinfectant the slides were examined to confirm the presence of the parasite eggs At ambient temperature 20 2 C each prepared slide was placed into an empty Petri dish and sufficient attemperated Contec ProChlor applied to immerse the slide Following a 10 minute exposure period each slide was removed and carefully rinsed with distilled water to remove the disinfectant leaving the eggs on the tape Immediately following rinsing each slide was placed into an empty Petri dish and sufficient hatch medium Dix 2004 attemperated at 37 2 C applied to immerse each slide The slides were incubated at 37 2 C for a minimum of 12 hours 216

Poster Presentations Assessment of results Following incubation the hatch medium was rinsed carefully with distilled water and the slides examined microscopically to assess the numbers of hatched and un hatched eggs present on the slide The results are expressed in terms of percentage eggs hatched Group 1 Untreated control no exposure to Hypochlorous acid Group 2 Distilled water control rinse instead of Hypochlorous acid Group 3 Contec Prochlor 10 minute exposure 217

Poster Presentations The of hatched eggs in the group treated with hypochlorous acid is calculated as a of the untreated control group 2 57 55 9 4 60 This equates to a 95 40 kill 10 minute exposure to 2000ppm Hypochlorous Acid 200 Conclusion Under the stated test conditions 2000ppm ready touse hypochlorous acid kills 95 4 of environmental Syphacia species eggs in 10 minutes Acknowledgements Huntingdon Life Sciences Study Director John N Carter Head Department of In Vitro Technologies Study Design and Implementation Kate Read MRCVS References Flynn R J 1973 Parasites of Laboratory Animals Ames Iowa State University Press 238 240 Dix J Astill J and Whelan G 2004 Laboratory Animals 38 11 16 218

December 2014 Animal Technology and Welfare Increasing useable surface area in standard mouse cages ANDREW NEWMAN1 and SABRINA BRANDO2 1 2 Biological Services College of Life Sciences University of Dundee Animal Concepts Zoom 1813 8225 KM Lelystad The Netherlands Corresponding author a newman dundee ac uk Winner of the LBS Serving Biotechnology Ltd Best Poster Award 2014 Abstract We have been investigating how an interactive environment can be developed in a laboratory caging system These ideas were tested on cages in our unit but could also be used in other caging systems We presented a poster at the Canadian Association for Laboratory Animal Science in Winnipeg Canada in 2013 We have continued to look at additional ideas on enrichment in relation to mouse caging While most of the ideas are more suitable for cages with a height greater than the recommended minimum 120mm there are still possibilities in incorporating them in a standard cage This poster will cover some of these enrichments Finding enrichment that is both physical and cognitive for an animal in the confines of a laboratory cage would help remove a sterile environment and give a more enriched life within the cage This poster shows some of the initial concepts and our attempts to turn these into a working reality Introduction The sizes of laboratory animal cages in particular rodent cages make activity within the cage difficult to achieve The following ideas are an attempt to give mice an active environment that will keep them in good physical condition to provide for cognitive stimulation and a more complex environment with more choices It is too early to say if these designs will help in reducing obesity significantly increase physical fitness and cognitive skills but they certainly help in allowing activity and variation within the cage Other than bedding and nesting materials the two most common enrichment items found within a mouse cage are plastic tubes tunnels and mouse houses both are normally placed on the floor and are passive in nature There are a few examples that allow the tunnel and mouse house to hang from the cage lid Active enrichment could also be different types of running wheels The ideas being put forward have an underlying goal of universal adaption to most mouse cages are easily kept clean and are durable Although these are prototypes and need refinement there is no reason why some of these cannot be used as they are There are two areas that are generally dead areas the centre of the cage and the area between the floor and cage top Directive 2010 63 EU requires that animals have enrichment Annex III section 3 3 b All animals shall be provided with space of sufficient complexity to allow expression of a wide range of normal behaviour They shall be given a degree of control and choice over their environment to reduce stress induced behaviour Establishments shall have appropriate enrichment techniques in place to extend the range of activities available to the animals and increase their coping activities including physical exercise foraging manipulative and cognitive activities as appropriate to the species Environmental enrichment in animal enclosures shall be adapted to the species and individual needs of the animal concerned The enrichment strategies in establishments shall be regularly reviewed and updated Method Cages Thoren USA 9 cage W170 D280 H120mm Thoren USA 2 cage W280 D280 H120mm Tecniplast Italy GR900 W300 D300 H200mm 219

Poster Presentations Materials as shown in photographs Figure 4 New test products Ring binders and various chains are easily obtained The T bar was specially made to our design Figure 1 Commonly used products for animal enrichment Figure 2 If a number of paper houses can be placed in a cage the mice will use these like a maze to climb and run through The mice find these paper houses light and easy to manipulate Figure 5 This shows a young mouse climbing through a very small ring They will do this several times in succession Figure 3 Unusual enrichment stimulus for texture and touch The aquarium plastic grass was not very practical but something more suitable and longer lasting could give a new stimulus This grass was completely chewed at the stem within 24 hours Figure 6 Ring binders connected in a form of a lattice for climbing through The cage tops are still able to be cleaned and stacked with these rings still attached 220

Poster Presentations Figure 7 A cage showing ring binders and a climbing frame fixed to the side of the cage to give extra space for activities Figure 8 An example of a climbing frame in a small cage This cage is not ideal as the height of the cage restricts climbing Figure 9 More climbing frames due to the height of this type of cage this works very well These frames also work if placed on the cage floor Figure 10 An elaborate tunnel Interconnected waste pipes can make various shapes More interesting than a short straight tube normally used this allows for exploring the tunnel system Figure 11 Tunnels set up in a small and large mouse cage Having multiple entrances allows mice more freedom to move if another mouse is in the tunnel Figure 12 A cage with ring binders and a tunnel This allows the complete use of the cage by mice 221

Poster Presentations Conclusion Figure 13 This large mouse cage small rat cage is ideal for endless enrichment products This photograph shows a perpendicular running wheel and a ladder for climbing Figure 14 A good example of ring binders being used Whole pelleted diet in a cage adds to enrichment as it allows the mice to manipulate the diet 1 Except for the T bar which was made specifically to our design all the other equipment came from stationer y suppliers hardware stores and pet shops to test out various ideas The ring binders have great potential due to their simplicity and ease of use various additions like chains and metal bars can be fixed on or between these rings for greater enhancement The only negative aspect we found was the noise these ring binders made when hitting the metal cage lid or the cage side The tunnels are used to a greater extent when a complex design is incorporated compared to a short straight short tube Mice will continuously enter and exit these tunnels 2 Bath plugs were tried as an alternative to the T bar as these looked to give more space within the cage but have the same effect viz climbing and balancing The metal plugs were more popular than the rubber plugs Being a heavier material the metal plug did not move as easily as the rubber plug A dedicated form of disk could easily be developed 3 Climbing frames have potential but need a height greater than 120mm to have any impact as adult mice can reach the cage top without any difficulty These frames can also give divisions with in the cage which the mouse would have to negotiate 4 A perpendicular running wheel was preferred to the disk wheel when they had the choice of both types 5 The aquarium grass was an attempt to look at texture and touch as an enrichment At present little progress have been made on this 6 The recommended minimum cage height for a mouse cage limits potential enrichment Mice will continuously climb and explore if given the appropriate environment We found the largest cage tested to be excellent for incorporating environmental enrichment References Figure 15 The bath plug disk is more space saving than the T bar This could be developed into something more practical with either a chain or solid bar attached to the disk 222 Balcomb J P 2006 Laboratory environments and rodents behavioural needs a review Laboratory Animals 2006 40 217 235 Hawkins P et al 2004 Rodent enrichment dilemmas the answers are out there Animal Technology and Welfare 2004 3 143 147 Newman A 2010 A simple improvement of rodent tunnels for environmental enrichment Animal Technology and Welfare 2010 9 57 58 Phillips S et al 2012 Environmental enrichment what works and do animals need it Animal Technology and Welfare 2012 11 49 51 Sztainberg Y et al 2010 An environmental enrichment model for mice Nature Protocols 2010 9 1535 1539 Published online 12 August 2010 Wurbel H et al 2007 Refinement of rodent research through environmental enrichment and systematic randomisation 2007 NC3Rs 9 www nc3rs org uk

December 2014 Animal Technology and Welfare Application of molecular technologies on non invasive samples to enhance health monitoring programmes of rodents in IVCs a comparison with a traditional approach ANGELA KERTON1 SAVVAS PIPERELIS1 MANDY THORPE1 C CLARKE1 FRANCISCO DIAZ1 K HENDERSON2 C CLIFFORD2 COLIN DUNN3 S TAFT M FOA3 and R NORRINGTON3 1 2 3 CBS Department South Kensington Campus Faculty of Medicine Imperial College London SW7 2AZ Charles River Wilmington MA USA2 Charles River UK Margate Kent UK Corresponding author a kerton imperial ac uk Introduction Methods Individually ventilated cage IVC systems are widely used to protect the health status of laboratory mice Meaningful sampling of the IVC housed population to establish the health status is challenging This study compares pairs of immunocompetent outbred mouse sentinels CRL CD1 exposed to soiled bedding on a weekly basis utilising laborator y methodologies typically described in FELASA health monitoring recommendations against non terminal sampling from pairs of immunodeficient and immunocompetent outbred mouse sentinels Crl CD1 Foxn1 nu nu nude and Crl CD1 Foxn1 nu using specified real time PCR tests and a serology panel The study was designed around ten single sided IVC racks consisting of 80 cages holding five mice per cage which had been routinely monitored by traditional methods managed as a unit in one facility spread over three rooms and involved two sample points each being submitted to two independent laboratories Sentinels aged three weeks on arrival followed by a 7 day acclimatisation period were exposed to 10 20 ml per transfer of soiled bedding over a 12 week period during routine cage cleaning Samples were taken from the animals on two separate occasions 12 weeks and 24 weeks post commencement of study Two different approaches for detection of agents were utilised i standard serology and bacteriology by submission of live animals Lab A and ii submission of faecal pellets for TaqMan OpenArray Digital PCR analysis from Crl CD1 Foxn1 nu nu sentinels and serum from Crl CD1 Foxn1 nu sentinels for analysis by MFIA Lab B To comply with the list of agents to be screened according to FELASA recommendations additional fresh faecal pellets were collected and pooled 10 1 in formalin as a media to enable Faecal Centrifugation Concentration FCC testing for the screening towards non pathogenic protozoa forming cysts For the same purpose and to potentially corroborate the results obtained via PCR only for the second screening serum was collected from each Crl CD1 Foxn1nu to be tested for a full list of agents via Multiplex Fluorescent Immunosorbent assay MFIA Blood was centrifuged 2000g for 10 minutes The serum 0 1 0 2 ml and faecal pellets sent for testing were shipped at 4 C The work reported here was aimed firstly at testing the efficiency of an immunodeficient Nu Nu sentinelbased health monitoring programme under field conditions in a multi user IVC experimental unit The rationale to use an immunodeficient mouse as a bedding sentinel is that it should not clear infection and may sustain a higher population of some infectious agents Therefore it should always shed and could be used at any time to provide reliable samples for PCR Secondly it was aimed at determining whether the immunocompetent Nu mouse because of its proximity with the infected nude becomes a superior contact sentinel for use in traditional health monitoring HM i e serology Thirdly a comparison of traditional and PCR based methods for routine screening was compared and collated over a six month time period 223

Poster Presentations Results The exposure of nude mice to soiled bedding with nonterminal sampling of those mice enabled the detection of Helicobacter species pinworms and various opportunistic bacteria In contrast the traditional exposure and detection methods were consistently negative at each time point Minimal clinical signs were recorded in any of the nude mice those noted were attributed to old age Room Room 13 Laboratory Room 15 23 Rack A 0 2 0 2 0 2 Rack B 0 2 0 2 0 2 Rack C 0 2 0 2 0 2 Rack D N A N A 0 2 A Standard Testing Procedures N number of live sentinel animals submitted Laboratory Staphylococcus Staphylococcus B Trialled aureus xylosus Figure 1 and 2 TaqMan OpenArray Digital PCR 0 1 Laboratory B Trialled Approach Aspiculuris Approach 1 pooled tetraptera 1 pooled sample of sample of 10 faecal 10 faecal pellets pellets from from sentinels sentinels has been has been tested for tested for each room each room Table 1 Comparative results for health screening after 12 week exposure time Figure 3 Crl CD1 Foxn1 nu nu and Crl CD1 Foxn1 nu sentinels at 6 months of age 224

Poster Presentations Room 15 23 0 2 Rack A Room 13 Laboratory Room 0 2 0 2 Testing of faecal pellets replaces transportation of live animals to commercial laboratories eliminating transpor t stress and demonstrating considerable refinement over current techniques A Standard Testing Rack B 0 2 0 2 0 2 Rack C 0 2 0 2 0 2 submitted Rack D N A N A 0 2 Procedures pos live sentinel animals Laboratory Rack A 0 1 Staphylococcus Helicobacter B Trialled Approach Rack B Rack C 0 1 0 1 xylosus Aspiculuris spp Klebsiella Rack D N A tetraptera oxytoca Rack A 0 1 Staphylococcus xylosus Table 2 Results for health screening Ethically the use of immunodeficient animals should always be balanced with the benefits that their use might generate and any unnecessary pain suffering or distress avoided and stopped if present For the purpose of this study comparing data between the two HM systems we were fortunate to conduct the study in a facility with extensive historical information of the presence of infectious agents and that were used as reference positive controls However we would not recommend using nude sentinels in any microbiological environment associated with morbidity and over t clinical disease as this would be incompatible with their welfare Acknowledgements We gratefully acknowledge the indispensable contributions from Katie Horan Ann Clarke and Ray Edgar CBS Depar tment Imperial College Hammersmith Campus and the Molecular Diagnostic Laboratory staff within Research Animal Diagnostic Services Charles River Conclusions These data potentially indicate that the use of a more targeted soiled bedding programme exposing nude mice combined with molecular diagnostic approaches may be more sensitive for the detection of communicable agents The use of the nude sentinel mouse exposed to soiled bedding and PRIA testing appears to have some advantages over exposing immunocompetent sentinels using traditional screening methods reliant on serological testing It allows the possibility to test non invasive samples from nude sentinel mice and to minimise the use of animals For the purpose of this trial during a six month period in the trialled approach only 20 animals have been utilised as sentinels while in the traditional approach 40 sentinels have been used This reduction would also apply to transportation of live animals For the purpose of the trialled approach no live animals needed to be transported The nude mice were closely monitored and examined throughout the study and presented with minimal health problems The clinical signs noted in 3 individual animals included reduced mobility hunched posture and some mild skin discoloration These were attributed to old age at the end of the study 7 months of age Molecular diagnostics also support refinement through the shipping of clinical non terminal samples rather than animals 225

Animal Technology and Welfare December 2014 You can t keep anything clean without getting something else dirty Care of the nude mouse as a sentinel strain in a non sterile IVC environment MANDY THORPE ANGELA KERTON and SAVVAS PIPERELIS CBS Department South Kensington Campus Faculty of Medicine Imperial College London SW7 2AZ Corresponding author s piperelis imperial ac uk Introduction IVC cage systems are widely used in many rodent research facilities Traditionally these units are monitored using immunocompetent sentinels exposed to dirty bedding and tested via methods including serology bacteriology and parasitology This screening method can be costly and requires the use of many animals Also because various factors may influence detection it is not always effective Data presented will summarise the strict housing and checking regimes health recording weight records and incidence of spontaneous disease These protocols have been reviewed and approved by our establishment s ethics committee It supports the theme of FELASA s Congress Better Science from Fewer Animals and the 3Rs as there is an overall reduction in animals used for routine health screening Figure 1 Crl CD1 Foxn1 nu nu and Crl CD1 Foxn1 nu 226 and avoidance of stress associated with live animal transport Our poster demonstrates how the use of immunocompetent sentinels is partially replaced by introducing immunodeficient sentinels where samples are tested via PCR Whilst there may not be any advantage with regards to the transmission to the sentinel cage there is an increased susceptibility of the sentinel to pathogens Methodology We focused on the use of the nude mouse as a sentinel housed in non sterile conditions over a 6 Figure 2 Individually ventilated cages IVC XJ cages hitemp cage

Poster Presentations month period in a large academic establishment All the sentinel animals were purchased from a UK Schedule 2 supplier at 3 weeks of age and were microbiologically defined in accordance with Federation of European Laboratory Animal Science Associations FELASA guidelines They were fed expanded rodent SDS RM1 diet Lillico Biotechnology Surrey UK and demineralised water ad libitum via 260mL polycarbonate bottles Allentown New Jersey USA Housing conditions within the facility were within Home Office Code of Practice recommendations Mice were housed in pairs within negative pressure individually ventilated cages IVC XJ Cages Hi Temp cage 535cm2 Allentown New Jersey USA at a pressure of 25 27 Pa and ventilated at a rate of 50 air changes per hour Bedding was provided grade 12 corncob with shredded paper as nesting material LBS Biotech and a variety environmental enrichment Procedures Ten single sided IVC racks were selected from the long term experimental holding at the facility to be set up with 2 sentinel cages and 1 control cage per rack One sentinel cage on each rack contained 2 immunocompetent week old Crl CD1 females which had been traditionally used in the facility for health screening The other sentinel cage and control cage on each rack contained Crl CD1 Foxn1nu One Nu Nu and One Nu 5 6 weeks old In terms of exposure of sentinels to dirty bedding each week an aliquot of dirty bedding form each cage of the rack 10 20 ml has been transferred into the cages of the sentinel animals during cage cleaning No dirty bedding was transferred into the control cages All animals were weighed on a weekly basis Data relating to mice body weights throughout the programme are presented in Tables 1 and 2 Results The exposure of immunodeficient mice to soiled bedding with non terminal sampling of those mice enabled the detection of Helicobacter species pinworms and various oppor tunistic bacteria In contrast the traditional exposure and detection methods were consistently negative at each time point Minimal clinical signs were recorded in any of the immunodeficient mice those noted were attributed to old age The nude mouse should not be able to clear infection and may sustain a higher population of some infectious agents Therefore it should always shed and could be used at any time to provide reliable samples for PCR The hypothesis that its heterozygous immunocompetent cage mate continuously exposed to higher levels of pathogens results in an improved detection rate was also investigated Table 1 Weights of Crl CD1 Foxn1 nu nu sentinel Table 2 Weights of Crl CD1 Foxn1 nu nu control The 3Rs Figure 3 Crl CD1 Foxn1 nu nu and Crl CD1 Foxn1 nu Testing of remote samples replaces transportation of live animals to commercial laboratories eliminating transpor t stress and demonstrating another considerable refinement over current techniques used The use of nude mouse demonstrated an increased susceptibility of sentinels to the infectious agents and samples were non terminal and non invasive 227

Poster Presentations The use of nude sentinels would significantly reduce the numbers of animals used per year in comparison to our current methods of screening Conclusion A stringent checking regime was instigated at the start of the study and the nude and nu mice were closely monitored and examined throughout and presented no major health problems despite being maintained under conventional husbandry conditions However as age progressed signs of discomfort became evident At the end of the study whilst waiting to obtain the results from the laboratory three out of the ten animals were examined for having reduced mobility hunched postures and some mild skin discoloration On veterinary examination the animals were bright and alert but showed reduced motility so were placed on wet diet and increased monitoring All animals returned to normal after a few days but continued to receive more intensive monitoring for future reference Strict humane end points outlined in the project licence were adhered to however based on the experience of the present experiment we would strongly recommend the replacement of the Crl CD1 Foxn1nu nu nu and nu sentinels at 7 months of age in order to avoid the occurrence of such problems Ethically the use of immunodeficient animals should always be balanced with the benefits that their use might generate and any unnecessary pain suffering or distress avoided and stopped if present Acknowledgements We gratefully acknowledge the indispensable contributions from Katie Horan Ann Clarke and Ray Edgar CBS Depar tment Imperial College Hammersmith Campus and the Molecular Diagnostic Laboratory staff within Research Animal Diagnostic Services Charles River 228

December 2014 Animal Technology and Welfare Refined techniques for acclimatising female macaques to restraint LEE REED TERRI JACKSON LAUREN DEAN and STUART BAKER Institute of Neuroscience Henry Wellcome Building for Neuroecology Newcastle University Framlington Place Newcastle upon Tyne NE2 4HH Corresponding author lee reed ncl ac uk Introduction Restraint of Rhesus macaques in Biomedical Research is often necessary for experimental requirements and user safety Taking advantage of the social and intelligent nature of Macaques can aid in acclimatising these animals to restraint devices By using Positive Response Training PRT techniques alongside Diazepam initial stress levels in the early training phase can be reduced Presented here is a refined method broken down into simple steps to Illustrate the methods and techniques employed This hand touching is then developed into a hand hold by only rewarding when initially a prolonged touch is given changing the reward time until this develops into a hold Step Two To reduce the initial stress levels of what can be a very stressful situation Diazepam up to 2mg kg is given over a 5 7 day period with decreasing doses The neck bar is applied after the conditioned hand hold with immediate reward for both hand hold and neck bar closure Step One Prior to any neck bar training macaques are brought into the laboratory for habituation to the new environment at this stage a fruit reward is given just for being in the laboratory Rhesus macaques are encouraged reach and touch user s hand when this occurs a reward is given reinforcing this behaviour Figure 2 Neck bar closed Step Three This step places less emphasis on the neck bar and more upon the comfort of the Macaque whilst in the restraint by beginning to stretch the time the Macaque is in the laboratory and beginning to develop any other interactions such as illustrated the removal of the front cage panel Figure 1 Macaque and trainer holding hands This step is often the best time to introduce or reintroduce any task that the NHP is going to be using 229

Poster Presentations Conclusion Using these techniques has allowed for neck bar training to be completed within 14 days and in the most successful cases as fast as 6 days Figure 3 Macaque in restraint cage with front panel removed Step Four This step focuses upon decreasing need for the hand hold and the Macaque feeling comfortable enough to place their head into the neck bar To do this firstly the need for hand holding at this point should be minimal with no reaction to the neck bar being noticed By simply releasing the hand as the neck bar goes on at decreasing intervals the Macaques will get to the point where no handholding is necessary and will go into the restraint as required on cue of the neck bar opening Figure 4 Macaque allowing neck bar to be fitted 230

December 2014 Animal Technology and Welfare Instructions to Authors Subjects considered for publication may include original articles technical notes and reviews pertaining to all aspects of animal science and technology management and education The Editorial Board wishes to offer particular encouragement to papers leading to improvements in environmental enrichment the general care and welfare of the animals used in particular those species and strains exhibiting harmful genetic defects and papers describing refinements in techniques a reduction in the number of animals that need to be used or alternatives to animal use Papers describing experimental procedures will only be accepted for publication if authors clearly state that the procedures conform to the prevailing principles and Codes of Practice of the Animals Scientific Procedures Act 1986 Papers submitted from outside the U K should state what legislation and or ethical approval the work has been carried out under In addition authors who describe surgical techniques with recovery should include details of post operative care and any analgesic therapy provided All submissions should follow the ARRIVE Animal Research Reporting of In Vivo Experiments guidelines Kilkenny C Browne WJ Cuthill IC Emerson M Altman DG 2010 Improving Bioscience Research Reporting The ARRIVE Guidelines for Repor ting Animal Research PLOS Biol 8 6 e1000412 doi 10 1371 journal pbio 1000412 The Editorial Board reser ves the right to seek independent advice on any aspect of the content of an article but the final decision on acceptance or rejection remains with the Board Submission Material submitted for publication will be considered provided that it is contributed exclusively to Animal Technology and becomes the property of the Institute of Animal Technology The relevant ar ticle must clearly indicate where photographs and or graphs are to be inserted Address for submission atw iat org uk Hard copy The original manuscript plus two copies should be sent to the address below together with a copy on disk CD or DVD All sheets should be typewritten on one side in double spacing and serially numbered Any photographs or graphs should be supplied as originals and conform to the format in 4 below Address for submission Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL No responsibility will be accepted for loss or damage to such articles Electronic files of submissions are required together with separate files of photographs and any graphics that appear in the manuscript Electronic submissions should be sent via email via atw iat org uk alternatively manuscript plus two copies may be sent as hard copy to the address below All sheets should be typewritten on one side in double spacing with 4 cm margins and serially numbered Additionally a copy on disk should be provided or sent by email via atw iat org uk Articles for submission should be sent to Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL No responsibility will be accepted for loss or damage to such articles Format Articles may be submitted either electronically or by hard copy as follows 1 The first sheet of the article should contain the following Electronic i the full title of the paper ii the initials and last name of the author s iii the full address of the depar tment s and institution s where the work was carried out iv the address for correspondence if different to above Articles should be submitted in Word format with double spacing to the lines and all pages serially numbered Any photographs or graphs must be submitted as separate files and conform to the format in point 4 below 2 For the remainder of the paper the text should be 231

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INDEX TO ADVERTISERS December 2014 Allentown Inc iii ARMIS R W Associates vi AVID plc Labtrac xv Bell Isolation Systems IFC Contec iv Harlan Laboratories viii Institute of Animal Technology xi xiii xvii IPS Product Supplies Ltd IBC LBS Biotech xiv Learning Curve Development Ltd v PFI Systems xvi Special Diets Services xii Sychem Ltd x Tecniplast UK OBC VetTech Solutions vii