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IAT Journal Animal Technology and Welfare G Mouse identification welfare issues G RSPCA farm animal welfare report G You are heroes G Further Felasa SECAL congress posters Official Journal of the Institute of Animal Technology and European Federation of Animal Technologists ISSN 1742 0385 Vol 13 No 1 April 2014

CONTENTS Vol 13 No 1 April 2014 Editorial Jas Barley Chair of the Editorial Board ix Mouse identification methods and potential welfare issues a survey of current practice in the UK Nur Mazlan Noelia L pez Salesansky Charlotte Burn and Dominic Wells 1 Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience Kelly Alves Bicalho and Ivanete Presot 11 PAPER SUMMARY TRANSLATIONS 19 TECH 2 TECH Animal technology at the National Institute for Medical Research a century of innovation Alan Palmer 27 You are heroes share your truth Cindy Buckmaster 35 ARRIVE Guidelines 38 Report of an RSPCA AHVLA meeting on the welfare of agricultural animals in research cattle goats pigs and sheep Penny Hawkins R Eddie Clutton Ngaire Dennison Mirjam Guesgen Matt Leach Finula Sharpe Hugh Simmons Adrian Smith John Webster and Ute Weyre 43 AS ET SPECIAL TRAVEL BURSARY 2013 ESSAYS What do you regard as the most important issues in optimising the care and welfare of laboratory animals Jan Bilton 57 POSTER PRESENTATIONS A refined method of restraint for dogs used in inhalation studies refinement in the acclimatisation procedure Simon Moore Mick Timothy and Steven Hawes 59 Drying off procedure for dairy cows Dean Cooper 62 3Rs Centre Utrecht Life Sciences Jan van der Valk Saskia Arndt Nelleke Verhave and Pim Rooymans 64 Systematic reviews and the Three Rs Judith van Luijk Rob de Vries Carlijn Hooijmans Kim Wever Marlies Leenaars and Merel Ritskes Hoitinga 66 Control of the estrous cycle in guinea pig Cavia porcellus Anne Gr goire A Allard E Huam nc S Le n R M Silva S Buff Marion Berard and T Joly 68 The impact of automation on cage wash operations Mary Robinson Tom Rodriguez Jr Elmer Banes Ronaldo Elenzano Suzanne Craig and Peggy Tinkey 70 Animal research in a global organisation advantages and challenges of a centralised oversight group Gill Fleetwood 72 Instructions to Authors 74 Index to Advertisers xx 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

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FIAT CBiol MSB Phil Ruddock MIAT RAnTech Ted Wills HonFIAT RAnTech Dorothy Woodnott FIAT Honorary Members John Frogley FIAT RAnTech Andrew 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 Ken Applebee OBE FIAT CBiol FSB RAnTech Honorary Secretary Wendy Steel BSc Hons FIAT RAnTech 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 Kate Burton MSc FIAT 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 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April 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 Writing this editorial just before Christmas in 2013 it seems very strange to think that by the time you read this first issue of Animal Technology and Welfare ATW of 2014 we will already be be a quarter of the way through the year and rapidly approaching IAT Congress 2014 During 2012 and 2013 the Council of the Institute of Animal Technology IAT met several times with younger representatives from the IAT Branches commonly described as the Youth members to discuss how the Institute can engage the interest of our younger members From the discussions arising both during these meetings and at others where members of the IAT Editorial Board have met with interested individual youth members it has become clear that changes in style and content would help younger members to maintain an interest in both IAT Bulletin and Animal Technology and Welfare as well as the IAT in general Obviously it is important to serve all our members and to remember that although the IAT is increasingly using electronic methods of communicating with our members the Bulletin in particular is still the primary means of providing members with information about IAT activities and developments as well as other aspects of our profession You will have noticed that the layout of the IAT Bulletin in 2014 has a new look which was one of the ideas that the youth members thought would encourage everyone to read the Bulletin The Editor Sarah Lane will be introducing further changes to increase the appeal of the publication ATW is also changing and I hope that you approve of the new style information pages at the front of this issue My thanks go to Sue Ojakowa from PRC Associates and Jon Clucas of Warwick Printing for their help in designing the new layout A further change in this volume of ATW is the formal adoption in our Instructions to Authors of the ARRIVE Animal Research Reporting In Vivo Experiments guidelines developed by the National Centre for the 3Rs NC3Rs and which are intended to improve the reporting of animal experiments Previous work by the NC3Rs showed that many publications reporting publicly funded animal research from the UK and US lacked key information on how the study was designed conducted and analysed which could limit their value in informing future scientific studies and policy The guidelines have been developed to improve standards of reporting and ensure that the data from animal experiments can be fully evaluated and utilised The guidelines are primarily aimed at scientists writing up their research for publication and for those who are involved in peer review Developed in consultation with the scientific community including researchers statisticians journal editors and funders the guidelines consist of a 20 point checklist of the essential information that should be included in publications reporting animal research During 2013 ATW has already introduced other small changes including the increase in the number of Tech 2 Tech articles adapted directly from Congress Platform presentations These allow ATW to provide ix

Editorial readers with the latest developments in Animal Technology rather than waiting several months for formal papers to be written following Congress However there is always a need for formal articles giving full details of new technology etc and I would encourage as many of you as possible to translate your work into a paper or article As it states in the Instructions to Authors published at the back of each issue 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 Of course a list of publications that you have authored or co authored also adds creditability to your CV One way to get your work published and win a cash prize together with a free registration for Congress is to submit an entry for the Andrew Blake Tribute Award which is given to the Animal Technologist judged to have made the most significant contribution to improving standards in laboratory animal welfare over the previous twelve months I am pleased to announce that the winner of the 2014 award is Natalie Edwards from the University of Cambridge with her paper Improving animal welfare for neurodegenerative mice which shows that a relatively simple change in where feed for the mice is placed makes significant improvement to their welfare This issue offers papers on Mouse identification methods and potential welfare issues from Nur Mazlan and his colleagues and from Brazil an article on Quality Management Systems implemented at an animal breeding facility In addition to the ARRIVE guidelines mentioned above our Tech 2 Tech contributions includes courtesy of Alan Palmer a look back over the past one hundred years at the changes in animal technology at the National Institute for Medical Research and a report from the RSPCA AHVLA meeting on the welfare of Agricultural Animals in Research I am also delighted to include another entry from the 2013 entries for the AS ET Special Travel Bursary posters from the last year s IAT Congress as well as more posters from FELASA SECAL Congress held in Barcelona This will appear in the August issue of ATW xi

April 2014 Animal Technology and Welfare Mouse identification methods and potential welfare issues a survey of current practice in the UK NUR MAZLAN1 NOELIA L PEZ SALESANSKY2 CHARLOTTE BURN2 and DOMINIC WELLS1 1 2 Department of Comparative Biomedical Sciences Royal Veterinary College Royal College Street London NW1 0TU Department of Production and Population Health Royal Veterinary College Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Corresponding author nmazlan rvc ac uk Summary Marking mice to identify individuals is routine practice in laboratory animal facilities but little is known about the current methods of choice or their perceived animal welfare logistical or experimental design consequences Therefore an online survey on mouse identification was sent to laborator y animal establishments throughout the UK The survey link was sent to 83 recipients generating 62 responses from 54 animal establishments Most establishments were academic 61 and over 50 of the responses were from unit managers and or Named Animal Care and Welfare Officers The two most commonly used identification methods were ear punch or ear notch 85 and marker pen application 63 The use of microchips had been discontinued by 37 of institutions Toe clip was considered to be severely stressful or and painful by 53 of the respondent while microchips 45 and tail tattoo 35 were regarded as being moderately stressful or and painful Ear punch or ear notch was the most commonly used method for tissue collection for genotyping Potential welfare issues associated with each identification method are discussed in the context of the survey results Keywords mouse identification laboratory animal welfare refinement standardisation husbandry Introduction Most biomedical research is carried out on rodents especially mice mice were used in 71 of the 3 8 million scientific procedures commenced in Great Britain in 2011 1 Since mice are usually housed in visually homogenous groups individual identification is often required A wide range of methods have been used to identify individual mice with some methods being more invasive in nature than others Permanent identification methods include ear notch ear punch ear tag toe clip tattoo and microchip Temporary identification can be achieved by the use of hair dyes fur trimming or non water soluble marker pens In general most permanent identification methods are invasive breaking the skin while most non permanent methods are usually non invasive Regardless of being invasive or non invasive all procedures involve restraint of the animal which is itself normally stressful 2 3 although it may be possible to modulate the degree of anxiety and stress through the use of alternative handling and restraint methods 4 Identification marking schemes are rarely included in the Methods sections of scientific publications but arguably they could be regarded as Welfare related assessments and interventions that were carried out before during or after an experiment which are suggested for inclusion by the ARRIVE guidelines 5 The invasive and or intrusive nature of the methods means they have the potential to differentially affect mouse welfare and are a possible source of variation that could affect experimental results Earlier in 2013 two working groups of the Federation of European Laborator y Animal Science Association FELASA have published separate reports on their sur vey findings and recommendations on animal identification and rodent genotyping The FELASA working group on animal identification found that ear notch clip 20 out of 42 responses and ear tag 15 out of 42 responses were the most used methods in the USA Canada and in Europe while toe clip and ear tattoo were the least used methods The group recommended using an identification method with minimal adverse effects on the animals while considering the type of research involved although the 1

Mouse identification methods and potential welfare issues a survey of current practice in the UK Methods Advantages Disadvantages Ear punch notch 1 Generally easy to perform costefficient12 2 Easy to read handling may not be necessary 3 Allows individual identification of maximally a few hundred mice 4 Provides tissue sample for genotyping2 7 12 13 19 1 Possibly painful13 2 Punched ear may induce aggression among cage mates13 3 Some strains reseal ear punches eg MRL MpJ 23 24 Toe clip distal phalanx removal 1 Markings are truly permanent19 2 Provides tissue sample for genotyping19 3 Allows early genotyping 3 to 7 day old pups14 18 1 Possibly painful19 23 2 Only to be done on mice before 14 days old18 19 3 Handling or and restraining may be needed to read markings Ear tag 1 Allows identification of a very high number of individual mice6 12 2 Quick and easy procedure6 3 Relatively inexpensive6 12 1 Possibly painful19 2 Restraint may be necessary to read tag 3 May not be a permanent method risk of losing tag6 12 19 4 Tag is a potential irritant6 12 Tattoo 1 Allows identification of a very high number of individual mice 2 Little risk of misidentification19 3 Footpad tattoo can be applied on mice of all ages12 14 19 1 Possibly painful19 in rats micro tattoo more painful than others9 2 Personnel must be trained6 12 14 3 Anaesthetics or analgesics may be necessary12 4 Ink may fade illegible with time12 Microchip 1 Allows identification of a very high number of individual mice6 2 Minimal identification errors compared to other methods6 12 19 3 Allow registration of data in computerised tracking systems27 28 1 Personnel must be trained for application chip positioning 2 Expensive6 28 29 3 Potentially causes discomfort distress14 28 tumours19 29 4 Handling or and restraining needed to read chip code Fur shave cut 1 Easy to apply no special skills training needed 6 2 Easy to read6 3 Less likely to be painful6 4 Low cost6 1 Temporary may only last for 14 days12 up to 3 weeks13 2 Need frequent handling to clip the hair19 3 Can only distinguish a limited number of mice6 4 Some shavers are noisy possibly stressful to mice Coat dyes bleach 1 Easy to apply no special skills training needed 2 Easy to read6 3 Less likely to be painful6 4 Low cost6 1 Temporary12 2 Need frequent handling to reapply dyes19 3 Can only distinguish a limited number of mice6 4 Potential toxicity6 13 19 Marker pen 1 Applicable to all ages6 2 Easy to apply no special skills training needed 3 Easy to read6 4 Less likely to be painful6 5 Low cost6 1 Temporary frequent remarking is necessary12 2 Potential adverse response to solvents in pens6 3 Aversive response to odour released from marker pen has been reported in rats15 Revolving pliers ear Lancet tail foot pad Micro tattoo system ear tail foot pad Electric tattoo equipment tail Table 1 Overview of the advantages and disadvantages of mouse identification and genotyping methods Relevant references are given in brackets where possible 2

Mouse identification methods and potential welfare issues a survey of current practice in the UK precise methods of choice were not named 6 From a survey covering 25 European countries the FELASA working group on rodent genotyping found that tail biopsy 121 out of 158 respondents was the most used method for sampling genotyping while ear punch notch 72 out of 158 respondents and ear tag 39 out of 158 respondents were the methods of choice for identifying genetically modified rodents That working group recommended using a method that is able to simultaneously identify an individual animal and provide tissue for genotyping 7 for making significant welfare improvements by choosing or modifying an identification method to minimise pain stress and other negative welfare consequences It is impor tant to note that improvement to laboratory animal welfare will often not just benefit the animal humane implication but can also benefit the scientific community scientific implication by promoting valid reliable and reproducible experimental data that are not being confounded by the element of pain and stress experienced by the animal 11 A literature review was carried out to find available information on mouse identification methods and their welfare consequences The search terms were rodent identification mouse identification identification methods marking methods genotyping microchip transponder toe clip tattoo ear tag ear notch ear punch and marker pen The related references cited within the selected literature were also reviewed A simplified over view of the advantages and disadvantages of each mouse identification and genotyping method are detailed in Table 1 In summary most articles on different mouse identification and or genotyping methods focused their investigation on the acute effects on mice the ease of performing each method reliability and durability of each method Also different institutions or even different researchers have their own set preference of mouse identification and genotyping method We conducted a survey on mouse identification to assess the current practice in the laborator y establishments throughout the UK and perceived animal welfare practical and scientific issues related to different identification methods To our knowledge the survey is the most comprehensive to date in terms of participation from one country provides novel information summarising perceptions and practice in mouse identification in UK animal units There has been only limited investigation of the welfare consequences of each method for identifying mice For example ear punching is a routine husbandry procedure but it is likely to cause stress during restraint and because it penetrates the sensitive tissues it may cause acute pain at the time of marking and potentially a degree of chronic pain afterwards Indeed mice vocalised more during ear notching 30 of 26 mice than a sham procedure 8 of 24 mice 8 There is also evidence in other mammals rats showed significantly greater mean arterial pressure during the period 1 16h after ear notching than after micro tattooing or eartattooing9 while ear tagged and ear notched piglets showed increases in pain related behaviours vocalisations salivary cortisol and blood lactate than controls 10 There is scope for refinement in marking methods For example in genetically modified mice a biopsy is needed for genotyping and it is possible to combine the biopsy with marking methods such as ear punch ear notch or toe clip Combining biopsy for genotyping with identification marking method would require only one potentially stressful event rather than two 7 Given the numbers of mice used in experimental procedures annually and the need for the majority of them to be unambiguously identified there is potential Methods An online survey was created using SurveyGizmo Online Survey Software Questionnaire Tool and the survey link was sent by email to a mailing list targeting facility managers and Named Animal Care and Welfare Officers NACWOs of laboratory animal establishments Other personnel who are routinely involved in handling laboratory animals such as technical staff scientists and the Named Veterinary Surgeon NVS could also take part on behalf of the facility manager or the NACWO Each response was anonymous unless the respondents chose to include their affiliation so we made it clear that we only needed one response from each animal establishment for the survey results to be meaningful We also promised to maintain the anonymity of individual institutions and individual respondents The survey comprised of 11 questions on mouse identification methods Supplementary Material 1 Aside from straightforward questions on the current practice of identification and genotyping there were also questions which required the respondents to rate stress or and pain three points from mild to severe and level of ease three points from quite hard and rather slow to very easy and quick associated with each identification method Respondents were also asked to name the best identification method for experimental standardisation and rate the criteria of an ideal identification method three points from being less important to very important The survey was carried out in two phases in the period of February to June 2012 Ethical approval for the survey was granted by the RVC Ethics and Welfare Committee URN 2012 0052H 3

Mouse identification methods and potential welfare issues a survey of current practice in the UK Results Survey coverage We obtained 62 survey responses from 54 animal units from all over the UK England 44 animal units Northern Ireland 1 animal unit Scotland 7 animal units and Wales 2 animal units Background of respondents Academic institutions made up the highest percentage 61 of establishment type surveyed followed by government scientific research institutions GSRI 17 pharmaceutical establishments 13 contract research organisations CRO and other types of establishments 4 each while 1 respondent chose not to give any affiliation details Figure 1a Figure 1c Respondent s age n 60 with each slice labelled as the age category followed by the number of respondents Mouse identification methods A range of different mouse identification methods were used in laboratory animal establishments throughout the UK The most commonly used methods were ear punch or ear notch 85 marker pen 63 microchip 31 and ear tag 22 Figure 2a Figure 1a Type of institution taking part in the survey n 54 GSRI government scientific research institution CRO contract research organisation Most of the respondents taking part in the survey were unit managers and NACWOs 55 and 48 out of the total number of 60 respondents respectively Figure 1b Most of the time the unit manager and the NACWO of a laboratory animal establishment were the same person 81 of NACWOs were also the unit manager Figure 1b Respondent s position n 60 NACWO named animal care and welfare officer NVS named veterinary surgeon Most respondents 67 were between 35 and 54 years old Figure 1c 50 of the respondents were females 43 were males while the remaining 7 chose not to include the gender information 4 Figure 2a Mouse identification methods used in UK animal units n 54 Figure 2b Discontinued identification methods n 30 56 of the establishments had discontinued the use of some identification methods due to different reasons described in Table 2 Among the methods that had been discontinued were microchip 37 ear tag 30 tattooing 23 and toe clip 17 Figure 2b

Mouse identification methods and potential welfare issues a survey of current practice in the UK Identification method Reasons for discontinuing Microchip Cost n 4 microchips kept moving slipping loss of microchips unnecessary excessive for animal welfare not needed anymore Ear tag Animal welfare n 3 not easy to identify at a glance difficult to identify after some time can be torn off tags fell out more stressful to animal front limb caught in ear tag and infection very likely to tear off if males fight used for specific reasons before Tattoo Welfare of animal n 2 unnecessary for such painful method too fiddly other less invasive equally reliable method available caused local inflammation deemed unsuitable for neonates by NACWO NVS Toe clip Excessive for animal welfare n 2 unnecessary not visually easy to identify Ear punch Difficult to carry out and read changed to microchip linked to database excessive for our needs and not easy to identify at a glance Marker pen Used only for short term study now use mostly black mice used for specific reasons before Hair dye Not permanent enough frequent reapplication needed took too long to apply other equally reliable methods are available Fur shave Impractical Bar code Unreliable attached using superglue to 1 day old pups when they sweated the bar codes came off Table 2 Reasons for discontinuing certain methods Each reason was suggested by only one respondent unless stated Most mice were identified at the age of two to four weeks 61 or between four to six weeks 15 Figure 2c or and pain another 25 stated that they did not know about the degree of stress or and pain while 22 rated tail tattoo as being severely stressful or and painful The highest percentage of respondents regarded the microchip as being moderately stressful or and painful 45 while another 38 rated it as a mild procedure Marker pen 82 ear punch or ear notch 70 hair dyes 63 fur shave or fur cut 67 and ear tag 41 were rated by most respondents as being only mildly stressful or and painful Figure 3a Figure 2c Mouse age during identification n 54 with each slice labelled as the age category followed by the number of respondents Perception of potential animal suffering personnel preference level of ease associated with each identification method and preferred identification method for standardisation The method perceived by respondents as causing the greatest harm to mouse welfare was toe clipping with 53 of the respondents rating it as being severe while 28 stated that they did not know about the degree of stress or and pain of a toe clip procedure and 19 rated the procedure as being moderately stressful or and painful 35 of the respondents regarded tail tattoo as causing moderate stress Figure 3a Identification methods according to animal stress or and pain level as perceived by respondents Ear punch or ear notch had the highest percentage of respondents rating it as being most preferred 57 followed by microchip 34 and marker pen 30 The methods which most respondents rated as being least preferred were toe clip 74 ear tag 71 and toe tattoo 64 Figure 3b 5

Mouse identification methods and potential welfare issues a survey of current practice in the UK 87 Also 75 of the respondents thought it was very important for an identification method to be long lasting and 71 of them thought ease of application was another very important criterion for an ideal identification method Figure 4 Figure 3b Identification methods according to respondent preference Identification methods rated as being very easy to carry out were marker pen 78 hair dyes 56 fur shave or fur cut 55 and ear punch or ear notch 52 All tattooing methods were regarded as being quite hard to carry out as 26 to 46 respondents gave this rating for each tattooing method Figure 3c Figure 4 Criteria of an ideal identification method The number of responses for each criterion is given in Supplementary Material 2 Genotyping The three most commonly used methods to collect DNA sample for genotyping genetically modified mice in the UK were ear punch or ear notch 85 tail snip 46 and blood sampling 22 Hair pluck and toe clip were also used by 4 each of the animal units taking part in the survey Figure 5a Figure 3c Identification methods according to their ease of application The number of responses for each method is given in Supplementary Material S2 A large percentage of the respondents listed microchip 76 and ear punch 76 as the best identification methods for standardisation Figure 3d Figure 5a Tissue collection methods for genotyping n 54 Figure 3d Perceived best identification method for standardisation n 62 Criteria of an ideal identification method The criteria rated as being very important by the most respondents were reliability 92 of respondents ease of reading the identification number or code achieved 89 and having minimal welfare concern 6 Figure 5b Using ear punch toe clip for both identification and genotyping purpose n 48 with each slice labelled as the method category followed by the number of respondents

Mouse identification methods and potential welfare issues a survey of current practice in the UK A high percentage 92 of animal units practising ear punch or toe clip to collect tissue samples for genotyping stated that they also utilised both methods for the purpose of identification Figure 5b In the three cases where ear punch was not used to satisfy both purposes respondents repor ted the following reasons genotyping was only done on future breeding stock or sometimes mice arrived already tagged or notched and researchers could not get genotyping results from ear notch sample obtained during identification so they performed a tail biopsy for genotyping When asked if they had found any disadvantages when attempting relatively non invasive sampling procedures hair pluck and mouth or rectum swab to obtain DNA samples for genotyping 12 out of 20 respondents reported that they found no disadvantages while the other eight reported they had found disadvantages Five respondents gave details on the disadvantages as listed hair pluck to collect DNA sample cannot serve as an identification method hair pluck is still invasive to animal and easy to contaminate some groups reported that their equipment was not sensitive enough to complete genotyping using samples obtained by noninvasive methods or they are afraid of crosscontamination hair sampling large number of mice resulted in contamination and they still need to be identified and mouth swab was not very good in giving clear genotyping results Discussion Looking at the survey results it appears that some identification methods were more preferred by animal technicians or researchers than others For example ear punch or ear notch was used as an identification method in about 85 of participating animal units Indeed ear punch or ear notch is a quick procedure which requires only simple tools and therefore has lower running costs than other identification methods such as tattooing using a tattooing machine or implantation of a microchip 12 Other methods such as fur shave or fur cut and tattooing were least used in the animal units surveyed Fur shaving is not permanent12 13 while tattooing requires specific equipment and sufficient training6 14 thus making them less favourable compared to other permanent identification methods Ear punch or ear notch which was the method of choice in most animal units was rated as a method which causes only mild stress or and pain by 70 of respondents putting it on a par with other non invasive identification methods such as fur shave or fur cut hair dyes and marker pens This suggests that most people who work with mice assume that there is very little stress or and pain experienced by mice during ear punch or ear notch despite some evidence suggesting the ear punch is a potentially painful procedure as indicated by increased mean arterial pressure in rats 9 and vocalisation8 This perception could be due to the fact that the procedure for ear punch or ear notch is very quick with little opportunity for handlers to notice any sign of stress or and pain Observing for the signs of stress or and pain after returning mice to their home cage following the procedure is not usually practiced and analgesia is not normally given Further research may be necessary to clarify whether or not ear punch or notch causes significant pain to mice Besides being non permanent non invasive techniques could be the identification methods that involve the least stress or and pain From the survey results it was evident that marking using marker pens which were perceived by 82 of respondents to be a mild procedure was practiced widely 63 in UK animal units However nothing is known about possible adverse effects of marker pen inks for mice which need to be investigated further given that rats have been shown to react in a complex manner Tail marked rats appeared bolder in an elevated plus maze and yet they showed more pronounced aversion related Harderian gland secretion chromodacryorrhoea in response to handling compared with unmarked cage mates and unmarked rats avoided open pens significantly more than closed pens in a choice test suggesting that the solvent odour is aversive to them 15 There is also the possibility for toxicity or chemicals entering mouse s body which may interfere with research results 6 Permanent identification methods such as toe clip and ear punch or ear notch will cause a variation in pain and stress levels due to variations in the handling duration number of painful events clips punches and or notches and the amount of tissue being removed according to their designated identification number On the other hand every animal may experience similar levels of pain and stress with other permanent identification methods such as ear tag and microchip So from this point of view ear tag or microchip might be a more preferable permanent identification method for experimental standardisation In agreement with the points mentioned above the survey results showed that most respondents had chosen microchip 76 and ear punch 76 as the best identification methods for standardisation The FELASA Working Group on animal identification considered metal ear tags used by 22 of respondents here as being the worst choice of identification method due to pain and distress as well as posing a risk for inducing various tissue reactions 6 However the literature has suggested that tissue reactions due to metal ear tags could arise from inaccurate placement of the tags or by using metal ear tags in a mouse strain known to be susceptible to squamous cancers 16 17 Whenever tissue samples are needed for genotyping genetically modified mice ear punch or ear notch would 7

Mouse identification methods and potential welfare issues a survey of current practice in the UK be the recommendation as performing one invasive procedure to satisfy two goals is a refinement in experimental procedures 7 unless less invasive procedures are possible such as a mouth swab for genotyping and marker pen for identification if these are indeed found to cause less stress The FELASA working group on genotyping recommended ear punch or ear notch as the method of choice starting from 14 days of age only when permanent identification and tissue for genotyping are needed 7 Currently ear punch or ear notch seems to be the method of choice for collecting tissue sample to genotype mice in the UK since 85 out of 54 animal units reported its use for genotyping In fact 90 of all units who perform ear punch or toe clip to genotype genetically modified mice reported that they utilise ear punch or toe clip as a means of identification too In comparison to our findings a survey carried out by the FELASA working group has found that 46 out of 149 respondents from 15 European countries including the UK reported using ear punch or ear notch for genotyping genetically modified mice weanlings or older 7 In our survey we found that only two out of 54 animal units performed a toe clip for genotyping purposes There was not a single animal unit who reported the use of toe clipping for mouse identification By looking at questions in which we asked the respondents to rate the procedure according to the level of stress or and pain it causes it was evident that respondents regarded toe clip and all tattooing methods as causing a higher level of stress or and pain than other methods Figure 3a Relatively they are also not easy to perform and would require a significant training period before one can master the skill and gain sufficient experience Although there are articles reporting that three to seven days old mice showed little reaction to toe clip and that the procedure did not significantly impair their grip strength motor abilities coordination and balance 14 18 the survey findings suggest that many do not regard the toe clip is as good as or even better than an ear punch or ear notch Toe clipping is still a controversial highly debated procedure in the UK The BVAAWF FRAME RSPCA UFAW Joint Working Group on Refinement recommended not to use toe clipping unless as an absolute last resort and that it should only be performed in mice below the age of two weeks old 13 On the other hand Norecopa s Norwegian Consensus Platform for Replacement Reduction and Refinement of animal experiments Board has stated that toe clipping should not be permitted even with the refinement described by the Nor wegian Animal Research Authority allowing only one toe to be clipped on each hind leg 19 However in the latest edition of the Guide for the Care and Use of Laboratory Animals by the National Research Council USA the clause on toe clipping has changed from toe clipping as a method of 8 identification should be used only when no other individual identification method is feasible and should only be performed on altricial neonates 20 to as a method of identification of small rodents toe clipping should be used only when no other individual identification method is feasible It may be the preferred method for neonatal mice up to 7 days of age as it appears to have few adverse effects on behaviour and wellbeing at this age Castelhano Carlos et al 2010 Schaefer et al 2010 especially if toe clipping and genotyping can be combined 21 The change suggests that toe clipping is now viewed in a different perspective after no scientific evidence of behavioural or motor impairment was found in two studies In their recent publications the FELASA Working Groups on animal identification and genotyping recommended distal phalanx removal toe clip for identification and genotyping in young pups approximately seven days old by removing only the most distal phalanx of one toe per paw 6 7 The survey results demonstrated that there was a high level of welfare awareness among animal care personnel as 87 of the respondents rated minimal welfare concern as a very important criterion of an ideal mouse identification method Also the use of several identification methods had been discontinued due to welfare concerns Table 2 Ten respondents reported that they preferred to use other equally reliable identification methods that are more welfare friendly or less invasive in nature Their concern for mouse welfare during identification gives a positive indication that they would be willing to improve their current practice if scientific evidence to support such a change on welfare grounds is presented There are certainly more questions that could have been added to the sur vey to make it more comprehensive but at that point of time we felt that the questions were sufficient to establish the basic information on the current practice of animal identification in the UK It would be useful to add a question on the number of mice kept in each facility and a few questions on the use of analgesic or anaesthetic during identification Although there is a possibility of anaesthesia being aversive 6 there is evidence that procedures such as ear tattooing in rabbits cause pain and application of EMLA cream prior to the procedure is effective in preventing pain associated with the procedure 22 Furthermore the BVAAWF FRAME RSPCA UFAW Joint Working Group on Refinement recommended the application of local anaesthetic spray prior to tail tattooing 13 Another plan for future work is to target specific groups such as animal unit staff researchers and NVS since the results from this survey comprised mostly of answers from unit managers and NACWO Although some animal unit staff NVS scientists and a deputy facility manager took part in the survey the number was fairly

Mouse identification methods and potential welfare issues a survey of current practice in the UK small and under represented which makes a fair comparison impossible In the future we would be interested to find out whether results from different groups would vary Since the sur vey the Federation of European Laborator y Animal Science Associations FELASA Working Groups has published two repor ts with recommendations on rodent identification and genotyping 6 7 It should be noted that these reports may subsequently have influenced identification marking in the UK and elsewhere Nevertheless the survey has indeed given some useful baseline information on mouse identification methods used in the UK and how they are perceived particularly by unit managers and NACWOs The welfare consequences of the commonly used identification methods have not been extensively studied so further research is required to compare the most commonly used mouse identification methods namely ear punch or ear notch marker pen microchip and ear tag Acknowledgements We would like to thank everyone who has given their valuable time and input to answer the survey on mouse identification carried out by the Royal Veterinar y College NHM was funded by a studentship from the Ministry of Higher Education Malaysia and Universiti Putra Malaysia References 21 22 23 24 25 26 27 Home Office Science Statistics of Scientific Procedures on Living Animals Great Britain 2011 http www homeof fice gov uk publications scienceresearch statistics research statistics other scienceresearch spanimals11 spanimals11 view Binar y 2012 accessed 21 January 2013 Cinelli P Rettich A Seifert B Burki K and Arras M 2007 Comparative analysis and physiological impact of different tissue biopsy methodologies used for the genotyping of laboratory mice Laboratory Animals Vol 41 No 2 174 184 Balcombe J P Barnard N D and Sandusky C 2004 Laboratory routines cause animal stress Contemporary Topics in Laboratory Animal Science Vol 43 No 6 4251 Hurst J L and West R S 2008 Taming anxiety in laboratory mice Nature Methods Vol 7 No 10 825 826 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 PLoS Biology Vol 8 No 6 e1000412 Dahlborn K Bugnon P Nevalainen T Raspa M Verbost P and Spangenberg E 2013 Report of the Federation of European Laborator y Animal Science Associations Working Group on animal identification Laboratory Animals Vol 47 No 1 2 11 Bonaparte D Cinelli P Douni E et al 2013 FELASA guidelines for the refinement of methods for genotyping genetically modified rodents A report of the Federation of European Laborator y Animal Science Associations 28 29 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 Working Group Laboratory Animals Vol 47 No 3 134145 Williams W O Riskin D K and Mott A K 2008 Ultrasonic sound as an indicator of acute pain in laboratory mice Journal for American Association of Laboratory Animal Science Vol 47 No 1 8 10 Kasanen I H Voipio H M Leskinen H Luodonpaa M and Nevalainen T O 2011 Comparison of ear tattoo ear notching and microtattoo in rats undergoing cardiovascular telemetry Laboratory Animals Vol 45 No 3 154 159 Leslie E Hern ndez Jover M Newman R and Holyoake P 2010 Assessment of acute pain experienced by piglets from ear tagging ear notching and intraperitoneal injectable transponders Applied Animal Behaviour Science Vol 127 No 3 4 86 95 Poole T 1997 Happy animals make good science Laboratory Animals Vol 31 116 124 Wang L 2005 A primer on rodent identification methods Lab Animal NY Vol 34 No 4 64 67 Robinson V Morton D B Anderson D et al 2003 Refinement and reduction in production of genetically modified mice Laboratory Animals Vol 37 Suppl 1 S1S49 Castelhano Carlos M J Sousa N Ohl F and Baumans V 2010 Identification methods in newborn C57BL 6 mice a developmental and behavioural evaluation Laboratory Animals Vol 44 No 2 88 103 Burn C C Deacon R M and Mason G J 2008 Marked for life Effects of early cage cleaning frequency delivery batch and identification tail marking on rat anxiety profiles Developmental Psychobiology Vol 50 No 3 266 277 Cover C E Keenan C M and Bettinger G E 1989 Ear tag induced Staphylococcus infection in mice Laboratory Animals Vol 23 No 3 229 233 Baron B W Langan G Huo D Baron J M and Montag A 2005 Squamous cell carcinomas of the skin at ear tag sites in aged FVB N mice Comparative Medicine Vol 55 No 3 231 235 Schaefer D C Asner I N Seifert B Burki K and Cinelli P 2010 Analysis of physiological and behavioural parameters in mice after toe clipping as newborns Laboratory Animals Vol 44 No 1 7 13 Norecopa Toe clipping in mice An evaluation of the method and alternatives http www norecopa no norecopa vedlegg Norecopa toeclip pdf 2008 accessed 26 July 2012 National Research Council USA 1996 Guide for the Care and Use of Laboratory Animals The National Academy Press Washington DC National Research Council USA 2011 Guide for the Care and Use of Laboratory Animals Eighth Edition The National Academies Press Washington DC Keating S C J Thomas A A Flecknell P A and Leach M C 2012 Evaluation of EMLA cream for preventing pain during tattooing of rabbits Changes in physiological behavioural and facial expression responses PLoS ONE Vol 7 No 9 e44437 Li X Gu W Masinde G et al 2001 Genetic control of the rate of wound healing in mice Heredity Vol 86 No 6 668 674 Rajnoch C Ferguson S Metcalfe A D Herrick S E Willis H S and Ferguson M W J 2003 Regeneration 9

Mouse identification methods and potential welfare issues a survey of current practice in the UK 25 26 27 28 29 10 of the ear after wounding in different mouse strains is dependent on the severity of wound trauma Developmental Dynamics Vol 226 No 2 388 397 Karsak M Gaffal E Date R et al 2007 Attenuation of allergic contact dermatitis through the endocannabinoid system Science Vol 316 No 5830 1494 1497 Kitagaki M Suwa T Yanagi M and Shiratori K 2003 Auricular chondritis in young ear tagged Crj CD SD IGS rats Laboratory Animals Vol 37 No 3 249 253 Kort W J Hekking Weijma J M Tenkate M T Sorm V and VanStrik R 1998 A microchip implant system as a method to determine body temperature of terminally ill rats and mice Laboratory Animals Vol 32 No 3 260269 Warn P A Brampton M W Sharp A et al 2003 Infrared body temperature measurement of mice as an early predictor of death in experimental fungal infections Laboratory Animals Vol 37 No 2 126 131 Le Calvez S Perron Lepage M F and Burnett R 2006 Subcutaneous microchip associated tumours in B6C3F1 mice A retrospective study to attempt to determine their histogenesis Experimental and Toxicologic Pathology Vol 57 No 4 255 265

April 2014 Animal Technology and Welfare Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience KELLY ALVES BICALHO1 2 and IVANETE PRESOT3 1 2 3 Laborat rio de Imunologia Celular e Molecular Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brazil Centro de Ci ncia Animal CCA Universidade Federal de Ouro Preto Ouro Preto MG Brazil Servi o de Qualidade Biosseguran a e Ambiente Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brazil Corresponding author bkelly cpqrr fiocruz br Summary This work was carried out at the animal breeding facility of the Ren Rachou Research Center CPqRR FIOCRUZ Belo Horizonte MG Brazil Quality Management System QMS are the rules and principles relating to day to day quality in organisations The aim of this study is to report QMS implementation at a breeding animal facility in Brazil To achieve the planned objectives a review of all the existing guidelines was first carried out which led to the choice of ISO 90011 and the requirements of the Association for Assessment and Accreditation of Laboratory Animal Care AAALAC 2 The following steps were taken to implement the system awareness of the top management definition of policy and quality objectives survey of critical points at the breeding animal facility defining structural work staff training and elaboration of documents A situational diagnosis allowed the identification of critical points such as inadequate control of animal production lack of animal health and genetics monitoring programmes deficiencies in ventilation monitoring programme systems of lighting temperature humidity and noise in the areas of animal breeding deficiencies in the routine disinfection in the animal facilities areas and sterilisation of materials and supplies lack of personnel access control deficiency of equipment control lack of animal welfare programme An action plan was established and corrective actions were taken The implementation of the QMS resulted in better planning organisation greater involvement and understanding of work processes improving service to users of laboratory animals improvement of the working environment health and welfare of both animals and people Keywords breeding animal facility management system implementation quality Introduction Biomedical research requires that experimental animal models are standardised as to their health genetics condition and appropriate animal facility environment Architectural installations with appropriate control of variables such as temperature humidity noise ventilation and lighting are necessar y for the standardisation of animal models 2 Healthy laboratory animals genetically and microbiologically controlled imply reliable and reproducible experimental results physical and psychological animal welfare reducing of the number of animals used in research teaching and testing financial resources optimisation and workers health and safety 2 4 5 6 7 Currently there are a growing number of organisations that adopt Quality Management System QMS which gives them better organisation increased productivity and more credibility thus ensuring the reliability of results The benefits of QMS in a breeding animal facility are easily perceived when an effective system has been implemented with a view to their suitability to the purposes and goals of the institution for example peer recognition standards in animal handling contribution to the 3Rs Replacement Reduction Refinement improving the quality of studies using laboratory animals increased laborator y animals users satisfaction greater efficiency as a result of the focus on prevention rather than correction 8 9 10 Although in recent years concrete efforts to improve infrastructure facilities and equipment of the laboratory animal facilities of public education and 11

Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience research in Brazil have been made through funds obtained from funding agencies and institutions themselves the quality of mice and rats laboratory produced in the most institutions is still below the standard required by international standards of modern laboratory animal science Therefore it is necessary to implement sustainable programmes to modernise animal facilities developed in conjunction with qualified professionals in the field of laboratory animal science seeking to adapt and improve not only the infrastructure but also the formation technical personnel for management and maintenance of colonies of laboratory animals 2 11 12 13 In this context the importance of implementing the QMS to promote continuous improvement of the breeding animal facilities processes is unquestionable This will contribute to animal facilities ability to achieve quality standard thus enabling the production and supply of laboratory animals Choice of guideline to be implemented So far there is no Brazilian or international standard to define quality requirements specific to laborator y animal facilities Therefore three QMS standards are quoted and they can be adapted to animal facilities 1 Good Laborator y Practices GLP 14 2 Quality Management System requirements ISO 9001 1 3 Requirements of the Association for Assessment and Accreditation of Laboratory Animal Care AAALAC 2 or specific recommendations of domestic and international associations for laborator y animal science associations such as Federation of European Laboratory Animal Science Associations FELASA 13 15 2 GLP14 is targeted for the study in order to guarantee the quality of the management including personnel and level of expertise facilities and equipment conditions in which the study is planned developed registered archived and reported The focus of GLP is the scientific process and in particular the study protocol The standard requires that management keeps a master list with all studies recently completed or underway and that are developed under the Standard Operating Procedures SOPs which must be periodically reviewed updated and any changes must be authorised Each unit that follows this standard must have a Quality Assurance Programme to ensure that studies are conducted according to GLP The Quality Assurance staff must do inspections to verify if the study plans and SOPs are being followed by the staff involved and whether they are in accordance with GLP requirements GLP is primarily based on the reliability of the results from laboratory studies and therefore mainly applicable to experimental animal facilities 2 GLP for animal facilities is intended to ensure the maintenance of quality and safety of 12 animals used in the laborator y that conducts biomedical and behavioural studies and trials for product testing 15 16 The second standard cited for laborator y animal facilities ISO 9001 2 may be applied to any product or service In the case of laboratory animal facilities this standard is basically applicable to breeding animal facilities assumed as production processes As for experimental animal facilities ISO 90012 criteria are very general and its application is reduced 11 The standard encourages a process approach in the management of an organisation It is divided into four interrelated blocks management responsibility resource management product realisation measurement analysis and improvement 1 ISO 90012 emphasises the impor tance of determining what customers require and expect and to improve continuously the system products and services in order to better meet their requirements After QMS implementation if the institution decides to apply for certification the system will be evaluated by the certifying body employed to carry out the auditing process It begins usually by an adequacy auditing critical analysis of the quality manual followed by onsite compliance auditing After cer tification maintenance audits will be six monthly or annually and recertification audits will be conducted every three years depending on the certifying body 1 The AAALAC requirements or specific recommendations of domestic and international associations for laboratory animal science e g FELASA focus is on specific procedures for the establishment management and animal control and their application is specific and complementar y to breeding and experimental animal facilities 1 The AAALAC requirements evaluate the organisation and animal handling procedures to ensure appropriate use safety and animal welfare considering the control of facilities procedures and techniques as well as the management of workers health and safety 1 An impor tant function of AAALAC is to provide an opportunity for self assessment of laboratory animal facilities which allows identification of strengths and weaknesses Management bodies of laboratory animal facilities who wish to apply for accreditation should prepare a dossier that describes the characteristics of the animal facility and its management processes The document is used as a guide for the auditing team to evaluate the programme and facilities The auditing team will compare the laboratory animal facilities practices with the principles of the Guide for the Care and Use of Laboratory Animals 3 The auditors visit the facilities interview the staff and if necessary they will require information about specific procedures or protocols A report will be prepared after the auditing which must be retained for analysis by the accreditation board The auditing body will make their

Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience recommendation as to whether or not the audited laborator y animal facilities should be accredited according to their findings1 Based on ISO 9001 and AAALAC standards we defined the internal QMS at the breeding animal facility of the Research Center Rene Rachou CPqRR Oswaldo Cruz Foundation The Oswaldo Cruz Foundation The Oswaldo Cruz Foundation Fiocruz is considered a multidisciplinary public health centre and its activities on the national and international level have become a world reference in the area of science and technology in health There are five sector programmes that it participates in following the objectives and guidelines of the federal government science technology and health innovation pharmaceutical assistance and strategic supplies betterments in health work and education sur veillance and prevention of risks associated with the production and consumption of consumer goods and ser vices and sur veillance prevention and control of diseases and harm 17 G top management awareness and definition of G G G G G quality policy survey of critical points at the animal facility definition of the work structure personnel training elaboration of documents process of production planning of production activities health monitoring programme genetic monitoring programme control environment personnel access control equipment control animal welfare programme The record was made using an appropriate spreadsheet following the 5W 1H method as proposed by Santos 18 It was prepared with these items definition of what will be done what when it will be done when who will who where it will be done where why will be done why and how will be done how The action plan progress was monitored by regular meetings as proposed by Falconi 19 IV Results The Rene Rachou Research Center Fiocruz Minas Gerais Brazil In 1970 during a series of changes in the Ministry of Health the Centre was incorporated with Fiocruz The mission of the Institute is to generate knowledge and tools for a better quality of life for the population regarding health care policies uniting effor ts of research technological development teaching and reference services The CPqRR develops activities in the field of parasitological and viral research including studies on schistosomiasis leishmaniasis Chagas disease malaria dengue virus yellow fever amongst others focussing on sur veillance diagnosis and vaccines The unit has breeding and experimental animal facilities in order to fulfil research projects that use animal models a Top management awareness and definition of quality policy The awareness of the senior management team was essential for the success of the QMS implementation and their commitment to the system by providing the necessar y suppor t for its development implementation and maintenance This was achieved through education by instruments such as specific project for QMS implementation lectures and seminars It is important to emphasise that the quality policy of the CPqRR was established and widely publicised in the Institution It is committed to promoting technical and scientific improvement seeking the quality of research and other services provided by the institution ensuring compliance with the QMS requirements and satisfaction of both internal and external customers The CPqRR enjoys national and international reputation and gives guidance for the Brazilian public health care system providing information and support for control measures of tropical diseases b Survey of critical points at the animal facility The objective of this paper is to report the process of implementing a QMS in a breeding animal facility illustrating all the stages involved in the process A site visit was made in order to understand work processes and together with animal facility staff and management to identify critical actions necessary to improve the process III Methods The following steps were taken to implement the QMS at the CPqRR breeding animal facility With the aim of facilitating the logic of the process implementation an action plan was elaborated in which the stages were defined The following critical points were identified inadequate control of the animal production lack of health monitoring programme lack of genetic monitoring programme deficiency in monitoring of ventilation systems lighting temperature relative humidity and noise at the animal breeding poor control in routine periodic disinfection of areas at animal facility and 13

Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience sterilisation of materials and supplies lack of access control of personnel absence or deficiency of equipment control lack of animal welfare programme c Definition of the work structure A work structure formed by a control group and action groups were created in order to manage quality The first was formed by quality and animal breeding facility management and where necessary by a laboratory animal technician representative Action groups were formed by management and staff of the animal breeding facility d Personnel training For participation and involvement of all employees in the QMS implementation was elaborated as a training plan in accordance with the level of people empowerment and activities they per form The objective was to train all professionals in the standards to be implemented The QMS training includes the training of new employees training in the workplace and periodic updating training Training in the workplace has been prepared considering technicians activities and it was complemented by discussions meetings training offered by the institution and through materials applied to tasks and species with which each person works Among the updating training it can be emphasised Update Course on Laboratory Animal Science for the laboratory animal facility s staff which included in its programme the following topics a Laboratory Animal Facility definition purpose and classification Installation and sanitar y barriers equipment materials and supplies Laboratory animals classification according to health and genetic status routine and operational procedures b handling of laboratory rodents c handling of mice and rats in micro isolators d handling of mice and rats in isolation e animal health and environmental monitoring f genetic monitoring of lineages g animal welfare h biosafety in breeding and experimental animal facilities bio exclusion and biocontainment i maintenance calibration and equipment qualification j materials cleaning cleaning disinfection and sterilisation k processes validation l preparation of solutions m quality management system e Elaboration of documents The following quality system documents were developed document control records control noncompliance control treatment of corrective and preventive actions internal audits and management review of the QMS Then we defined the standard operating procedures SOPs specific to the animal facility such as cleaning up and disinfection of the breeding animal facility personnel access to breeding animal facility disinfections and sterilisation of materials and supplies treatment of cleaning utensils from Clean Area Technicians behaviour in the Clean 14 Area supply of animals derivatives and final inspection The flow char ts of the production processes at the breeding animal facility were drawn up allowing the visualisation of the production activities inspection test and critical points which guided the need for procedures monitoring and process control Finally we developed the Organisation Manual the Quality Manual and Biosafety Manual of the breeding animal facility f Process of production Planning of production activities To facilitate the planning of the production activities it was elaborated the SOP for animals prediction where it should be established the semi annual animals prediction by laboratory animals users according to species strain sex weight and age Based on these data the animal production in the breeding animal facility will be scheduled Health monitoring programme A health monitoring programme was implemented in accordance with the recommendations of FELASA20 21 to assess the animals health status at the breeding animal facility Parasitological necropsy serological microbiological and molecular methods are used to detect rodents major ectoparasites endoparasites bacterial and viral infections Genetic monitoring programme Genetic control was initiated through the organisation of inbred C57BL 6 and BALB c transgenic and knockouts mice colonies establishing production and foundation colonies Animals of the same colour of coat were not created in the same room Genetics certification of isogenic strains as well as genetically modified animals transgenics and knockouts is conducted annually by appropriate laboratory methods Control environment Before the activities control of the sanitisation process was necessary to implement the standardisation in periodic cleaning and disinfection routine of the breeding animal facility areas materials and supplies The environmental monitoring programme is being implemented to control the variables that can impact the quality of laboratory animals produced by the breeding animal facility The programme includes a assessment of procedures for sanitation and sterilisation of caging micro isolators supplies and rooms b assessment of supplies water food and bedding c assessment of environmental conditions temperature humidity airflow lighting and noise and d pest control The quality assurance programme in activities for the sterilisation proposed for CPqRR breeding animal facility was developed according to literature recommendations22 with some modifications and it

Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience consists of four steps equipment control exposure control load control and records The equipment control is performed by mechanical indicators time temperature and pressure calibration of measuring instruments and preventive maintenance of autoclaves use of biological indicator after maintenance and realisation of the Bowie Dick test to detect the presence of residual air in pre vacuum autoclaves The exposure control is accomplished by the use of external and internal indicators The external such as adhesive tapes aims to distinguish processed from unprocessed material The internal indicators are positioned within each package to be sterilised in a place of difficult access of the sterilising agent Regarding the control of the load this is performed weekly or every ten cycles by the use of biological indicators spores of Bacillus stearothermophillus which is the most important test that ensures the quality of sterilisation Autoclaves certification are per formed periodically The documentation of the process to ensure the effectiveness of sterilisation includes records such as item identification batch number of the load cycle number operator time and temperature of the cycle biological indicator results date of sterilisation and expiration date The next step to be implemented will be the evaluation of sanitisation using contact plates Rodac Plate in caging racks and other equipment followed by incubation and colony counting The biosafety area control also will be done periodically collecting samples with sterile swabs in critical points of the clean area such as corners of walls and baseboards lamps door handles shelves walls diffusers of air ducts and others The contact plates also will be used for evaluation of surfaces and room floors of the breeding animal facility The food and bedding used at the breeding animal facility are purchased from qualified suppliers In the process of food acquisition analysis a report is presented indicating nutritional values microbiological and chemical levels within acceptable limits Microbiological and physical chemical assessment of potable water is done periodically In order to improve environmental conditions there is fundamental periodic monitoring of heating ventilation and air conditioning HVAC and lighting Daily monitoring of maximum and minimum temperature and relative humidity is performed by thermo hygrometer in the morning and afternoon Air changes and airflow in the rooms are assessed periodically by means of instruments suitable for this purpose Light timers are used to provide the desired photoperiod The next steps will be periodic evaluation of the differential pressure gradient in the environment by the use of pressure gauges assessment of the light intensity by illuminometer and noise levels to ensure the levels are in accordance with those recommended The pest control programme is conducted weekly by specialised firm hired by the institution The use of insecticides is regulated within the breeding animal facility and there is no direct application in animal rooms Personnel access control Considering the impact of personnel access rules in biosafety areas were inserted into the breeding animal facility routine measures for access control within the different areas of the animal facility The access control in biosafety area was defined in SOP which is used in personnel training programmes including hands and forearms asepsis use of sanitising showers garment and behaviour rules within the facility among other activities Equipment control An equipment control programme was set up including calibration and maintenance of equipment In the equipment collection managing was essential to identify each one as well as archiving the manufacturer s instruction The main actions established were preventive maintenance of autoclaves and cage washer semi annual validation of exchange stations and calibration of pressure gauges thermometers and manovacuometers of the autoclaves In addition SOPs were defined for use of all equipment in the animal facility use and maintenance of ventilated racks cage wash and autoclaves operation Animal welfare programme Zootechny health and animal welfare maintenance in animal facilities must be accompanied by guidance and supervision of a veterinarian These professionals have obligation to provide researchers and other people involved in the work sector guidance on the sensitivity of the animal pain and other sensations that can reach them as well as zootechnical conditions to maintain the same in ideal conditions with regard to their biological characteristics 23 At the CPqRR breeding animal facility the handling has been refined through procedures that minimise the suffering and promote animal welfare and update information is widely available to employees A proposed animal welfare programme for the breeding animal facility includes environmental enrichment with the use within the cages of nontoxic plastic objects that allow adequate sanitisation such as pieces of polyvinyl chloride PVC pipe spinning wheel and igloos acting as burrows for protection addition of nontoxic wood shavings or cellulose in the form of small bundles that allow preparation of bedding or nest 2 The action plan showing the principal steps and actions taken is presented in Table 1 15

Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience Steps Main actions Top management awareness and definition of quality policy Top management awareness was achieved through education by instruments such as specific project for QMS implementation lectures and seminars The quality policy was established by management Survey of critical points at the animal facility Critical points were performed after the site visit and understanding of the work processes along with management and the animal facility staff Definition of the work structure A control group and action groups were created in order to manage quality Personnel training QMS and specific training for all laboratory animals professionals Elaboration of documents Verifying if the creation of a document was really necessary choosing an appropriate author standardising a document to serve as a model for the creation of others Process of production in the breeding animal facility Establish the health monitoring programme genetic monitoring programme control environment personnel access control equipment control animal welfare programme Table 1 Action plan for Quality Management System QMS implementation at breeding animal facility V Discussion When we talk about quality mainly in public institutions it is necessary to think about resistance to change fear insecurity involvement participation since the human element is essential towards the success of any programme or management system Our great allies to success are awareness communication information and effective training of employees Once employees understand QMS benefits they become involved and determined and active par ticipants in its implementation The QMS implementation at the CPqRR breeding animal facility was possible by the commitment from the top management which appointed those responsible for carrying out the activity The joint work of the quality management and breeding animal facility staff was instrumental in the successful QMS implementation The situational diagnosis of the breeding animal facility to survey the critical points and the elaboration of the action plan allowed turning the PDCA Plan Do Check and Act to achieve continuous improvement of processes Staff training programme within the different areas of the breeding animal facility results in benefits to animals science and institution 23 The training programme of the CPqRR expanded the employees knowledge about the risks to human health animal and environmental inherent to activity that each one develops and allowed the participation and involvement of everyone in the QMS implementation process Fostering the development of people is a strategic issue since they are responsible for promoting necessary changes in an institution and for making decisions that guide towards the future The training programme was conducted in a design that rescued the trainee as action subject 16 considering the contributions from him as a thinking and social person Only in this way the training is considered an instrument of remaking and it will contribute effectively to improvement of services 24 The success of any quality programme is dependent on a documentation system that plans and defines how the activities will be made The quality manual is the strategic level document that shows the quality policy objectives and guidelines of the QMS It is a reference to how the system works by linking with other documents 1 In addition the manual is a tool for internal communication with clients and auditors Since the procedures are documents that detail the guidelines in the manual to comply with established policies SOPs were extremely impor tant in the routines standardisation facilitating the understanding and suppor ting the improvement of activities by employees 15 10 They were valuable tools in personnel training and evaluation system in internal audits Quality records are documents attesting the quality management system and they are maintained to demonstrate that the activities were carried out as planned according to the specified requirements 1 10 They demonstrate the QMS adequacy Once the documentation was drawn up by people involved in the process teamwork was encouraged and this fact provided perceptions of activities duplication communication failures important activities of the process that were not being performed Thus the joint preparation of the documentation generated commitment and subsequently activities conducted safely and efficiently The environmental monitoring programme has allowed detection and immediate correction of deficiencies in the physical structure devices and procedures ensuring quality care of animals Environmental contamination

Quality Management System implementation at breeding animal facility the CPqRR FIOCRUZ experience compromises animals health 5 so special attention was given to employees who work in clean area where people can be considered an important source of contamination 15 17 Uniforms and suitable individual protection equipment for restricted use on the work area reduced the risk of contamination and allergies Signalling hygiene and safety work procedures at the animal facility were elaborated in order to warn employees against hazards and accidents and ensure workers safety and health Furthermore they restrict the access to breeding animal facilities areas Use of animals in research minimising pain suffering and distress and promoting their welfare is as important as the goal of obtaining experimental results 25 7 This is important for ethical reasons for good science for economic reasons and to meet the legal principles advocated by the Brazilian and international legislation to protect the use of experimental animal models The QMS implementation has resulted in greater involvement and understanding of the processes by employees planning organisation and refinement of the work process improving service to users of laboratory animals improvement of the working environment health and welfare of employees and animals alignment of the breeding animal facility management model with the concept of sound management as recommended by the World Health Organisation 10 11 12 13 144 15 16 17 18 References 21 22 23 24 25 26 27 28 29 Associa o Brasileira de Normas T cnicas ABNT 2008 ISO 9001 Sistema de Gest o da Qualidade requisitos ABNT Rio de Janeiro Committee for the Update of the Guide for the Care and Use of Laboratory Animals National Research Council 2010 Guide for the Care and Use of Laboratory Animals 8th ed National Academy Press Washington Bicalho K A Ara jo F T M Rocha R S and Carvalho O S 2007 Sanitary profile in mice and rat colonies in laboratory animal houses in Minas Gerais I endo e ectoparasites Arquivo Brasileiro de Medicina Veterin ria e Zootecnia 59 1478 1484 Carty A J 2008 Opportunistic infections of mice and rats Jacoby and Lindsey revisited ILAR Journal 49 272276 Bicalho K A Presot I M Carvalho O S and Rocha R S 2006 Implanta o do Sistema de Gest o da Qualidade em Biot rio de Produ o Parte II final Controle de Contamina o 81 35 38 Massironi S M G 2009 Padr o Gen tico In Cuidados e Manejo de Animais de Laborat rio Lapchik VBV Mattaraia VGM KO GM Eds Atheneu S o Paulo 385 390 Poole T 1997 Happy animals make good science Laboratory Animals 31 116 124 Bicalho K A Presot I M Carvalho O S and Rocha R S 2005 Implanta o do Sistema de Gest o da Qualidade em Biot rio de Produ o Parte I de II Controle de Contamina o 80 33 36 Bicalho K A Presot I M Carvalho O S and Rocha R S 2006 Implanta o do Sistema de Gest o da 19 20 21 22 23 24 25 Qualidade em Biot rio de Produ o Parte II final Controle de Contamina o 81 35 38 Rosemberg F J 2007 Sistemas de gest o da qualidade em biot rios de cria o e de experimenta o In A ci ncia entre bichos e grilos reflex es e a es da biosseguran a com animais Cardoso TAO Navarro MBMA Orgs Hucitec S o Paulo Gilioli R Andrade L A G Passos L A C Silva F A Rodrigues D M and Guaraldo A M A 2000 Parasite survey in mouse and rat colonies of Brazilian laboratory animal houses kept under differents sanitary barrier conditions Arquivo Brasileiro de Medicina Veterin ria e Zootecnia 52 33 37 Gilioli R Sakurada J K Andrade L A Kraft V Meyer B and Rangel H A 1996 Virus infection in rat and mouse colonies reared in Brazilian animal facilities Laboratory Animal Science 46 582 584 Silva A C 1994 Trinta anos em apoio pesquisa e ao desenvolvimento Funda o de Amparo pesquisa do Estado de S o Paulo Edusp S o Paulo M hler M and Nicklas W 2004 Health Monitoring In The Laboratory Mouse Hedrich H Bullock G Petrusz P eds Elsevier Academic Press Hannover 449 462 Majerowicz J 2008 Boas Pr ticas em Biot rios e Biosseguran a Interci ncia Rio de Janeiro Molinaro E M Majerowicz J and Valle S 2008 Biosseguran a em biot rios Interci ncia Rio de Janeiro Brasil Minist rio da S ude Funda o Oswaldo Cruz 2009 Relat rio de Gest o 2009 Rio de Janeiro Fiocruz Dispon vel em Acessed in 01 Mar 2012 Santos M B 2007 Mudan as Organizacionais T cnicas e m todos para a inova o Lastro Belo Horizonte Falconi V C 1994 Gerenciamento da Rotina do trabalho do dia a dia UFMG Belo Horizonte Nicklas W Baneux P Boot R Decelle T Deeny A A Fumanelli M and Illgen Wilcke B FELASA Federation of European Laboratory Animal Science Associations Working Group on Health Monitoring of Rodent and Rabbit Colonies 2002 Recommendations for the health monitoring of rodent and rabbit colonies in breeding and experimental units Laboratory Animals 36 20 42 Nicklas W Deeny A Diercks P Gobbi A IllgenWilcke B and Seidelin M 2010 FELASA guidelines for the accreditation of health monitoring programs and testing laboratories involved in health monitoring Laboratory Animals 39 43 48 Young M 1998 Controlling and improving the sterilization process using biological and Chemical Indicators Infection Control Today s l s n Voipio H M Baneux P Gomez de Segura I A Hau J and Wolfensohn S 2008 Joint Working Group on Veterinary Care Guidelines for the veterinary care of laboratory animals report of the FELASA ECLAM ESLAV Joint Working Group on Veterinary Care Laboratory Animals 42 1 11 Filho H R P 2007 A valoriza o do capital humano no ambiente empresarial Banas Qualidade XVII 32 38 Baumans V 2005 Environmental enrichment for laboratory rodents and rabbits requirements of rodents rabbits and research ILAR Journal 46 162 170 17

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April 2014 Animal Technology and Welfare PAPER SUMMARY TRANSLATIONS INHALTVERZEICHNIS Methoden der identifikation von m usen und potentielle tierschutzprobleme ein berblick gegenw gertiger Praktiken im vereinigten k ngreich NUR MAZLAN1 NOELLA L PEZ SALESANSKY2 CHARLOTTE BURN2 und DOMINIC WELLS1 1 2 Fakult t f r vergleichende biomedizinische Wissenschaften K nigliche Hochschule f r Tiermedizin Royal College Street London NW1 0TU Fakult t f r Betriebs und Bev lkerungsgesundheit K nigliche Hochschule f r Tiermedizin Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Entsprechender autor nmazlan rvc ac uk berblick Das Markieren von M usen zur Identifizierung einzelner Tiere ist bliche Praxis in Labortiereinrichtungen aber es ist nur wenig ber die momentanen verwendeten Methoden oder die angenommene artgerechte Tierhaltung bekannt ber die Folgen von logistischem oder experimentellem Design Daher wurde eine Online Umfrage zur Identifikation von M usen an Labortiereinrichtungen im gesamten Vereinigten K nigreich geschickt Der Umfrage Link wurde an 83 Empf nger gesendet und erbrachte 62 Antworten von 54 Tiereinrichtungen Bei den meisten Einrichtungen handelte es sich um akademische 61 und ber 50 der Antworten kamen von Bereichsleitern und oder erw hnten Tierpflege und Tierhaltungsangestellte Die am h ufigsten verwendeten Identifikationsmethoden waren Ohrstanzen oder Ohrzwicke 85 und Tintenmarkierung 63 Die Verwendung von Mikrochips wurde von 37 der Einrichtungen eingestellt Fu rasten wurden von 53 der Beantwortenden als extrem stressverursachend oder und schmerzhaft bezeichnet w hrend Mikrochips 45 und Schwanzt towierungen 35 als m ig stressverursachend oder und schmerzhaft eingesch tzt wurden Ohrstanzen oder Ohrzwicke waren die am h ufigsten verwendeten Methoden f r die Gewebeentnahme zur Gentypisierung M gliche Probleme hinsichtlich artgerechter Tierhaltung in Bezug auf Identifikationsmethoden werden im Kontext der Umfrageergebnisse diskutiert Schl sselw rter Identifikation von M usen Artgerechte Tierhaltung von Labortieren Verfeinerung Standardisierung Haltung 19

Paper Summary Translations Umsetzung eines Qualit tsmanagementsystems in einer Tierzuchteinrichtung die CPqRR FIOCRUZErfahrung KELLY ALVES BICALHO1 2 und IVANETE PRESOT3 1 2 3 Laborat rio de Imunologia Celular e Molecular Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brazil Centro de Ci ncia Animal CCA Universidade Federal de Ouro Preto Ouro Preto MG Brazil Servi o de Qualidade Biosseguran a e Ambiente Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brazil Entsprechender autor bkelly cpqrr fiocruz br bersicht Diese Arbeit wurde an der Tierzuchteinrichtung des Ren Rachou Forschungszentrums CPqRR FIOCRUZ Belo Horizonte MG Brasilien durchgef hrt Qualit tsmanagementsysteme QMS beziehen sich auf die Regeln und Prinzipien t glicher Qualit t in Organisationen Das Ziel dieser Studie ist es ber den Einsatz von QMS an einer Tierzuchteinrichtung in Brasilien zu berichten Um die geplanten Zielsetzungen zu erreichen wurde zuerst ein berblick ber alle existierenden Richtlinien ausgef hrt die zur Wahl von ISO 9001 und den Anforderungen der Gesellschaft f r Bewertung und Zulassung der Pflege von Labortieren AAALAC Die folgenden Ma nahmen wurden f r den Einsatz des Systems ergriffen Bewusstsein des Spitzenmanagements Definition von Strategie und Qualit tszielen berblick ber kritische Punkte bei der Zuchttiereinrichtung Bestimmung struktureller Arbeit Mitarbeiterausbildung und Ausarbeitung von Dokumenten Eine situationsbezogene Diagnose f hrte zur Identifizierung folgender kritischer Punkte unangemessene Kontrolle der Tierzucht mangelnde Tiergesundheit und genetische Untersuchungsprogramme fehlende Bel ftungskontrolle Belichtungssysteme Temperatur Feuchtigkeit und L rm in den Tierzuchtbereichen mangelnde Routinedesinfektion in den Tiereinrichtungen sowie mangelnde Sterilisation von Materialien und Zubeh r Des weiteren mangelte es an Kontrollen des Personalzutritts und der Ausr stung sowie an Tierschutzprogrammen Ein Aktionsplan wurde erstellt und korrigierende Ma nahmen ergriffen Die Umsetzung von QMS resultierte in besserer Planung Organisation gr erer Einbindung und Verst ndnis von Arbeitsprozessen indem der Service f r Nutzer von Labortieren verbessert wurde Verbesserung des Arbeitsklimas der Gesundheit und des Wohlergehens von Tieren und Menschen Schl sselw rter Tierzuchteinrichtung Qualit tsmanagmentsystem Umsetzung 20

April 2014 Animal Technology and Welfare CONTENU DE LA REVUE M thodes d identification de la souris et probl mes potentiels li s au bien tre une tude sur les pratiques actuelles au Royaume Uni NUR MAZLAN1 NOELIA L PEZ SALESANSKY2 CHARLOTTE BURN2 et DOMINIC WELLS1 1 2 D partement des sciences biom dicales comparatives Royal Veterinary College Royal College Street Londres NW1 0TU D partement de production et de la sant de la population Royal Veterinary College Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Auteur correspondant nmazlan rvc ac uk R sum Le marquage des souris pour identifier les individus est une pratique courante dans les animaleries de laboratoire mais on sait relativement peu de choses sur les m thodes actuelles de choix ou sur le bien tre apparent des animaux et les cons quences de la conception logistique ou exp rimentale Par cons quent une enqu te en ligne sur l identification de la souris a t envoy e aux animaleries de laboratoire dans tout le Royaume Uni Le lien pour acc der l enqu te a t envoy 83 destinataires g n rant 62 r ponses de la part de 54 animaleries La plupart de ces tablissements taient des universit s 61 et plus de 50 des r ponses provenaient de directeurs de sites et ou des agents charg s des soins et du bien tre des animaux Les deux m thodes d identification les plus fr quemment utilis es sont la perforation de l oreille ou l entaille l oreille 85 et une application au crayon marqueur 63 L utilisation des micropuces a t abandonn e par 37 des tablissements L amputation de l orteil est consid r e comme tant extr mement stressante ou et douloureuse par 53 des r pondants tandis que les micropuces 45 et le tatouage de la queue 35 sont consid r es comme tant mod r ment stressants ou et douloureux La perforation de l oreille ou l entaille l oreille sont les m thodes les plus couramment utilis es pour le pr l vement de tissus des fins de g notypage Les probl mes potentiels li s au bien tre associ s chaque m thode d identification sont examin s dans le contexte des r sultats de l tude Mots cl s Identification de la souris bien tre des animaux de laboratoire am lioration standardisation levage 21

Paper Summary Translations Mise en uvre d un syst me de gestion de la qualit dans un tablissement d levage animalier l exp rience CPqRR FIOCRUZ KELLY ALVES BICALHO1 2 et IVANETE PRESOT3 1 2 3 Laborat rio de Imunologia Celular e Molecular Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Br sil Centro de Ci ncia Animal CCA Universidade Federal de Ouro Preto Ouro Preto MG Br sil Servi o de Qualidade Biosseguran a e Ambiente Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Br sil Auteur correspondant bkelly cpqrr fiocruz br R sum Ces travaux ont t effectu s au sein de l tablissement d levage animalier du centre de recherche Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Br sil Les syst mes de gestion de la qualit SGQ sont les r gles et les principes relatifs la qualit dans les organisations au quotidien L objectif de cette tude est de produire un rapport sur la mise en uvre d un SGQ dans un centre d levage animalier au Br sil Pour atteindre les objectifs pr vus un examen de toutes les directives existantes a d abord t effectu ce qui a conduit au choix de la norme ISO 9001 et aux exigences de l Association pour l valuation et l accr ditation du traitement des animaux de laboratoire AAALAC Les mesures suivantes ont t prises pour mettre en uvre le syst me La prise de conscience des hauts responsables la d finition de la politique et des objectifs de qualit une enqu te sur les points critiques de l tablissement d levage animalier un travail structurel d cisif la formation du personnel et l laboration de documents Un diagnostic situationnel a permis l identification de points critiques tels que un contr le insuffisant de la production animale un manque de programme de suivi sur la sant animale et la g n tique des d ficiences dans le programme de contr le de la ventilation les syst mes d clairage la temp rature l humidit et le bruit dans les domaines d levage animalier des insuffisances dans la d sinfection de routine des installations d levage animalier et dans la st rilisation du mat riel et des fournitures un manque de contr le de l acc s du personnel un manque de contr le des quipements le manque d un programme pour le bien tre des animaux Un plan d action a t tabli et des mesures correctives ont t prises La mise en uvre du SGQ a abouti une am lioration de la planification de l organisation une plus grande participation et une meilleure compr hension des processus de travail am liorer le service fourni aux utilisateurs d animaux de laboratoire une am lioration de l environnement de travail de la sant et du bien tre des animaux et des personnes Mots cl s Centre levage Animalier Syst me Gestion Qualit Mise en uvre 22

April 2014 Animal Technology and Welfare INDICE DE LA REVISTA M todos de identificaci n de ratones y problemas probables en el bienestar una encuesta sobre las pr cticas actuales en el Reino Unido NUR MAZLAN1 NOELIA L PEZ SALESANSKY2 CHARLOTTE BURN2 y DOMINIC WELLS1 1 2 Department of Comparative Biomedical Sciences Royal Veterinary College Royal College Street Londres NW1 0TU Department of Production and Population Health Royal Veterinary College Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Autor correspondiente nmazlan rvc ac uk Resumen Marcar a ratones para identificarlos es una pr ctica rutinaria en las instalaciones de animales de laboratorio pero se sabe poco sobre los m todos actuales de elecci n o las consecuencias del dise o experimental o log stico o de bienestar animal percibidas Por tanto se envi una encuesta en l nea sobre identificaci n de ratones a varios centros de animales de laboratorio del Reino Unido El enlace con la encuesta se envi a 83 destinatarios generando 63 respuestas de 54 centros de animales de laboratorio La mayor a de los centros eran acad micos 61 y m s del 50 de las respuestas eran de jefes de departamentos y o encargados del bienestar y el cuidado animal Los dos m todos de identificaci n m s utilizados eran la punci n o perforaci n de oreja 85 o la marca con rotulador 63 El uso de microchips ha sido suspendido por un 37 de las instituciones El clip en la pata se consider que era demasiado estresante y o doloroso por un 53 de los participantes mientras que los microchips 45 y las marcas en la cola 35 se consideraban que eran menos estresantes y o dolorosos La punci n o perforaci n de oreja era el m todo m s utilizado para la recogida de tejidos para la genotipificaci n En el contexto de los resultados de la encuesta se presentan los posibles problemas sobre bienestar asociados con cada m todo de identificaci n Palabras clave Identificaci n de ratones bienestar de animales de laboratorio perfeccionamiento estandarizaci n cr a 23

Paper Summary Translations Implementaci n del sistema de gesti n de la calidad en instalaciones de cr a de animales la experiencia CPqRR FIOCRUZ KELLY ALVES BICALHO1 2 y IVANETE PRESOT3 1 2 3 Laborat rio de Imunologia Celular e Molecular Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brasil Centro de Ci ncia Animal CCA Universidade Federal de Ouro Preto Ouro Preto MG Brasil Servi o de Qualidade Biosseguran a e Ambiente Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brasil Autor correspondiente bkelly cpqrr fiocruz br Resumen Este proyecto se llev a cabo en la instalaci n de cr a de animales del Centro de Investigaci n Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brasil El Sistema de Gesti n de la Calidad QMS es la norma y los principios relacionados con la calidad cotidiana en las organizaciones El objetivo de este estudio es informar sobre la implementaci n de QMS en instalaciones de cr a de animales de Brasil Para conseguir los objetivos planeados primero se llev a cabo una revisi n de todas las directrices existentes que llevaron a la elecci n de ISO 9001 y los requisitos de la Asociaci n para la Evaluaci n y Acreditaci n sobre el Cuidado de Animales de Laboratorio AAALAC Los siguientes pasos se llevaron a cabo para implementar el sistema concienciaci n de los principales directivos definici n de la pol tica y los objetivos de calidad encuesta sobre los puntos cr ticos en las instalaciones de cr a de animales definici n del trabajo estructural formaci n del personal y elaboraci n de documentos Un diagn stico de la situaci n permiti identificar los puntos cr ticos como Un control inadecuado de la producci n animal falta de salud en animales y de programas de control gen tico deficiencias en los programas de control de la ventilaci n sistemas de iluminaci n temperatura humedad y ruido en las zonas de cr a de animales deficiencias en la desinfecci n rutinaria en las instalaciones para animales y la esterilizaci n de materiales y suministros falta de control de acceso de personal deficiencia en el control de equipos falta de programas de bienestar animal Se estableci un plan de acci n y se llevaron a cabo acciones correctivas La implementaci n del QMS result en una mejor planificaci n organizaci n una mayor participaci n y comprensi n de los procesos laborales una mejora en el servicio a los usuarios de animales de laboratorio mejora del entorno de trabajo y de la salud y bienestar de los animales y las personas Palabras clave Instalaciones de cr a de animales Sistema de Gesti n de la Calidad Implementaci n 24

April 2014 Animal Technology and Welfare INDICE DELLA REVISTA Metodi di identificazione dei topi e potenziali rischi connessi alla salute un sondaggio sulle pratiche adottate attualmente nel Regno Unito NUR MAZLAN1 NOELIA L PEZ SALESANSKY2 CHARLOTTE BURN2 e DOMINIC WELLS1 1 2 Department of Comparative Biomedical Sciences Royal Veterinary College Royal College Street Londra NW1 0TU Department of Production and Population Health Royal Veterinary College Hawkshead Lane North Mymms Hatfield Hertfordshire AL9 7TA Autore corrispondente nmazlan rvc ac uk Sintesi La marcatura sui topi una pratica svolta abitualmente nei laboratori con sperimentazione animale tuttavia sono poco noti i metodi attuali di selezione o le presunte conseguenze sulla salute delle specie dovute alla progettazione logistica o sperimentale Per tale ragione stata inviata un indagine on line sull identificazione dei topi a tutte le aziende che utilizzano animali da laboratorio nel Regno Unito Il link dell indagine stato inviato a 83 destinatari e ha ottenuto 62 risposte da parte di 54 aziende animali La maggior parte delle aziende erano universitarie 61 e oltre il 50 delle risposte proveniva da responsabili di unit operative e o da operatori addetti alla cura e alla salute degli animali I due metodi di identificazione pi adoperati risultavano la perforazione o la recisione auricolare 85 e la marcatura con penna 63 Il 37 delle istituzioni aveva interrotto l applicazione dei microchip L asportazione delle dita delle zampe era considerata estremamente violenta e o dolorosa dal 53 degli intervistati mentre i microchip 45 e il tatuaggio sulla coda 35 abbastanza violenti e o dolorosi La perforazione o la recisione auricolare risultavano i metodi pi adoperati per prelevare i tessuti ai fini della genotipizzazione I potenziali rischi sulla salute connessi a ciascun metodo di identificazione adoperato vengono trattati nell ambito dei risultati del sondaggio Parole chiave Identificazione dei topi salute degli animali da laboratorio ridefinizione normalizzazione zootecnia 25

Paper Summary Translations Attuazione del Quality Management System Sistema di gestione della qualit nelle strutture per l allevamento di animali l esperienza del CPqRR FIOCRUZ KELLY ALVES BICALHO1 2 e IVANETE PRESOT3 1 2 3 Laborat rio de Imunologia Celular e Molecular Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brasile Centro de Ci ncia Animal CCA Universidade Federal de Ouro Preto Ouro Preto MG Brasile Servi o de Qualidade Biosseguran a e Ambiente Centro de Pesquisas Ren Rachou CPqRR FIOCRUZ Belo Horizonte MG Brasile Autore corrispondente bkelly cpqrr fiocruz br Sintesi Questo lavoro stato svolto presso la struttura per l allevamento di animali Ren Rachou Research Center CPqRR FIOCRUZ Belo Horizonte MG Brasile Il Quality Management System QMS un complesso di regole e principi relativi alla qualit su base giornaliera nelle organizzazioni Lo scopo di tale studio riferire l attuazione del QMS in una struttura per allevamento di animali in Brasile Al fine di raggiungere gli obiettivi prestabiliti stata dapprima operata una revisione di tutte le linee guida esistenti che ha portato a selezionare la ISO 9001 e i requisiti dell Association for Assessment and Accreditation of Laboratory Animal Care AAALAC Per attuare il sistema sono state adottate le seguenti misure conoscenza dei dirigenti definizione degli obiettivi della politica e della qualit sondaggio sui punti critici presso le strutture per l allevamento di animali definizione del lavoro strutturale formazione del team ed elaborazione dei documenti Una diagnosi situazionale ha consentito di identificare i punti critici come ad esempio controllo inadeguato della produzione animale mancanza di programmi di controllo genetico e sanitario degli animali insufficienza di programmi di controllo dell aerazione dei sistemi di illuminazione temperatura umidit e rumore nelle aree di allevamento degli animali insufficienza di disinfezioni regolari nelle strutture adibite agli animali e sterilizzazione dei materiali e dei rifornimenti mancanza di controllo di accesso del personale insufficienza del controllo dei dispositivi mancanza di un programma per la salute degli animali stato istituito un piano di azione e sono state intraprese azioni correttive L attuazione del QMS ha favorito il miglioramento della pianificazione e dell organizzazione un maggior coinvolgimento e comprensione dei processi lavorativi un miglior servizio per il personale che opera con gli animali da laboratorio un migliore ambiente di lavoro migliori sanit e salute sia degli animali che degli individui Parole chiave Struttura per l allevamento di animali Quality Management System attuazione 26

April 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 Animal technology at the National Institute for Medical Research a century of innovation ALAN PALMER Biological Services National Institite for Medical Research The Ridgeway Mill Hill London NW7 1AA Corresponding author apalmer nimr mrc ac uk Based on an IAT LAVA Congress 2013 Platform Presentation Summary Understanding the past to appreciate the present A century of animal science and technology at the MRC NIMR As the MRC celebrates its centenar y year this presentation takes a look back at the history of animal use at the National Institute for Medical Research The development of key principles techniques and design that improved animal welfare standards and helped shape the modern animal technology industry along with the important scientific discoveries made using a range of species will be described Particular attention will be paid from the 1940s through to the 1970 s regarded by some as the golden age of animal technology and science this period saw some major advances in caging systems breeding techniques laborator y animal nutrition animal technician training and education the Laboratory Animals Bureau and animal house design to name just a few Insights into these developments and the key people who helped drive forward advances in animal welfare will be presented providing a fascinating account into how life in the animal house has changed over the past 100 years Keywords National Institute for Medical Research centenary advances animal welfare Introduction The modern barrier maintained animal facility is controlled with computer monitored environmental conditions and staffed by highly trained technologists caring for animals housed in state of the art caging systems However it was not always like this and as part of MRC National Institute for Medical Research Centenary celebrations this presentation recounts a collection of articles anecdotes and pictures that describe what life was like in the Animal House and how certain technologies and practices evolved 27

Tech 2 Tech During the late 1940s the whole of the animal science and technology industry was beginning to rapidly develop and standardisation of feeding maintenance colony management breeding and supply was taking shape In response the Medical Research Council MRC set up the Laboratory Animals Bureau LAB in 1947 later to become the Laboratory Animals Centre LAC in 1958 The main function of the LAB was to facilitate the standardisation of the supply of animals for research However during the post war years 1945 1950s building materials and labour were prioritised for rebuilding of war damage and new towns and with funds in short supply it was not uncommon for research animals to be housed in wooden sheds where disease spread easily Up until then many laboratory animals were sourced from the commercial pet trade usually with inferior stock which often resorted to poorly funded researchers having to resort to buying large quantities of mice regardless of health and condition the research industry was being supplied with inferior animals by unscrupulous breeders as the prime stock could be sold as pets for more money It was therefore essential for Laboratory Animal welfare to take priority not only to improve the standards of health and welfare for all species but to also supply a good standard animal for research Barrier Maintained Animal Facilities The MRC National Institute for Medical Research NIMR has a long history of pioneering the use of barrier maintained animal facilities since it first recognised in the early 1920s the importance of keeping research animals at the Rhodes Farm facility free from any chance infection The initial innovation was the introduction of a bathing house that all researchers and care staff would use to wash in and change into protective rubber clothing before entering the dog compound Figure 2 A miscellany of huts and sheds that housed many of the animals at NIMR until early 1960s NIMR Director Sir Peter Medawar referred to this as shanty town Mill Hill Sir Peter Medawar made it his priority when taking directorship of NIMR in 1962 to modernise all the animal units at NIMR a project that was initiated by Sir Charles Harrington in the early 1960s Sir Peter Medawar was adamant that strictly controlled purpose built animal facilities was important to help reduce pathogenic burden affecting animal welfare and experimental results The sound proof dog unit was the first of the new units to be completed and was opened in 1966 there being a very good reason why it had to be priority and completely sound proofed Figure 1 Animal attendant wearing protective rubber clothing standing at the entrance to the dog compound bathing house 28 Figure 3 The new dog facility incorporated kennels indoor exercise passage outdoor exercise yard bathing room and it also boasted a state of the art hospital standard operating theatre

Tech 2 Tech Figure 5 6 Pictures of the old units the cats were kept in racks of cages for stock and littering down but there was also a rotation system in place allowing the cats out for exercise Figure 4 Bath time in the new dog facility Beagles and greyhounds were both housed in the facility the beagles being mainly used for Diabetes research and the greyhounds which were ex racing dogs used for Osteoporosis research Due to an increasing demand the dogs had been moved from their original wooden hut to a block of kennels on the 3rd floor of the south west wing in the main building where they also had an outside run This proved highly entertaining for the local youths after closing time at the local public house as they would stand on the road close to the building at this time there was no perimeter security fencing barking and whipping the dogs up into a frenzy of noise that was the source of many local complaints It was therefore considered essential to design the new unit to contain all noise by the use of soundproofing that would absorb the noise while also making it comfortable for staff and dogs Apparently after moving the dogs from the main building into the new sound proof unit the local inebriated youths would still bark but although they got no reaction several weeks passed before all human barking ceased The next priority was to build a modern cat facility as originally the cats were kept in two wooden huts one for breeding and one for holding In 1967 the new modern cat unit was built in the valley which incorporated stud male pens breeding room and rearing pens as well as offices and staff showers rest room over two floors During its highest period of activity the new cat units were producing over 1750 cats per year approximately half of these would be for internal use and the rest would be made available commercially to other research institutions The demand for a good quality research cat was increasing not only at NIMR by the Physiology Figures 7 8 The new NIMR cat facility circa 1969 29

Tech 2 Tech department which required 500 per year over 3 kilos in weight but also across the whole research industry and the Laboratory Animals Centre wished to take the MRC and the rest of the research industry out of the infamous cat market as many cats were being supplied by unscrupulous dealers with animals rife in parasites that was causing a national concern There was a substantial economic cost to running the cat unit but Doug Short MBE FIAT the original Chief Animal Technician at NIMR insisted it solved the greater ethical problem of where and how to source quality cats and also provide them with the best possible welfare Specific Pathogen Free SPF Unit Construction of the SPF unit started in 1969 and was completed commissioned and ready for occupation in December 1972 The original stock animals were Caesarean rederived germ free rats and mice that were maintained in pressure isolators in the Gnotobiotics unit and later contaminated with suitable identified gut flora Rabbits and Guinea pigs were introduced a few years later By late 1973 the SPF unit was producing over 10 000 mice and 2 500 rats per month and supplying nearly all the institutes rodent requirements The efficiency of the unit is reflected by the fact that the death rate among the 50 000 mice and 8000 rats housed was less than 0 1 compared to over 25 in the stocks previously housed in conventional units the elimination of infantile diarrhoea in mice being the major contributory factor It is interesting to note that when the NIMR director Sir Peter Medawar first proposed the building of the SPF unit in the 1960s he faced opposition from certain elements of the scientific community for proposing that in the future all major biological research institutes reestablish their animal colonies on a specific pathogen free basis and that the use of pure inbred strains and first generation F1 hybrids would be the research models of choice It is well known that many within the scientific community still believed that the use of these animals was in some way unnatural or artificial and would give misleading results and not be representative of the characteristics of mammalian life A quote by Sir Peter Medawar sums up his foresight into what is today regarded as standard The provision of SPF specific pathogen free animals is not to be thought of as a great romantic adventure in the world of biological research but as something that will in a few years time be as commonplace as the provision of inbred strains is today 30 Automation and feeding the rabbits Another popular research animal at NIMR during the 1960 1980s was the rabbit which were to be given a new home in the soon to be developed SPF unit However the original rabbitry is still very interesting for two reasons The first was the development of a new pelleted rabbit diet in 1959 the previous rabbit diet was known as Diet181 which was developed by H M Bruce and A S Parkes in 1945 and was very successful However some of the ingredients of Diet18 in particular linseed cake meal and bone meal were prone to becoming rancid if stored for any length of time The number of rabbits being used was increasing rapidly and it was impractical to order the amounts of the diet needed as fresh regular orders and if ordered in bulk it went rancid before it could be used This problem was also being reported elsewhere in the industry with animals becoming lethargic and out ofcondition when fed diet18 that had been kept in storage As a solution for this problem Doug Short and Len Gammage NIMR Senior Animal Technician developed a new pelleted diet with a high protein content with human food grade ingredients less prone to deterioration and with the help of a nutritionist the new diet was trialled successfully with animals quickly improving condition and breeding per formance improving The new diet was supplemented with hay for rabbits and could also be used for Guinea pigs when supplemented with hay and green stuffs This new diet was to become known as SG1 after Short and Gammage and was eventually made available commercially and the formula used in the industry for over 2 decades The only problem with the SG1 diet was that the animals found it so palatable ordinary stock animals became fat as they would not stop eating it when fed via a conventional hopper system and it was thought that feeding little and often was too time consuming for the technicians After a visit to a local poultry farm where he saw a chicken battery with an automatic feeding system Doug Short being the visionary that he was decided that the way for ward in the animal house was automation Thanks to the help of the NIMR engineering department an automatic chicken battery was converted into a fully automated feeding and watering system for the rabbit facility In this converted chicken battery the food hopper would go around a track on the outside of the rabbit cages 672 cages in total and stop for short intervals at each cage this would then be followed a few minutes later

Tech 2 Tech Figures 9 12 Automated rabbit feeding watering and cleaning system by the water trough on the same track and re filled from the mains water supply after triggering a valve Not being content with just automated feeding and watering Doug Shorts modifications also incorporated fully automated cleaning the rabbit cages having a grid bottom floor that allowed urine and faeces to drop through onto a plate glass shelves that were bonded together to run the length of the rack and a squeegee was attached to the hoppers that pushed the rabbit excreta along the glass shelf and emptied it into a drain where it was flushed into the sewers Some mice were also kept in large glass bowls similar to goldfish bowls which was standard practice in the industry Until the years following World War 2 WW2 mice and rats were fed this diet of wet mash which would be made by the animal care staff incorporating pretty much anything they could get their hands on from local food suppliers and the works canteen kitchen waste supplemented where possible with dried milk and egg products and margarine there was no provision for water bottles and the mash would be changed daily This system had no adverse effect on the animals breeding or temperament and the rabbits could not gorge themselves on the food constantly It also reduced labour considerably and odours Rodent cage design and standardisation of animal feeds at MRC NIMR When the Small Animals Department at Rhodes Farm Mill Hill was built in the early 1920s the rats and mice were kept in wooden boxes made by an on site carpenter These original wooden boxes had no food hopper or water bottle holder as the animals were fed a diet of wet mash only dried cubed feed was not yet being commercially available Figure 13 Replica of an early wooden mouse cage Figures 14 15 The first NIMR metal cages 31

Tech 2 Tech and fed in rations to lactating mums first followed by pregnant breeders and young weaners with the stock animals being fed what was left However post WW2 the necessity to change because of food shortages and lack of manpower changing the soiled wet mash daily was seen as both time consuming and wasteful The wet mash also soiled the animals cages and bedding so was far from ideal from a welfare perspective This led to a purpose built rodent box designed in the late 1940s by NIMR Animal Technicians and then Senior Technician Doug Short with a bottle and food hopper These cages were still made on site as required and were seen as a revelation in terms of animal welfare as the animals could be checked regularly without removing lids and fed watered daily with a minimum of fuss and no wet mash to soil the cages The wooden rodent boxes had limited life as the animals were quite adept at gnawing their way out so new metal caging was designed and implemented and by 1930 both the Hampstead laboratories and Rhodes Farm small animals department were now using metal rodent cages as standard These cages were still made on site as cages made commercially were still not readily available food and water hoppers were not incorporated into the design as the animals were still being fed the diet of wet mash However by 1945 Sir Alan Parkes and Hilda Bruce had started working on standardisation of the animal feeds by developing compound diets in a cubed and pelleted form that would be suitable for all standard small laboratory animals In 1949 and after nearly five years of research at NIMR a diet called 41B went into commercial production as the first complete unsupplemented dried rodent diet for mice and rats and was used in the same formula industry wide for over 3 decades The new standard dried and cubed rodent diets now meant provision of separate water was a necessity as was an easier method of feeding the diet As metal was becoming easier to source post WW2 cages were being made out of zinc the original zinc boxes just had a per forated lid which made for a very cold and damp environment in the winter for the animals These boxes were adapted for inclusion of a small diet hopper and water bottle holder The first water bottles used were actually ink bottles with a small rubber bung and spout This also caused the problem during the winter months of the water bottles freezing as the animals were kept in huts with little or no environmental control heating Figure 16 A modern style rodent cage circa 1950s Standardisation of animal supply and training The MRC had been breeding and supplying other research institutes for some time but were unable to meet demand so they set up a system of regulation and an accreditation scheme for commercial breeders to the research industry This accreditation scheme was developed at NIMR and based on the knowledge expertise and experience of the animal care team and senior researchers Sir Alan Parkes and Hilda Bruce The accreditation scheme was initiated for regulating laborator y animal supply nationally Commercial breeders were invited to apply for accreditation and most breeders of repute did so The accreditation scheme demanded strict regulations such as stock must be raised primarily for research use breeding stock must be self contained strict standards of hygiene should be implemented annual inspections will be carried out The Laboratory Animals Centre published the Mouse Newsletter until the mid 1970s detailing all mouse strains available to the researcher and also maintained 15 of the commonest laboratory mouse strains at NIMR which it supplied to institutes and commercial 32

Tech 2 Tech breeders with a nucleus of stock together with instructions on how to breed using the traffic light system in order to maintain genetic integrity The LAB initiated annual conferences for exchanges of information for animal technicians and scientists on all aspects of laboratory animal care and welfare but there was also a spontaneous movement amongst animal technicians nationwide that resulted in the formation of a new technical organisation The Animal Technicians Association ATA at the 1950 LAB conference this new organisation with Sir Alan Parkes as its first president would put the provision of training for animal technicians as a priority and put an end to the belief which had prevailed for too long that the work of the animal technician was an unskilled occupation Figure 17 List of papers printed in the 1st Journal of the Animal Technicians Association by the President Dr A S Parkes FRS Although Sir Alan Parkes and Hilda Bruce set the foundations for the standardisation of animal technology industry with their work on diets and breeding NIMR animal technicians led by Doug Short and the Animal Technicians Association ATA later to become the IAT Institute of Animal Technology 33

Tech 2 Tech spearheaded the development of modern standards and pioneered many advances that would not only improve standards at NIMR but also help establish the standard for the whole industry 1962 Publication of the first concise textbook of Laborator y Animal Technology based on the ATA education syllabus with 24 chapters covering everything from animal handling to the law all but 6 were written by NIMR or LAC people The late 1950s and early 1960s also saw the publication of many important books following the formation of the LAB and ATA as there became an ever increasing demand for the sharing of knowledge 1959 publication of Russell Burch book The Principles of Humane Experimental Technique which introduced the concept of the 3Rs Replacement Refinement and Reduction and they credit the MRC s LAB for carrying out the 1st systematic survey of animal use in the UK and the work carried out by the ATA for improving the efficiency of animal technicians and encouraging communication So the animal technology industry was by now a modern forward thinking profession with the skilled animal technician regarded as an important asset to biological research and a quote by Dr Lane Petter director of LAC in 1962 sums up the advances perfectly 1961 Following the 3Rs book the MRC s LAC published the 1st practical guide for commercial breeders and those involved in their care the MRC LAC accreditation scheme developed at NIMR formed the nucleus for this book Conclusions Animal Technologists and care staff across the whole Biomedical industry have been responsible for some ground breaking advances in laboratory animal science and welfare in the last few decades This is just a small snapshot of advances and innovations made in the past 100 years at MRC NIMR that have contributed towards improved laboratory animal welfare across the whole industry advances and innovations driven forward largely by animal care staff working alongside researchers and veterinarians Undoubtedly a similar story could be told for many establishments across the country References 1 34 Bruce H M and Parkes A S 1946 Feeding and breeding of laboratory animals growth and maintenance of rabbits without fresh green food Journal of Hygiene Cambridge Vol 44 pp 501 507

April 2014 Animal Technology and Welfare You are heroes share your truth CINDY BUCKMASTER Centre for Comparative Medicine Baylor College of Medicine One Baylor Plaza BCM 145 Houston Texas 77030 USA Corresponding author buckmast bcm ed Presented at the IAT LAVA Congress 2013 and reprinted with kind permission of AALAS Laboratory Animal Science Professional Magazine Our work is a labour of love for people and animals How did we become villains I spend a lot of time thinking about the public s misunderstanding of animalbased research and I spend even more time thinking about the truth of it all The public demands cures and safe consumer products Animal based research is still necessary to meet those demands Arguments to the contrary are based in guilt not fact We are villains to people who have blocked out the facts because they do not want to accept the true cost of their demands They blame us for the reality we created together and they want to argue rather than make things right Restoring balance Before we can get to that point we have to dismantle the misperceptions that have been planted in the minds and hearts of the public about the people who work with animals in research Nearly half of our fellow Americans do not support animal based research Some of them do not think it s really necessary for biomedical progress and most of them believe that animals mean nothing to us and we abuse them for profit They view us as monsters and are horrified sickened and enraged by our depravity Our attempts to correct these vulgar distortions of the truth have not had the impact we would like because we ve been battling an emotional crisis with reason Most of our outreach efforts have been focussed on educating the public about the necessity of animalbased research for improving and saving human and animal lives This has been helpful but it is not what people really want to know What they really want us to tell them is that they have been lied to about how we feel about our animals They want to know that we value and care about them They want to know that we treat them with compassion and respect And they need to know that our work is a labour of love for animals and people The selfless nature of this love is extraordinary and uncommon We are now so lost in the details of our arguments that we can t see the truth that joins us we all love our animals and we would all rather they were not necessary for reliable biomedical progress For now they are Arguing about whether animal based research is right or wrong does nothing to change that fact and it does nothing for our animals It may be more realistic for us to qualify our thoughts and actions as loving and not loving When we do our best to be loving under all circumstances we can find common ground and then we can create the change we feel so strongly about together 35

Tech 2 Tech We are heroes for love and compassion and our truth is the key to restoring balance in the hearts and minds of the public The activist community has filled people s minds with grotesque disturbing and distor ted representations of animals in biomedical research This loop of hideous and desperate images is cued to play over and over by every pamphlet magazine letter television commercial and billboard poster prepared by the activists for this purpose Taking it public Let us begin with one of our least favourite questions So what do you do Many of us avoid answering this question because we do not know how to phrase our truth in a way that will elicit interest and understanding and convey our extraordinary compassion for all living beings In the dialogue displayed below I propose some responses for you to consider that may help guide your conversation toward the truth of our work P1 your new acquaintance P2 you Unlocking mysteries with compassion The laboratory animal science community can change this now In fact no one else can do this as quickly and powerfully as we can because no one else can tell our stories When we talk about our work with animals people see the joy and pain in our eyes and they feel the truth in our hearts They come to understand that we love our animals and that we do what we do because we love people too They begin to appreciate the demanding and heartbreaking nature of our work and they become grateful that we are here for our animals and for them and their loved ones This has been my experience with everyone who has asked me about my work I know that some of you reading this article have had the same experience in casual conversations with friends family and strangers It is time now for those who have remained silent to join us in sharing our truth with the public but you need to do so carefully and I have some suggestions to help you get started Guiding Your Conversation P1 So what do you do P2 I care for those animals who care for us all P1 What do you mean P2 Do you know anyone being treated for an illness or chronic disease Any friends or relatives P1 Oh Are you a researcher P2 Is the treatment helping P2 No Researchers study diseases and develop treatments My job is to give research animals the best quality of life I can while they are with us P1 It seems so P1 How do you do that P2 Well I care for those animals who make that possible for all of our loved ones including our pets P2 By providing for their physical and emotional needs with compassion and respect P1 Sure At this point your new acquaintance will probably want to know more about the details of your work and you should be as transparent as you comfortably can The first few lines of your ensuing conversation are the 36

Tech 2 Tech most critical and every word used has to be chosen carefully The activists use a variety of words to cue up the loop of distorted images I mentioned earlier If you use one of those words this loop will play in the mind of the person you are speaking with and they will shut you out Some examples of words to avoid are included below Make your words count Words that may be perceived to objectify animals should be avoided because they confirm the activists assertion that animals are meaningless things to us Do not refer to any of your animals as the animal or it in your conversation Use your animals names when you share your story and refer to them as yours That is how you think of them and it evokes the accountability you feel for your animals and the quality of their lives Similarly avoid saying that research is done on animals or that they are used for research These are words used commonly to describe tasks done with things not living beings Instead refer to your promising work with animals and the hope that research with them brings to millions of people and animals suffering with serious and life threatening illnesses Avoid using the word experiment The activists use this word repeatedly in their propaganda against research because it exploits the representation of poorly conceived mad science that popular horror movies and books have etched in our minds Biomedical progress relies upon carefully designed studies of nature that allow us to improve the quality of human and animal lives This is loving not mad Choosing Your Words Along these same lines avoid saying any of the other words used commonly in activist propaganda like torture and vivisection Do not say them ever even to disagree with them They are cues Instead we have plenty of experiences to share that convey our professionalism and compassion Talk about your relationships with your animals and how you address their species typical needs with environmental enrichment Talk about your animals endearing behaviours and how you work with them through positive reinforcement training to cooperate in studies Be sure to point out that in doing so we shape the quality of lives today and tomorrow because outstanding animal care leads to outstanding biomedical discovery Effecting change Our stories will revise the images playing in the collective mind of the public and begin to unite us in the reality of our unrelenting demands for cures public safety and convenience Animal based research is still necessary for the intense progress we require We are all accountable for this reality There is no us and them We need solutions not arguments In addition we need viable and reliable non animal alternatives that will allow us to continue to produce the answers and conveniences we will never stop demanding And we need to focus the rest of our collective energy and resources on providing those animals still necessary for our needs with the best quality of life we can provide as we work to replace them I am hopeful that we can all get on the same side of that truth because we all love our animals AVOID USE Use of animals Work with study Research on Research with Experiments Studies The animal it Ours yours theirs Their words Your words 37

Animal Technology and Welfare April 2014 The ARRIVE Guidelines Animal Research Reporting of In Vivo Experiments Adapted from the NC3Rs website Introduction The guidelines are NOT intended to The ARRIVE Animal Research Reporting of In Vivo Experiments guidelines were developed in collaboration with researchers statisticians and journal editors as part of an NC3Rs initiative to improve the design analysis and reporting of research using animals maximising information published and minimising unnecessary studies The objective of the ARRIVE guidelines is to maximise the amount of high quality data from a minimum number of experiments to avoid unnecessary animal use Promote uniformity stifle creativity or encourage authors to adhere rigidly to all items in the checklist Some of the items may not apply to all studies and some items can be presented as tables figure legends or flow diagrams e g the numbers of animals treated assessed and analysed The guidelines consist of a 20 item checklist of the key information that should be included in a manuscript Title Abstract Introduction Methods Results and Discussion to ensure comprehensive and transparent reporting The checklist contains the minimum amount of information to ensure that a study can be reviewed analysed and repeated The guidelines were published in the online journal PLOS Biology1 in June 2010 and are currently endorsed by scientific journals major funding bodies and learned societies and have been adopted by Animal Technology and Welfare ATW as the standard for all future submissions of formal papers The guidelines are intended to Improve reporting of research using animals Guide authors as to the essential information to include in a manuscript and not be absolutely prescriptive Be flexible to accommodate reporting a wide range of research areas and experimental protocols Promote reproducible transparent accurate comprehensive concise logically ordered well written manuscripts Improve the communication of the research findings to the broader scientific community 38 Be a guide for study design and conduct However some items on the checklist such as randomisation blinding and using comparator groups may be useful when planning experiments as their use will reduce the risk of bias and increase the robustness of the research Who are the guidelines aimed at G G G G novice and experienced authors journal editors peer reviewers funding bodies What kind of research areas do the guidelines apply to The guidelines will be most appropriate for comparative studies where two or more groups of experimental animals are being compared often one or more of the groups may be considered as a control They apply also to studies comparing different drug doses or for example where a single animal is used as its own control within subject experiment Most of the recommendations also apply to studies that do not have a control group The guidelines are suitable for any area of bioscience research where animals are used How might these guidelines be used The guidelines provide a checklist for those preparing or reviewing a manuscript intended for publication

Tech 2 Tech The ARRIVE Guidelines Checklist Animal Research Reporting In Vivo Experiments Item Recommendation Title 1 Provide as accurate and concise a description of the content of the article as possible Abstract 2 Provide an accurate summary of the background research objectives including details of the species or strain of animal used key methods principal findings and conclusions of the study Background 3 a Include sufficient scientific background including relevant references to previous work to understand the motivation and context for the study and explain the experimental approach and rationale b Explain how and why the animal species and model being used can address the scientific objectives and where appropriate the study s relevance to human biology Objectives 4 Clearly describe the primary and any secondary objectives of the study or specific hypotheses being tested Ethical statement 5 Indicate the nature of the ethical review permissions relevant licences e g Animal Scientific Procedures Act 1986 and national or institutional guidelines for the care and use of animals that cover the research Study design 6 Section paragraph For each experiment give brief details of the study design including a The number of experimental and control groups b Any steps taken to minimise the effects of subjective bias when allocating animals to treatment e g randomisation procedure and when assessing results e g if done describe who was blinded and when c The experimental unit e g a single animal group or cage of animals INTRODUCTION METHODS A time line diagram or flow chart can be useful to illustrate how complex study designs were carried out 39

Tech 2 Tech Experimental procedures 7 For each experiment and each experiment group including controls provide precise details of all procedures carried out For example a How e g drug formulation and dose site and route of administration anaesthesia and analgesia used including monitoring surgical procedure method of euthanasia Provide details of any specialist equipment used including supplier s b When e g time of day c Where e g home cage laboratory water maze d Why e g rationale for choice of specific anaesthetic route of administration drug dose used Experimental animals 8 a Provide details of the animals used including species strain sex developmental stage e g mean or median age plus age range and weight e g mean or median weight plus weight range b Provide further relevant information such as the source of animals international strain nomenclature genetic modification status e g knock out or transgenic genotype health immune status drug or test na ve previous procedures etc Housing and husbandry 9 Provide details of a Housing type of facility e g specific pathogen free SPF type of cage or housing bedding material number of cage companions tank shape and material etc for fish b Husbandry conditions e g breeding programme light dark cycle temperature quality of water etc for fish type of food access to food and water environmental enrichment c Welfare related assessments and interventions that were carried out prior to during or after the experiment Sample size 10 a Specify the total number of animals used in each experiment and the number of animals in each experimental group b Explain how the number of animals was arrived at Provide details of any sample size calculation used c Indicate the number of independent replications of each experiment if relevant Allocating animals to experimental 11 a Give full details of how animals were allocated to experimental groups including randomisation or matching if done b Describe the order in which the animals in the different experimental groups were treated and assessed Experimental outcomes 12 Clearly define the primary and secondary experimental outcomes assessed e g cell death molecular markers behavioural changes 40

Tech 2 Tech Statistical methods 13 a Provide details of the statistical methods used for each analysis b Specify the unit of analysis for each dataset e g single animal group of animals single neuron c Describe any methods used to assess whether the data met the assumptions of the statistical approach Baseline data 14 For each experimental group report relevant characteristics and health status of animals e g weight microbiological status and drug or test na ve prior to treatment or testing This information can often be tabulated Numbers analysed 15 a Report the number of animals in each group included in each analysis Report absolute numbers e g 10 20 not 50 2 b If any animals or data were not included in the analysis explain why Outcomes and estimation 16 Report the results for each analysis carried out with a measure of precision e g standard error or confidence interval Adverse events 17 a Give details of all important adverse events in each experimental group b Describe any modifications to the experimental protocols made to reduce adverse events Interpretation scientific implications 18 a Interpret the results taking into account the study objectives and hypotheses current theory and other relevant studies in the literature b Comment on the study limitations including any potential sources of bias any limitations of the animal model and the imprecision associated with the results 2 c Describe any implications of your experimental methods or findings for the replacement refinement or reduction the 3Rs of the use of animals in research Generalisability translation 19 Comment on whether and how the findings of this study are likely to translate to other species or systems including any relevance to human biology Funding 20 List all funding sources including grant number and the role of the funder s in the study RESULTS DISCUSSION Funding The reporting guidelines project was funded by the National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs 41

Tech 2 Tech Acknowledgements The NC3Rs gratefully acknowledges the expertise and advice that all the contributors have given to developing the guidelines We would par ticularly like to acknowledge the contribution of the NC3Rs Reporting Guidelines Working Group We would also like to thank NC3Rs grant holders the Medical Research Council Biotechnology and Biological Sciences Research Council Wellcome Trust Parkinson s Disease Society British Heart Foundation and their grant holders and funding committee members who provided feedback on the guidelines Further information www nc3rs org uk ARRIVE enquiries nc3rs org uk References 1 2 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 PLOS Biol 8 6 e1000412 doi 10 1371 journal pbio 1000412 Schulz K F Altman D G Moher D the CONSORT Group 2010 CONSORT 2010 Statement updated guidelines for reporting parallel group randomised trials BMJ 340 c332 42

April 2014 Animal Technology and Welfare Report of an RSPCA AHVLA meeting on the welfare of agricultural animals in research cattle goats pigs and sheep PENNY HAWKINS1 R EDDIE CLUTTON2 NGAIRE DENNISON3 MIRJAM GUESGEN4 MATT LEACH5 FINULA SHARPE6 HUGH SIMMONS7 ADRIAN J SMITH8 JOHN WEBSTER9 and UTE WEYER7 1 2 3 4 5 6 7 8 9 Research Animals Department RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Royal Dick School of Veterinary Studies University of Edinburgh Easter Bush Campus Midlothian EH25 9RG Home Office Animals in Science Regulation Unit PO Box 6779 Dundee DD1 9WW Institute of Veterinary Animal and Biomedical Sciences Massey University Private Bag 11 222 Palmerston North 4442 New Zealand Pain Animal Welfare Science PAWS Group School of Agriculture Food Rural Development Newcastle University Newcastle Upon Tyne NE1 7RU HLS Woolley Road Alconbury Huntingdon Cambridgeshire PE28 4HS AHVLA Animal Sciences Unit AHVLA Weybridge Addlestone Surrey KT15 3NB Norecopa c o Norwegian Veterinary Institute P O Box 750 Sentrum 0106 Oslo Norway Old Sock Cottage Mudford Sock Yeovil Somerset BA22 8EA Corresponding author penny hawkins rspca org uk Introduction The Royal Society for the Prevention of Cruelty to Animals RSPCA and the Animal Health and Veterinary Laboratories Agency AHVLA jointly convened a meeting to bring together animal technologists researchers veterinarians and students with an interest in the welfare of cattle goats pig and sheep used in research and testing for a programme of talks and discussion sessions The meeting held in June 2013 addressed a range of topics including cognition and emotion in agricultural animals welfare in containment systems positive reinforcement training pain assessment and alleviation and assessing actual severity It was initiated as a follow up to an international meeting on harmonisation of the care and use of agricultural animals in research held by Norecopa the Nor wegian consensus platform for alternatives in September 2012 Norecopa a national consensusplatform for the 3Rs Adrian J Smith Norecopa In 1999 following an initiative at the 3rd World Congress on Alternatives and Animal Use in the Life Sciences in Bologna the same year a European organisation was established with the purpose of bringing together all four stakeholders interested in animal research regulators academia industry and animal welfare organisations This organisation known as ecopa European Consensus Platform for Alternatives www ecopa eu offers membership and support to national platforms which follow the same principles Currently there are 14 national platforms in Europe that are approved by ecopa Nor way s national consensus platform Norecopa www norecopa no was established in 2007 Institutions and individuals can become members of Norecopa and most Norwegian research institutions have done so Norecopa signals its commitment to all 3Rs of Russell Burch by describing itself as a national platform for replacement reduction and refinement of animal experiments Inspired by the 2004 FELASA meeting in Nantes entitled Internationalisation and Harmonisation of Laboratory Animal Care and Use Issues Norecopa has arranged a series of international consensus meetings on the care and use of animals in research These meetings have so far covered fish in 2005 2009 wildlife in 2008 and agricultural animals in 2012 43

Tech 2 Tech Norecopa wishes these meetings to be a lasting resource for the international research community so each meeting has its own website http www norecopa no sider tekst asp side 21 where all the presentations are available together with links to guidelines on the care and use of the species in question In addition participants write a consensus document describing the challenges to implementing the 3Rs which were identified during the meeting These consensus documents have been used actively by Norecopa and others to initiate tasks designed to address these challenges Norecopa has also established email based discussion forums following these meetings but interest in these has so far been low the reasoned interpretation of sensation and experience can help to improve welfare by teaching the animal to cope However it may in some circumstances increase the degree of suffering when the coping mechanisms fail Never theless suffering is an emotional state it is a fallacy to assume that the greater the apparent cognitive ability of a species still less its phenotypic similarity to humans the greater its capacity to suffer There is no good argument to suggest that the capacity of a chimpanzee to suffer is any greater than that of a dog or pig 2 Norecopa welcomed the RSPCA AHVLA meeting as an initiative which should help to advance efforts to increase implementation of the 3Rs in the care and use of agricultural animals Agricultural animals in research cognition emotions and ethics Emotion Security Fear Pleasure The consensus document resulting from the 2012 meeting on agricultural animals included comprehensive recommendations and a list of tasks relating to the implementation of the 3Rs when using agricultural animals in research and testing These included further meetings and discussion fora on 3Rs topics for a range of stakeholders There was considerable support for this among UK researchers animal technologists and veterinarians so delegates from the RSPCA and AHVLA went on to convene a meeting based in the UK in June 2013 Positive aspect Negative aspect Comfort Pain Satiety Appetite Hunger Joy Hope Despair Anxiety Table 1 Aspects of different animal emotions I also contend that sentient animals do not simply live in the present Primitive sensations like hunger or pain lead a domestic animal to hope that a meal will be provided on time or fear that they may receive a beating An animal with the capacity for hope will also have the capacity for despair 3 How then should we apply these concepts of sentience to our approach to the care of agricultural animals especially those used in research where we are obliged to weigh any harms to the animals against benefits to the society of humans and other animals I suggest the following rules of engagement John Webster University of Bristol Agricultural animals used in research are considered to be sentient Definitions of sentience vary as does the nature of sentience itself within the animal kingdom However within the homeothermic classes of mammals and birds I stand by my definition of sentience as feelings that matter 1 Sensations resulting from environmental stimuli motivate the sentient animal to actions that are designed to avoid suffering and promote a satisfactory emotional state If the actions are successful the animal learns to cope If they fail then the animal is likely to suffer Failure to cope may occur either when the stresses are too severe complex or prolonged or when the animal is constrained in such a way that they are unable to take constructive action necessary to relieve the stress The primary motivation for sentient animals including humans is emotion the need to feel good avoid suffering and ignore the things that do not matter see Table 1 for some examples of animal emotions and their positive and negative aspects Cognitive ability or 44 1 Harms should be assessed not only in terms of their short term effects on the physical state of the animal but also in terms of their longer term effects on emotional state e g anxiety learned helplessness 2 The day to day husbandry of the animal must not only be sufficient to meet physical needs for nutrition comfort etc but should also wherever possible provide the resources necessary to allow the animal to achieve a satisfactory emotional state through their own actions 3 Species selection must be made on a case by case basis where the benefits are assessed by weighing the scientific evidence relating to the predictivity of the animal model against the harm that may accrue to the animals both from the scientific procedures and from their lifetime experience within the research environment 4 The capacity of an animal to experience suffering must be defined in terms of their own sentience not their status in human society It is unethical to promote agricultural animals e g minipigs as more acceptable than dogs or monkeys simply on

Tech 2 Tech the basis of public identification of the pig as a food animal the dog as a companion animal and primates as those species closest to man Box 1 The Five Freedoms 1 Freedom from hunger and thirst 2 Freedom from discomfort Welfare challenges in high disease containment research Hugh Simmons AHVLA and NADIR Network of Animal Disease Infectiology Research Facilities 3 Freedom from pain injury and disease 4 Freedom to behave normally 5 Freedom from fear and distress Disease causing infectious agents must be contained if they are i exotic and contagious to animals ii infectious to humans iii genetically modified or a combination of two or all three of these factors These three different containment requirements are covered by three different pieces of legislation in the UK which specify four different levels of containment in each piece of legislation There is some variation in the detail of the containment required at each level The highest are Categories 3 and 4 Category 3 applies to pathogens such as Classical Swine Fever Blue Tongue Virus and Tuberculosis TB while Category 4 applies to Foot and Mouth Disease Rabies and High Pathogenicity Avian Influenza Freedom from hunger and thirst Fitted troughs drinkers and water bowls are often found in Category 3 buildings just as in low security facilities However Category 4 buildings are more challenging due to the greater stringency on how waste can be removed from the room or building In a lot of designs virtually all the waste has to be washed down the drain to the ETP This means a complete diet has to be offered without long stem fibre for ruminants which reduces normal rumination function As there is no suitable alternative source of long stem fibre studies are planned so as to minimise the duration of category 4 housing and animals are fed a diet with high content of fibre but not long stem These disease containment legislations define key elements of building design and management necessary to ensure containment such as air handling units with high efficiency particulate air HEPA filters effluent treatment plant ETP on the drainage system sterilisation or incineration of waste and changing facilities where staff change into the required personal protective clothing and equipment PPE These factors can impose restrictions on the kind of housing and environment that can be provided for the animals for example long stem fibre roughage and bedding may block ETPs or PPE may affect interactions between animals and staff Once such highly specialised facilities have been built it can be difficult or impossible to update them to take account of some new developments in housing and care due to their construction and complexity Freedom from discomfort The risk of environmental discomfort should be low as building management systems within containment facilities keep temperature and relative humidity within appropriate limits With respect to physical comfort animals need somewhere comfortable to rest which can present problems if it is not possible to provide conventional bedding materials If animals are held on non slip flooring without bedding they may be uncomfortable and can develop sores or bursae at pressure points during longer experiments One way in which animal comfort is assessed is to evaluate how long they spend lying on different materials and then provide the preferred material unless there are significant veterinary or scientific constraints Considering all of this undertaking research using farm animals into diseases that require high containment can present challenges that must be overcome to ensure animal welfare Currently all the European organisations with these types of facilities for farmed animals including AHVLA are members of NADIR This is a FP7 project funded by the European Union with the aim of sharing good practice in a number of areas including animal welfare http www nadir project eu Some of the approaches used by NADIR and its members to improve animal welfare are set out below using the Five Freedoms as an analysis tool Box 1 If some discomfort is unavoidable the aim is to address this within the experimental design so that non infectious phases are conducted outside keeping the time in containment to a minimum Freedom from pain injury and disease The nature of the infectious agent used and design of the experiment will also both be critical to the animal s freedom from pain injury or disease In common with animals kept in low security accommodation at AHVLA a Health Plan is implemented for each animal to eliminate any health problems that are not related to the study It is particularly important that animals are in good health before going into high containment not only to minimise any effects on welfare of the experimental procedures and any husbandr y 45

Tech 2 Tech constraints but also due to the logistical problems of treating any intercurrent disease in high containment facilities This would have a potentially catastrophic impact on the validity of the study especially as high containment experiments are generally done on small numbers of animals It is obviously impossible for animals to be free from pain or disease when they are used in disease research so refinement is critical The impact of disease can be reduced by implementing an effective health and welfare monitoring protocol that includes humane endpoints Staff contact is unfortunately limited to defined periods when animals are in higher categories of containment but CCTV can be used for continuous obser vations Although this cannot substitute for interactions with an empathetic and experienced observer or extra monitoring for animals on critical points of severe protocols CCTV can help to maintain surveillance and can also provide footage that can be sampled and reviewed The welfare score charts used at AHVLA are always tailored to individual projects and approved by the institutional ethics committee Humane endpoints are refined using predictive indicators wherever possible For example clinical signs and disease progression vary between strains of classical swine fever but real time analysis of white blood cell WBC and platelet counts have enabled predictive endpoints to be established Animals are now humanely killed when the WBC and platelet counts fall so that they do not have to experience the full progression of the disease Other measures to reduce suffering and refine procedures at AHVLA include G G G G G G pre start meetings for researchers veterinarians animal technologists and care staff to ensure that all are aware of critical indicators and decision points telephone contact list for out of hours if euthanasia is required procedures timed so that the phase with the greatest potential to reach end point will occur early or mid week samples taken early in the morning so that results will be available that day enabling better decision making with respect to endpoints experienced staff are responsible for animal monitoring and care for groups at greatest risk e g unvaccinated controls use of microchips to monitor body temperature which reduces handling Freedom to behave normally The legal minimum space allowances for animals in high containment are no different from those for animals in conventional housing However containment 46 systems can have an impact on the kind of environment that can be offered to the animals and consequently on their freedom to behave normally Environmental enrichment is possible in containment building designs and should be tailored to the species being housed e g long fibre foodstuffs and social groups are very important for ruminants straw and other materials or objects to manipulate are good for groups of pigs reducing bullying and injuries within the group Freedom from fear and distress A key concept here is facilitating emotional resilience i e providing a set of conditions that allow emotional adaptation to different forms of adversity at different times in the animal s life Ways of fostering emotional resilience include allowing acclimatisation periods to the housing system before animals are used and very importantly training animals before the experiment including giving food rewards Care is also taken to select individuals and breeds with suitable temperaments as not all breeds are suited to high containment This is also a human health and safety issue when working with large animals in contained spaces In addition to the Five Freedoms at AHVLA we also sum up our approach as Four Ps Positive cultural attitude towards identifying any issues or problems The facility user group holds both pre start and wash up meetings before and after studies for open discussion and learning for future studies Proactive ongoing development of welfare score sheets continually refining humane endpoints with the goals of moving to predictive endpoints as well as providing comfort and environmental enrichment Preventative measures to directly or indirectly reduce suffering such as building design experimental design developing emotional resilience in animals Productive approach acknowledging that better welfare and better science go hand in hand Clicker training in minipigs at Huntingdon Life Sciences Finula Sharpe HLS Note much of this presentation has previously been published in Animal Technology and Welfare see Arblaster 2010 4 Introduction Handling minipigs throughout long term scientific procedures can pose both welfare and practical problems Mature minipigs can weigh 30 kg and

Tech 2 Tech animals of this size and strength are difficult for the majority of animal technologists to lift or carry safely which can result in stress to the animal and physical strain for the handler to walk into the dosing trolley by the end of the 2 week acclimatisation period while others were more cautious about taking treats from the trainer s hand and took a little longer Clicker training has therefore been introduced as a method of avoiding stressful carr ying and sling restraint for minipigs undergoing dosing during dermal studies 5 The aim of the clicker training programme at HLS was to train 52 minipigs to walk voluntarily from their pen into a dosing trolley stand for the dosing procedure and walk back to their pen once it had been completed To ensure consistency in the training method each technologist involved was given training and a set of instructions to follow in addition to the six stages of training Method The training programme used operant conditioning in which the minipigs were trained to form positive associations between certain behaviours and fruit rewards such as pieces of apple and grape The minipigs began training at the age of 4 weeks and each animal was trained individually in an empty pen for a session lasting approximately 5 minutes with the handler seated on the pen floor Animals were taken through 6 stages of training listed below which gradually progressed towards walking into a dosing trolley If at any point a minipig was showing signs of confusion then the session would be stopped and training taken back to the previous stage to reestablish the desired behaviour Stage 1 Click of clicker each time the minipig takes a treat from the trainer s hand Stage 2 Treat and simultaneous click of the clicker each time the minipig moves towards a target Stage 3 Treat click each time they touch the target Stage 4 Treat click each time they follow the target around the pen Stage 5 Treat click each time they follow the target out of the pen and back in again Stage 6 Treat click each time they follow the target into the dosing trolley and out again Progress was recorded on a daily tick sheet which highlighted for example if the animal touched the target followed the target around the pen or was nervous of the handler This enabled us to ascertain whether or not animals were ready to move onto the next stage of training Training sessions were carried out daily for 2 weeks before the start of the study Outcome The initial aim of training all minipigs to walk voluntarily into the dosing trolley was achieved by the third week of the study This was aided by animals having access to their daily food ration while standing in the dosing trolley as it provided a distraction from the dosing procedure Some minipigs were sufficiently confident Clicker training minipigs it is important that the minipig is alone in the pen during each training session G only reward the minipig when they perform the desired behaviour G the moment a minipig per forms the desired behaviour they should be rewarded immediately with a simultaneous treat and click G it is important to have a positive attitude at all times and not get frustrated if the animal doesn t perform what is required they should be ignored until they can focus on the trainer again G if the minipig becomes confused or unfocused training should be stopped and taken back to a safe phase that the animal is more familiar with G if the animal becomes scared and runs away while training outside the pen calmly walk do not run after them and bring them back with the target and a reward G before training animals to walk into the dosing trolley raise and lower the trolley outside the pen so the minipig can get used to the sound G once trained the minipigs will not need the target any more as they will be attracted by their daily food ration in the trolley G Conclusion The aim of the clicker training programme was achieved and this positive behaviour continued throughout the remainder of the study While the age of the minipigs was a significant issue it was an unavoidable factor due to the constraints of the study protocol which required animals to be 6 weeks of age at the start of the study Clicker training did also serve a purpose in allowing the minipigs to become more familiar with handlers outside of normal husbandry procedures Clicker training minipigs provides a significant improvement in the dermal dosing procedures used at Huntingdon Life Sciences as it removes the need to handle the minipigs and therefore reduces stress to the animals Using positive reinforcement as a method of refining dermal dosing procedures therefore improves animal welfare It also reduces physical strain placed on the animal technologists involved Clicker training could also be used for other studies using minipigs where animals need to be moved from the animal unit to a dosing room 47

Tech 2 Tech Pain assessment and alleviation in agricultural species used in research R Eddie Clutton University of Edinburgh Introduction Non avian agricultural species cattle sheep pigs and goats are used for a number of different purposes in research6 Two examples of research fields where pain may be caused are i the investigation of legal though potentially painful husbandr y operations e g castration and ii experimental surgery with recovery e g organ transplantation Pain assessment and alleviation are clearly very important in both of these contexts When applying a harm benefit assessment it may be possible to justify inadequate pain management in i for example to demonstrate that a husbandry procedure is a welfare issue or to evaluate potential techniques for pain relief However inadequate pain management can never be justified in the case of ii Sheep goats and pigs have also been used to try to study human pain In accepting the validity of these species as human pain models the scientific community must accept that these species feel pain7 and act accordingly the provision of effective analgesia when these species undergoing painful procedures is a matter of justice as well as an act of refinement The extensive use of pigs in biomedical research is constantly justified on the basis of their physiological and anatomical similarity to human beings so it is surprising that with few exceptions the literature devoted to laboratory pig welfare is notable by its absence of references to pain recognition and treatment Pain recognition Acute pain behaviours resulting from husbandr y procedures in young agricultural animals are well recognised Little is known about similar behaviours after experimental surgery and it may be imprudent to extrapolate information because age and breed are confounding factors Pain evaluation is also complicated because in general agricultural species show less obvious to the human eye pain behaviours than horses or carnivores Behaviours may also be counterintuitive to humans for example sheep may display obsessive eating behaviour when in pain but this may be interpreted as animals feeling well The fact that some agricultural animals are less likely to mount fight or flight reactions may account for their popularity as large animal models along with legallycondoned speciesism Unfortunately all of this means that attempts to categorise pain behaviours in order to assess pain 48 severity may be misleading Indeed no validated nor widely accepted pain scoring system currently exists for farm animal species undergoing procedures that are unrelated to husbandry8 On a more positive note if animal technologists are familiar with individual animals undergoing potentially noxious procedures they can be better able to detect subtle behavioural changes that indicate the presence of pain and initiate suitable treatment It can be time consuming to build up this level of trust but empathetic staff play an important role in ensuring that animals are given the benefit of the doubt with respect to experiencing pain and resources should be made available for this Pain alleviation Information on analgesic drugs in agricultural species including in the scientific literature is extremely limited beyond those used in painful production disorders e g mastitis This may be because there has been no perceived need for analgesia if pain behaviours are subdued or because legislation drafted to protect human food supplies limits commercial incentives for the pharmaceutical industry to fund pain studies in food animals Determining the efficacy of ad hoc analgesic therapy is complicated by the limited pain behavioural repertoire in the species involved However the literature is gradually increasing and attitudes are changing Strategies being increasingly employed as par t of anaesthetic techniques to minimise an animal s post operative discomfort are pre emptive analgesia the administration of analgesics before surgical injury is created polymodal pain therapy the administration of analgesics from several therapeutic classes at relatively low doses such that the combined effect is adequate analgesia but a reduced risk of side effects par tial intravenous anaesthesia in which analgesic drugs are used at higher doses in order to contribute to the state of anaesthesia and thus reduce the requirement for general anaesthetic and prolonged post operative analgesia which involves anti inflammatory medication being given until the surgical wound has healed All four strategies may be combined within a given anaesthetic technique Conclusion The dearth of information on pain recognition and treatment in agricultural animals relative to companion and other laboratory species prompts consideration of a major limitation on their use in potentially noxious recovery experiments There is no justification for withholding analgesia or some kind of pain relief technique when conducting potentially painful procedures on agricultural animals

Tech 2 Tech Assessing pain using the Lamb Grimace Scale LGS Mirjam Guesgen1 and Matt Leach2 1 Institute of Veterinary Animal and Biomedical Sciences Massey University New Zealand and 2 School of Agriculture Food Rural Development Newcastle University Pain in animals is of considerable public concern Within biomedical research pain is of additional concern because it not only compromises animal welfare but can also have implications for the validity of scientific results In order to address this and alleviate pain it is essential to assess its severity and duration effectively New behaviour based methods for assessing pain are providing evidence for a relationship between pain and behaviour and providing additional tools for pain management For example facial expressions are routinely used to assess pain in humans particularly those who are nonverbal and it has also been shown that human observers tend to focus on animals faces when assessing pain This led to studies of pain faces in non human animals and the use of facial expressions as indicators of pain is now being developed for several species such as rodents9 10 11 and rabbits 12 In these studies analysis of video stills has enabled the development of species specific Grimace Scales which have been shown to be accurate reliable and valid measures of pain A further benefit is that evaluation studies can be done using video footage of animals already undergoing scientific or husbandry procedures for other purposes thereby gaining information that can help to reduce suffering but without causing additional harms The aim of this study was to identify whether lambs produce noticeable changes in facial expressions when experiencing pain and if so to develop and validate a Grimace Scale that incorporates these expressions 1 2 First the LGS was defined using five so called action units orbital tightening mouth changes nose changes cheek flattening and ear changes Each one is scored on a 3 point scale 0 not present 1 moderately present and 3 obviously present Figure 1 depicts the Rabbit Grimace Scale12 to give an impression of the appearance of animal facial action units the Lamb Grimace Scale will be published at a later date The LGS was then trialled using still images of the faces of restrained Romney X lambs from video footage taken before and after sham docking or routine taildocking with a rubber ring in an on farm situation Images were randomly selected and scored for four of the action units not ear position as lambs were restrained by four treatment blind observers The results showed that in this instance the LGS had high reliability within and between observers but low accuracy i e people were scoring consistently but there was a high proportion of false positives This was likely due to restraint stress which can also influence facial expressions In a follow up study video sequences of the faces of unrestrained lambs were taken before and after taildocking or sham docking and scored by treatment blind observers as above with ear position also included This time reliability was also high within and between observers and accuracy was significantly improved with fewer false positives In summary we found that the LGS changes in response to tail docking and that this can be reliably coded by observers However as LGS is also affected by restraint further validation is required to see whether it is possible to identify those action units that are associated with pain and with distress This study demonstrates that lambs change a number of their facial features when experiencing pain and that these features can be accurately and reliably coded and quantified It is hoped that the LGS can go on to become a routine tool for pain and welfare assessment 3 Lifetime experiences and actual severity Figure 1 Facial action unit in a rabbit orbital tightening Ngaire Dennison Animals in Scientific Procedures Inspectorate ASPI Legend The eyelid is partially or completely closed and the globes may also be drawn in toward the head so that they protrude less Score 0 not present 1 moderately present 2 obviously present the eye closure reduces the visibility of the eye by more than half 12 The UK Animals Scientific Procedures Act 1986 ASPA requires that a project licence to carry out a programme of scientific work using living animals may only be granted where there has been a favourable evaluation of the proposed project This evaluation must include a harm benefit analysis of the programme to assess 49

Tech 2 Tech whether the harms that would be caused to animals in terms of pain suffering and distress could be justified by the expected outcome taking into account ethical considerations and the expected benefits to human beings animals and the environment Part of the prospective assessment of harm requires that the series of procedures that an animal undergoes during an experiment be classified as non recovery mild moderate or severe with respect to suffering that may occur In addition due to the changes in the ASPA required to implement the recently enacted European Directive 2010 63 EU there is now a requirement for the assessment and reporting of the actual severity of the regulated procedures There are many benefits associated with assessing and reporting actual severity Table 1 but with the proviso that judgements have to be suitably informed objective and consistent Actual severity assessment should G G G G ensure that what actually happened to the animal is recorded and understood provide evidence based information for future prospective harm benefit assessments facilitate effective retrospective review identify areas where the application of the Three Rs should be focused G provide feedback to help refine ongoing studies G provide input for thematic reviews G G lead to statistics that will better reflect the actual harms to animals improve transparency and understanding of the impact of science on animals Table 1 Benefits of actual severity assessment The law requires that the assessment of actual severity be made by a competent person and as with the prospective classification classified as non recovery mild moderate or severe but in the case of actual severity it must be based on the actual impact on the individual animal A number of sources of information should be used to determine the experience of the individual including day to day obser vations of behaviours and clinical signs observed throughout the procedure This requires good planning team work and training as well as appropriate choices of indicator and recording systems Training including ensuring appropriate attitudes is critically important for staff working with farm animals as these animals can be perceived by those without the appropriate education as stoic due to lack of understanding of their behaviour as discussed above or simply as production animals somehow less deserving of consideration than companion animal species 50 Further guidance on all of the aspects of assessing actual severity outlined in the paragraph above has been produced by the European Commission 13 14 which applies across all species but the examples provided are of limited help to those working with farm animals for two main reasons First the worked examples are in rodent and rabbit models and it is important to consider cases of relevance to large animals to identify any species or model related difficulties in assessing severity for work involving larger animals Second the examples tend to relate to the classification of a single procedure whereas farmed animals are often on study for a long time and are more likely to undergo a series of experimental techniques Re use of farm animals in further scientific procedures for a second and separate purpose may be requested for a number of reasons including reduction of overall animal numbers avoidance of the requirement for na ve animals for surgery or for bio security reasons There may be an additional financial element to considerations if significant resource has been expended on training or otherwise preparing the animals There are understandable concerns in the scientific and welfare communities about achieving compliance with the legal requirement for assessment of actual severity especially in terms of ensuring accurately and consistency and also with respect to how other potential causes of discomfort pain or stress within the life time experience of the animal e g transport should be factored in People are often anxious that they will get it wrong and may simply opt for reporting the prospective severity limit which is not helpful because it i provides little advantage over the previous system of reporting and ii provides no information about studies that are more or less severe than expected Other concerns relate to the time taken to do the assessment for each animal but if the assessment is done properly the benefits in Table 2 should outweigh this and confusion about what is meant by consistency in assessments It is important to recognise that the level of actual suffering may differ from the prospective judgement and also may vary between different animals undergoing exactly the same procedures depending on the individual s responses to these That is consistency is about everyone scoring each animal in the same way it is not about expecting the same model to give the same results between or even necessarily within establishments In the UK any such re use must be authorised in advance by the Secretary of State The fact that the animal has been used before must be included within the harm benefit assessment the actual severity of the previous procedures must be assessed and checked against defined criteria and a veterinar y surgeon with knowledge of the lifetime experience of the animal has to advise that the animal s general state of health and wellbeing have been fully restored following the previous use

Tech 2 Tech So how to achieve this Besides making full use of the available guidance from the European Commission and the forthcoming guidance from the Home Office in the UK it is a good idea to communicate with and consult others with relevant expertise Within the establishment this could include animal technologists the Named Animal Care and Welfare Officer NACWO veterinarians the Named Information Officer NIO researchers and internal animal care and use committees including the Animal Welfare and Ethical Review Body AWERB Externally useful contacts could be members of the Home Office Inspectorate HOI or scientists and technicians from other groups doing similar work What people were doing to assess suffering Day to day welfare was mainly assessed by animal technologists whom delegates also believed to be best at the task see Figure 2 Most delegates 31 recorded observations using clinical record sheets that were mostly free text whereas fewer 10 used numerical or binar y score sheets three used electronic data logging systems and 11 reported that the recording system depended on the species and experimental protocol In a session with speciesspecific breakout groups delegates listed the indictors that they commonly used to assess both good and suboptimal welfare Appendix 1 Communication with the local Home Office Inspector HOI is absolutely essential if the severity limit of the protocol is found to have been exceeded e g if an animal on a study with a moderate severity limit has experienced severe suffering Some concerns have been expressed that this would automatically and in itself be non compliant with the requirements of ASPA However provided each case is promptly reported to the HOI as required by standard condition 18 of the project licence the user should normally still be compliant with the Act Condition 18 would only be breached if the HOI was not notified Discussion session The rest of the meeting was devoted to a discussion session in which delegates exchanged views on the way in which they assessed severity and the kind of resources they felt might be useful to help achieve good standards of objectivity and consistency An overview of the outcomes is presented below note that TurningPoint was used to survey the audience so the numbers of responses will vary Audience demographics The audience of around 70 delegates comprised mostly animal technologists 28 of whom 12 were NACWOs also 14 researchers 9 veterinarians several members of AWERBs NIOs Named Training and Competency Officers NTCOs and two HOIs Most people 36 worked in government research or diagnostic facilities with fewer employed within academia 19 and industry 10 Delegates worked with pigs 44 people sheep 34 cattle 31 poultry 23 and goats 10 The majority of attendees 53 had attended because they felt there were welfare challenges associated with farm animal use whereas 27 people felt reasonably happy with the situation but wanted to ensure they were updated For non UK readers the NACWO is a senior animal technologist responsible for inter alia ensuring day to day care of the animals and the NIO is responsible for ensuring that staff have access to relevant information specific to the species at the establishment Figure 2 Delegate poll who assesses animal welfare day to day and who does it best Legend Delegates could vote for up to three options for who assesses welfare and one for who does it best With respect to the latter question 7 answered it depends Figure 3 Delegate poll are the Directive and or the documents produced by the EC and its Expert Working Groups of any help with respect to assessing the welfare of agricultural animals Legend The figures refer to the number of delegates voting for each option 51

Tech 2 Tech The requirement to record and report actual severity Delegates were asked whether their facility was ready to take on the task of assessing actual severity 19 said yes 28 said they were working on it and 17 said no Most people were entirely or at least partly satisfied with the training they had received relating to welfare assessment 24 and 22 respectively as opposed to 15 who were not satisfied but only 12 of 67 delegates who answered 18 see Figure 3 had read the guidance from the EC on severity assessment and found it useful Discussion on a potential forum for large animal users A straight question was put to the audience would you use a large animal network such as a discussion forum or virtual actual meetings if one were available Fifty four delegates answered yes as opposed to twelve who replied that they would not The potential benefits of a UK forum are set out in Figure 4 There was a fairly equal preference amongst delegates for an online discussion forum and meetings and 22 people felt that the IAT could do more for those working with large animals Clearly any forum would have to be hosted by an appropriate body and the audience were given a number of options to vote for a scientific welfare organisation 18 votes a leading agricultural research facility 15 the NC3Rs 11 a body like IAT or LASA 10 other 8 or the authorities 1 Figure 4 Delegate poll what would you want from a UK forum Legend The figures refer to the number of delegates voting for each option While there was support for a UK forum of some kind at the meeting Norecopa s attempt to set up a similar multinational forum was met with limited interest Further thought and consultation will be necessary as to the kind of format that could be supported by relevant bodies The outcome of this discussion will be passed on to IAT LASA the NC3Rs the UK Animals in Science Committee and the Home Office The RSPCA and AHVLA are also exploring the potential for another joint meeting at the time of writing 52 Overall action points The following action points are suggested on the basis of the talks and discussions on the day Be prepared to accept and recognise emotional states in agricultural animals challenge assumptions to the contrary Consider how a positive emotional state might be encouraged for the animals in your care taking into account as appropriate factors such as selection of species strain and individual the potential for refining housing care procedures and the way in which they are conducted recognising and assessing desirable behaviours that indicate positive wellbeing Review large animal housing and care with reference to the Five Freedoms and AHVLA s Four Ps above including animals housed under all biosecurity levels If animals in containment do not have suitable housing and husbandry refinements identify any obstacles and work to overcome these Consider whether there is the potential to use positive reinforcement training within any procedures either to obviate restraint or provide stimulation for the animals Make no assumptions about the nature and level of pain an animal may be experiencing on the basis of their behaviour alone Keep up with the literature and current thinking on the assessment of pain and suffering including new approaches such as Grimace Scales Challenge any requirements to withhold analgesia or some other appropriate pain management technique Review the guidance on actual severity assessment produced by the European Commission and relevant national regulator e g the UK Home Office seek further advice and clarification if needed and feedback to the authors if the guidance is not useful at your facility If you have any thoughts relating to a discussion forum for those working with agricultural animals or ideas for further meetings on agricultural animal use contact research animals rspca org uk Acknowledgements Thank you to all of the delegates and speakers for their support and enthusiasm for the meeting and for the useful and interesting discussions Many thanks also to AHVLA staff for their very welcome assistance which helped to make the day such a success References 21 22 Webster J 2005 Animal Welfare Limping towards Eden Blackwell Oxford Webster J Bollen P Grimm H and Jennings M 2010 Ethical implications of using the minipig in regulatory toxicology studies Journal of Pharmacological and Toxicological Methods 62 160 166

Tech 2 Tech 23 24 25 26 27 28 29 Webster J 2011 Anthropomorphism and zoomorphism useful fallacies Animal Welfare 20 29 36 Arblaster F 2010 Clicker training in minipigs Animal Technology and Welfare 9 115 118 Bertelsen T Nielsen T C Lund G and Gade L P 2008 The use of clicker training in minipigs Ellegaard Newsletter 30 6 7 http minipigs dk fileadmin filer Newsletters Newsletter_30 pdf last viewed 30 October 2013 Norecopa 2012 Consensus Statement Harmonisation of the Care and Use of Agricultural Animals in Research http www norecopa no norecopa vedlegg Consensusstatement agricultural animals pdf last viewed 30 October 2013 Morton D B and Griffiths P H M 1985 Guidelines on the recognition of pain distress and discomfort in experimental animals and an hypothesis for assessment Veterinary Record 116 431 436 Molony V and Kent J E 1997 Assessment of acute pain in farm animals using behavioural and physiological measurements Journal of Animal Science 75 266 272 Langford D Bailey A Chanda M Clarke S Drummond T Echols S Glick S Ingrao J KlassenRoss T LaCroix Fralish M Matsumiya L Sorge R Sotocinal S Tabaka J Wong D van den Maagdenberg A Ferrari M Craig K and Mogil J 2010 Coding of facial expressions of pain in the laboratory mouse Nature Methods 7 447 449 10 11 12 13 14 Sotocinal S Sorge R Tuttle A Marton L Wieskopf J Mapplebeck J C S Wei P Zhan S Zhang S McDougall J J King O D and Mogil J S 2011 The Rat Grimace Scale A partially automated method for quantifying pain in the laboratory rat via facial expressions Molecular Pain 7 55 http www ncbi nlm nih gov pmc articles PMC316360 2 last viewed 30 October 2013 Leach M C Klaus K Miller A L Scotto di Perrotolo M Sotocinal S G and Flecknell P A 2012 The assessment of post vasectomy pain in mice using behaviour and the mouse grimace scale PLoS ONE 7 4 e35656 Keating S C J Thomas A A Flecknell P A and Leach M C 2012 Evaluation of EMLA cream for preventing pain during tattooing of rabbits Changes in physiological behavioural and facial expression responses PLoS ONE 7 9 e44437 European Commission 2012 Working Document on a Severity Assessment Framework http ec europa eu environment chemicals lab_animals interpretation_en htm last viewed 30 October 2013 European Commission 2013 Examples to Illustrate the Process of Severity Classification Day to day Assessment and Actual Severity Assessment http ec europa eu environment chemicals lab_animal s interpretation_en htm last viewed 30 October 2013 Appendix 1 Commonly used indicators of good and suboptimal welfare in cattle pigs and sheep These indicators were listed by breakout groups addressing single species They have been organised into the categories set out by the EC Working Group on severity assessment13 with the intention of providing examples of indicators that may be helpful when reviewing welfare assessment for cattle pigs and sheep The procedure specific indicators are usually tailored to particular procedures species breeds where appropriate and stages of development but the examples in the tables below apply to a range of different procedure types for each species Good welfare cattle High level categories Areas to focus on when observing animals Specific indicators to monitor Appearance Body condition Good body condition although this can depend on the breed e g Holstein Fresians typically appear to have lower condition scores than other breeds so other indicators should be considered in conjunction with body scores for these Coat and skin condition Shiny coat Environment Enclosure environment including any litter nesting material enrichment items Normal faecal consistency Interest in novel objects curiosity Behaviours Social interaction Socialisation and group interaction including playfulness and social grooming Good response to stockperson Posture and mobility Good posture Lies down easily Other Chewing cud Coming forward and vocalising for food Alert 53

Tech 2 Tech Good welfare pigs High level categories Areas to focus on when observing animals Specific indicators to monitor Appearance Eyes Bright eyes Environment Enclosure environment including any litter nesting material enrichment items Foraging using toys playing with low grunting Inquisitive curious e g when person enters the room in response to noise new objects or change in environment Rooting exploring with nose and mouth contact Behaviours Social interaction Interacting with other pigs snorting and running together Positive responses to stimulation Making eye contact nose up Posture and mobility Bouncing pop corning Relaxed posture Other Tail wagging when happy Vocalising before feeding interest in food Nest building with new bedding bouncing into new beds Barking when excited or anticipating Quiet vocalisations e g soft grunt when greeting Scratching dry skin Good welfare sheep High level categories Areas to focus on when observing animals Specific indicators to monitor Appearance Body condition Good health Body functions Food water intake Good appetite Behaviours Social interaction Socialisation with humans Normal interactions with other sheep Other Expression of normal range of behaviours Suboptimal or poor welfare cattle High level categories Areas to focus on when observing animals Specific indicators to monitor Appearance Body condition Poor body condition although this can depend on the breed see above Eyes Sunken eyes Wide open eyes Other Bloat this is an immediate cause for concern Respiration Rapid heavy breathing Food water intake Depressed appetite Body functions 54

Tech 2 Tech Other Increased heart rate Decreased lactation Environment Enclosure environment including any litter nesting material enrichment items Changes in defaecation Behaviours Social interaction Isolation this is a cause for concern Posture and mobility Changes in posture movement or gait Reluctance to rise ot increased lying Other Escape attempts Agitation Specific vocalisations associated with pain or fear Specific tail movements associated with fear Pawing the ground Decreased rumination in adult including dropping the cud Grinding teeth this is a cause for concern These are identified on the basis of the individual project its potential adverse effects and expected indicators of these Sensitivity to touch Restlessness Depression Decreased maintenance behaviours and social grooming General malaise head and ears down inappetance Difficulty in changing position Hunched posture Dehydration skin pinch test Salivation Tail flicking Aggression including head thrashing kicking For infection studies respiratory changes mastitis gastrointestinal signs e g scour bloat dropped cud blood in faeces Procedurespecific indicators Suboptimal or poor welfare pigs High level categories Areas to focus on when observing animals Specific indicators to monitor Appearance Body condition Poor body condition Body functions Food water intake Poor food consumption or reduced drinking Body temperature Problems thermoregulating e g panting blotchy skin skin colour changes Environment Enclosure environment including any litter nesting Diarrhoea Reduced interest in nesting material or toys not curious or even neophobic Behaviours Social interaction No eye contact do not come to pen door Isolation from group Nervous of other pigs Undesirable behaviours Fighting tail biting Bar biting or chewing e g surfaces nipple drinkers tails ears 55

Tech 2 Tech Posture and mobility Other Procedurespecific indicators Posture tail position and carriage ear position Reduced foraging Quiet with reduced vocalisation or barking when startled angry or afraid These are identified on the basis of the individual project its potential adverse effects and expected indicators of these Dog sitting Apathy Restlessness e g postoperatively shows that animal is uncomfortable Laboured respiration after surgery Bark or squeal reaction to acute pain e g intramuscular injection Changes in skin colour could be stress or problem thermoregulating Regurgitation Persistent vocalisation is a serious concern Suboptimal or poor welfare sheep High level categories Areas to focus on when observing animals Specific indicators to monitor Appearance Body condition Poor body condition score Coat and skin condition Poor fleece or skin condition Other Ear position Respiration Panting Food water intake Inappetance Other Disease prone Social interaction Isolation from other sheep Undesirable behaviours Stereotypies Posture and mobility Changes in gait or lameness Prolonged lying down Other Vocalisation Body functions Behaviours Procedurespecific indicators 56 Aggression Behaviour signs of fear Unresponsiveness

April 2014 Animal Technology and Welfare AS ET SPECIAL TRAVEL BURSARY 2013 ESSAYS What do you regard as the most important issues in optimising the care and welfare of laboratory animals 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 Entry from the AS ET Special Travel Bursary 2013 As an Animal Technologist who has worked in the industry for nine years the care and welfare of laboratory animals is something that is always in my mind There are many factors that contribute towards optimising the care of animals in science from law and legislation all the way through to the practical aspects such as the provision of environmental enrichment Speaking as an animal technologist I have chosen the two most important issues that I feel are crucial for the animals in our care Due to the way we run experimental studies and the way that laboratory animals are housed and managed species used in research are inevitably totally reliant on technologists for all their needs from the provision of a suitable housing environment to ventilation veterinary attention water and diet Therefore as laboratory animals are so reliant on others for all their needs the first issue that I regard as crucial in optimising their care and welfare is the staff Therefore conscientious and obser vant animal technologists are one of the most valuable assets a laboratory can have A good technologist needs to have empathy while at the same time understanding the scientific needs of a study Hiring the right technologists is difficult in some cases due to the low entry wages which often results in many new technologists being school leavers who perhaps initially do not see laboratory animal work as a long time career In my experience within the UK staff turnaround can be quite high which can make it more difficult to maintain a high standard of care However many school leavers who fall into this line of work do develop into valuable key members of staff if given the right opportunities I think comprehensive training programmes and clear paths of progression and opportunity can help to attract and retain members of staff who see caring for animals in science as a rewarding long term career Working towards recognised qualifications such as IAT diplomas can also help to retain staff as well as expanding their skill sets Once working in an animal facility technologists should receive in depth training in all aspects of animal care from husbandry and handling to animal behaviour and procedural work where applicable should always be taught and carried out to the highest standards possible Animal Technologists should be observant and vigilant and be confident in rapidly recognising any signs of stress pain and disease They need to display empathy and also know to recognise relevant end points including when action needs to be taken if an animal is suffering Technologists must also be familiar with all current legislation that affects their work and importantly should feel able to come forward if they see something 57

AS ET Special Travel Bursary 2013 essays happening within the workplace that they do not believe to be professional or that is impeding the care or welfare of the animals In the UK every unit must have a Named Veterinary Surgeon NVS and Named Animal Care and Welfare Officer NACWO and to optimise the welfare of animals in their care technologists should always know who these people are or their equivalents and the channels though which they could be contacted I believe that to be a good animal technologist it is essential to be an animal lover However other key traits that a good animal technologist must possess are a clear understanding of why you are doing what you do and also to be able to see things from a research point of view Inevitably in this industry animal technologists will at times see things they do not like but understanding the research that is being carried out and the advances that are being made makes these times easier The second issue that I consider important in the care and welfare of laboratory animals is related to the environments that we create for the animals in our care specifically to allow animals to behave as naturally as possible Allowing animals to exhibit their natural behaviours as much as is practical is very important for their welfare It is beneficial to the animals themselves the staff that care for them and crucially it results in the collection of better quality scientific data as happier and unstressed animals are healthier animals Understandably sometimes the experiments and scientific protocols can affect the way that animals are cared for and the environment in which they are housed but wherever possible we should try and make their environment as similar to their natural one as we can without impeding daily tasks health checks dosing etc Whenever feasible and appropriate to the species animals should be group housed as this is one of the best forms of enrichment Being able to groom play and curl up with a companion can relieve stress and be of great comfort to laboratory animals Lonely animals of social species can quickly become depressed anxious and stressed sometimes even to the point of displaying stereotypical behaviour or self mutilation and consequently become of little use as scientific models Due to this I think group housing should be used wherever possible It is always important however to observe group housed animals closely for any signs of bullying or fighting and if practical provide an area where cage companions can avoid social contact if they choose All animals should be housed in conditions appropriate for their needs in terms of both their microenvironment and macroenvironment Temperature light humidity and noise levels in their macroenvironment should be 58 the optimum for their species and their microenvironment should be of an adequate size to enable plenty of movement exercise and the displaying of natural behaviours for example different levels in cages to enable animals to gain access to higher levels if that is where they are most comfortable Larger species should have access to areas where they can run stretch and exercise wherever possible experimental protocols permitting Large play pens that can be used for set periods of time in the day are a good example of this Nesting material in relevant forms is a great source of environmental enrichment and enables nest building and provides somewhere to hide However it is important to balance factors and although bedding and nesting material should always be given it is also important that animals can be checked thoroughly every day and viewed easily to allow any problems to be identified as soon as possible Environmental enrichment in the form of houses domes beds etc is beneficial in encouraging behaviours such as nest building climbing and playing However it is essential that these items do not interfere with the ability for animals to still be easily accessible and observations performed Chew blocks treats tubes and other loose toys should be used for as well as providing health benefits such as maintaining teeth through gnawing they also make an animal s environment more stimulating and encourage play and environmental enrichment also has the benefit of reducing abnormal repetitive behaviours such as barmouthing back flipping circling and barbering which are often seen in singly housed animals or where there is little or no stimulation Adapting feeding methods to more closely resemble an animal s natural pattern is also a useful enrichment tool Rodents for example are foragers and are kept interested and stimulated when provided with forage material rather than only in a food hopper although this is not always possible e g in studies where food consumption is recorded However there are many cases where a sprinkling of seeds or other forage on the cage floor is permissible and makes a significant difference to the rodent s quality of life and welfare Wherever possible it seems that the benefits of environmental enrichment far outweigh any negatives and so should be used wherever the scientific data will not be affected by its use In conclusion I know from experience having welltrained dedicated and attentive animal technologists undoubtedly optimises the care of laboratory animals Also that providing living conditions that enable laboratory animals regardless of species to exhibit natural behaviours whenever possible while still enabling good science as well as being practical hugely enhances an animal s welfare and quality of life

April 2014 Animal Technology and Welfare POSTER PRESENTATIONS Originally presented at The 12th FELASA SECAL Congress 2013 and reprinted with the permission of both organisations and the authors A refined method of restraint for dogs used in inhalation studies refinement in the acclimatisation procedure SIMON MOORE MICK TIMOTHY and STEVEN HAWES Huntingdon Life Sciences Huntingdon Cambridgeshire PE28 4HS Corresponding author moores ukorg huntingdon com Abstract Huntingdon Life Sciences has pioneered the use of a minimal restraint method for dogs for over 10 years This poster provides information on the fur ther refinement of this methodology and presents a revision of the acclimatisation procedure to improve animal welfare compliance and study results The introduction of the facemask and air to the exposure system are both implemented earlier in the 14 day acclimatisation period However this is offset by a more gradual increase in the total time spent being acclimatised These refinements have resulted in increased animal compliance and overall study conduct Introduction For more than a decade Huntingdon Life Sciences has pioneered a new type of minimal restraint for dog inhalation studies Figure 1 rather than using the historical and restrictive sling method and as a result this refinement has been widely replicated The evolution of the method is outlined in the poster titled A refined method of restraint for dogs used in inhalation studies M Hussain and A Leach 2006 1 Fur ther improvements to animal welfare were presented in another follow up poster by R Kaleta V Knauss S Cracknell A Leach M Hussain and C Hardy 2006 2 where fleece bedding and jackets were Figure 1 Dogs on the exposure table using the minimal restraint system used to provide warmth during longer exposure durations or when test articles are known to depress the body temperature Figure 2 The inhalation route is selected when it is the intended clinical route of administration of pharmaceutical products In dogs inhalation dosing is achieved in the majority of studies by exposing the animals to an atmosphere containing the test substance by means of a facemask Facemask exposure is normally used for longer duration exposures at relatively high 59

Poster Presentations concentrations It involves restraint of the animal for a period of up to 6 hours and necessitates the fitting of a malleable rubber mask held over the nose and mouth by a leather muzzle Original training programme The original sham dosing regimen evolved many years ago and was the result of numerous inhalation studies It is presented in Table 1 and was based on our knowledge at that time Day Total time of restraint mins Mask fitted in minutes for each exposure time mins 15 Figure 2 Dogs on the exposure table with fleece bedding and jackets The guiding principles in animal research today are the 3Rs reduction refinement and replacement In animal research refinement refers to method improvements that reduce possible stress or discomfort to the animals involved It also encompasses measures to improve animals overall well being and environment This poster describes the refinement of the acclimatisation phase of inhalation exposure to improve animal welfare compliance and overall study conduct which were driven by specific technologist observations made over years of conducting this procedure Basic technique The basic technique used is fully outlined in the poster by M Hussain and A Leach 2006 1 To summarise dogs are placed in a simple and snug body harness During each day of the training period the dogs are walked using standard leads from the housing units to the dosing room This helps to build up a relationship between handler and dogs It also serves to teach the dogs that when they are being walked on leads they will be going to the dosing room and will know what to expect This routine serves to make the dogs happier and more cooperative The harness is fastened to a restraining lead which is attached to a metal pole at the rear of the exposure bench Most dogs will become excited during the first few times they are placed in the harness Repetition calms them down so that they may be restrained for extended periods in the harness The minimal tethering combination of the harness and lead gives the animals a full range of movement this enables acceptance of the exposure system procedure whilst allowing interaction between dogs being exposed The success of this technique lies in the training of the dogs to accept the restraint method 60 1 2 3 4 5 6 7 8 9 10 11 12 13 14 60 120 15 60 120 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 30 30 45 45 60 60 60 60 60 60 60 60 15 30 30 34 60 75 90 120 120 120 120 120 120 120 0 0 0 0 0 0 15 15 15 15 15 15 15 15 0 0 0 0 0 0 15 30 45 60 60 60 60 60 0 0 0 0 0 0 15 30 60 90 120 120 120 120 With air introduction NO NO NO NO NO NO NO NO NO NO YES YES YES YES Table 1 Original training programme for acclimating dogs to the restraint system Revised training programme Although the original training programme described in Table 1 had been amended and gradually improved over a number of years we felt there were still enhancements to be made The revised procedure is presented in Table 2 with the changes to Table 1 highlighted The number of sham occasions would still take 14 days prior to the first test article exposure as this has proved to be optimum However we felt that the animals were originally restrained on the benches for too many occasions before the facemasks were introduced When the animals were first placed in the dosing suite it was not unusual for them to display signs of hyperactivity or exploratory behaviour which encouraged chewing of expensive body harnesses and restraining leads When the facemasks were originally introduced observations suggested this did not allow staff a sufficient number of occasions to condition the animals and so minimise hyperactivity or exploratory behaviour and chewing of equipment whilst still ensuring that the high standards we demand for study conduct particularly for the longer inhalation dosing regimens were upheld Equally we felt that the introduction of air was too late in the original acclimatisation procedure and too close to the first compound exposure for full benefit of the

Poster Presentations regimen to be achieved so we decided to introduce air at an earlier stage in the process The introduction of facemasks and connection of air has been brought forward to day 2 Day Total time of restraint mins Mask fitted in minutes for each exposure time mins 15 1 2 3 4 5 6 7 8 9 10 11 12 13 14 60 120 15 60 120 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 30 60 60 60 60 60 60 60 60 60 15 15 15 15 30 60 90 120 120 120 120 120 120 120 0 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 30 60 60 60 60 60 60 60 60 60 60 15 15 15 30 30 60 90 120 120 120 120 120 120 120 With air introduction NO YES YES YES YES YES YES YES YES YES YES YES YES YES Table 2 Original training programme for acclimating dogs to the restraint system This earlier introduction of the mask and air was compensated by maintaining the length of periods of restraint to 15 minutes for either 2 or 3 days instead of daily increments from that point of exposure lasting 60 or 120 minutes respectively The exposure periods increasing gradually to the proposed duration Implementation As anticipated the dogs soon adapted to the revised procedure and by the end of the 14 day training period the animals were even more compliant than the original acclimatisation procedure The animals continue to lie down next to each other and continue to fall asleep This revised and refined training programme has improved study conduct and results To date we have conducted over 40 studies utilising the revised acclimatisation procedure with study duration ranging from single exposures to 52 week chronic studies using periods of restraint of up to 6 hours References 1 2 Hussain M and Leach A 2006 A refined method of restraint for dogs used in inhalation studies Kaleta R Knauss V Cracknell S Leach A Hussain M and Hardy C 2006 A refined method of restraint for dogs used in inhalation studies System adaptation and improvement 61

Animal Technology and Welfare April 2014 Drying off procedure for dairy cows DEAN COOPER Huntingdon Life Sciences Huntingdon Cambridgeshire PE28 4HS Corresponding author cooperd ukorg huntingdon com Introduction Dairy cows are often used on studies at Huntingdon Life Sciences and frequently milk samples will need to be obtained and analysed as part of the study investigations However sometimes the animals are required to be dry before dosing can commence The term dry refers to an animal that is no longer producing a milk yield to feed its young The following procedure will give guidance on how to dry off a cow for studies which require this Method Naturally a cow s lactation cycle lasts approximately 1 year The cow will begin lactating just before calving The yield will gradually rise in line with the growth of the calf hitting a peak at around 40 60 days after calving Production will then steadily decline over the next 250 or so days 310 in total after calving The cow will then become naturally dry for the remaining 50 days before the cycle begins again Cow calves Production peaks at 40 60 days To manually dry off a cow record the amount of milk given For the purposes of Table 1 this is 100 The amount of concentrate given at this time is also 100 Day 1 Day Day Day Day Day Day Day 2 3 4 5 6 7 8 commence drying off by halving food collect 75 of milk yield before stopping 50 food 50 milk 25 food 25 milk no food 10 milk no food maintain at 3 4L no food maintain at approx 2L no food final milk 1L or less no food dry Table 1 Reducing milking occasions to one session per day Steady decline for 250 days Cows naturally dry to prepare for next cycle Despite this natural cycle it is possible to alter a cow s lactation period The cow will continue to give a yield as long as the milk is being used While this yield is being maintained a large amount of concentrate will also be given to allow the animal to produce the milk These two factors dictate the yield given By reducing or removing these the cow will give less milk Therefore to reduce the yield reduce the amount of milk taken and the amount of concentrate given Although it is possible to dry a cow off whilst maintaining a large concentrate ration the food will not be used for milk 62 production and will result in the cow putting on weight To avoid this reduce the food ration in relation to the reduction of milk being taken Hay grass or grass nuts can be given to maintain the animal s health Figure 1 The above schedule can be amended as necessary depending on the amount of milk being given For animals giving less than 10L a day begin the process of Day 3 Once the cow has been successfully dried off an intra mammary injection of suitable antibiotic must be given to combat the onset of mastitis The antibiotic will be advised by the on site Veterinary Department and the injection given in accordance with LAA DOS 014 The successful administration of the anti biotic will be the point at which the cow is considered dry

Poster Presentations Procedure for increasing milk yield in dairy cows A cow s milk production is dependent on several factors The main factor is the stage of the lactation cycle the cow is in Production peaks at 40 60 days post calving and steadily declines over the following 210 days However it is possible to increase a cow s yield Although the volumes achieved may not be as high as those seen at peak production the gradual decline can be abated by implementing some simple techniques The route to the milking parlour can also play a large role in increasing milk yield The route should be as simple as possible with minimal distractions If the cow has only one route in which to go the level of adrenaline released will be decreased The release of adrenaline interrupts the oxytocin based milk let down response meaning less milk will be given Studies have also shown that on high production milking units the playing of gentle background music can increase yield by up to 10 Classical music is the preferred choice due to its calm and slow tempo Researchers at the University of Leicester believe the calming influence of the music reduces stress in much the same way it does for humans The familiarity of background noise also blocks or dulls sudden noises which may otherwise startle the animals In addition to this talking to the cow in a gentle way is also believed to help increase yield Again it is thought familiarity and a calm environment reduce the stress on the animal decreasing adrenaline output and allowing the oxytocin hormone to promote milk production Figure 2 Milk production in cows throughout lactation cycle The simplest way of increasing a cow s yield and slowing the decline over time is to increase milking to three times a day instead of the usual two By increasing the occasions the cow is exposed to more of the hormones that stimulate milk secretion in the mammary gland The milk produced also contains an inhibitor that has a negative feedback control on production A more frequent removal of this inhibitor therefore results in higher production Another factor to improve yield is ensuring the cow is being milked in a calm and comfortable environment Factors which can be detrimental to this include slippery floors leading up to the parlour poor handling techniques poor housekeeping in and round the parlour and bad ventilation Signs of discomfort include kicking at the milking cluster defecating in the milking stall and reluctance to enter stall By incorporating some small measures milk production can be increased These can be as simple as hanging a fly catcher near to the parlour to prevent a build up of flies during milking and the use of desk fans or similar to increase ventilation in and around the stalls Conclusion In summary it is not possible to change the natural lactation cycle of a cow However it is possible using the techniques given above to interrupt prolong or reduce the length of the cycle depending on the outcome desired It is impor tant to feed the correct amount of concentrate to a lactating cow when milking The food consumed will be used for milk production which means a greater amount of concentrate will result in a greater yield By feeding at the same time as milking it is believed to prolong and increase the release of the hormone oxytocin It also increases milk flow which in turn decreases milking duration which places less stress on the animal 63

Animal Technology and Welfare April 2014 3Rs Centre Utrecht Life Sciences JAN VAN DER VALK1 SASKIA ARNDT NELLEKE VERHAVE and PIM ROOYMANS 3Rs Centre ULS NKCA Department of Animals in Science and Society Faculty of Veterinary Medicine Utrecht University Yalelaan 2 NL 3584 CM Utrecht The Netherlands Corresponding author j vandervalk uu nl 3Rs Centre ULS The 3Rs Centre Utrecht Life Sciences provides information on and stimulates the development acceptance and implementation of 3Rs methods within Utrecht Life Sciences ULS every research objective Therefore we need to focus also on reducing the number of animals and refining experimental methods to reduce suffering Implementing the 3Rs in our protocols and research can only be done if we are aware of their existence No unless Animal experiments are not allowed in the Netherlands unless no alternative is available Therefore the use of the 3Rs is obligatory by law 1 A non animal method to replace an animal experiment is not yet available for Figure 3 Cage enrichment an example of refinement minimising animal distress 3Rs expertise Figure 1 Cell and tissue culture an example of replacement Figure 2 Computer programmes an example of methods that reduce animal use 64 The 3Rs Centre ULS facilitates the communication on initiatives within and outside the ULS that increase the use of 3Rs Within the ULS there are already several research and education projects focussed on the 3Rs Figure 4 Education best practice videos reduction

Poster Presentations The 3Rs Centre ULS identifies these initiatives and communicates about them also outside the ULS In addition it advises on new 3R applications and on their use in research and education The centre collaborates with other 3Rs centres and the Netherlands Knowledge Centre on Alternatives NKCA 3Rs Centre for you Strategy G G identify and advise on possible cooperation between public private partners also non ULS partners identify and communicate on funding opportunities Animal experimentation G G G advising Animal ethics committees involved in new developments e g consequences on the new EU directive on animal experimentation advising researchers and educators on 3R issues in animal experiments Education G G G G G course on Laboratory Animal Science postgraduate course on Laboratory Animal Science 1 year in Dutch only MSc Course on Animal Welfare 18 months Veterinary Master Responsible use experimental animals lectures on the 3Rs by invitation Cooperation G G G G G chair on animal welfare and laboratory animal science chair on alternatives to animal experiments chair on alternatives to animal experiments for toxicological risk assessment animal welfare officers communication departments Communication G G G G regular newsletter website http www uu nl 3RsCentreULS email 3RsCentreULS uu nl twitter 3VCentrumULS References 1 Wet op de dierproeven experimentation Dutch Act on animal 65

Animal Technology and Welfare April 2014 Systematic reviews and the Three Rs JUDITH VAN LUIJK ROB DE VRIES CARLIJN HOOIJMANS KIM WEVER MARLIES LEENAARS and MEREL RITSKES HOITINGA SYRCLE Radboud University Nijmegen Medical Centre Post 231 SYRCLE CDL PO Box 9101 6500 HB Nijmegen The Netherlands Corresponding author J vanLuijk cdl umcn nl Summary Replacement Reduction Refinement the 3Rs are the leading principles in Laboratory Animal Science LAS for a scientifically and ethically sound use of animals in research Although the implementation of 3Rs methodologies in research is obligatory according to the new EU Directive 2010 63 the search for relevant 3Rs methods and eventually their implementation can be ver y challenging The Systematic Review SR approach may be part of the solution here Main differences between the two types of review Narrative review Systematic review Common in animal based research Common in clinical research No or general review question Specific and clear review question Subjective method Evidence based method Study selection not specified Transparent study selection Introduction No attempts to avoid bias Risk of bias analyses When planning and designing a new animal experiment the 3Rs principle needs to be applied The 3Rs stand for Replacement Reduction and Refinement of animal use in Research A literature review can be conducted to answer a number of questions in the planning stage e g Is an animal experiment necessary if so which animal model is most suitable And what evidence is already available No combined data analysis Meta analysis of data Systematic Review A Systematic Review SR is a literature review focussed on a research question that tries to identify appraise select and synthesise all primary animal studies relevant to that question Unfortunately while it can provide many advantages to enhance implementation of the 3Rs principle in research this methodology is not yet common in animal based research Three Rs principle Replacement Reduction and Refinement according to Article 4 of the EU Directive 2010 63 A scientifically satisfactory method or testing strategy not entailing the use of live animals Ensure that the number of animals used in projects is reduced to a minimum without compromising the objectives of the project Ensure refinement in order to eliminate or reduce to the minimum of any possible pain suffering distress of lasting harm to the animals 66 Table 1 Main differences between the two types of review Comprehensive search The comprehensive search approach of the SR methodology can be used to find more evidence based answers compared to the conduct of narrative review Performing a comprehensive search of animal studies seems a ver y promising approach to implement information from previously per formed animal experiment and the 3Rs all together One of the advantages of this approach is that it provides structure to the search process and makes it transparent This transparency is also key in a fair ethical evaluation of the animal experimental design which is also one of the topics addressed in the EU Directive 2010 63 Challenges Searching for specific 3Rs information can be very challenging due to the scattered information over many 3Rs databases and the different required strategies to search them Additionally various studies have shown that poor repor ting and publication bias are still a concern when conducting a full SR of animals and require serious attention from the research community

Poster Presentations Advantages of Systematic Review of animal studies As Table 1 shows there are many differences between a narrative and a systematic review Once a comprehensive search is conducted it becomes easier to if appropriate and desired pursue with the conduct of a full SR A full SR has the following advantages Gain new insights and determine knowledge gaps Better interpretation of all relevant published studies Prevent unnecessary duplication of animal experiments Up to date overview within a specific research topic Provide valuable information for an evidence based study design taking the 3Rs into account Improve transparency of the review process Stimulate multi disciplinary cooperation 67

Animal Technology and Welfare April 2014 Control of the estrous cycle in guinea pig Cavia porcellus A GR GOIRE A ALLARD E HUAM Nc S LEON R M SILVA S BUFF M BERARD and T JOLY 1 2 3 4 Institut Fran ais d Etudes Andines UMIFRE17 CNRS MAEE Lima 18 Peru Universit de Lyon VetAgroSup IsaraLyon Unit ICE Cryobio 69 243 Lyon France CIETE Ministry of Agriculture La Molina National Agrarian University Lima Peru Institut Pasteur Animalerie Centrale 75724 Paris France Corresponding author anne gregoire gmail com Abstract The aim of this work was to look for a simple method to obtain synchronised ovulation in Guinea pigs under farming conditions while respecting animal welfare The luteolytic activity of three different prostaglandins F2 PGF2 analogs D cloprostenol D L cloprostenol and Luprostiol and a daily treatment with oral progestagen Altrenogest were tested successively at different stages of the estrous cycle on the same group of females during a period of 8 months The estrous cycle length was not modified by the administration of prostaglandins whatever the stage of the estrous cycle when the treatment was initiated Our results led us to reject the use of PGF2 analogues to induce practical synchronisation of the estrus in this species In all the females n 29 given 15 days with Altrenogest 0 1 mL p o once a day ovulation occurred 4 43 0 13 days Figure 1 Traditional Guinea pig breeding in the Andes 68 after the end of the treatment Altrenogest treatment was followed by mating No negative impact of the treatment on the pregnancy rates delivery rates and litter sizes were observed This standard method of guinea pig estrus synchronisation is less stressful towards the animals compared to techniques using progesterone tubing Materials and methods 44 multiparous females from the Maria Marcela farm Puente Piedra Peru were used They were between 18 to 24 months old weighed between 1 to 1 5 kg and showed normal estrous cycles The animals were housed under farming conditions groups of 5 females living together on vegetal litter and were fed on commercial pellets and tap water ad libitum Figure 2 Oral treatment

Poster Presentations 4 treatments were tested 3 used PGF2 analogs to evaluate their luteolytic activity D Cloprostenol D LCloprostenol et Luprostiol and one used an oral progestagen Altrenogest to block ovulation The vaginal opening of the Guinea pigs was detected by daily observation The day of ovulation was further determined by smears performed every 12 hours G after mating the fertility of the females that had been synchronised with Altrenogest was comparable to that of females mated naturally pregnancy rate of 79 birth rate of 66 mean size of the litters of 3 09 0 27 pups Conclusion and perspectives These results validate the first essential step of the development of a simple and reliable method of cryopreservation of guinea pigs lines The results of this work are currently used to secure guinea pigs lines with high agronomic and economic value in Peru and two unique laboratory lines used at the Institut Pasteur as animal models for the study of tuberculosis Figure 3 Daily inspection for signs of heat Results The mean length of the Guinea pigs estrous cycle was 16 17 0 21 days 83 cycles were studied The treatments using PGF2 did not modify the length of the cycles P 0 05 which were respectively 15 76 031 17 32 0 42 and 16 17 0 36 days under D Cloprostenol D LCloprostenol and Luprositol On the contrary the treatment with Altrenogest synchronised ovulation G the estrous cycle was prolonged as long as Altrenogest was given G 100 of the treated females ovulated 4 43 0 13 days after the treatment was stopped Altrenogest N u m b e r o f f e m a l e s Interval treatment ovulation days 69

Animal Technology and Welfare April 2014 The impact of automation on cage wash operations MARY ROBINSON TOM RODRIGUEZ Jr ELMER BANES RONALDO ELENZANO SUZANNE CRAIG and PEGGY TINKEY Department of Veterinary Medicine and Surgery University of Texas MD Anderson Cancer Center 1515 Holcombe Blvd Houston TX 77030 USA Corresponding author MARobinson2 mdanderson org Introduction G G G G G many institutions face an ever increasing rodent population in facilities advances in rodent housing equipment allow a greater number of cages to be handled by the same number of care staff likewise automation in the cagewash facility has been touted to be the solution to some ergonomic and logistical issues surrounding the processing of the large number of dirty cages generated in facilities with high rodent populations to assist with the job of scraping and dumping over 20 000 cages each week we installed a robot on the dirty side of cagewash in two facilities in 2012 an assessment was performed to determine the impact of the robot on cage wash operations number and type of injuries reported by staff since robot use started whether any improvements to the process had been made to increase number of cages processed Results G G G more cages were processed per day using robot manual maximum 2210 per day and robot maximum 2800 per day downtime was more likely to be caused by problems with either the bedding collector or the tunnel washer than with the robot itself downtime was between 2 and 24 hours in duration Goal To assess whether use of the robot results in more efficient cagewash operations by increasing the number of cages processed and decreasing cage wash downtime Methods Supervisors at each facility were asked to record the following data for a 2 week period G G G G G Number of cages processed by robot daily Number of cages processed manually daily Frequency of downtime in using robot Reason for downtime in using robot Length of downtime In addition supervisors were asked length of time personnel had to attend to the robot per hour length of time personnel spent ensuring proper cage stacking on trolleys 70 Figure 1 Dirty side cage wash work flow pre robot G G G two personnel one assigned to dump and scrape cages one assigned to load cages onto conveyor belt of tunnel washer

Poster Presentations checking and restacking cages on the trolley takes 1 to 7 minutes personnel have fewer strains of upper body compared with manual dumping using trolleys to stack cages for autoclaving is less efficient 640 per load vs 744 per load but is more ergonomic as trolleys are easier to push cleaning the bedding collection pan twice daily instead of once weekly decreased downtime due to clogging by 75 at one location Conclusions Using the dirty side cage wash robot G G G G G increased daily cage dumping by 25 with one fewer staff member per location allowed reassignment of staff member to other tasks decreased repetitive motion strains in staff robot has excellent reliability after initial programming and troubleshooting downtime more likely caused by problems in other cage wash equipment than robot Acknowledgements Figure 2 Dirty side cage wash work flow using robot G G G one person loads trolleys into queue and handles robot controls robot dumps and scrapes cages and loads onto conveyor belt of tunnel washer same person removes empty trolleys and attends to robot The authors wish to thank Claro Mordeno Joe Mitra Freddie Santiago Marvin Bernardino Noran Urbanec Kristin Pegram Raymond Blanco Rosauro Mauleon Rolando Digan Maria Tiangco Melanice Gibson and Adrian Wells for assistance in collecting data This research is supported in part by the NIH through the MD Anderson s Cancer Center Support Grant CA016672 Figure 3 Number of cages dumped by robot and personnel at both locations N Nor th Campus S South Campus attending to robot takes 1 5 minutes per 2 trolleys processed 71

Animal Technology and Welfare April 2014 Animal research in a global organisation advantages and challenges of a centralised oversight group GILL FLEETWOOD GlaxoSmithKline Office of Animal Welfare Ethics and Strategy Gunnels Wood Road Stevenage Hertfordshire SG1 2NY Corresponding author Gill 2 Fleetwood gsk com Introduction GlaxoSmithKline is an international organization which carries out animal research in many countries Regulations governing animal care and use var y throughout the world the laws reflecting the differing cultures of regions and countries GSK wishes the care for our animals to be equivalent irrespective of where in the world research is conducted on our behalf We address this by having G G G an overarching policy on animal care and use core principles that must be met for all GSK animal studies irrespective of location a centralised group independent of the operational responsibilities husbandry care study design and support to lead the company vision for responsible animal research Office of Animal Welfare Ethics and Strategy OAWES GSK core principles G G G G G G G G G G access to species appropriate food and water access to species specific housing including appropriate temperature and humidity levels access to humane care and a programme of veterinary care animal housing that minimises the development of abnormal behaviours adherence to principles of replacement reduction and refinement in the design of in vivo or ex vivo studies review of study design and purpose by institutional ethical review panel commitment to minimising pain and distress to the animal during in vivo and ex vivo studies work is performed by staff trained to conduct the procedures for which they are responsible training is documented and verified processes are in place to minimise animal use Applies for all studies irrespective of their location or whether they are carried out within company facilities or performed externally on our behalf Advantages G G G G G G G head of group is the GSK Risk Owner for Animals demonstrates GSK commitment to animal welfare provides single point of contact for all issues related to animal research common principles for animal care and welfare worldwide mechanism to facilitate identification and dissemination of improved practice between business units independent of medicine discovery and development organisation provides consistent oversight throughout GSK Challenges G G 72 OAWES is a small group distributed worldwide defining the role of OAWES and communicating where we can add value to the business

Poster Presentations G G G G G G complexity of GSK organisation worldwide distribution of diverse operations Different regulations Different culture combining governance role with leading strategy balancing time spent determining longer term strategy with that required to ensure continuity of everyday business linking aspirations of senior management with experience of scientists carrying out studies retaining links with day to day business Animal research strategy Animal Research Strategy Introduce scientific peer review of animal model strategies Map animal model portfolio and define Challenge areas of opportunity G G Education G completed an analysis of regulatory expectations and training needs in US and EU G initiated development of global systems for training delivery and documentation as well as registering training needs G three workstreams focusing on people trainers assessors trainees materials training standards and assessments IT system e learn training and competency records Aim is to train and assess through a matrix of competent experts G Budget secured to accelerate provision of training materials from external providers Advocacy G Data Examine in vivo data sets Investigated approaches to standardise data in the discovery space Invest Strategically invest in alternatives Identify programs to progress to proof of concept without animals Share G G Investigate the feasibility of sharing precompetitive animal model data Establish partnerships with others interested in sharing data in this area G Our commitment Rigorously challenge the need for animal studies and work to minimise the impact on animal welfare by investing in the development of alternative studies and sharing animal based data from the GSK Corporate Responsibility Report 2012 Quality G G worldwide risk oversight committee launched 4Q2012 Animal Welfare and Quality Council accountable people in all areas that commission animal research established independent Quality Assurance Assessment AQA of animal work both internally and externally Specific assessments ethical review process China global Animal and regulator y records permissions and licences global Level 1 process Animal welfare programmes associated with 225 proposed external collaborations influenced the transposition of EU Animals Directive into national legislation contributed to UK and European discussions on continuity of supply maintaining public support for animal research publicising pharmaceutical industry progress in implementing 3Rs partnered with GSK Corporate Responsibility and Global Communications teams to formulate animal research related sections of Corporate social responsibility report external policy statements set up new methods to communicate with those interested in animal research within GSK e g animals in Research internal website animal Welfare Community of Practice talk at GSK induction for new staff Ethical review G G Achievements for 2012 GLPQA review of AQA processes showed processes considered effective in assessing compliance and quality no significant findings contributed to Internal Audit of GSK China G G provided secretariat for US IACUCs and UK Animal Welfare Body conducted analysis of the GSK animal model review process against regulatory requirements UK US engaged with GSK ethical review groups in France Spain Belgium and China initiated review of operation and memberships of GSK ethical review committees to streamline interaction with Animal Welfare and Quality Council 73

Animal Technology and Welfare April 2014 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 74 2 For the remainder of the paper the text should be

Instructions to Authors clear and concise and where appropriate sub divided under the following headings i ii iii iv v vi vii Summary Introduction Methods Results Discussion Acknowledgements References 3 Measurements should be given in metric units see The use of S I Units 1969 British Standards Institution publication and spelling should follow that of the Oxford English Dictionary Abbreviations must be defined in full at their first appearance in the text The 24 hour clock should be used for times Words to appear in italic type should be underlined Designation of inbred strains should be in accordance with the International Index of Laboratory Animals 6th edition compiled edited and published by M W Festing 1993 4 Photographs should have clear and well contrasted tone values and be in colour All illustrations charts e g histograms and graphs and photographs should be submitted separately and bear on the reverse side the author s name a number corresponding to the order in which it appears in the text e g Figure 1 and an arrow pointing to the top Journals Surname and initials of author s date title of article Name of journal in full volume number first and last page numbers e g Saigeman S 1998 Environmental enhancement of cats what why how Animal Technology Vol 49 No 3 145 154 Books Surname and initials of author s date title of book Name of publisher Town of publisher e g Flecknell P A 1987 Laborator y Animal Anaesthesia Academic Press London Chapter from a multi author book Surname and initials of chapter author s date title of chapter In title of book surname and initials of book editors Name of publisher Town of publisher first and last page numbers of chapter e g Gregory J A 1985 Principles of Animal Husbandry In Laboratory Animals An Introduction for Experimenters Second Edition Tuffrey A A John Wiley Sons Ltd Chichester 87 105 Papers accepted for publication but not yet published should be included in the list of references followed by in press Papers in preparation personal communications and unpublished observations should be referred to as such in the text only Illustrations charts and photographs supplied on disk should be in JPEG TIFF or EPS formats and have a resolution of no less than 300dpi Content The captions for illustrations charts and photographs should be typed in double spacing in numerical order on a separate sheet of paper Papers describing procedures involving the use of animals should always include full details of the animals and husbandry conditions used These would be as follows 5 References Only essential references should be included Authors are responsible for verifying them against the original source material ATW uses the Vancouver referencing system references should be identified in the text by superscript Arabic numbers e g 12 after any punctuation and numbered and listed at the end of the paper in the order of when they are first cited in the text Automatic numbering should be avoided References should include the names and initials of up to six authors If there are more than six authors only the first three should be named followed by et al Publications for which no author is apparent may be attributed to the organisation from which they originate Simply omit the name of the author for anonymous journal articles avoid using Anonymous References should be set out as follows Animals Species Breed or strain Sex Age and weight at start of procedure Genetic status inbred outbred hybrid mutant Source Microbiological status conventional specified pathogen free define which pathogens animals are free from gnotobiotic define which microorganisms are present Quarantine or acclimatisation period Husbandry during procedure Type of housing material size cage type if relevant Number of animals per cage or unit Bedding type quality any pretreatment 75

Instructions to Authors Type of system conventional barrier ventilated rack isolator Environmental temperature C range Relative Humidity range Lighting natural artificial state hours of light and dark Ventilation number of air changes per hour Period of acclimatisation before start of procedure Feed type composition any pretreatment amount frequency Water type quality any pretreatment amount frequency Scientific procedure Number of animals and any pretreatment Time of day of procedure s Quantity and frequency of any samples Statistics Tests used should be named Reprints Free reprints are no longer provided but the ATW Editorial Board are happy to provide PDF files of articles after publication Use of these files is subject to Copyright restrictions 76

INDEX TO ADVERTISERS April 2014 3Rs LAB xx AAALAC Institute of Animal Technology xvii Allentown Inc iv Allentown Inc xvi ARMIS R W Associates xiii AVID Plc xiv Bell Isolation Systems v Charles River Laboratories IFC Harlan Laboratories viii Institute of Animal Technology x IPS Product Supplies Ltd IBC LBS vii PFI Systems xviii Special Diets Services vi Surrey Diagnostics xv xix Sychem Ltd xii Tecniplast UK OBC VetTech Solutions iv xx