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IAT Journal Animal Technology and Welfare INCLUDES G Andrew Blake Tribute Award winner G An early look at the Revised ASPA regulations G IAT Education past present future G 2012 Winter Meeting posters Official Journal of the Institute of Animal Technology and European Federation of Animal Technologists ISSN 1742 0385 Vol 12 No 1 April 2013

OFFICERS President Dr Robin Lovell Badge FRS Animal Technology and Welfare Immediate Past President Professor Sir Richard Gardner MA PhD FSB HonFIAT FRS Vice Presidents Stephen Barnett BA MSc CBiol FSB RAnTech John Bleby TD JP DVetMed DLAS CBiol FSB MRCVS Brian Cass CBE Miles Carroll PhD Gerald Clough BSc PhD EurBiol CBiol MSB SFZSL Paul Flecknell MA Vet MB PhD DLAS DipLECVA MRCVS Barbara Mortimer BVetMed DLAS MRCVS Judy MacArthur Clark CBE BVMS DLAS CBiol FSB MRCVS Fiona McEwen BSc BVM S MSc MRCVS Tim Morris BVetMed PhD DipACLAM DipECLAM CBiol FSB CertLAS MRCVS Jos Orellana BVSc MSc Clive Page PhD BSc Sophie Petit Zeman PhD Gail Thompson RLATG Robert Weichbrod PhD RLATG Sheila Whitehead BVMS MSc CertLAS MRCVS Lord Robert Winston FMedSci DSc FRCOG FRCP FRCS Ed FSB Life Members Roger Francis MSC FIAT RAnTech Pete Gerson MSc FIAT RAnTech John Gregory BSc Hons FIAT CBiol FSB RAnTech Patrick Hayes FIAT DipBA RAnTech John Kelly FIAT Robert Kemp FIAT Hon RAnTech Keith Millican FIAT CBiol MSB Phil Ruddock MIAT RAnTech Ted Wills HonFIAT RAnTech Dorothy Woodnott FIAT Honorary Members John Frogley FIAT RAnTech John Lesley FIAT RAnTech Ronald Raymond FIAT RAnTech Peter Russell FIAT RAnTech Ray Thatcher FIAT RAnTech Members of Council Ken Applebee Jas Barley Kate Burton Charlie Chambers Steven Cubitt Andy Cunningham Glyn Fisher Carol Fox Cathy Godfrey Alan Graham John Gregory Patrick Hayes Linda Horan Andy Jackson Elaine Kirkum Adele Kitching Sarah Lane Norman Mortell Steve Owen Wendy Steel Allan Thornhill Lynda Westall Debbi Young Members of Executive Committee Council Officers Chair Steve Owen FIAT RAnTech Vol 12 No 1 April 2013 CONTENTS Editorial Jas Barley Chair of the Editorial Board ix Body condition scoring for Laboratory Zebrafish Carole Wilson Karen Dunford Carly Nichols Heather Callaway Jenna Hakkesteeg and Matthew Wicks 1 An effective method for terrestrial arthropod euthanasia Neil Bennie Christopher Loaring Mikaella Bennia and Steven Trim 9 An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive Tim Betts 15 PAPER SUMMARY TRANSLATIONS 21 TECH 2 TECH IAT Education past present and future Kenneth Applebee Animal Management Software making it work for you 41 Alison Hopkins Report on the PTLLS Preparing to Teach in the Lifelong Learning Sector course Alan Palmer 45 Report of the 2012 RSPCA UFAW Rodent Welfare Group meeting Penny Hawkins Secretary Jess Gimpel Andrew Rice Dominic Wells Judi Latcham Kate Heath Mark Gardiner Trevor Wattam Elliot Lilley Maggy Jennings and Robert Hubrecht 49 Immediate Past Chair Ken Applebee FIAT CBiol FSB RAnTech POSTER PRESENTATIONS The use of contrast agents to enhance soft tissue imaging using micro CT in laboratory rodents Jordi Tremoleda and Willy Gsell Honorary Secretary Wendy Steel BSc Hons FIAT RAnTech Tumours models core Biological Research Unit Paul Mackin Lisa Young and Steven Kupczak Honorary Treasurer Glyn Fisher FIAT RAnTech 59 63 Let the fish decide How group size gender and 69 dominance affect preference for environmental enrichment in zebrafish Paul Schroeder Soffia Jones Iain Young and Lynne Sneddon Assistant Treasurer Charlie Chambers MIAT RAnTech A review of German animal research applications from 2010 to identify which anaesthetic and analgesic regimens are used in experiments involving rodents Kathrin Herrmann and Paul Flecknell 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 continued on page ii 73 Non surgical embryo transfer with the NSET TM device is a 3Rs refinement technique that reduces stress in CD 1 mice Kendra Steele Barbara Stone James Hester Brett Spear and Angelika Faith Goodin Chair Board of Educational Policy Ken Applebee FIAT CBiol FSB RAnTech 75 Instructions to Authors 77 i

BRANCH SECRETARIES 2013 Officers continued from page i 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 Aberdeen Ms Donna Wallace Rowett Research Institute Greenburb Road Bucksburn Aberdeen AB21 9BJ Cambridge Ms Fran Flack Wellcome Trust Sanger Institute Wellcome Trust Genome Campus Hinxton Cambridgeshire CB10 1SA EFAT Representatives Charlie Chambers MIAT RAnTech Kate Burton MSc FIAT RAnTech Cheshire Ms Julie Humphreys Block 53 Mereside Astrazeneca Alderley Edge Nr Macclesfield Cheshire SK10 4TG Council Website Coordinator Allan Thornhill FIAT RAnTech Edinburgh Ms Janice Young MRC Human Genetics Unit Western General Hospital Crewe Road Edinburgh EH4 2XU IAT INFORMATION Hertfordshire Essex Ms Hazel Broom GSK Safety Assessment Toxicology Support Park Road Ware Hertfordshire SG12 0DP 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 Huntingdon Suffolk Norfolk Ms Pam Stott Huntingdon Life Sciences Woolley Road Alconbury Huntingdon Cambridgeshire PE28 4HS Board of Educational Policy Ken Applebee Chair Steven Cubitt Secretary Carol Fox CPD Officer Sarah Lane Ireland Mr Jason Radford Biological Service Unit University College Cork Eire Board of Moderators Cathy Godfrey Chair Glyn Fisher Secretary Moderators Gary Childs Joanna Cruden Nicky Gent Linda Horan Sue McHugh Kent Ms Fiona Jameson Pfizer Ltd PO 155 PGRD Sandwich Kent CT13 9NJ Communications Group Norman Mortell Chair Kate Burton Linda Horan Elaine Kirkum Allan Thornhill Lynda Westall London Ms Karen Robinson BSU Rayne 4th Floor Lambeth Wing St Thomas Hospital London SE1 1UL Registration and Accreditation Board Ken Applebee Charlie Chambers Chair Gerald Clough Carol Fox 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 Midlands Mr Ian Fielding Tecniplast UK BCM Box 3058 London WC1N 3XX North East England Mathew Sanderson Centre for Cardiovascular Metabolic Research Wolfson Building University of Hull Hull HU6 7RX Oxford Mr Adrian Woodhouse Prosidion Windrush Court Transport Way Watlington Road Cowley Oxford OX4 6LT 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 Surrey Hampshire Sussex Ms Lesley Hughes c o Building 200 ASU Veterinary Laboratories Agency Woodham Lane New Haw Addlestone Surrey KT15 3NB West Middlesex Mrs Wendy Steel BSU Royal Veterinary College Hawkshead Lane North Mymms Hertfordshire AL9 77A Although every effort is made to ensure that no inaccurate or misleading data opinion or statement appear in the journal the Institute of Animal Technology wish to expound that the data and opinions appearing in the articles poster presentations and advertisements in ATW are the responsibility of the contributor and advertiser concerned Accordingly the IAT Editor and their agents accept no liability whatsoever for the consequences of any such inaccurate or misleading data opinion statement or advertisement being published Furthermore the opinions expressed in the journal do not necessarily reflect those of the Editor or the Institute of Animal Technology West of Scotland Ms Linda Horan Biological Procedures Unit University of Strathclyde SIBS Building c o Phys and Pharm 27 Taylor Street Glasgow G4 ONR 2013 Institute of Animal Technology All rights reserved No part of this publication may be reproduced without permission from the publisher Western Counties Mrs Elaine Kirkum Lillico Biotechnology PO Box 431 RH6 0UW ii

April 2013 Animal Technology and Welfare THE INSTITUTE OF ANIMAL TECHNOLOGY ETHICAL STATEMENT IN THE CONDUCT OF THEIR PROFESSIONAL DUTIES ANIMAL TECHNOLOGISTS HAVE A MORAL AND LEGAL OBLIGATION AT ALL TIMES TO PROMOTE AND SAFEGUARD THE WELFARE OF ANIMALS IN THEIR CARE AND TO RECOGNISE THAT GOOD LABORATORY ANIMAL WELFARE IS AN ESSENTIAL COMPONENT OF GOOD LABORATORY ANIMAL TECHNOLOGY AND SCIENCE Editorial Jas Barley Chair of the Editorial Board Welcome to the first issue of Animal Technology and Welfare for 2013 Firstly I need to highlight a correction to the email for correspondence that appeared in Volume 11 No 3 published in December 2012 for the Tech 2 Tech article No Guts no Glory this has now changed to gbgcre gmail com The modifications to the Animals Scientific Procedures Act 1986 to enable the transposition of the EU Directive 2010 63 EU have now been published and we welcome the early insight as to how the modifications will affect the work of UK Animal technologists provided by Tim Betts paper An Early Overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive The ATW Editorial Board hopes to include further offerings on how things have changed with the transposition of the Directive into UK and European legislation I am regularly being informed of the value that many of our readers both within the UK and overseas find in Animal Technology and Welfare both as a source of information and as a teaching aid especially in those countries that do not enjoy the benefits of formal education for animal technologists such as that provided by the Institute of Animal Technologists IAT It is therefore with great pleasure that we are including a transcript of one of the presentations from the John Bleby 80th Birthday symposium held in 2012 given by Ken Applebee on IAT Education Past Present and Future John Bleby has been a long time supporter of Animal Technologists in general and in particular recognised the contribution that knowledgeable AT s make to research Although the information included in the presentation has all been presented to IAT members and our Stakeholders in the past albeit in various stages as things changed and progressed this presentation was probably the first time the complete story to date had been explained As a means of developing our educational role we are delighted that the IAT Council has approved an initiative to increase contributions from European Animal Technologists by accepting papers in their native language with translation into English being arranged by the Editorial Board All we will need from potential authors is an abstract in English so that we can assess the potential of the paper in terms of interest for our readers Please note at the present time this is only available to technologists however we of course welcome contributions from any of our academic readers as well but trust that they would be able to supply a full manuscript in English In addition to an insight into the modified legislation we are pleased to include the 2012 Andrew Blake Tribute Award winning submission The Andrew Blake Award 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 and the 2012 Award goes to Carole Wilson and the team at University College London for their submission Body Condition Scoring for Laboratory Zebrafish Other submissions for the Award are also included in this issue We are also pleased to have had made available to us several of the posters exhibited at the Laboratory Animal Science Association LASA 2012 Winter Meeting The majority of the posters relate to the 3Rs and I commend them to you topics include the use of a Non surgical Embryo Transfer device NSET as a means of refinement the preferences of fish for varying types of environmental enrichment and a review of the use of anaesthetics and analgesics in German research applications ix

April 2013 Animal Technology and Welfare Body condition scoring for Laboratory Zebrafish CAROLE WILSON KAREN DUNFORD CARLY NICHOLS HEATHER CALLAWAY JENNA HAKKESTEEG and MATTHEW WICKS Division of Biosciences University College London Gower Street London WC1E 6BT Corresponding author carole wilson ucl ac uk Winner of the 2012 Andrew Blake Tribute Award Summary Zebrafish are a rapidly expanding model in biomedical research The numbers reported in Home Office returns do not always show the true numbers of Zebrafish actually held in facilities as much work is conducted on embryonic forms that do not need to be included The true husbandry requirements of Zebrafish remain elusive much work is still required to identify their actual needs This lack of information is reflected in many unstandardised husbandry methods the true impact upon some scientific disciplines is now beginning to be understood Currently there are no standardised methods for health monitoring of Zebrafish In this paper we describe how we have developed and described a tank side body condition scoring system which may also be called the traffic light system We created a pilot study to trial this system using 45 volunteer participants from a variety of backgrounds We asked them to score fish in various different conditions without any training and then again after receiving limited training to try to identify whether the participants responses could be standardised From our trial we determined that using Body Condition Scoring is a way of standardising health monitoring and how such a system could be implemented Although the idea of Body Conditioning Scoring for a Zebrafish facility does require further work we believe that it has many advantages over current methods being used It can standardise basic health monitoring within a facility and complement other health monitoring processes such as sentinel water quality and biofilm screening Introduction Zebrafish Danio rerio are fast becoming the animal model of choice for a range of scientific disciplines beyond the more traditional developmental biology1 2 and large scale mutagenic screens3 4 The disciplines now using Zebrafish embryos include drug and small molecule screening5 6 and adults for tissue regeneration studies7 8 and cancer research9 10 Although Zebrafish have become an increasingly popular model Home Office statistics show fish in general represented 15 of all returns in 201111 this does not tell the whole story much work is done on embryonic forms so colony size can be much larger than is suggested by Home Office returns The speed and growth rate of scientific interest in Zebrafish may have also have outpaced the speed at which husbandry methodology has advanced There is a lack of general information about the husbandry requirements of Zebrafish although studies have been made of them in the wild12 13 information about their true husbandry requirements remains elusive14 This has led to a variety of various husbandry protocols being developed personal observation and few if any truly standardised methods used in the husbandr y of Zebrafish One way of introducing elements of standardisation across the facility could be to introduce a body condition scoring system This paper describes a pilot study of a body condition scoring system devised and trialled by UCL fish facility staff Scoring systems are broadly used in many areas and allow for monitoring of various aspects of animals health and behaviour In agriculture body condition scoring systems are common for assessing farm animal health primarily cattle and sheep using body condition as an indicator For example in dairy herds they can be used as an indicator of body fat and potential milk yield15 as well as general health16 in sheep as a measure of body fat17 and in horses using estimated weight as an indicator of body condition18 19 In laboratory mammalian species scoring systems have been developed for a variety of different purposes from those used to assess welfare and health under a variety of circumstances to those which aid scientific research more directly such as assessment of chimeric mice20 Body conditioning scoring systems have been developed as non invasive methods for scoring in both rats21 and mice22 and are used to assess health and general welfare especially when body weight may not 1

Body condition scoring for Laboratory Zebrafish be applicable such as in cancer tumour models Scoring systems have been developed to assess and monitor depth of anaesthesia in laboratory species usually determined by respiration rate heart rate blink reflexes and various pinch reflexes More recently systems have been developed to determine pain by facial expression grimace scales in both mice23 and rabbits24 Although there has been use of scales to both classify and monitor aspects of fish husbandry and welfare both within the scientific community for example guidance on the severity classification of procedures of fish has been published25 and in fisheries ecology26 and more general aquaculture the use of body condition indices is relatively common27 28 In monitoring any aspect of Zebrafish health and welfare the use of body condition scoring is rare and not well developed or utilised Although condition factor indices measuring growth and mass have been used in assessing Zebrafish condition29 this type of body condition scoring is inappropriate for large scale use as weighing and measuring the fish would be a substantial stressor to the fish Additionally the large numbers of fish housed in many facilities would also make this extremely labour intensive and practically impossible Other types of body condition scoring used for mammalian species may also be difficult to translate directly to fish as they may involve handling animals15 17 In developing our body conditioning scoring system we have tried to address some of the problems presented by other systems for body condition scoring and recognise some of the difficulties presented by body conditioning scoring of Zebrafish We have developed a body conditioning scoring system that is non stressful to the fish can be performed at the same time as the daily checks required by The Animal Scientific Procedures Act 1986 is easy to learn and introduces a standardised method All Zebrafish were held in either 3 or 10 litre tanks on recirculating systems at no more than 5 fish per litre and each tank has 6 water changes per hour The fish water is conditioned from reverse osmosis water with marine salts added back The water is changed 6 times per hour The water parameters are temperature 28 C 1 C PH 7 0 5 conductivity 450 600 S ammonia 0mg litre nitrites 0 0 3mg litre and nitrates 0 25mg per litre Dissolved gases and water hardness are not routinely measured The fish are fed combinations of a dr y microencapsulated diet and Instar I artemia two three times daily It should be noted that it is rare to find such extreme examples of ill health within any Zebrafish population and we have calculated that the examples we have shown represent less than 0 01 of our total fish population at any given time that could be classified with a body condition score of BSC2 amber or 3 red The traffic light Body Conditioning Scoring System for Zebrafish The traffic light BCS body condition scoring system comprised of 4 stages Table 1 and Figure 1 These 4 stages grade various aspects of both fish behaviour and general body condition The four stages indicate various stages of health decline of health that maybe obser ved in a general population of Zebrafish especially if the population is large30 BCS1 Black Fish presenting these symptoms have little swimming movement little gill movement If these are isolated cases these fish should be removed from the systems if there are high numbers of fish presenting these signs further immediate action is necessary as this may indicate a water quality issue Method This trial comprised of two parts the development of a traffic light system where various stages of body condition were graded and a second part when we assessed the suitability of the method where different groups of volunteers were asked to use the system to score the body condition of different fish All work was carried out in accordance to the Animals Scientific Procedures Act 1986 no work was carried out under a specific project licence as this study involves a variation of standard husbandry procedures The fish used for this trial were from the UCL fish facility general adult population and healthy adults were representative of a mixed population of sex ages and genetic status from a total of approximately 50 000 Zebrafish Danio rerio The microbiological status of these fish was untested 2 Figure 1 a Body Conditioning Score 1 Black

Body condition scoring for Laboratory Zebrafish BSC2 Red Fish presenting these symptoms should be removed from the system as many of these conditions can represent underlying infectious disease such as mycobacteria31 and Pseudoloma neurophilia32 Other explanations may be genetic water quality issues and age related33 Body Conditioning Score 2 Remove immediately Club tail tail deformity Emaciated wasted low body to head ration Lesion Cataracts Tumour Dropsy scales puff outwards Scoliosis and lordosis curvature along spine Scale and pigmentation loss BCS3 Amber I Amber II Representing both over and under conditioned fish Fish presenting these symptoms require monitoring as none of the conditions are necessarily serious in themselves and maybe the result of genetics If listing and gasping are not in isolated fish this is a serious problem as it is an indication of gas saturation34 within the water and must be acted on immediately BSC4 Green The vast majority of any fish Zebrafish population should fall into this category Healthy Zebrafish should have good symmetric body shape complete fin complement not showing any signs of distress when swimming or breathing and it should be easy to determine the sexes through body shape Male body shape is more bullet like female is more rounded especially in the abdomen Body Conditioning Score 4 Looks like Well conditioned Sleek body Consistent pattern and colour Sexes visible Popeye protruding eye Severely eggbound Also includes Reversed orientation swimming at an extreme angle Decayed fins Figure 1 b Body Conditioning Score 2 Red Body Conditioning Score 3 Monitor Also includes Gasping Missing dorsal or pectoral fin Figure 1 d Body Conditioning Score 4 Green Figure 1 c Body Conditioning Score 3 Amber Table 1 The different stages of the Body Condition Scoring system traffic light system 3

Body condition scoring for Laboratory Zebrafish The Trial of the traffic light Body Conditioning Scoring System Six fish representing at least one fish representing each of the stages were selected from the main facility After the trial all fish were culled in accordance with the Animals Scientific Procedures Act 1986 Schedule 1 All fish staged at either black or red would have been culled and all other fish were surplus to scientific requirements and were due to be culled by a Schedule 1 method Body Conditioning Score 1 All volunteer participants were ranked according to previous knowledge of fish and animal husbandry We determined four different categories within the volunteer participants Category 1 Fish technicians responsible for the daily care of fish and researchers whose primary work is done with Zebrafish and have responsibilities under Animals Scientific Procedures Act 1986 Body Conditioning Score 2 Looks like Dead immobile white grey colour missing body parts Dying laying on bottom of tank not moving when provoked Looks like Emaciated wasted body to head ratio Tumour Reversed orientation swimming at an extreme angle Decayed fins Scale and pigmentation loss Scoliosis lordosis curved back Popeye protruding eye Dropsy scales puff out Body Conditioning Score 3 Body Conditioning Score 4 Looks like Thin Gasping Listing Missing operculum Missing dorsal or pectoral fin Looks like Well conditioned Sleek body Consistent pattern and colour Sexes visible Figure 2 Body Condition Scoring sheet used to train participants 4

Body condition scoring for Laboratory Zebrafish Category 2 Students who see the fish regularly but neither conduct research nor per form husbandr y duties Category 3 Experienced animal technologists who work with mammalian species but do not work with fish Category 4 Participants who had neither knowledge of Zebrafish as a research model nor of the husbandry of any animal used in research Five individual tanks housing the fish were arranged one was coded BCS1 black dead one was coded BCS4 green healthy and the remainder were variously coded BCS2 red and BCS3 amber and had various different conditions as stated and shown in Table 1 Figure 2 The individual tanks were transparent and housed individual fish in order for the participant to view the fish completely before assigning it a BCS traffic light colour A 10 litre tank was also set up containing either one or two fish from categories BCS1 black BCS3 amber various stages of ill health and eight to ten BSC4 green normal healthy fish for the participants to view Each participant was asked to give a BCS traffic light colour score The fish in the individual tanks were given a BSC traffic light score The scoring for the 10 litre tank had to be assigned according to the BSC traffic light score of the least healthy fish not an average score or a score based on the majority of BCS4 green fish but a score that could inform someone else what the worst scenario was within the tank Each participant was asked to score all the tanks initially according to their own experience and knowledge of fish no help was given at this stage Once complete the participant was given a short amount of training 510 minutes shown a BCS traffic light scoring sheet Figure 2 with the various levels stages explained including examples of diseases ill health with accompanying pictures Then with the aide memoire of the BCS traffic light scoring system they were asked to rescore all tanks Zebrafish in a research setting Group 2 Participants not familiar with Zebrafish in a research setting Using the two groups Figure 3 we found that at 64 of group 1 fish technicians and research workers using Zebrafish were able to correctly score the fish before training and 74 were then able to correctly score fish after training The combined categories of 2 4 as group 2 were able to correctly score fish in 53 of all cases prior to training and increased to 63 after training Interestingly a breakdown of the categories within group 2 Figure 4 showed a significant rise in the categor y 2 par ticipants students familiar with Zebrafish but not conducting either husbandry or research ability to correctly score the Zebrafish rose from 56 prior to training to 84 after training Conversely category 4 participants ability to score the fish after training dropped slightly 57 to 53 although this is not significant data not shown A one way ANOVA was performed on the data A P value of 0 039 for the data generated before training indicates that there is a significant difference between the groups A P value of 0 042 for the data generated after training indicates that there is still a significant difference between the groups Figure 3 The difference in results between group 1 and group 2 Results In total 45 volunteers participated in the trial over the course of nine different events The number of participants from each of the 4 different categories was as follows Category Category Category Category 1 2 3 4 21 5 7 12 We then decided to combine categories 2 4 to analyse some of the results to give us two groups Group 1 Participants familiar and working with Figure 4 The differences in results between the 3 categories in group 2 5

Body condition scoring for Laboratory Zebrafish Discussion Although this is a very preliminary pilot trial the results show that even with a very limited amount of training some degree of standardisation from a visual inspection can be achieved It is unsurprising those participants with the most experience working with Zebrafish could identify and score the most accurately What is more surprising is that students who were used to looking at Zebrafish but had no research or husbandry experience appeared to be able to score the different stages of health the most accurately after training suggesting that what may be required prior to any training is for people to spend some time generally familiarising themselves to the species The failure of participants in category 4 to improve scoring after training also suggests this as well as a need to improve and lengthen the time spent training We recognise that more work needs to be done to fine tune the different stages in the traffic light system which we believe can be achieved by greater range of images in and longer more extensive periods of training We recognised that participants from all categories had problems identifying single fish with a status other than BSC4 green in a tank of otherwise healthy fish Fish displaying distended abdomens egg bound or tumours or displaying dropsy were also not obvious to all participants This will be a problem and require further work to overcome as all participants were looking at 5 tanks in total however in a real life situation the number of tanks examined could be in the thousands This is not a perfect system some things would need additional training and or explanation for example gasping at top on tank by many fish may indicate gas bubble disease gas super saturation and a problem with the systems pushing in gas under pressure to the tanks this is an emergency and not to be treated as amber BCS3 which would only be seen in individual fish Since large number of individuals housed together may make it difficult or impossible to pinpoint individual conditions this may be a better method for identifying conditions that affect larger numbers of fish or at least for conditions that have a profound phenotype It would be less successful for those conditions which have subtle expressions As living fish are always in water it may also make visualisation of health status more difficult and this could also be impeded by non handling something more likely to happen with a mammalian species neurophilia32 It can also be used as an indicator of age through the effects of both fecundity and fertility and used as a marker of possible ill health that have a genetic link for example it may be possible to track health problems in individual lines strains using a BCS traffic light system35 A BCS traffic light system can be used to monitor fish housed singularly or those housed in groups therefore it would make monitoring of individual tanks and groups of fish possible It is important to remember that the health and welfare of each individual fish is important irrespective of how many fish a facility may hold overall A BCS traffic light system would be beneficial to animal technologists who may not be familiar with fish health and welfare and for new staff as well as a useful tool in explaining decline or lack of condition to other interested parties A BCS traffic light system has advantages over some other condition scoring methods e g using K factors29 as length and weight may vary from facility to facility biased by the lack of standardised conditions such as diets and water quality Furthermore BCS is a more rapid and consistent system causing no stresses to the fish and is a better indicator of health status As the trend for using Zebrafish in research increases and the size and holding capacity of Zebrafish facilities correspondingly increases to the point where facilities hold thousands perhaps hundreds of thousands of Zebrafish a health monitoring and scoring system such as we suggest here is likely to become an additional part of any Zebrafish health screening and monitoring process Acknowledgements We would like to thank all of the volunteers who took the time and participated in the trial References 21 22 23 24 However a BCS traffic light system does have several advantages in monitoring fish health and welfare Not only can it allow standardisation of monitoring fish it can also enable tracking and observations of specific conditions for example it may help to identify outbreaks of infectious disease in fish colonies and Pseudoloma especially mycobacterium31 6 25 26 Kimmel C B et al 1995 Stages of Embr yonicDevelopment of the Zebrafish Developmental Dynamics 203 3 p 253 310 Kimmel C B Kane D A and Ho R K 1991 Lineage Specification during Early Embryonic Development of the Zebrafish Cell Cell Interactions in Early Development p 203 225 Mullins M C et al 1994 Large Scale Mutagenesis in the Zebrafish in Search of Genes Controlling Development in a Vertebrate Current Biology 4 3 p 189 202 Haffter P et al 1996 The identification of genes with unique and essential functions in the development of the zebrafish Danio rerio Development 123 p 1 36 Crawford A D Esguerra C V and de Witte P A M 2008 Fishing for drugs from nature Zebrafish as a technology platform for natural product discovery Planta Medica 74 6 p 624 632 Tan J L and Zon L I 2011 Chemical Screening in

Body condition scoring for Laboratory Zebrafish 27 28 29 10 11 12 13 14 15 16 17 18 19 20 21 22 Zebrafish for Novel Biological and Therapeutic Discovery in Zebrafish Disease Models and Chemical Screens 3rd Edition 105 p 493 516 Poss K D 2007 Getting to the heart of regeneration in zebrafish Semin Cell Dev Biol 18 1 p 36 45 Jopling C et al 2010 Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation Nature 464 7288 p 606 9 Etchin J Kanki J P and Look A T 2011 Zebrafish as a Model for the Study of Human Cancer in Zebrafish Disease Models and Chemical Screens 3rd Edition 2011 105 p 309 337 Amatruda J F et al 2002 Zebrafish as a cancer model system Cancer Cell 1 3 p 229 231 Home Office 2012 Statistics of Scientific Procedures 2011 Home Office Editor 2012 The Stationery Office Limited London Engeszer R E et al 2007 Zebrafish in the Wild A Review of Natural History and New Notes from the Field Zebrafish 4 1 p 21 U126 Spence R et al 2007 Diet growth and recruitment of wild Zebrafish in Bangladesh Journal of Fish Biology 71 1 p 304 309 Lawrence C 2007 The husbandry of zebrafish Danio rerio A review Aquaculture 269 1 4 p 1 20 Edmonson A J et al 1989 A Body Condition Scoring Chart for Holstein Dairy Cows Journal of Dairy Science 72 1 p 68 78 Pankaj P K Mishra A and Jain R 2008 Body Condition Scoring and Its Application in Dairy Cattle Management Veterinary Practitioner 9 2 p 171 175 Russel A J F Doney J M and Gunn R G 1969 Subjective Assessment of Body Fat in Live Sheep Journal of Agricultural Science 72 p 451 Carroll C L and Huntington P J 1988 Body Condition Scoring and Weight Estimation of Horses Equine Veterinary Journal 20 1 p 41 45 Kimball C 2008 Equine body condition scoring Veterinary Technician 29 9 p 548 White G et al 2011 Development of a photographic reference series for the scoring of chimeras Animal Technology and Welfare 10 3 p 229 223 Hickman D L et al 2008 Characterizing a Body Condition Scoring Technique and Scale for Rats Journal of the American Association for Laborator y Animal Science 47 5 p 110 110 Ullman Cullere M H and Foltz C J 1999 Body condition scoring A rapid and accurate method for assessing health status in mice Laboratory Animal Science 49 3 p 319 323 23 24 25 26 27 28 29 30 31 32 33 34 35 Langford D J et al 2010 Coding of facial expressions of pain in the laboratory mouse Nat Methods 7 6 p 447 9 Keating S C J et al 2012 Evaluation of EMLA Cream for Preventing Pain during Tattooing of Rabbits Changes in Physiological Behavioural and Facial Expression Responses Plos One 7 9 Hawkins P et al 2011 Guidance on the severity classification of scientific procedures involving fish report of a Working Group appointed by the Norwegian Consensus Platform for the Replacement Reduction and Refinement of animal experiments Norecopa Laboratory Animals 45 4 p 219 224 Bolger T and Connolly P L 1989 The Selection of Suitable Indexes for the Measurement and Analysis of Fish Condition Journal of Fish Biology 34 2 p 171 182 Lambert Y and Dutil J D 1997 Can simple condition indices be used to monitor and quantify seasonal changes in the energy reserves of cod Gadus morhua Canadian Journal of Fisheries and Aquatic Sciences 54 p 104 112 Gaylord T G and Gatlin D M 2000 Assessment of compensator y growth in channel catfish Ictalurus punctatus R and associated changes in body condition indices Journal of the World Aquaculture Society 31 3 p 326 336 Siccardi A J et al 2009 Growth and Survival of Zebrafish Danio rerio Fed Different Commercial and Laboratory Diets Zebrafish 6 3 p 275 280 Lawrence C 2011 Advances in Zebrafish husbandry and management Methods Cell Biol 104 p 429 51 Kent M L et al 2004 Mycobacteriosis in zebrafish Danio rerio research facilities Comparative Biochemistr y and Physiology C Toxicology Pharmacology 138 3 p 383 390 Kent M L and Bishop Stewart J K 2003 Transmission and tissue distribution of Pseudoloma neurophilia Microsporidia of Zebrafish Danio rerio Hamilton Journal of Fish Diseases 26 7 p 423 426 Kishi S et al 2003 The zebrafish as a vertebrate model of functional aging and very gradual senescence Experimental Gerontology 38 7 p 777 786 Matthews J L 2004 Common diseases of laboratory Zebrafish Zebrafish 2nd Edition Genetics Genomics and Informatics 77 p 617 643 Gerhard G S et al 2002 Life spans and senescent phenotypes in two strains of Zebrafish Danio rerio Experimental Gerontology 37 8 9 p 1055 1068 7

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April 2013 Animal Technology and Welfare An effective method for terrestrial arthropod euthanasia NEIL BENNIE CHRISTOPHER LOARING MIKAELLA BENNIE and STEVEN TRIM Venomtech Limited Kent Enterprise Hub University of Kent Canterbury Kent CT2 7NJ UK Correspondence dh323 cam ac uk Reproduced and adapted with permission of the Company of Biologists from the Journal of Experimental Biology 2012 215 4237 4241 A 2012 Andrew Blake Tribute Award submission Summary As scientific understanding of invertebrate life increases so does the concern for how to end that life in an effective way that minimises potential suffering and is also safe for those carrying out the procedure There is increasing debate on the most appropriate euthanasia methods for invertebrates as their use in experimental research and zoological institutions grows Their popularity as pet species has also led to an increase in the need for greater veterinary understanding Through the use of a local injection of potassium chloride KCl initially developed for use in American lobsters this paper describes a safe and effective method for euthanasia in terrestrial invertebrates Initial work focused on empirically determining the dose for cockroaches which was then extrapolated to other arthropod species For this method of euthanasia we propose the term targeted hyperkalosis to describe death through terminal depolarisation of the thoracic ganglia as a result of high potassium concentration Key words inver tebrate anaesthesia potassium chloride KCl targeted hyperkalosis Introduction A good death is an important part of a good life for all animals in captivity and a lack of effective euthanasia methods for invertebrates has meant that the practice is not in tune with the theory This gap in knowledge has gained significant attention recently Cooper 2011 Murray 2012 as moral concern grows over just what constitutes suffering in these animals Suitable euthanasia methods need to be not only effective and conducted in a way that causes minimal pain but also relatively simple to perform acceptable to the person conducting the procedure American Veterinary Medical Association 2007 as well as compatible with any research involved Battison et al Battison et al 2000 published a new method of euthanasia for the American lobster Homarus americanus using injection of a saturated solution of potassium chloride KCl This method caused immediate immobilisation and death through circulator y arrest in approximately 1 min and therefore fits the criteria for a euthanasia procedure Currently anaesthesia followed by immersion in fixative such as 70 ethanol has been cited as a preferred method Pizzi 2012 but it is inappropriate for microbiology Cooper 2011 or RNA extraction Freezing is often suggested by private and professional keepers but this compromises histological examination and is increasingly regarded to be inhumane Pizzi 2012 without prior anaesthesia to a depth at which recovery is impossible before death from freezing This study extrapolates the work performed by Battison et al Battison et al 2000 to a range of terrestrial arthropod phyla Our aim is to produce a universal method of arthropod euthanasia that is not only more effective than current methods but is quicker more economical and importantly for transcriptomic and proteomic studies preserves tissue quality Animal nervous systems contain ion channels permeable to potassium and sodium which respectively control the flow of these ions across the cell membrane and subsequently the electrical potential In the field of electrophysiology it is well known that neurones at a resting potential maintain a high intracellular potassium concentration and low intracellular sodium concentration relative to the extracellular environment Addition of extracellular potassium ions K in the form of KCl immediately depolarises the cell through neutralisation of the potassium concentration gradient across the cell membrane This resulting change in potential voltage across the cell membrane causes opening of voltage gated sodium channels Thus 9

An effective method for terrestrial arthropod euthanasia excess extracellular KCl causes exaggerated sodium influx that depolarises the cell and becomes toxic to it The excess K remaining in the extracellular environment also prevents repolarisation Takahashi et al 1999 Therefore application of excess K around the neurones of the thoracic ganglia in the form of KCl abolishes the neural input and results in circulatory collapse and then death Evolutionarily conser ved across a wide range of arthropod orders the arthropod ventral nerve cord with some exceptions is positioned on the ventral midline and exhibits a chain of serial ganglia The anterior ganglia are the largest and have been referred to as the invertebrate brain Gullan and Cranston 1994 but their main role is processing the large amount of sensory input from the head Therefore destruction of the brain in invertebrates principally affects the input of sensory information from the eyes and antenna but not from the rest of the body Consequently a method of euthanasia targeting the ganglia that control the vital functions would be a more appropriate way to successfully terminate arthropod life Humans are not the first species to use K ions as pharmacological tools scorpions have evolved to utilise K ions Parabuthus species possess potent venom peptides that act on many ion channels including K channels Yet they have also evolved a prevenom with a high concentration of K ions of approximately 80mmoll 1 Inceoglu et al 2003 A study on the effect of this prevenom on two insect species Trichoplusia ni and Sarcophagia bullata concluded it is an energy efficient way of prey capture through paralysis Inceoglu et al 2003 We demonstrate that if K is delivered directly to the thoracic ganglia its effects are rapid and avoid generating potential nociceptive action potentials Assessment of proposed injection volume Prior to any dosing studies six giant cockroach nymphs Blaberus giganteus Linnaeus 1758 were anaesthetised in the manner described below and allowed to recover in fresh air to record a baseline recovery time Ten nymphs with a mean s d mass of 3 25 1 03g were anaesthetised individually in a 670ml anaesthesia chamber Reed et al 2011 that delivered carbon dioxide CO2 at 0 5 lmin 1 through a low range flow metre Harvard Apparatus Edenbridge UK Once anaesthetised nymphs were abdominally injected with sterile Ringers solution at 100lg 1 body mass allowed to recover and then monitored for signs of unusual behaviour and or illness The time taken for full induction i e abolishment of the righting reflex and any movement were recorded along with the recovery time The time to full recovery was defined as time to the return of the righting reflex This was important as it enabled us to establish when recovery was expected Development of the euthanasia method Rather than performing a full dose response curve three dose groups were selected to assess the initial efficacy of KCl as a method of euthanasia this was to We hereby propose the term targeted hyperkalosis to describe the euthanasia of invertebrates through local injection of K to deliberately depolarise the thoracic ganglia and bring about rapid death The direct derivation of targeted hyperkalosis is a local highpotassium state that accurately describes the method Materials and methods All animals used in this study were euthanised to provide tissue for other research projects and as such the development of a suitable method of euthanasia was required Although the species involved are not currently legally protected they were maintained under the ethos of the UK Animals Scientific Procedures Act 1986 A SP A 86 All animals were acclimatised for 2 weeks to confirm health and nutritional status and were weighed during total anaesthesia 10 Figure 1 Blattodea injection site The first arthodial membrane red arrow used for euthanasia by targeted hyperkalosis in cockroaches Blaberous giganteus order Blattodea

An effective method for terrestrial arthropod euthanasia reduce the number of animals receiving a toxic but nonfatal dose As a suitable euthanasia protocol needs to be effective for every animal every time standard toxicological parameters such as LD50 lethal dose in 50 of population and LD90 lethal dose in 90 of population were not deemed necessary for this study KCl was dissolved at 300mg ml 1 in sterile water which avoids the need for an incubation step as conducted by Battison et al Battison et al 2000 and eliminates the risk of precipitation if the solution is kept at cooler temperatures The solution was sterile filtered through a 0 2m filter Merck Millipore Watford Hertfordshire UK and aliquoted for storage in 1 5ml centrifuge tubes Fisher Scientific Loughborough Leicestershire UK Blaberus giganteus were anaesthetised as previously described and injected with 30 mg ml 1 KCl into the first leg sinus via the arthrodial membrane Fig 1 using a 25G needle and 1ml syringe Fisher Scientific Doses assessed were 10 lg 1 N 5 animals 2 72 1 26g 50lg 1 N 5 animals 2 92 0 71g and 100l g 1 N 10 animals 2 49 0 42g Double the number of animals was used in the higher dose group to confirm a 100 fatality rate ventral midline at the junction between the first leg plate and adjacent ventral plate Fig 3 Acanthoscurria cordubensis and Hadogenes troglodytes were selected based on the requirement for their venom glands Acanthoscurria cordubensis were dosed centrally via the sternum Fig 4 whereas H troglodytes were injected into the junction between the second leg coxa and the sternum rostrally to the genital operculum Fig 5 Results A rising concentration of CO2 was effective at reversibly anaesthetising and therefore immobilising all species used during this study Individuals from several insect species particularly those from the orders Blattodea and Phasmidae were observed to vomit during anaesthesia and occasional exaggerated limb movements were noted Little other evidence of distress was observed Assessment of proposed injection volume Injections for Gr yllus bimaculatus and Locusta migratoria were administered in the first leg sinus via the arthrodial membrane In Hierodula membranacea the injection was via the arthrodial membrane but in the second leg sinus Initial baseline data indicated that anaesthesia induction for B giganteus took 1min 28s 6s for full anaesthesia with recovery taking 1min 41s 13s After injection of 10 v w of body mass of sterile Ringers solution whilst under anaesthesia animals recovered in 1min 46s 9s No unusual behaviour was noted in the 48h observation time post injection and subsequently no further observations were made The E calcarata nymph given 100lg 1 sterile Ringers solution into the ventral thoracic cavity recovered after 6min and showed no detectable behavioural differences up to and beyond the 48h observation time This demonstrates that the large volume alone 10 v w of body mass does not cause any noticeable effect on survival or behaviour Myriapoda species Scolopendra polymorpha and Narceus americanus were injected into the joint between the second and third segment along the ventral midline Fig 2 Phasmids were injected into the Battison et al Battison et al 2000 used ultrasonography to confirm circulatory arrest in H americanus in this study the authors relied upon irreversible cessation of movement and sensation Extrapolation of the euthanasia method to other species In order to determine the effectiveness of KCl euthanasia in different terrestrial invertebrates we tested the method in eight other invertebrate species from eight orders Table 1 Species Common name Life stage N Mass g Dose v w Gryllus bimaculatus De Geer 1773 African field cricket Adult 6 0 97 0 22 10 Locusta migratoria Linnaeus 1758 Migratory locust Adult 6 1 62 0 29 10 Hierodula membranacea Burmeister 1838 Giant Asian mantid Nymph 5 0 39 0 06 10 Scolopendra polymorpha Wood 1861 Sonoran desert centipede Juvenile 5 0 32 0 2 20 Narceus americanus Palisot de Beauvois 1817 Florida millipede Juvenile 7 2 87 0 81 10 Eurycantha calcarata Lucas 1869 Giant spiny stick insect Nymph 8 2 00 0 89 10 Acanthoscurria cordubensis Thorell 1894 Sub adult 10 18 68 3 21 0 5 Hadogenes troglodytes Peters 1861 Flat rock scorpion Juvenile 9 5 08 2 59 10 Masses are presented as means s d Table 1 KCI dosing for euthanasia of test species 11

An effective method for terrestrial arthropod euthanasia which was defined as no movement or recovery during a 24 hour period Doses of 1 and 5 v w KCl in B giganteus were non fatal 5 v w caused marked local paralysis of forelimbs and antennae which persisted for over 1min 10 v w of KCl caused instant paralysis and inward contraction of the limbs as well as total abolishment of all nociceptive responses and nonrecovery within 24 hours Therefore 10 v w was identified as the 100 effective dose for follow up studies Results of extrapolation of euthanasia method to other species In G bimaculatus 10 v w was immediately effective in all but one individual However this was a suspected inaccurate dosing because for such small species a 33 G needle is required to limit leakage and maintain accuracy For L migratoria H membranacea and E calcarata 10 v w KCl euthanasia was effective in all animals The only unusual observation made was the forceful autotomisation of a rear leg by one L migratoria The Myriapoda unique body plan presented a challenge as nerve ganglia and heart tissue are duplicated throughout its length this most likely brought about the results observed The N americanus responded to Figure 2 Chilopoda injection site Ventral midline injection site red arrow between segments two and three for centipedes Solopendra polymorpha order Chilopoda 12 10 v w KCl with a wave of paralysis moving anterior to posterior with a maximum latency to death of 13s Over half of the study group four animals were deemed dead before removal of the needle The passage of paralysis anterior to posterior was slower in S polymorpha but after a single dose of 10 v w KCl none of the study group had died within 10min Immediate paralysis of the anterior segments was evident in all animals while the posterior three segments remained active in all animals A second dose of 10 v w KCl was required and resulted in immediate cessation of all movement As such a 20 v w KCl final dose was effective in all the S polymorpha tested and was therefore proposed as the protocol for the Chilopoda class of Arthropoda The 10 v w KCl dose in the H troglodytes caused immediate paralysis and death in all animals signified by contraction of the limbs and ablation of responses to noxious stimuli limb crush Initial studies on the Theraphosidae revealed that injection of 10 v w into the central nerve ganglion was not possible due to volume limitations and a substantial back pressure was observed in the syringe Although it was not possible at this time to record haemolymph pressure in Figure 3 Phasmidae injection site Ventral midline injection site red arrow for Eurycantha calcarata order Phasmidae

An effective method for terrestrial arthropod euthanasia the Theraphosidae prosoma and the authors are unaware of any articles on the subject reducing the dose to 0 5 v w and leaving the syringe in place for 10s resolved the issue Thus 0 5 v w can be injected into the central sternum as this is technically easier than reaching the ganglia through an ar throdial membrane grisella Fabricius 1794 Lepidoptera hover fly unknown species Diptera and several beetles Pachnoda marginata Drury 1773 Smaragdesthes africana oertzeni Kolbe 1895 and Dicronorrhina derbyana conradsi Kolbe 1909 Coleoptera For euthanasia of the small insects a 10l glass syringe Hamilton Company Reno NV USA was used with a 25G needle The data presented here are summarised in Table 2 as a list of proposed euthanasia protocols listed by insect order For heavily armoured arthropods such as the crustaceans the thoracic ganglia can be reached through the arthroidial membrane sinus Battison et al 2000 For terrestrial arthropods the ventral midline should be used where possible Figure 4 Theraphosidae injection site Central sternum injection site red arrow for Acanthoscurria cordubensis order Theraphosidae Injection of 0 5 v w administered centrally via the sternum was effective in ablating the nervous system and caused death in all animals Injection of 0 5 v w into the prosoma ganglia is terminal and nonrecoverable in Theraphosidae haemolymph can be collected via cardiac puncture but only for approximately 1min as this appears to be when the circulation stops During development of this protocol we discovered that direct intra cardiac delivery of 1 v w KCl is also an effective euthanasia method for Theraphosidae spiders but it does not appear to work for Araneomorphae spiders data not shown Thus the proposed spider protocol is 0 5 v w delivered directly to the prosoma ganglia Other species tested Representatives of other insect orders can also be euthanised with 10 v w KCl administered directly via injection to the thoracic ganglia through the sternal membrane between the forelegs in the ventral midline Adult species tested were the lesser wax moth Achroia Figure 5 Scorpion injection site Dosing site red arrow for Hadogenes troglodytes order Scorpiones Discussion The data presented here demonstrate that KCl dosing is a rapid and effective method of euthanasia in the arthropod species tested With training this method is easy to perform and causes immediate death which allows rapid tissue collection for experimental transcriptomic and proteomic studies An understanding of the species specific neural anatomy is crucial to performing this technique As such it is important that this is understood prior to working with such animals Before obtaining vertebrates for scientific study it is vital that staff can identify potential suffering and are proficient in carrying out euthanasia This is detailed in the A SP A 86 and we feel that this approach should be adopted for invertebrates as well KCl is an ideal agent for euthanasia as it is cheap effective doesn t require any special storage has a very long shelf life and is safe to use KCl is compatible with a wider range of 13

An effective method for terrestrial arthropod euthanasia Order Common name Injection site Dose v w Blattodea Cockroaches Between first pair of legs 10 Phasmidae Stick insects Between first pair of legs 10 Diplopoda Millipedes Between second and third segments 10 Chilopoda Centipedes Between second and third segments 20 Orthoptera Grasshoppers Between first pair of legs 10 Aranae Spiders Mid sternum 0 5 Scorpiones Scorpions Between second pair of legs 10 Mantodea Mantids Between first pair of legs 10 Coleoptera Beetles Between first pair of legs 10 Diptera Flies Between first pair of legs 10 Table 2 Summary of euthanasia dose protocols pathological and research investigations than any of the current protocols such as transcriptomics and proteomics with the exception of primary neuronal culture due to the nature of action However there are still situations where KCl is impractical such as for very small species e g Drosophila sp or when culling large numbers of invertebrates where tissue samples are not required In these instances anaesthesia followed by a confirmator y procedure such as immersion in fixative or rapid freezing should be performed but only as long as the anaesthesia is deep enough so death occurs before recovery is possible List of symbols and abbreviations A SP A 86 Animals Scientific Procedures Act 1986 CO2 carbon dioxide KCl potassium chloride LD50 lethal dose in 50 of population LD90 lethal dose in 90 of population v w volume by weight Acknowledgements Thank you to Dr Carol Trim for critical review Funding This research was funded internally by Venomtech and therefore received no specific grant from any funding agency in the public commercial or not for profit sectors 14 References American Veterinary Medical Association 2007 AVMA guidelines on euthanasia 2007 Available at https www avma org KB Policies Documents euthanasia pdf Battison A MacMillan R MacKenzie A Rose P Cawthorn R and Horney B 2000 Use of injectable potassium chloride for euthanasia of American lobsters Homarus americanus Comp Med 50 545 550 Cooper J E 2011 Anaesthesia analgesia and euthanasia of invertebrates ILAR J 52 196 204 Gullan P J and Cranston P S 1994 The nervous system and coordination In The Insects An Outline of Entomology pp 61 65 London Chapman and Hall Inceoglu B Lango J Jing J Chen L Doymaz F Pessah I N and Hammock B D 2003 One scorpion two venoms prevenom of Parabuthus transvaalicus acts as an alternative type of venom with distinct mechanism of action Proc Natl Acad Sci USA 100 922 927 Murray M J 2012 Euthanasia In Invertebrate Medicine ed G A Lewbart and I A Ames pp 441 443 Oxford Blackwell Publishing Pizzi R 2012 Spiders In Invertebrate Medicine ed G A Lewbart and I A Ames pp 187 222 Oxford Blackwell Publishing Reed S Lock M and Trim S 2011 Safe laboratory management for arachnids of medical importance Anim Technol Welf 10 85 90 Takahashi S Shibata M and Fukuuchi Y 1999 Role of sodium ion influx in depolarization induced neuronal cell death by high KCl or veratridine Eur J Pharmacol 372 297304

April 2013 Animal Technology and Welfare An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive TIM BETTS Consultant to Charles River UK Ltd Manston Road Margate Kent CT9 4LT Introduction The Home Office released in early November 2012 a consolidated version of ASPA as modified by the regulations put in place following European Directive 2010 63 EU with the proviso that the document should not be taken as a definitive statement of law that must await its passage through parliament later in 2102 It should however be expected to make the deadline of January 1st 2013 for implementation This paper attempts to simplify some of the less clear material that has been added and remind readers that we must wait until well into 2013 before the new Guidance on the Act is released and we can then start to settle into the new regime The purpose of this brief overview is to summarise for Animal Care Staff a group recognised specifically by the European Directive the main changes that will occur and how our organisations will deal with the new legislation as it beds into our system Protected animal The definition remains unaltered except for the addition of cephalopods A protected animal for the purposes of this Act means any living vertebrate other than man and any living cephalopod Note there is no geographical limitation or specification in this definition so it applies to all vertebrate and cephalopods out as far as our 12 mile sea limit and does not limit the remit to animals held in laboratories The animal shall be regarded as continuing to live until the permanent cessation of circulation or the destruction of its brain Remember this is not the either or it appears to prove death after a Schedule 1 kill for example you must be certain that no blood can reach the brain so that neurological tissue will rapidly die from hypoxia you cannot rely solely on what might appear to be the physical destruction of the brain The regulated procedure This definition appears to have undergone some changes but in practice the outcome will remain more or less the same A regulated procedure for the purposes of this Act means any procedure applied to a protected animal for a qualifying purpose which may have the effect of causing the animal a level of pain suffering distress or lasting harm equivalent to or higher than that caused by the introduction of a needle in accordance with good veterinary practice The novelty here is the term for a qualifying purpose which brings in a term we are all familiar with it is applied for an experimental or other scientific procedure and in addition which will be welcomed by many it is applied for an educational purpose However how are you to judge whether this threshold level of pain has been reached I do not think you will need to make that judgement as the crucial part of the definition lies in what is defined as pain suffering and distress and I suspect there will be no change here because Inspectors will rely on their standard definition which as far as I can tell remains as follows Commencement of protection For mammals birds and reptiles as eggs two thirds of the gestation or incubation period for the relevant species has elapsed and for other animals fish amphibia when they become capable of independent feeding Pain suffering distress and lasting harm encompass any material disturbance to normal health physical mental and social well being of the animal disease injury and physiological or psychological discomfort whether immediately or in the longer term There is a necessity to explain the term living and the old definition has been retained The licensee should also be aware that section 2A stipulates that a regulated procedure has been 15

An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive performed on an animal not yet reaching the maturity of a protected animal but it is intended to keep the animal alive until that stage of maturity is reached and it is expected that the definition above can be applied to the outcome The regulated procedure also covers breeding animals that have been subjected to genetic manipulation if the outcome is likely to be as described above The killing of protected animals is a regulated procedure unless they are killed by an appropriate method under Schedule 1 The new legislation is clearer on what is not a regulated procedure a non experimental agricultural practices b non experimental clinical veterinary practice c practices undertaken for the purpose of recognised animal husbandry d procedures under taken under the Veterinar y Medicines Regulations 2011 e ringing tagging or marking an animal for identification provided that it causes no more than only momentary pain or distress and no lasting harm persons dealing with those animals are adequately educated and trained and are supervised until they have demonstrated the requisite competence TCO e a person to be responsible for ensuring that the conditions of the licence are complied with could be licence holder or deputy With Home Office agreement a person may fulfil more than one of these positions The NACWO or NVS should if they have concerns about the health or welfare of any animals kept on the premises notify the personal licensee in charge of the animal or if that is not possible they should take steps to ensure the animal is cared for or if necessary killed The Establishment licence remains in force until revoked but the Home Office reserves the right to vary the licence if they believe circumstances require it Personal licence Change of wording in this section but overall no significant change with one exception the personal licence is no longer tied to an establishment and in theory may be used anywhere in the UK or Europe provided the licence is linked to an active project licence Principles of replacement reduction and refinement No person shall personally apply a regulated procedure to an animal unless The principles of the 3Rs are even more prominent than in the previous legislation featuring in the overview of the Act and in each of the licensing sections a they hold a personal licence for described procedures and animals b the procedures and animals are par t of a recognised project c the place where the procedures are carried out has an Establishment licence detailed in the project licence d the licensee must be endorsed by the Training and Competency Officer e the licensee must be competent to apply the procedures within the conditions stipulated Establishment licence Licensing is still at three levels but we now have a more appropriate title for the licensing of the workplace The 2C licence taking its name from the section of the European Directive which deals with where animals are held and procedures are performed will seamlessly replace the Certificate of Designation It is in this licence that we see the most innovative changes The Licence Holder Certificate Holder as was must nominate five people to fulfil specific roles within the Institution a a person to be responsible for overseeing the welfare and care of the animals kept at the place specified in the licence NACWO b a veterinary surgeon with expertise in laboratory animal medicine or other suitably qualified person fish amphibia reptiles cephalopods to provide advice on the welfare and treatment of those animals NVS c a person responsible for ensuring that the persons dealing with those animals have access to any information they need about the species concerned d a person to be responsible for ensuring that the 16 Project licence As before the project licence specifies a programme of work using specified regulated procedures to animals of specified descriptions at a specified place that place must have a 2C licence A different place may be specified if there is a scientific justification acceptable to the Secretary of State PODE The application for a project licence is broadly similar to what we are used to although there is now a timetable for the outcome of the licence application to be made known to the applicant within 40 working days this can be extended under some circumstances The project licence holder will have to accept responsibility for the overall implementation of the

An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive programme of work and ensure that it is carried out in compliance with the conditions stated on the licence this includes ensuring that severity limits are adhered to and that retrospective assessment may need to be made of the actual response the animals made to the treatment given The five year option for the life of a project remains and as before the licence terminates on the death of the project licence holder Compliance Where it appears to the Secretary of State Home Office Inspector that the holder of a licence under this Act is failing or has failed to comply with a condition of the licence or a provision of the Act then a compliance notice will be served which a specifies the condition or provision that has not been complied with b specifies the action to be taken by the licence holder to ensure the failure is not continued or repeated c specifies the action to be taken to eliminate or reduce any consequences of failure d requires the holder to take that action within a specified time e explains the effect of the prescribed action such as revocation of the licence The licence may under certain circumstances be suspended revoked or varied A licensee whose licence is suspended revoked or varied without their agreement is entitled to make written or oral representations to a person appointed for that purpose who on completion of their investigation will write a report to Secretary of State a copy of which will be provided to the licence holder concerned Re use Animals that have been used for regulated procedures must not be used again unless The severity conditions of the first procedure have been included in a project licence A veterinary surgeon with knowledge of the animal s lifetime experience has advised that the animal s general state of health and well being have been fully restored following the application of the previous procedures The further procedure must be covered by a project licence and should not exceed non recovery mild or moderate Euthanasia and Schedule 1 A person must not intentionally kill a protected animal unless it is done in a place specified in a section 2C licence and the person uses an appropriate method and is registered as competent to kill animals of recorded species by specific methods as listed on a register held by the establishment Schedule 1 at the time of writing has undergone little change except that the animals may be exposed to sedatives or anaesthesia prior to euthanasia providing that this does not result in more distress than would otherwise occur As in the current table the only method approved under all circumstances is the appropriate overdose of anaesthetic and whichever of the five techniques listed are used an animal cannot be left until death is confirmed by permanent cessation of the circulation destruction of the brain dislocation of the neck exsanguination the onset of rigor mortis or the instantaneous destruction of the body in a macerator Schedule 2 Schedule 2 remains the same with the addition of the amphibians Xenopus laevis and tropicalis Rana temporaria and pipiens and most impor tantly Zebrafish Licence conditions Conditions of section 2C licence Establishment Licence 1 The licence holder shall ensure that the regulated procedures are carried out in a manner consistent with the principles of the 3Rs 2 There must be a register of people who are competent to kill protected animals and each registered person must be adequately educated and trained in both the species to be killed and the methods of killing to be used This register must be made available to the Inspectorate and the Secretary of State 3 The licence holder must notify the Secretary of State of any proposed change in a the full name of the Holder b the full name and qualifications of the named person responsible for compliance c the full name and qualifications of the named animal care and welfare officer d the full name and qualifications of the named veterinary surgeon e the full name and qualifications of the named information officer f the full name and qualifications of the named training and competency officer g the areas on the schedule of premises and the class of use they are put to h the types of protected animals to be held or used in regulated activities 4 a all protected animals must be provided with 17

An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive 5 6 7 8 18 adequate care and housing appropriate to their type or species b restrictions on an animal s ability to satisfy its physiological and ethological needs must be kept to a minimum c unless authorised in a project licence husbandry conditions should be appropriate for the animals housed d the licence holder shall ensure that all installations and equipment function correctly to give reliable results and cause the least possible distress to the fewest number animals possible any defects detected should be eliminated as soon as possible e the health and well being of the animals and the environmental conditions in which they are kept shall be checked at least once a day by a competent person and any deficits discovered should be eliminated as quickly as possible f the holder shall ensure that conditions of transport are appropriate for the animal s health and well being g unless specified in a project licence the standards of care and accommodation as set out in the Codes of Practice must be met The licence holder must ensure that the establishment is appropriately staffed at all times to ensure the well being of the protected animals This requires adequate education and training under supervision until they can demonstrate the requisite competence The licence holder is required to establish and maintain an Animal Welfare and Ethical Review Body consisting of a NACWO a NVS a Project licence holder and any other such persons as may be specified in Guidance to be issued by the Secretary of State The AWERB shall as a minimum carry out the following tasks advise staff dealing with animals on welfare matters and on the application of the 3Rs review internal operational processes with regard to monitoring reporting and checking welfare issues follow the development and outcome of projects identifying fur ther refinements and reductions in animal numbers where possible They should also advise on the appropriate social housing of animals and advise on re homing schemes where these are appropriate A record must be kept of such advice for a minimum of three years Limitations on breeding non human primates from animals not bred in captivity Records to be kept at the licensed establishment a Number and species of animals bred acquired supplied and used in procedures or discharged from the Act b The origin of the animals and if they were bred for use in procedures c Dates on which the animals were acquired supplied or discharged d From whom the animals were acquired e The name and address of the recipient of the animals f The number and species of animals which died or were killed in each establishment For the animals that die the cause of death shall when known be noted g If the licence authorises the use of regulated procedures then the projects for which the animals are used shall be recorded These records listed above shall be kept for 5 years and be submitted to the Secretary of State or Home Office Inspector if requested 9 The licence holder shall ensure that an individual history file be kept for every dog cat or non human primate held stating its date of birth if it was bred for research any relevant reproductive veterinary or social information about the animal together with the procedures it has undergone When the animal dies or is set free or is transferred to another place the file must be kept for at least three years and if requested be submitted made available to the Secretary of State or a Home Office Inspector 10 Any unmarked cat dog or non human primate must be marked permanent individual identification before weaning or as soon after weaning as possible in a manner that causes the least possible pain 11 Inspectors shall have access at all reasonable times to the parts of the establishment where animals are housed used or bred 12 The areas of the establishment that have been approved cannot be changed without approval of the Secretary of State 13 Unless there is formal approval from the Secretary of State the establishment may only keep the protected animals specified in the licence in the areas specified on the licence 14 Health records of the animals kept bred or used at the establishment are to be maintained under the supervision of the NVS and made available to the Inspectorate or the Secretary of State on request 15 The licence holder shall nominate and be responsible for the performance of the named persons listed in section 3 above 16 If for any reasons the named persons mentioned are not available arrangements must be made to provide adequate for all the animals 17 Security to be maintained to prevent animal escapes and intrusion by unauthorised persons 18 Quarantine and acclimatisation facilities shall be provided and used appropriately 19 Adequate precautions against fire shall be maintained 20 The licence holder shall take all reasonable steps to prevent the per formance of unauthorised procedures mistakes 21 The licence holder must liaise effectively with all the named persons and licensees

An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive 22 A project licence holder s death has to be notified to the Secretary of State within seven days of the death and the 2C licence holder must assume responsibility for the project during the ensuing 28 days until the licence is revoked or re submitted 23 Animals kept alive following the completion of regulated procedures must come under the supervision of the NVS or other qualified person unless the animal is moved to another establishment or its re use is authorised by the Secretary of State 24 These conditions must be made available for consultation by all licensees and named persons 25 The licence remains the property of the Secretary of State and must be surrendered on request Many of the above conditions are essentially the same as before but the additional material covers the new named people and has higher requirements for record keeping supervision and animal husbandry Personal licence conditions Many of these conditions remain little changed and some of the new ones appear very repetitive the old condition 10 which stipulated that the licensee had to defer to the Home Office Inspector if in the Inspector s opinion the animal was suffering and should be killed appears to have been dropped The licence application form has been greatly simplified and the personal licence is no longer restricted to the establishment from which the application was made It is probable that some of the conditions below will receive some clarification when the Guidance Notes are published later in 2013 The personal licence will reflect general aspects of research activity without being too specific details of procedures as at present will be found on the project licence 1 In exercising his or her responsibilities the licence holder will at all times act in a manner consistent with the principles of the 3Rs 2 The licence holder is entrusted with the primary responsibility for the welfare of the animals on which they perform regulated procedures 3 The holder must not apply a regulated procedure to an animal if it causes severe pain suffering or distress that is likely to be long lasting and cannot be alleviated 4 The holder must not apply a procedure unless precautions have been taken to prevent or reduce to a minimum any pain suffering distress or discomfort that may be caused 5 Where a procedure is applied to an animal the holder must ensure that any pain suffering distress or lasting harm that is being caused is stopped 6 Where a procedure has been applied and caused pain suffering or distress the holder must take steps to ameliorate the pain suffering or distress 7 The holder shall ensure that where a procedure has been applied that death as an end point is avoided as far as possible and is replaced by an early humane end point 8 In all circumstances where an animal has been subjected to a regulated procedure and is in severe pain suffering or distress which is likely to be long lasting and cannot be ameliorated then the licence holder must ensure that it is immediately killed in accordance with section 15A in effect Schedule 1 9 The licence holder may apply a regulated procedure without the use of anaesthetics only if they are satisfied that the procedure will not inflict serious injuries capable of causing severe pain or if the anaesthetic will in itself be more traumatic to the animal than the intended procedure or if it would frustrate the purpose of the procedure 10 When anaesthesia either local or general is used it shall be of sufficient depth to prevent the animal from being aware of pain arising during the procedure 11 Once the anaesthetic has worn off the licence holder must ensure that pain relieving methods are applied to prevent the break through of post procedural pain 12 The holder must use analgesia or another appropriate method to ensure that pain suffering and distress are kept to a minimum 13 It is the responsibility of the personal licence holder to notify the project licence holder if it appears that the severity limit of the procedure or any adverse effects described in the project licence look likely to be exceeded 14 The licence holder must ensure that suitable arrangements are made for the care and welfare of the animals for which they are responsible during any period when they themselves will not be in attendance 15 The licence holder shall ensure that whenever necessary veterinary advice and treatment are obtained for the animals in their care 16 The licence holder shall ensure that all cages pens or other enclosures are clearly labelled so as to enable an Inspector NVS or NACWO to identify the Project Licence the project licence protocol the start date of the protocol and the Personal licensee involved 17 The personal licensee may not perform procedures un supervised until they have achieved a level of competence for the technique which has been signed off by the TCO or other authorised person 18 The licence holder may delegate to assistants under their control tasks which form an integral part of the regulated procedures but do not require technical knowledge or skill Such tasks will be illustrated in the Guidance Notes when published 19 The licence holder must take all reasonable steps 19

An early overview of the Animals Scientific Procedures Act 1986 as modified by the European Directive to ensure that they have the appropriate personal and project licence authority before commencing a procedure 20 The licence holder shall maintain a record of all animals on which procedures have been carried out including details of super vision and declarations of competence as appropriate The records shall be kept for at least 5 years and submitted to an Inspector or the Secretary of State on request 21 The licence holder must give any necessar y assistance to Inspectors carrying out their visits 22 The licence remains the property of the Secretary of State and shall be surrendered on request Licence applications will in future be assessed by licensing staff at the Home Office and will only be referred to Inspectors if the application involves the use of Neuromuscular blocking agents or falls into a categor y regarded as more complex Other wise applications will fall into five categories A Minor minimally invasive procedures not requiring sedation analgesia or anaesthesia B Minor minimally invasive procedures involving sedation analgesia or brief anaesthesia this will include minor surgical procedures under brief terminal anaesthesia C Surgical procedures involving general anaesthesia with recovery D Use of neuromuscular blocking agents E Procedures conducted for education and training Project licence required Application for the above licences will depend on the submission of the current modular training certificates which will be tweaked to accommodate the European Modular system The licence application will have to present a declaration by the TCO that supervision will be provided until competence is demonstrated and recorded 20 Project licence The project licence conditions follow more closely those already in existence covering the conditions in the personal licence by way of close super vision of licensees and ensuring that severity limits are complied with and that accurate records are kept and reported on where required Summary Most people working under the Act will not notice a great deal of difference to their everyday activities it would seem that supervision will be more rigorous and record keeping will become more important The bureaucratic nature of recording the progress of competence through practice appears to be more onerous and the Animal Welfare and Ethical Review Body will under most circumstances be able to move into its slightly enhanced role without any major changes required Individuals may see less of their Inspector than they used to unless of course their Establishment scores highly on the risk assessment carried out on their activities and attitudes There is a greater emphasis on animal welfare and it would seem highly likely that given current trends welfare charts will need to be kept to enable the accurate reporting of retrospective assessments of severity in at least some projects Good examples of mild moderate severe and non recovery have been published and no doubt other examples will follow and the Guidance Notes will no doubt make clear some of those areas that require more detailed interpretation The interpretations and opinions expressed in this article are those of the author and do not imply the agreement of either the Institute of Animal Technology The Home Office or Charles River UK Ltd

April 2013 Animal Technology and Welfare PAPER SUMMARY TRANSLATIONS INHALTVERZEICHNIS Body Condition Scoring K rperkonditionsbeurteilung von Zebrafischen im Labor CAROLE WILSON KAREN DUNFORD CARLY NICHOLS HEATHER CALLAWAY JENNA HAKKESTEEG und MATTHEW WICKS Division of Biosciences University College London Gower Street London WC1E 6BT Korrespondierende Autorin carole wilson ucl ac uk Gewinner des Andrew Blake Tribute Award 2012 Abstract Die wahren Erfordernisse artgerechter Haltung von Zebrafischen sind bisher kaum erforscht es ist noch viel Arbeit n tig um ihre wahren Bed rfnisse zu identifizieren Dieser Mangel an Informationen spiegelt sich in vielen nichtstandardisierten Versuchsmethoden wider und man versteht inzwischen langsam die tats chlichen Auswirkungen auf verschiedene wissenschaftliche Disziplinen Es existieren derzeit keine standardisier ten Methoden zur Gesundheits berwachung von Zebrafischen In diesem Dokument erl utern wir wie wir ein tankwandseitiges Body Condition Scoring System entwickelt und beschrieben haben das auch als Ampelsystem bezeichnet werden kann Wir erstellten eine Pilotstudie mit 45 freiwilligen Teilnehmern aus verschiedensten Bereichen Wir baten sie um Beurteilung von Fischen unterschiedlicher Kondition ohne jegliches Training und sodann erneut nach Erhalt begrenzten Trainings um so zu versuchen zu ermitteln ob eine Standardisierung der Antworten der Probanden m glich sei Bei unserem Versuch stellten wir fest dass Body Condition Scoring eine Methode standardisierter Gesundheits berwachung ist und wie solch ein System realisiert werden k nnte Auch wenn das Konzept des Body Conditioning Scoring f r eine Zebrafischanlage weiterer Erforschung bedarf glauben wir dass es viele Vorteile gegen ber aktuell verwendeten Methoden besitzt Es kann zur Standardisierung grundlegender Gesundheits berwachung in einer Anlage dienen und andere Gesundheits berwachungsverfahren wie Sentinel Wasserqualit t und Biofilm Screening erg nzen 21

Paper Summary Translations Eine effektive Methode zur Euthanasie terrestrischer Arthropoden NEIL BENNIE CHRISTOPHER LOARING MIKAELLA BENNIE und STEVEN TRIM Venomtech Limited Kent Enterprise Hub University of Kent Canterbury Kent CT2 7NJ Korrespondierender Autor s trim venomtech co uk Nachdruck mit Genehmigung der Company of Biologists aus dem Journal of Experimental Biology 2012 215 4237 4241 Ein f r den Andrew Blake Tribute 2012 eingereichter Beitrag Abstract Mit zunehmendem wissenschaftlichen Verst ndnis ber das Leben von Wirbellosen verst rkt sich auch die Suche nach M glichkeiten einer effektiven Beendigung dieses Lebens die potenzielles Leiden minimiert und gleichzeitig jenen Sicherheit bietet die dieses Verfahren durchf hren Angesichts der wachsenden Nutzung von Wirbellosen in experimentellen Forschungs und zoologischen Einrichtungen wird die Debatte ber die f r Wirbellose am besten geeigneten Euthanasiemethoden intensiver gef hrt Ihre Beliebtheit als Haustiere macht zudem ein gr eres veterin risches Verst ndnis erforderlich Ausgehend von der urspr nglich f r den amerikanischen Hummer entwickelten lokalen Injektion von Kaliumchlorid KCl beschreibt dieses Dokument eine sichere und effektive Methode zur Euthanasie wirbelloser Landtiere Die Arbeit konzentrierte sich zun chst auf die empirische Bestimmung der Dosis f r Kakerlaken die dann f r andere Arthropodenarten extrapoliert wurde Wir schlagen f r diese Methode der Euthanasie den Begriff gezielte Hyperkalosis vor um den Tod durch terminale Depolarisation der Thoraxganglien als Ergebnis hoher Kaliumkonzentration zu beschreiben 22

Paper Summary Translations Ein erster berblick ber den durch die EU Richtlinie ge nderten Animals Scientific Procedures Act 1986 TIM BETTS Berater f r Charles River UK Ltd Manston Road Margate Kent CT9 4LT Abstract Einf hrung Anfang November 2012 ver ffentlichte das Innenministerium eine konsolidierte Version des Animal Scientific Procedures Act 1986 der durch die im Zusammenhang mit der EU Richtlinie 2010 63 EU erlassenen Verordnungen ge ndert wurde mit der Ma gabe dass das Dokument nicht als endg ltige Rechtsvorschrift aufzufassen und seine Verabschiedung durch das Parlament im Verlauf dieses Jahres abzuwarten ist Es ist jedoch zu erwarten dass die f r die Durchf hrung geltende Frist bis 1 Januar 2013 eingehalten wird Dieses Dokument unternimmt den Versuch einer vereinfachten Darstellung des mehr oder weniger eindeutigen Materials das hinzugef gt wurde und erinnert die Leser daran dass wir bis weit ins Jahr 2013 hinein warten m ssen ehe die neue Anleitung zum Gesetz ver ffentlicht wird und wir uns dann in das neue Regelwerk hineinarbeiten k nnen Zweck dieses kurzen berblicks ist es f r Animal Care Staff eine konkret durch die EU Richtlinie anerkannte Gruppe eine Zusammenfassung ber die wesentlichen eintretenden nderungen und ber die Art und Weise zu geben wie unsere Organisationen mit der neuen sich in unser System integrierenden Gesetzgebung umgehen 23

Animal Technology and Welfare April 2013 CONTENU DE LA REVUE valuation de l tat corporel des poissons z bres de laboratoire CAROLE WILSON KAREN DUNFORD CARLY NICHOLS HEATHER CALLAWAY JENNA HAKKESTEEG et MATTHEW WICKS D partement des BioSciences University College London Gower Street Londres WC1E 6BT Auteur ressource carole wilson ucl ac uk Gagnant du prix hommage M Andrew Blake d cern en 2012 R sum Les besoins r els en mati re d levage des poissons z bres restent ind finissables de nombreuses tudes sont encore n cessaires afin de pouvoir identifier leurs v ritables besoins Ce manque d information se retrouve dans de nombreuses m thodes d levage non normalis es et son impact r el sur certaines disciplines scientifiques est d sormais de plus en plus souvent pris en compte Il n existe actuellement aucune m thode normalis e permettant de surveiller la sant des poissons z bres Dans le pr sent document nous d crivons la fa on dont nous avons labor et d fini un syst me d valuation de l tat corporel des poissons z bres se trouvant dans l aquarium galement appel la m thode du feu de circulation Afin de tester cette m thode nous avons cr une tude pilote et avons fait appel 45 participants volontaires issus de milieux divers Nous leur avons demand de noter les poissons dans diverses conditions sans n avoir re u aucune formation Apr s qu ils aient re u une formation limit e nous leur avons demand de proc der nouveau ces notations afin de pouvoir d terminer si oui ou non les r ponses des participants pouvaient tre uniformis es Suite cet essai nous avons constat que l utilisation du syst me d valuation de l tat corporel permettait d uniformiser le syst me de surveillance de la sant des poissons et nous avons identifi le meilleur moyen de mettre en pratique ce syst me Bien que des tudes compl mentaires soient n cessaires dans le cadre du syst me d valuation de l tat corporel pouvant tre appliqu une installation h bergeant des poissons z bres nous pensons que ce syst me poss de plus d avantages que les m thodes actuellement utilis es Il permet d uniformiser les syst mes basiques de surveillance de la sant mis en place au sein d une installation et de compl ter d autres processus de surveillance de la sant tels que les examens li s aux sentinelles la qualit de l eau et aux films biologiques 24

Paper Summary Translations M thode efficace d euthanasie des arthropodes terrestres NEIL BENNIE CHRISTOPHER LOARING MIKAELLA BENNIE et STEVEN TRIM Venomtech Limited Kent Enterprise Hub Universit de Kent Canterbury Kent CT2 7NJ Auteur ressource s trim venomtech co uk Reproduit avec la permission de l association The Company of Biologists et extrait de la revue de Biologie Exp rimentale 215 4237 4241 2012 Dossier soumis en 2012 pour le prix hommage M Andrew Blake R sum Au moment o les connaissances scientifiques sur la vie invert br e augmentent il en est de m me pour les inqui tudes concernant l identification d une m thode permettant de mettre fin cette vie d une mani re qui soit efficace tout en minimisant les souffrances possibles et en garantissant un niveau de s curit lev pour les personnes r alisant cette proc dure Le d bat s intensifie sur les m thodes d euthanasie les plus appropri es pour les invert br s car ceux ci sont de plus en plus utilis s dans le cadre des recherches exp rimentales et au sein des institutions zoologiques Leur popularit en tant qu animaux familiers a entra n un besoin accru de l largissement des connaissances v t rinaires En faisant r f rence l utilisation d une injection locale de chlorure de potassium KCI d velopp e l origine pour tre utilis e sur le homard d Am rique le pr sent document d crit une m thode d euthanasie des invert br s terrestres s re et efficace Les premi res tudes taient ax es sur la d termination empirique de la dose administr e aux cafards Les r sultats de ces travaux ont ensuite t extrapol s d autres esp ces d arthropodes Dans le cadre de cette m thode d euthanasie nous proposons le terme hyperkali mie cibl e pour d crire le d c s suite une d polarisation terminale des ganglions thoraciques engendr e par une forte concentration de potassium 25

Paper Summary Translations Premier aper u de la loi sur les Animaux Proc dures Scientifiques de 1986 telle que modifi e par la directive europ enne TIM BETTS Consultant chez Charles River UK Ltd Manston Road Margate Kent CT9 4LT R sum Introduction Au d but du mois de novembre 2012 le minist re de l Int rieur Home Office a publi une version consolid e de la loi sur les Animaux Proc dures Scientifiques de 1986 telle que modifi e par les r glements mis en place suite la directive europ enne 2010 63 EU sous r serve que ce document ne soit pas consid r comme une d claration de droit d finitive puisqu il doit faire l objet d un examen au parlement un peu plus tard dans l ann e Il est toutefois pr vu qu il entre en vigueur au plus tard le 1er janvier 2013 Le pr sent document a pour objectif de simplifier quelques uns des l ments les plus ambigus qui ont t rajout s et tente de rappeler aux lecteurs que nous devrons patienter pendant une bonne partie de l ann e 2013 avant que les nouvelles directives de cette loi ne soient publi es C est ensuite seulement que nous pourrons commencer nous habituer au nouveau r gime L objectif de ce bref aper u est de r sumer pour le personnel responsable des soins aux animaux un groupe reconnu sp cifiquement par la directive europ enne les principaux changements qui seront op r s et la fa on dont nos organisations appliqueront la nouvelle l gislation lorsqu elle entrera en vigueur au sein de notre syst me 26

April 2013 Animal Technology and Welfare INDICE DE LA REVISTA Calificaci n de la condici n corporal para el pez cebra de laboratorio CAROLE WILSON KAREN DUNFORD CARLY NICHOLS HEATHER CALLAWAY JENNA HAKKESTEEG y MATTHEW WICKS Division of Biosciences University College London Gower Street Londres WC1E 6BT Autor correspondiente carole wilson ucl ac uk Ganadora del Premio Homenaje Andrew Blake 2012 Resumen Los verdaderos requisitos de cr a del pez cebra siguen siendo imprecisos es necesario mucho m s trabajo para identificar sus verdaderas necesidades Esta falta de informaci n se refleja en muchos m todos de cr a no estandarizados el verdadero impacto en algunas disciplinas cient ficas empieza ahora a ser comprendido En la actualidad no existen m todos estandarizados para el seguimiento del estado del pez cebra En este estudio explicamos c mo hemos desarrollado y descrito un sistema de calificaci n de la condici n corporal en acuarios que tambi n puede llamarse sistema sem foro Creamos un estudio piloto para probar este sistema utilizando a 45 voluntarios con distintos historiales Les pedimos que calificaran a peces en distintas condiciones primero sin haber recibido ninguna formaci n y luego de nuevo tras recibir una peque a formaci n para tratar de identificar si las respuestas de los participantes pod an estandarizarse Gracias a nuestra prueba pudimos determinar que la utilizaci n de calificaci n de la condici n corporal es una forma de estandarizar el seguimiento sanitario y determinar tambi n c mo se puede implementar dicho sistema A pesar de que la idea de Calificaci n de la condici n corporal para una instalaci n con peces cebra no requiere m s trabajo creemos que tiene muchas ventajas respecto a los m todos actuales en uso Puede estandarizar un seguimiento sanitario b sico en una instalaci n y complementar otros procesos de seguimiento sanitario como centinela la calidad del agua y las pruebas de pel cula biol gica 27

Paper Summary Translations Un m todo eficaz para la eutanasia de artr podos terrestres NEIL BENNIE CHRISTOPHER LOARING MIKAELLA BENNIE y STEVEN TRIM Venomtech Limited Kent Enterprise Hub University of Kent Canterbury Kent CT2 7NJ Autor correspondiente s trim venomtech co uk Reproducido con el permiso de Company of Biologists del Journal of Experimental Biology 2012 215 4237 4241 Presentaci n para el Premio Homenaje Andrew Blake 2012 Resumen A media que el conocimiento cient fico sobre la vida de los invertebrados va aumentando tambi n lo hace la preocupaci n sobre c mo poner fin a dicha vida de un modo eficaz que minimice el sufrimiento potencial y que sea segura para aquellos que lleven a cabo este procedimiento Hay un debate en aumento sobre los m todos de eutanasia m s apropiados para los invertebrados a medida que su uso en la investigaci n experimental y los centros zool gicos crece Su popularidad como animales de compa a ha dado lugar a la necesidad de un mayor conocimiento veterinario Mediante el uso de una inyecci n local de cloruro de potasio KCI inicialmente desarrollada para su uso en la langosta norteamericana esta investigaci n describe un m todo eficaz y seguro para llevar a cabo la eutanasia en los invertebrados terrestres El trabajo inicial se centr en determinar emp ricamente la dosis para cucarachas que luego se extrapol a otras especies de artr podos Para este m todo de eutanasia proponemos el t rmino hipercalosis objeto para describir la muerte mediante una despolarizaci n terminal de los ganglios tor cicos como resultado de una alta concentraci n de potasio 28

Paper Summary Translations Primer resumen de la Ley sobre animales Procedimientos cient ficos de 1986 modificada por la Directiva Europea TIM BETTS Asesor de Charles River UK Ltd Manston Road Margate Kent CT9 4LT Resumen Introducci n El Ministerio del Interior present a principios de noviembre de 2012 una versi n consolidada de la Ley sobre animales Procedimientos cient ficos de 1986 modificada por las regulaciones estipuladas por la Directiva Europea 2010 63 UE con la estipulaci n de que el susodicho documento no se considere una declaraci n definitiva de la ley ya que deber ser aprobado por el parlamento a finales de a o No obstante se espera que entre en vigor para el 1 de enero de 2013 Este estudio trata de simplificar las secciones menos claras del material que ha sido agregado y recuerda a los lectores que deben esperar hasta 2013 cuando las nuevas orientaciones sobre la Ley entren en vigor y entonces podamos empezar a implementar el nuevo r gimen El objetivo de este documento es resumir para Animal Care Staff grupo reconocido espec ficamente por la Directiva Europea los cambios principales que se llevar n a cabo y el modo en que nuestras organizaciones tratar n la nueva legislaci n a medida que empiece a entrar en nuestro sistema 29

Animal Technology and Welfare April 2013 INDICE DELLA REVISTA Punteggio della condizione corporea dei pesci zebra da laboratorio CAROLE WILSON KAREN DUNFORD CARLY NICHOLS HEATHER CALLAWAY JENNA HAKKESTEEG e MATTHEW WICKS Divisione di Bioscienze University College London Gower Street Londra WC1E 6BT Autore Corrispondente carole wilson ucl ac uk Vincitore del premio Andrew Blake Tribute Award 2012 Abstract I veri requisiti zootecnici dei pesci zebra continuano a essere sfuggenti e bisogna adoperarsi ulteriormente per identificare le loro vere e proprie esigenze Questa mancanza di informazioni evidente in molti metodi zootecnici non standardizzati e si sta cominciando a comprendere il vero impatto su alcune discipline scientifiche Attualmente non esiste alcun metodo standardizzato di stima dello stato fisiologico dei pesci zebra In questa relazione descriviamo come abbiamo sviluppato e descritto un sistema di valutazione della condizione corporea dall esterno di una pesciera a cui ci si pu riferire anche come sistema a semaforo Abbiamo ideato uno studio pilota per collaudare tale sistema rivolgendoci a 45 partecipanti volontari provenienti da background diversi Abbiamo chiesto loro di assegnare un punteggio ai pesci in svariate condizioni differenti senza sottoporli ad alcun training e nuovamente dopo aver ricevuto un training limitato per cercare di stabilire se era possibile standardizzare le risposte dei partecipanti Il trial effettuato ci ha permesso di determinare che l uso del Body Condition Score un metodo di standardizzazione della valutazione dello stato di salute e come pu essere implementato tale sistema Bench l idea del Body Condition Score per un ambiente di pesci zebra non richieda ulteriore ricerca siamo convinti che presenti molti vantaggi rispetto ai metodi utilizzati attualmente Pu standardizzare la stima dello stato fisiologico di base in ambito a una struttura e fare da complemento ad altri processi di monitoraggio dello stato di salute quali eventi sentinelIa qualit dell acqua e screening della formazione di biofilm 30

Paper Summary Translations Un metodo efficace di eutanasia in artropodi terrestri NEIL BENNIE CHRISTOPHER LOARING MIKAELLA BENNIE e STEVEN TRIM Venomtech Limited Kent Enterprise Hub University of Kent Canterbury Kent CT2 7NJ Autore Corrispondente s trim venomtech co uk Riprodotto su concessione della Company of Biologists dal Journal of Experimental Biology 2012 215 4237 4241 Candidato al premio Andrew Blake Tribute Award 2012 Abstract Con il progredire della comprensione scientifica della vita degli invertebrati aumenta anche la preoccupazione su come terminare tale vita in una maniera efficace che riduca al minimo le potenziali sofferenze e sia sicura per coloro che eseguono la procedura Si discute in modo crescente dei metodi eutanasiaci pi appropriati per gli invertebrati dal momento che il loro utilizzo in continua crescita negli istituti zoologici e nella ricerca sperimentale La loro popolarit come specie animale ha portato anche al diffondersi della necessit di un accresciuta comprensione della medicina veterinaria Facendo ricorso a un iniezione locale di cloruro di potassio KCl sviluppata inizialmente per essere utilizzata negli astici americani questa relazione descrive un metodo sicuro ed efficace di eutanasia negli invertebrati terrestri I lavori iniziali erano incentrati sul calcolo empirico della dose per gli scarafaggi poi estrapolati ad altre specie antropode Per questo metodo eutanasiaco proponiamo il termine targeted hyperkalosis per descrivere la morte attraverso la depolarizzazione del terminale dei gangli toracici come conseguenza di un alta concentrazione di potassio 31

Paper Summary Translations Panoramica introduttiva dell Animals Scientific Procedures Act del 1986 come emendato dalla Direttiva europea TIM BETTS Consulente di Charles River UK Ltd Manston Road Margate Kent CT9 4LT Abstract Introduzione All inizio di novembre 2012 il Ministero degli Interni ha rilasciato una versione consolidata dell Animal Scientific Procedures Act Legge sugli animali Procedure scientifiche del 1986 cos come emendato dalle norme vigenti in seguito alla Direttiva europea 2010 63 UE a condizione che il documento non pronunci definitivamente il diritto e che si attenda il varo da parte del Parlamento durante il corso di quest anno Si prevede comunque che la messa in atto verr effettuata entro la scadenza dell 1 gennaio 2013 Questa relazione mira a semplificare alcuni dei materiali meno palesi che sono stati aggiunti e a ricordare ai lettori che bisogna attendere fino al tardo 2013 prima che vengano pubblicate le nuove Linee guida che chiariscono i contenuti dell Act e che possiamo cominciare ad adattarci al nuovo regime Lo scopo di questa breve panoramica fornire all Animal Care Staff un gruppo specificamente riconosciuto dalla Direttiva europea un compendio dei principali cambiamenti che verranno apportati e come le nostre organizzazioni gestiranno la nuova legislazione in fase di subentro nel nostro sistema 32

April 2013 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 IAT Education past present and future KEN APPLEBEE Chair Board of Educational Policy Institute of Animal Technology Presented at the John Bleby 80th Birthday Symposium October 2012 Formal education of Animal Technicians began in 1950 with the founding of the Animal Technicians Association ATA later to become the Institute of Animal Technicians and then Technology IAT The IAT developed three levels of examination which were linked to professional membership and were G Preliminar y subsequently also known Intermediate and Certificate G Associateship subsequently also known Membership MIAT G Fellowship FIAT as as By far the biggest change in the syllabuses was the increase in the amount of background subjects being taught and examined particularly the amount of science required and this trend has continued to the present day However these syllabuses were written in such a way that there was no clear indication as the level or depth of knowledge required see topic 2 example below what would be examined to and no clear learning outcomes which was in common with the vast majority of other vocational qualifications at the time As can be seen from the Fellowship Examination syllabus examples below plate 1 a broad technical and husbandry knowledge base was required covering a range of species from mice to farm animals It can be seen from the reproductions of the 1958 IAT Fellowship examination syllabus and the 2004 version plate 2 during the first 50 years there was little change in subject topics except in one area However standards continually increased both in quality of teaching examination the depth of knowledge required and perhaps most importantly the students 33

Tech 2 Tech Plate 1 1958 Fellowship examination syllabus 34

Tech 2 Tech SYLLABUS FOR THE FELLOW LEVEL EXAMINATIONS Circa 2004 TABLE OF CONTENTS Plate 2 IAT Fellowship examination syllabus 2004 Fellowship Syllabus 2004 Topic 2 CARE AND HUSBANDRY OF ANIMALS WITH SPECIAL NEEDS This unit allows the learner to apply appropriate techniques for the maintenance of less common laborator y animals and animals housed under specialised experimental conditions Understands the methods used for housing maintaining and breeding as appropriate one named example from each of the following groups Less Common Species G a rodent such as a Chinese hamster or a wild rodent G a large bird other than chicken including artificial insemination if appropriate e g duck turkey goose G a medium size bird e g pigeon quail G a small cage bird e g budgerigar canary G a reptile G an amphibian G a fish G a cephalopod G an insect a named example of your choice G another invertebrate Animals with Complex Husbandry Requirements G gamma or X irradiated G mutant e g obese mice G immunodeficient animals e g SCID or athymic mice Up until the late 1980s the majority of courses were delivered in a formal setting at Colleges of Further Education with employers granting day release Tutors reserved the right to refuse to enter students into the IAT examinations even to the point of writing to an employer explaining that this was a likely outcome if students failed to meet college s expectations This would certainly be most unlikely to occur today especially as students increasingly turn to litigation Until the 1990 s the Fellowship examination took the form of two three hour theory papers one in the morning and the second the afternoon of the same day Until the introduction of the BSc and then MSc in Animal Technology in the early 1990s the Fellowship was considered the academic pinnacle and those who could attend college the course was generally extremely enjoyable and was delivered by experts Those that were successful were able to apply for the most senior positions normally managing the larger animal facilities However for many candidates sitting two written papers was a test of endurance and was not so much an examination of intelligence or even knowledge but one mainly of memory Successful candidates were then called for an Oral and Practical sections including Animal Handling and Sexing For those passing the Fellowship examination it was almost a rite of passage more a survival test than an examination However for the majority who were unsuccessful it became a source of resentment and frustration in some cases blighting their professional careers During the 1970 and 1980s public sector national pay scales e g the Whitley Council scales were based primarily on qualifications Those holding IAT qualifications including the Associateship or Membership and Fellowship would expect a higher rate of pay than those without While gaining the Fellowship probably did not help me attain my present post it did enable me to gain my first management role at the Royal College of Surgeons as John Cooper told me one day I employed you because you were young and had the FIAT IAT Examinations 1950 2000 There is no doubt that the IAT examinations from the 1950s up until the early 2000s enjoyed approximately 50 years of success and that every animal technologist both past and present owes an immense gratitude to those who went before and left something for others to build on During the late 1990s and early 2000 rumblings of discontent surfaced from both candidates and course providers concerning the difficulty and lack of transparency of obtaining a pass at all three levels of examination At the IAT 2004 AGM Steve Barnett proposed that IAT Council examine the structure of their examinations and their pass rates which was then seconded by Jas Barley and carried 35

Tech 2 Tech The then IAT Chair of Council Bob Kemp instructed the Board of Education to investigate this request and repor t back to Council within the next six months It can be seen from the following tables that not only were pass rates highly variable and often inflated by resits but also that fewer people were attempting the exams especially the Fellowship 0 1 2 34 0 1 2 So what had changed in the 2000s Science Students 36 The explosion of Genetic Modification technology which in turn has led to single specie mouse facilities with high levels of biosecurity and the failure of the examination to reflect this and other changes in science and technology made it difficult to show that the syllabuses were relevant to the industry of the day Employers no longer wanted a Jack of all Trades but needed specialists Many did not want to go to college certainly not in the evening or their own time Due to the changes in secondar y education many students did not have the skills to answer questions in long essay format although for many nonacademically minded students this was not a new problem Increase of graduates in the job market who were not interested in learning about species they did not work with such as farm animals or cephalopods Instead students wanted knowledge of subjects relevant to the emerging technologies and relevant to what they actually did in the workplace Computers Social Networking Internet students wanted ever ything readily available Not used to failing well not everything Again the changes in education generally had meant that students were now used to having any effor t acknowledged by some level of qualification however low For IAT examinations it was either a section pass or fail and some took failing badly Employers Less willing to grant day release Education to be focused on what happens in their workplace attaining skills leading to competence Employing more and more graduates Losing confidence in the relevance of IAT qualifications Employers also aware of the significant changes in secondary education Education Few college courses running IAT qualifications Availability of on line learning Qualifications taught in Modules or Units which meant that students gained some acknowledgement of learning regardless of whether they completed the full award Modular qualifications were also more flexible and fitted modern life styles meaning that students taking career breaks could come back to education without having to start again from the beginning Syllabuses based on learning outcomes Assessments more formative with more phase testing of topics during the learning process Educationalists were now setting the framework for vocational training and the traditional professional bodies were being criticised for not reflecting modern educational practices IAT qualifications NOT on the National Qualification Framework NQF which meant difficulty in comparing them with other qualifications Although not directly related during the 1990s and early 2000s when animal rights extremism was at its

Tech 2 Tech most intimidating It was apparent to many that the whole industry had an introverted and depressed atmosphere which may well have also contributed towards the critical view of the IAT s education system Whatever the reasons it was obvious to IAT Council that if we were not to lose our examination system the Institute had to change its whole educational structure and fast At the Animal Technician Education Training Stakeholders Meeting Royal Society on Friday 27th May 2005 this was indicative of a number of similar opinions expressed Felt that is was good that root and branch approach is being suggested and that tinkering around the edges was not now sufficient Report of the Education Review Group1 Council tasked a small group from Council to perform a review as to how Animal Technologist education needed to change and develop to equip our members in the 21st Century Institute representation to both the ABPI and MRC confirmed that a thorough review of animal technology education and training was required not only to respond to new technical skills required in the workplace but also to take account of modern educational practices and the educational abilities of school leavers In July 2005 IAT Council accepted the Educational Review Group Report despite strong objections from some members of the Board of Education who felt that adapting the current system would be sufficient To implement the report findings Bob Kemp took over as Chair of the Board of Education for one year and Brian Lowe would be approached to act as the IAT Educational Consultant Council wanted the new FE qualifications to be up and running within 18 months which was a formidable task but was felt to be achievable IAT Council set up a Syllabus Review Group SRG who were tasked to produce a syllabus based on a large part of the old syllabus but using learning outcomes and to be placed on the QCA framework In October 2006 the IAT met with representatives of the Qualifications Curriculum Authority QCA to discuss the requirements so the IAT s new level 2 3 qualifications could be allowed onto the National Qualification Framework NQF allowing the IAT to become a recognised awarding body The meeting was very positive and QCA representatives were very confident that provided the IAT put in place the appropriate systems that to them seemed to be a straight forward application The appropriate systems included the production of a full 200 page Prospectus which included a business case for the need for the qualifications IAT organisational structures syllabuses and educational policies and procedures and was submitted to the QCA on 8th December 2006 Following our submission the QCA consulted various exper ts in the field and following a few minor amendments the IAT received an email dated 25th June 2007 that stated This is confirmation that the IAT application to become an awarding body was put to the Regulators Accreditation Group yesterday and was approved This means that you are now a recognised awarding body The IAT as a QCA now Ofqual Awarding Body2 and the level 2 and 3 qualifications on the NQF3 now agreed to the following G The IAT as an awarding body organisation would no longer act as a provider of courses for which it awarded qualifications G Board of Moderators was formed to help to ensure standards are met and quality assess the providers 37

Tech 2 Tech Levels 2 and 3 FE Syllabus Module Part Award content The syllabus is divided into units this enables students to gain credits as they progress through the course Table 1 In addition relevant previous qualifications were also eligible for credits Clear guidance on assessment criteria was also given and coupled with learning outcomes students would know why they were failing to achieve required standards If a student was unable to complete the full award it was possible for them to gain recognition for the modules or new units they did achieve4 Table 2 Example of Learning Outcomes and Grading criteria Full award diploma Table 1 List of IAT units and qualifications Plate 3 Example of the full certificate 38

Tech 2 Tech The IAT has been consistent over many years and its longevity of its FE vocational qualifications is in direct comparison to many other shor t lived vocational qualifications IAT Qualifications 1950s 2006 IAT qualifications have consistently been formed of three levels and employers and technicians knew what they represented G IAT Intermediate Preliminary Husbandry G IAT Associateship Membership G IAT Fellowship IAT FE Qualifications 2006 to date 2006 to 2010 NQF G IAT First Certificate in Animal Husbandry Level 2 G IAT First Diploma in Animal Technology Level 2 G IAT National Certificate in Animal Technology Level 3 2010 onwards QCF G IAT Diploma in Laboratory Animal Husbandry Level 2 G IAT Diploma in Laboratory Animal Science and Technology Level 2 G IAT Diploma in Laboratory Animal Science and Technology Level 3 IAT Higher Education HE The need for graduate training has also been recognised by the In vivo sciences in the UK report5 Degreelevel animal technologists are becoming more common in CROs Contract Research Organisations and large pharmaceutical companies who have taken the strategic decision to pay higher salaries for technologists capable of fast tracking into managerial positions The IAT believes evidence shows that the academic sector has a need for senior technologists with a similar level of education and experience Since 2008 the IAT have been in discussions with the Open University and University of Manchester but due to a variety of reasons including student fees these negotiations failed IAT HE Certificate in Animal Technology Level 4 IAT Higher Certificate in Animal Technology developed by the Institute of Animal Technology IAT and accredited with Middlesex University took its first cohort of students in October 2008 and has been running successfully since The reason for its long term viability is that the course delivery is based around block release and on line learning and support This coupled with generous Industr y support and the enthusiasm of the students has enabled the IAT to keep costs down and lessen the need for students to be away from their place of work The IAT has since 1990 provided HE courses on animal technology and below is the list G BSc in Animal Technology 1990 2003 G Post Grad Dip 1994 1999 G MSc in Animal Technology These HE courses were popular and of a high quality however although reasonably priced even for the time they were London based day release so not only do employers lose 40 days working time costs were increased by travel and subsistence Many employers were unwilling to fund this aspect and supplementary funding was not easily available although some students did obtain bursaries from the IAT This resulted in the inability to sustain sufficient student numbers for long term viability B Sc in Laboratory Animal Science Technology The IAT are very aware that for animal technologists to attain their ultimate potential and to compete for the most senior posts they need to be educated to HE Level 6 IAT HE Diploma in Animal Technology Level 5 IAT Higher Diploma in Animal Technology developed by the Institute of Animal Technology IAT and accredited with Middlesex University took the first cohort of students in October 2012 The cost is 650 per module approx one third of the fees of most BSc degrees Despite this competitive price some employers still seem reluctant to fully fund their staff s fees To help enable animal technologists to gain access to HE Courses by the provision of bursaries the IAT helped to establish the charity Animals in Science Education6 The modular structure of the Certificate and Diploma allows people not wishing to study for the full awards to study individual modules as part of their Continuing Professional Development CPD requirements 39

Tech 2 Tech Overall structure of the HE Diploma Programme Module Level Applied communication skills for science Supervisory management Control and identification of disease Biological science 4 4 4 4 Animal law and ethics Experimental design Reproduction and Genetic Alteration Toxicology Qualification Higher Education Certificate in Animal Technology 5 5 5 5 Higher Education Diploma in Animal Technology Students may also take individual modules as CPD Table 3 Structure of current HE programme IAT BSc Laboratory Animal Science Technology Level 6 Negotiations with Middlesex University are ongoing and progressing well but will take another year for the IAT to gain HE institutional approval As a validated degree the Institute will continue to retain control of its qualifications and so will be consistent with previous standards The fees will be approximately 750 per module which equates to 9k for the full six year degree programme The first cohort of students should be enrolled from academic year 2014 and the four Level 6 modules will be G G G G Zero to Hero Over the past 60 years IAT Education and Examinations have enabled young technologists often school leavers who failed to gain any qualifications through school to develop their careers by gaining qualifications that are recognised and respected internationally The current educational system now in place will enable an easier movement of technologists through the various levels of the biomedical industry to thus fulfil their true potential while continuing to ensure the highest standards of animal care and welfare for animals used in science References 1 Animal Facility Design Project Management The physiology of pain and stress Project Future Educational Activities The Institute is currently working with the Medical Research Council MRC to develop Animal Technology apprenticeships As part of this we are also developing in conjunction with the Cambridge University Training College courses where Animal Technology subjects will be taught in a full time school environment 2 3 4 5 Ultimately the Institute sincerely hopes that graduate Animal Technologists will also have the opportunity to study for a Masters in Animal Technology and ultimately a PhD The Institute would now be extremely grateful for a period in which we can allow our education provisions to establish themselves so we would like to send a heartfelt plea to the various regulatory bodies please no more changes 40 6 Reformulate the Institute Examination and Qualification System Report of the IAT Educational Review Group 2005 Applebee Author Barley Gregory Kemp Ofqual Awarding Organisations http www ofqual gov uk how we regulate becoming arecognised awarding organisation Ofqual Register of Regulated Qualifications http register ofqual gov uk Qualification recognition Number RN5252 Applebee K A Cubitt S and Lowe B 2008 New modular flexible and transferable qualifications for animal technologists Proceedings of the Tenth FELASA Symposium 11 14 June 2007 16 19 Association of the British Pharmaceutical Industry and Biosciences Federation In vivo sciences in the UK sustaining the supply of skills in the 21st century October 2007 http www abpi org uk Details asp ProductID 325 Animals in Science Education Trust AS ET http www animalsinscienceeducationtrust org uk bursary htm

April 2013 Animal Technology and Welfare Animal management software making it work for you ALISON HOPKINS MIAT RAnTech a tune Software ag Julius Reiber Strasse 15 64293 Darmstadt Germany Correspondence ahopkins a tune com Summary Every year we are expected to produce more with less money and to achieve this in an environment of changing regulation 2013 is a particularly significant year for the animal management industry as we face the introduction of the new EU regulations and increasingly tight financial constraints Animal management software is available to help achieve the objectives of the new regulations and at the same time improve welfare and streamline our operations in line with the 3Rs Systems which meet the current regulator y requirements and also simplify the operation of facilities are available but how do we know what we should be looking for This article aims to G Look at the consequences of the new regulations and the impact on data management G Offer some guidelines to consider when selecting animal management software G Introduce tick lab as an example of appropriate data management software facilities are seeing the use of more transgenic animals greater numbers of breeding lines and enhanced communication with the scientists Many animal facilities recover their costs through recharging for animals and services and it is crucial that data used to make decisions is accurate and up to date Data management communication and collaboration are becoming more critical than in the past The environment that we have evolved is often inefficient and complex to manage The requirements from the new regulations and the drivers for continuous cost reduction will increase the pressure to have a system which is fit for purpose Data management is the development execution and supervision of plans policies programs and practices that control protect deliver and enhance the value of data and information assets 1 Compliance with the regulations will require a consistent management process and all facilities except perhaps the smallest and simplest operations will find it possible but difficult to comply efficiently using their current assortment of paper systems home grown spreadsheets and databases Introduction The new EU regulations are not in themselves so different from the current Animal Scientific Procedures Act 1986 but they are enforcing some changes to the way in which we record our data and indeed to which data we need to record Two key areas of focus are those of training and competency and the ability to capture retrospective severity for Home Office reporting Both of these areas for many will require a significant review of current practices and considerations for the best way to capture and record such information In today s economic climate cost is a key driver Research institutes are increasingly becoming a lot more aware of the cost of time and the efficiencies needed within processes to reduce outgoings Animal Traditional data management tools Conventionally animal facilities have used pen and paper to capture data Over time it has become apparent that the labour intensity of such a recording method is no longer practical or ideal within a research establishment particularly when it comes to retrieving data for reporting purposes A development from pen and paper was to record information in Excel spreadsheets These provide a reporting capability and as a readily available desktop tool have spread rapidly within organisations However Excel sheets are still single user and are missing the appropriate security and audit capabilities In the endeavour for efficiency and greater control many solutions became available within animal 41

Tech 2 Tech facilities A variety of in house programs and software solutions were developed for animal ordering material management animal stock management task allocation breeding and colony management health management and environmental monitoring These systems are rarely well integrated and their inability to reuse data and communicate together can create a tangle of inconsistency with a high cost of ownership Animal management software promises to help but it is important to choose the right one Animal Management Software AMS should be designed to provide an efficient and compliant infrastructure to manage the animal facility By providing a consistent view of the data and integration of that data to deliver the required Home Office reporting AMS enables managers to achieve compliance and at the same time to reduce the cost of operating the facility Guidelines to consider when selecting an animal management solution The system should G Fulfil the legal and operational requirements and be supported and future proofed to meet any potential requirements Regulatory changes are determined by law but the structures of organisations vary which means that the animal management system must be configurable to the unique requirements of each establishment G Demonstrate all the necessary functionality to meet animal management requirements G Allow the sharing and reuse of information so that information is entered only once as opposed to multiple times This creates consistency in data recording and saves time G Utilise drop down menus so tasks can be completed with minimum clicks G Have effective search capabilities to instantly narrow down data being viewed and quickly retrieve desired animal records G Be able to link to other systems already in place G Effectively suppor t your organisation s effor ts towards the 3Rs Reduce A comprehensive stock management system allows researchers to at a glance see the numbers of animals already in use against their project including those being held in stock before placing any additional orders reducing numbers of animals used through enhanced awareness Replace The ability to record retrospective 42 G G G G severity within an animal management system highlights changes allowing experiments to be reevaluated and re considered going forwards Refine A system should have the functionality to contain stipulations as to anaesthesia or analgesia surgery and non surgery procedures and treatments for certain species on certain projects It should also use a standardisation of terminology and procedures to increase transparency and the avoidance of misunderstanding par ticularly when communicating the nomenclature of transgenic strains Any good system should demonstrate an auditing workflow in which the veterinarians or the internal ethics committee can be involved This reliably ensures the least invasive methods are chosen and encourages comparison to the current science and research standards Be easy to use easy to learn and easy to implement with a high acceptance Provide functionality and data where you need it e g in the animal room lab and office Allow customised reporting so that all information entered into the system can be retrieved according to your chosen criteria Support mobile use tick lab One example for a fully integrated solution is tick lab from a tune Some functional highlights of tick lab include G G G G G G G G G G G Colony management Task management Health management HO data recording Reporting Training and competency Configurable cage cards Compatibility with RFID and bar code technology Ability to upload attachments Configurable system components Mobile use Conclusion With an investment in an effective animal management solution the research industry can reduce its overall costs and adequately meet the requirements that the new EU regulations impose Animal management software is available to help ease the burden of change and at the same time improve welfare and streamline our operations to save time and money The technology for Animal Management Systems has

Tech 2 Tech developed significantly in the last 10 years The software is mature and available as off the shelf products which can be configured for each customer The current regulatory and financial constraints are forcing many organisations to look closely at how they manage their data A well planned and executed AMS project will help to simplify record keeping in the unit and provide a platform for future growth Businesses are demanding better and more efficient ways of handling the volumes of data that are an integral part of our daily routine Many options are available for today s forward thinking professionals The only option not being explored is doing things the same old way We must ensure that when we consider options for technological development we consider those that will offer us the greatest future growth and continuous development with full support to meet regulatory requirements With the enforcement of the new regulations there is a need to change We are here to help References 1 Wikipedia http en wikipedia org wiki Data_ management http www alnmag com article integrated datamanagement lab animal research page 0 0 www a tune com http www a tune com fileadmin files documents tick_lab US a tune_EU_Directive_2010_63_EN_01 pdf a tune information leaflets or fur ther information is available upon request from ahopkins a tune com 43

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April 2013 Animal Technology and Welfare Report on the PTLLS Preparing to teach in the lifelong learning sector course organised by the IAT ALAN PALMER MRC National Institute for Medical Research The Ridgeway Mill Hill London NW7 1AA Correspondence apalmer nimr mrc ac uk Introduction The Institute of Animal Technology IAT recently organised the PTLLS training course for those in the industry requiring initial teacher training to carry out assessment and deliver courses training in the workplace The IAT qualifications include some level 2 units that can be assessed by academic or competency routes the competency based units can be assessed within the workplace by a RAnTech The new Animal Technology Apprenticeship Programme also includes competency based assessments in order to facilitate this in 2012 13 the IAT are kindly providing Preparing to Teach in the Lifelong Learning Sector PTLLS City and Guilds 6302 free of charge to registered Animal Technologists The course I attended was run over a four day period The following is a summary of each of the four days activities Day 1 The course began at 9am straight after a much needed coffee and the tutor Chris from course providers Andragogic Education introduced himself and detailed the basic aims and course objectives the ground rules and explained the purpose of the resource packs we were all given Although most of us had a chance of chatting and introducing ourselves over coffee before the course started the first activity of the day was to do the group introductions Each of the six delegates were asked to find out some basic facts about the person sitting next to them over the next few minutes and then present this to the rest of the group this was a good ice breaker exercise that was informal and fun and really did put everyone at ease and although from initially looking through the course pack we knew there was going to be a lot of group activity it certainly helped set the scene in promoting a friendly and informal class environment The rest of the day was spent exploring different ways of teaching using pedagogic methods where the teacher controls directs manages and suggests and andragogic methods the teacher facilitates a collaborative learning environment and the benefits of using andragogy to empower learners to learn for themselves Using a range of activities and group discussions we also explored the four main learning styles activists reflectors theorists pragmatists and Bloom s Taxonomy of Learning cognitive head attitude heart skills hands This prepared us in how to plan lessons that accommodate all learning styles and domains of learning this all sounds very confusing but is extremely enlightening and using the various classroom activities to explore the principles it all made perfect sense We also learned a lot about our own learning styles how we process information and more importantly retain it With the group being representative of all learning styles this helped us put into perspective the importance of planning a good learning experience for a variety of learners needs Day 2 The morning started with a review of what was covered in day 1 with a group discussion and then completing a short quiz to reinforce our learning We then started on the next topic which was motivation theory we watched a short film Herzberg s motivation theory 45

Tech 2 Tech and discussed the principles behind Frederick Herzberg s motivation theory how this can be applied into the workplace in the context of developing and empowering staff with the incentive to achieve After lunch was when the real work started The entire group working in pairs had to plan prepare and deliver a 15 minute peer teaching session to the rest of the class The topic for the lesson was Advantages Disadvantages of using various teaching aids each pair was given one i e audio visual flipchart handouts etc teaching aid to form the main topic of their lesson although all forms of teaching aids could be used in delivering the lesson Maxine my lesson partner and myself had to plan and deliver a 15 minute teaching session on the advantages and disadvantages of using a flipchart as a teaching aid We soon realised that the actual lesson content topic was not important it was how we incorporated all that we understood so far about the different learning styles into the lesson At the end of the lessons all the groups reviewed each other s sessions and we had feedback from the tutor this really helped to reinforce our learning so far and would be vitally important in what would be in store for us in the remaining days Day 3 As before we kicked off with a review of the previous day s activities and what we had learned so far plus some tips and advice in preparing for our final day s peer teaching session this would be a 30 minute lesson on any topic we liked as long as it incorporated everything we had learned so far We then discussed qualifications assessment practice and quality assurance mechanisms Again using group activities and exercises we reviewed what makes a national qualification Including maintenance of standards and quality assurance assessment of student learning and the advantages and disadvantages of using different assessment practices and the requirements of good assessment practice During these group activities we looked at and reviewed examples of various assessments practices and how various techniques can relate to the different learning domains Day 3 also incorporated another 15 minute peer teaching session on various topics as a practice for the final day s teaching session Jon my lesson partner this time and myself were to deliver an interactive session on the pros and cons of distance learning As before the topic was not the most important factor it was how we used different teaching strategies to benefit all the learning styles After lunch we had some time to start preparing for our peer teaching session the following day This would involve preparing a complete lesson plan developing lesson structure and content and also a resource pack The lesson had to encompass all that we had learned so far so we had to produce a structured lesson plan 46 demonstrating an andragogic and pedagogic approach that would be inclusive for all four learning styles addressed Blooms 3 learning domains and would be relevant interesting engaging challenging and have clear aims and objectives A tall order but the previous 3 days had prepared us well and the tutor was at hand to give advice and guide us through the process Day 4 This was the big day when all that we had learned would be put to the test Lesson plans and resource packs were to be handed in to the course tutor for evaluation and we all had a time slot when we were to deliver our lesson to the rest of the group while also being recorded on video This all sounds very nerve racking but as mentioned already the class activities had made us all bond in a nice informal learning environment So even though there were plenty of sweaty hands we were also quite fired up and ready to get on with it The final day passed in a blur with everyone giving interesting and well thought out lessons we all passed and learned a lot of new skills in the process As I said previously the lesson subject was not important it was how it was structured and delivered that was the key factor and we certainly had a variety of different subjects ranging from Making a sugar rose to Extracting venom from a Komodo Dragon

Tech 2 Tech After each teaching session the group all completed a peer assessment sheet of the lesson which was handed to the tutor Once these had been completed we had a debriefing chat with the tutor who gave us his feedback and reviewed the peer assessment forms with us this gave us a chance to talk through all aspects of the lesson and in particular what the strong points were and any areas that could be developed further We were given all feedback forms and a copy of the video recording of our session to take away On conclusion of the classroom sessions there are still 4 assignments that we need to complete to demonstrate to the assessor and City Guilds an understanding of the underpinning educational theory These are fully explained by the tutor and in our resource pack we are also given support from the course providers until all are completed and we can claim the full qualification In summary the skills and knowledge gained during the course was very enlightening and has certainly made me re think various in house training activities The course was very enjoyable and having an informal and activity andragogic based approach really made the whole learning process ver y enjoyable I would recommend this to anyone who trains teaches or assesses as part of their job it really will make you think about what you are doing in a positive way Both you and your candidates will benefit 47

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April 2013 Animal Technology and Welfare Report of the 2012 RSPCA UFAW Rodent Welfare Group meeting PENNY HAWKINS SECRETARY 1 JESS GIMPEL2 ANDREW SC RICE3 DOMINIC J WELLS4 JUDI LATCHAM5 KATE HEATH5 MARK GARDINER6 TREVOR WATTAM5 ELLIOT LILLEY1 MAGGY JENNINGS1 and ROBERT HUBRECHT8 1 2 3 4 5 6 7 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Centro de Investigaciones M dicas Pontificia Universidad Cat lica de Chile Santiago Chile Imperial College London South Kensington Campus London SW7 2AZ Comparative Biomedical Sciences The Royal Veterinary College Royal College Street London NW1 0TU GlaxoSmithKline Gunnels Wood Road Stevenage Hertfordshire SG1 2NY Mary Lyon Centre MRC Harwell Harwell Science and Innovation Campus Oxfordshire OX11 0RD UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Corresponding author penny hawkins rspca org uk Introduction Experimental animal use in Chile The RSPCA UFAW Rodent Welfare Group holds a oneday meeting every autumn so that its members can discuss current welfare research exchange views on rodent welfare issues and share experiences of the implementation of the 3Rs of replacement reduction and refinement with respect to rodent use A key aim of the Group is to encourage people to think about the whole lifetime experience of laborator y rodents ensuring that every potential negative impact on their wellbeing is reviewed and minimised Jess Gimpel Pontificia Universidad Cat lica de Chile The 2012 meeting addressed a range of topics including rodent use in Chile caring for aged mice reducing stress during blood sampling the welfare impact of different identification methods and implementing the Three Rs in antibody generation The meeting also discussed refinement and translatability issues with respect to selecting the appropriate age of rodent for in vivo studies and to rodent studies of neuropathic pain The day ended with a focus on refining severe substantial procedures with talks on an RSPCA initiative to reduce severe suffering and a practical example of refinements for SOD 1 mice a genetically altered strain used to study neurodegenerative disease Chile has a population of 17 million people and over 60 universities Nearly a third of these have medical faculties half have schools of veterinary medicine and there are a similar proportion of biology pharmacology and biochemistry schools Most of these entities do at least part of their research using animals and some of them base most of their scientific output on animal experiments There are also bio companies and nonprofit research organisations that use animal models About 15 registered rodent vivaria exist but it is still common practice for some researchers to have a small private animal colony Currently there is only one vivarium in Chile accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International AAALAC It achieved accreditation this year and there is no other animal facility applying for certification at the time of writing even though there are at least three major projects in the country that are setting up state of the art animal facilities Until recently Chile had one regulation relating to animal cruelty in its Penal Code that also had to be applied to scientific procedures using animals However this was a generic law and so was not a very satisfactory way of regulating research using laboratory 49

Tech 2 Tech animals An Animal Protection Law was sent to Parliament but then sat there for over 10 years While the law was discussed scientists and animal care professionals sought a way to implement voluntary standards specific to experimental animals in order to make progress in this area In 2004 a Standard based on and modified from ISO 10993 2 2006 Biological evaluation of medical devices Part 2 Animal Welfare Requirements was proposed the Chilean Norm NCh 2856 2 It was regarded as an important step at the time however there are still no vivaria currently certified according to that norm in the country In 2009 the Chilean Animal Protection Law 20 380 was finally promulgated This regulates all animal use including research It defines what animal experiments are where these may be performed and who can conduct them An important step is that the law forbids the use of animals for school demonstrations The law also required that a National Bioethics Committee which would draft all the regulations be established 60 days after its publication Three years later this committee is still not constituted hence the law albeit now official is still not ready to be properly implemented with regard to animal procedures Fortunately things have moved forward despite the lack of a proper legal backup This is mainly due to scientists training overseas international collaborations and journal editorial requirements of bioethics approval before research can be published A major incentive has also been provided by Conicyt the main national research funding agency www conicyt cl which introduced a requirement for an ethical review process to apply to all projects that include animal use This year Conicyt has also started to apply procedures for retrospective reviews and project audits Therefore institutions have started to set up their committees and review procedures In addition more vivaria have started to hire veterinarians and animal technologists and professionals working in animal facilities have formed a national association ASOCHICAL which meets monthly and organises courses with national and international speakers www asochical cl These have been very successful and demonstrate both the needs and interests of those involved in experimental animal care and use to learn and to do things right The future looks promising although we still have a long way to go There is still a great need for training both at veterinary and technical level as well as more awareness of scientists that good animal care is essential not only for the animals but also for their research to be sound Action points G Continue to make efforts to increase awareness among researchers of the importance of raising standards of animal care G Collaborate in suppor ting good practice when implementing regulations controlling experimental animal care and use 50 G Increase training opportunities for all those involved in animal research and testing especially animal technologists with particular emphasis on providing resources in Spanish Neuropathic pain Refinement and enhancing the clinical relevance of rodent in vivo models Andrew Rice Imperial College London Neuropathic pain is a type of chronic pain that is caused by a lesion or disease affecting the sensory nervous system1 Following the initial trauma or disease which damages the nervous system neuropathic pain may occur in the absence of any noxious stimulus Neuropathic pain has no apparent biological function but is usually both severe and chronic and with a population prevalence of about 7 and few effective therapies it presents an area of therapeutic need Although a number of novel treatment approaches have been developed in recent years in general their analgesic efficacy can be limited and their effectiveness varies between patients As a result there has been considerable effort to develop new drugs to alleviate neuropathic pain These efforts include animal use in studies of peripheral nerve injury which have helped to identify some putative paingenerating mechanisms as potential drug targets However in terms of predictive validity 2 these animal studies have limitations with respect to forward translation predicting the efficacy of new agents in clinical trials because they are not good at screening out compounds that go on to fail due to lack of efficacy In fact nearly all the drugs currently in use to alleviate neuropathic pain were initially developed for other conditions or discovered by chance There are also a number of ways in which the standards of experimental design and reporting of animal studies of neuropathic pain can be improved see references 1 and 2 One critical generic issue which pain shares with other therapeutic areas is the need to refine experimental design to minimise the impact of experimental bias in overestimation of treatment efficacy e g concealed allocation sample size calculation obser ver and analyser blinding reporting of withdrawals and drop outs and clear stating of pre determined inclusion and exclusion criteria etc 1 4 Furthermore animal studies should be reported in a format which includes all relevant information and allows the reader to easily ascertain 1 2 This produces perceptions of phenomena such as touch temperature body position proprioception and pain The ability of a model to correlate with or predict a future performance or variable

Tech 2 Tech the methodological rigour with which an experiment was conducted1 2 5 It is obviously critically important to use the most appropriate study designs and protocols for in vivo research otherwise animals will be wasted which is a serious ethical issue There are a number of new approaches to neuropathic pain studies that aim to both reduce animal suffering and improve translatability For example neuropathic pain in humans is not only caused by physical trauma but can be associated with a variety of different pathologies Traditionally animal models of neuropathic pain are of peripheral nerve trauma whereas human clinical trials tend to be conducted in painful polyneuropathy e g diabetic neuropathy or post herpetic neuralgia chronic pain complicating shingles In order to correct this dichotomy between the animal and human clinical trials an increasing number of animal models of neuropathic pain are now emerging which better reflect the range of clinical conditions that can provoke neuropathic pain Another relevant consideration is the type of clinical signs that are used in animal studies to assess neuropathic pain and the efficacy of any potential therapies Of course an emotional experience like pain can be self reported in humans but can never be directly measured in rodents we can only infer the presence or absence of pain from other usually behavioural observations Traditional outcome measures such as limb withdrawal to sensory stimuli e g a mechanical stimulus heat or cold are not especially translatable to the human condition in which there are other comorbidities such as anxiety emotional disturbance and interference with the normal circadian rhythm However natural behaviours that are ethologically important in the world of the rat can provide more useful outcome measures For example rodents in pain may display behaviours which decrease their exposure to predation risk for example in open field tests they may perform more risk assessment behaviours such as rearing and spend less time in the centre and more time in contact with the walls thigmotaxis These behaviours have also been used to pre clinically evaluate potential anxiolytic drugs Other behaviours such as spontaneous burrowing a sort of house keeping behaviour in which rodents dig into gravel or food pellet filled tubes in the home cage Figure 1 is also reduced in association with a number of pathologies including neuropathic pain6 Using outcome measures that are more relevant to the animal can not only help to improve translatability but can also provide more sensitive indicators of suffering for example behaviours such as burrowing may be altered before there are any clinical signs of suffering that are apparent to the human observer This approach could thus also be used to guide refinements such as implementing earlier humane endpoints Figure 1 Mouse in a burrowing tube Photo credit Image from Deacon R Assessing Burrowing Nest Construction and Hoarding in Mice J Vis Exp 59 e2607 DOI 10 3791 2607 2012 Action points G For researchers review the kind of outcome measures used to assess pain within pain studies especially neuropathic pain Would there be scope to change to more of a natural behaviour approach G Consider whether the approach to creating the animal model of neuropathic pain could be refined G Use a rigorous Good Laboratory Practice approach to experimental design and conduct in an effort to minimise experimental bias Report experiments which used animals according to the ARRIVE guidelines5 G For animal technologists consider whether the more animal centred outcome measures could be adapted to help assess suffering in other species and study types You might like to bring the topic to your local Ethical Review Process now the Animal Welfare and Ethical Review Body for discussion Survey of the welfare impact of different identification methods Dominic J Wells and Nur Mazlan Royal Veterinary College A number of different methods exist for identifying mice including noting natural markings ear punching and notching ear tags microchips tattooing clipping or dyeing fur and toe clipping These have varying degrees of invasiveness and toe clipping is the most controversial and is widely regarded as being of special 51

Tech 2 Tech concern due to the potential for both pain and loss of function It is accepted that the method with the least possible impact on the animal should be used in each case but there has been only limited investigation of the welfare consequences associated with different methods Given the huge number of mice used in experimental procedures annually and the need for the majority of them to be unambiguously identified the choice of identification method for mice can have a significant impact on animal welfare In the first half of 2012 we performed an on line survey of current identification practices in animal facilities in the UK receiving 60 responses out of a possible 83 Facilities that replied included academic institutions government research institutes pharmaceutical companies and contract research organisations Respondents were mostly facility managers or senior animal technologists Named Animal Care and Welfare Officers Ear punching or notching was the most common permanent identification system and marker pens were the most common temporary system A number of institutions had ceased to use certain methods including microchips ear tags tattoos and toe clip Table 1 Mice were most commonly marked Discontinued methods Reasons given Microchip Cost welfare concerns loss of chips Ear tag Welfare concerns not reliable hard to read after some time not easy to identify at a glance Tattooing Unnecessary too fiddly caused local inflammation other less invasive methods are available Toe clip Unnecessary welfare concerns not easy to identify visually without handling Marker pen Only for short term studies cannot be used on black mice Ear punch Difficult to do and read changed to microchip database linked not easy to identify at a glance Hair dyes Not permanent impractical other reliable methods are available Fur shave Impractical Bar coding Not reliable Table 1 Discontinued mouse identification methods with reasons Just over half the responding establishments reported making a decision to stop using one or more identification methods These are listed in the table with the most commonly discontinued method first microchip 25 of establishments that responded and the least appearing last bar coding under 5 52 between 2 and 4 weeks of age 61 with only 7 marked at under 2 weeks old The main criteria for selecting a method were reliability minimal animal welfare concern and ease of reading The high use of ear punch notch may reflect the current use of this method as the preferred technique for obtaining DNA samples for genotyping genetically altered mice although tail biopsy is still commonly used Toe clipping i e amputation of the distal phalanx was not favoured due to concerns about animal welfare and was used by under 5 of establishments for example in cases where it was felt that the practice could be justified due to a risk of a lethal phenotype that resulted in the death of the animal in the first days of life We are currently further investigating the welfare impact of the different methods of identification It is necessar y to consider each technique from the animal s perspective as even apparently innocuous methods can be stressful For example temporary identification can be achieved by the use of hair dyes or indelible marker pens This appears to be harmless although the solvent does appear to stress rats7 Impor tantly these and other procedures involve capture and restraint of the animal which is itself stressful and may actually be the major stressor associated with many identification techniques8 However it may be possible to reduce restraint stress by refining capture and handling for example by catching in the home cage tunnel or in cupped hands9 or by avoiding scruffing when using marker pens Reviewing the whole process from the animal s point of view in this way is a useful approach to refining identification and one that we encourage Action points G All identification methods apar t from natural markings will cause a degree of stress so keep a watching brief of new ideas to refine them G In general use the least invasive possible technique but also take account of the duration of the project or breeding programme Repeated application in the long term might actually cause more suffering than a more invasive one off identification procedure G Staff experience and exper tise have major influences on the animal s experience so ensure that training is adequate for those applying identification techniques G If genotyping is required consider combining biopsy and identification procedures e g using ear notch or punch material for genotyping Making the right choice of age for rodent in vivo studies Judi Latcham GlaxoSmithKline The choice of animals used in efficacy studies is a

Tech 2 Tech critical factor in determining data quality Criteria such as choice of species strain and sex are generally embedded in the decision making process However with some exceptions e g drug efficacy studies for neurodegenerative diseases the age of rodent used in efficacy studies is generally restricted to the early phase of their lifespan e g when the animals are about six to eight weeks old or at a set weight The absence of a clear scientific rationale for using this age range raises concerns about both the optimisation of rodent models for efficacy and their relevance to the clinical setting It also has ethical and economic implications if animals past the desired age are wasted because they are no longer considered to be suitable GSK has set up a project that aims to scientifically evaluate the effect of rodent age on data quality in efficacy studies and also aims to raise awareness about the choice of age in rodent models used in research Although most strains of mice are sexually mature at 35 days old relatively rapid maturational growth continues for most biological processes and structures until animals are about three months old This means that mature adulthood in the mouse lies between three and six months of age Mice are middle aged between 10 and 14 months and considered old between 18 and 24 months3 These life stages should be taken into account as part of good experimental design For example the immune system of the mouse does not reach maturity until adulthood An immunological study of the T dependent IgG response in CD1 mice found that the response was significantly less variable in 16 week old animals than in 7 week olds I Holyer personal communication Results such as these are of extreme importance and have had a significant effect when deciding which compounds to progress There are also implications for the Three Rs and the culture of care for example G Greater awareness of the importance of study design G More critical scrutiny of statements such as we have always done it this way G Reductions in animal numbers for example due to reduced variability and better quality data G Enhanced clinical relevance thus reducing wastage The outcome of the GSK project will be included in a comprehensive set of guidelines for the design of high quality animal studies A group co ordinated by the UK National Centre for the Three Rs NC3Rs will continue 3 See http research jax org faculty harrison ger1vLifespan1 html last viewed 10 January 2013 to collect data throughout 2013 to bring the project to a conclusion Action points G Raise awareness among colleagues of new thinking about mouse life stages G Consider and factor in the stage of maturity of mice during study design ensure that the correct age is selected G Challenge study protocols that state they will use mature mice aged 35 days Refining blood sampling in the rat Kate Heath GlaxoSmithKline As par t of the process of drug research and development blood samples are sometimes needed from rats immediately post dose Historically this was done via the caudal vein after whole body warming for approximately ten minutes in a cabinet at 39 C The rats were then restrained during blood sampling to avoid injury and ensure an effective procedure but the slanted table restraint technique used at the time required two operators was stressful to the animals and so was unpopular with staff There was an obvious need to refine this procedure and so the decision was made to find an alternative approach We found that another establishment was using a refined method of restraint for less stressful blood sampling from the jugular vein which led to a series of training visits in which we gained competence in the new technique Correct animal handling is the key to success in this method and it took 24 hours training in total to learn to restrain the rat s forepaws and make a finger stool to support the head Once animal handling and restraint were assessed as competent we moved on to learning the sampling procedure Training began with a butterfly or winged needle with flexible line Vygon UK Swindon which ensured that the hand was kept steady A second person was required to do the sampling in the initial stages and we were then able to move on to performing the technique without an assistant by restraining the rat with one hand and sampling with the other We were finally assessed for competency in successful sampling on both sides so that these could be alternated in the case of multiple sampling There was some scepticism about the feasibility of the new approach among staff when we brought this back to the facility so we began by training two volunteer members of staff who were especially experienced in blood sampling with a benchmark for competence of about 20 successful samples Once we started to implement the new technique at our 53

Tech 2 Tech establishment we reviewed its benefits over the method we had used previously We found that the manual restraint and jugular sampling route had a number of benefits for example the rats were calmer during sampling and showed no signs of distress afterwards the operator found it less stressful because the rats tolerated the procedure better and it was also faster to perform about one minute as opposed to three to four minutes for table restraint Stress to the rats was further reduced because there was no need for whole body warming or restraint apparatus We also made some changes of our own to the protocol Shaving was discontinued as it risked clipper abrasions without providing any health benefits It was also found that nitrile gloves made it more difficult to handle large rats confidently so we changed to powder free Aloe Vera latex gloves with a textured grip low protein 50 g g beaded cuff 4 5 mm thickness Sterile water was also used to part the fur instead of alcohol Using the new method we can sample blood immediately after dosing which provides scientific benefits as well as most importantly reducing stress to the animals4 We believe that opening our minds and embracing new ideas can really improve the welfare of laboratory animals and we have gone on to introduce this technique for other studies in Toxicology Support Safety Assessment Action points G Revisit routine protocols for procedures such as blood sampling and consider whether these could be further refined G Explore the potential to use expertise at other establishments when researching refinements and learning new techniques G Always review and evaluate refinements to ensure that animals are benefitting and staff are comfor table with new techniques and feel competent to conduct them 4 Note All animal studies were ethically reviewed and carried out in accordance with the Animals Scientific Procedures Act 1986 and the GSK Policy on the Care Welfare and Treatment of Animals Caring for aged mice and assessing their welfare Mark Gardiner Mary Lyon Centre MRC Harwell People are living longer so diseases associated with the ageing process are consequently placing increasing social and financial burdens on society During the past three years a new project has been initiated at MRC Harwell which focuses on diseases related to ageing 54 with the aim of improving quality of life in old age rather than increasing longevity This project has two parts i to comprehensively phenotype a selection of common inbred mouse strains at specific ages and ii to use large scale chemical mutagenesis to generate lines of mutant mice These mice are being screened throughout their lives for diseases that are prevalent in ageing populations such as diabetes sensor y deterioration neurodegeneration and osteoporosis10 As our animals are living for much longer than the average laboratory mouse we need to be able to care for them appropriately This includes recognising the difference between a healthy aged animal and one with welfare concerns or imminent health issues We designed tailored welfare assessment protocols in consultation with senior animal technologists researchers and the veterinarian using the Mouse Welfare Terms database of clinical signs www mousewelfareterms org In addition to monitoring the appearance and progression of age related ailments the welfare assessment system has also been used to chart the changing appearance of healthy ageing animals We have collected profiles demonstrating how body condition coat appearance and weight all change during the life of a mouse and how this varies between sexes and strains This has allowed us to refine our welfare assessments and to ensure the best scientific outcome from these animals whilst providing the most effective care for them Some of the phenomena observed as mice age include G Effects of dominance behaviours can become more marked as the animals age Mice of social strains are housed in groups of five because of the welfare benefits associated with housing social animals together but a degree of dominance is inevitable Over time this may be evidenced by different body weights between female cage mates as they age This need not necessarily be a welfare problem but animals are carefully monitored so that judgements can be made on a case by case basis G Barbering in which fur or whiskers are pulled out by the barbered animal or one or more other animals usually in a repetitive pattern may also occur more as animals get older Groups are observed closely if signs of barbering occur and any sore patches are treated topically Sometimes it is appropriate to remove the barber and animals are humanely killed if their sores do not resolve G As opposed to barbering natural hair thinning is commonly seen in older mice and we do not now see this as a welfare concern provided that the skin is unaffected Figure 2 Like humans aged mice can lose hair colour for example black mice may go grey or ginger

Tech 2 Tech Animals previously housed in groups are not left alone if their numbers have been reduced due to barbering or other humane endpoints they are used in terminal procedures or for tissues More aggressive strains such as BALB c are no longer used because we find they generally have to be singly housed and this would be for long periods of time in ageing studies It is very important that care is taken to review the animals behaviour and needs throughout their lives especially during the ageing process and that enough time is allocated to the care of ageing mice and this is built into research programmes Figure 2 Thinning coat on a healthy aged mouse Photo credit Mary Lyon Centre MRC Harwell G Weight increases or decreases may also be observed as animals get older Larger mice are handled carefully as for a heavily pregnant female and their weight is taken into account if there is a requirement to anaesthetise them A general endpoint of 15 weight loss is in place for aged animals unless there is scientific justification for maintaining them in which case a maximum weight loss of 20 may be deemed acceptable G Eye abnormalities such as swelling exophthalmia and very cloudy eyes are sometimes observed as mice age These are generally very rapid in onset and affect one eye only and animals with these kinds of significant eye problem are humanely killed Cataracts in which the cornea becomes opaque and there is a blue tinge to the eye but no swelling are monitored closely and animals euthanased if there are signs of distress or the condition progresses to become more severe as described above G As with many other species aged mice are prone to external growths such as warts or abscesses Abscesses are generally due to Staphylococcus infection and if they do not appear to be painful or interfering with movement may be treated by lancing using a local anaesthetic cream to control discomfort or pain There are no treatments for warts and other growths so animals with these are closely monitored to see whether there are any signs of discomfort or interference with feeding drinking normal posture or movement in which case the animal is humanely killed Animals with abscesses are also euthanased if the above applies or if the abscess does not clear up following treatment G Malocclusion of the teeth can become more prevalent in aged mice or within the mutagenesis programme If appropriate this can be treated by trimming the teeth using a new design of clipper we have found that is more effective and less stressful for the mice and providing wet mash If the animal begins to lose weight and there is no improvement they are euthanased Action points G Consider how long animals live in your establishment are any of them aged and if so are their needs being catered for G If experimental protocols for social animals of any age involve individuals being permanently removed review whether provisions are in place to ensure that animals are not left alone G Look at the Mouse Welfare Terms database and consider using its approach to observing and describing mice Evolution of antibody generation at GSK Trevor Wattam GlaxoSmithKline GlaxoSmithKline generates monoclonal and polyclonal antibodies as tools for research reagents for clinical assays and as therapeutic reagents for the treatment of disease The Antibody Generation Group AGG within GlaxoSmithKline has always focussed on the generation of high quality antibodies with strong emphasis on creating and maintaining a good culture of care when using animals Our goals are to use the minimum number of animals with the minimum number of immunisation procedures per animal causing minimum distress We also aim to maximise use of animal tissues The general culture of care also includes making empirical observations on the outcome of projects and discussions of these between the AGG relevant project teams and the Laboratory Animal Science Team all of which communicate regularly This has led to changes in immunisation strategies over successive project licences and refinement of the antibody generation process The current process can be summarised in these three steps 1 An Antibody Generation Proposal in which a request for the generation of an antibody is presented to the AGG This must include the type of antibody its target specificity and end user applications The proposal is discussed to confirm that the antibody 55

Tech 2 Tech reagent is suitable for its application and that valid and valuable research data would be obtained The AGG also checks for pre existing reagents both internally and externally and no new antibodies are generated if pre existing sources are found 2 An Expression Strategy Meeting including the requestor the AGG and others with relevant exper tise e g in gene cloning or protein purification This meeting discusses the target antigen type immunisation strategy screening methodology reagents required for immunisation and screening and the time line to generate the antibody Immunisations do not start until all reagents and screens are in place 3 The immunisation protocol GSK has a default protocol with respect to immunisation sites doses and timescale This includes refinements such as low immunogen doses and careful selection of adjuvants resulting in reduced adverse effects better serum titres and fewer immunisations Freund s complete and incomplete adjuvants are no longer used Discontinuing the use of 1 inch 23 gauge needles and replacing these with 0 5 inch 26 gauge needles has improved precision when immunising as well as reducing discomfort to the animal This process has led to 50 reductions in animal group sizes from four to two animals for all immunisations The refinements that have been implemented in immunogen preparation and immunogen delivery have allowed the AGG to maintain its success rate whilst performing all immunisation procedures under a mild protocol severity Adverse events are ver y rare occurring in less than 1 of immunisations However we believe that refinement never stops and aim to implement more improvements to the immunisation protocol that we hope will replace some animal use and further reduce the number of immunisations necessary These include DNA immunisation methodologies new adjuvants or combinations of adjuvants targeted immunisation approaches and complementary in vitro antibody generation techniques Action points G Ensure that there is a process in place for continually applying the Three Rs to routine procedures such as antibody generation G Make sure that all relevant staff and groups communicate with one another and share information on the Three Rs with respect to antibody generation with other establishments the RSPCA The Research Animals Department has recently increased its efforts to develop and promote ways of avoiding or reducing severe suffering We aim to identify G the kinds of procedures that can cause severe suffering G the factors that combine to make the level of suffering severe such as pain anxiety or long lasting procedures G the purpose of severe procedures for example some vaccine tests or studies of painful or distressing disorders G any perceived or actual obstacles to reducing suffering or avoiding these procedures and most importantly G what can be done to overcome these We are currently visiting a wide range of establishments to discuss the project collect case studies from those who have successfully avoided or reduced severe suffering and encourage people to pass on good practice in publications through discussion with colleagues and at scientific meetings It is a particularly good time to tackle this issue as the need for better recognition and assessment of animal suffering par ticularly cumulative suffering is increasingly recognised and the new Animals Scientific Procedures Act will require assessment of the actual severity of procedures with reporting in the annual Home Office statistics The RSPCA s work on severe suffering comprises three broad strands The first is identifying those procedures that have the potential to cause severe suffering and setting up a series of expert working groups operating in a similar way to the Joint Working Groups on Refinement to develop and promote refinements to these This work also builds on previous projects looking at recognising pain suffering and distress in laboratory animals and welfare assessment protocols We are aware that many potentially severe procedures have already been refined so that they cause less suffering but much of this important work has not been effectively published or disseminated With this in mind the second strand of the project is to collect examples of severe procedures that have been refined so as to i help disseminate these and encourage wider uptake of the methodologies used and ii see whether there are any common approaches that might be applied to refine other severe procedures Refining severe procedures Elliot Lilley RSPCA Research Animals Department Ending severe substantial suffering is a top priority for 56 Thirdly we are encouraging local ethical review processes to consider a stretch objective Could our establishment achieve an end to severe suffering either by refinement of existing approaches

Tech 2 Tech or by avoiding the use of such procedures What are the opportunities and challenges associated with achieving this Husbandry G G This is an on going area of work for the RSPCA and we welcome invitations to visit establishments examples of further case studies or suggestions for topics or procedures that could be addressed Please contact elliot lilley rspca org uk G G Mice are genotyped before weaning and then placed directly into their experimental groups Male and female mice are housed away from each other to avoid stress due to olfactory exposure Gloves are changed and work surfaces cleaned between the handling of male and female mice Some nesting material is carried over from the soiled to the clean cage when cage changing males as this reduces aggression Refinements for SOD1 mice Dominic J Wells and Hannah Kaneb Royal Veterinary College The SOD1G93A mouse is a frequently used model of amyotrophic lateral sclerosis ALS a form of motor neuron disease ALS is a rapidly progressive fatal disease commonly diagnosed at roughly 50 years of age following which survival time is commonly just a further two to four years Some 90 of cases are sporadic and the rest are familial Approximately 20 of familial cases are caused by mutations in the Cu Zn superoxide dismutase SOD1 gene Current treatments have only a very limited impact on survival and so the SOD1G93A mouse is commonly used to test potential therapies The progression of motor neuron loss and development of clinical signs in these mice is very predictable Treatments can be applied at various stages but arguably the most relevant studies for translation to the clinic are those in which the treatment starts at the time of early clinical signs and delays the progression of the disease This unfortunately means that the mice will exhibit progressive clinical signs during these studies and can ultimately experience severe suffering We take this very seriously and have set up a Standard Operating Procedure SOP for survival studies with SOD1 mice with the aim of alleviating symptoms and refining housing husbandry and endpoints so as to minimise suffering All investigators and animal technologists have a copy of the SOP which is summarised in Table 2 Some of the refinements are good practice for any type of study but are especially impor tant when striving to ameliorate severe procedures We are aware that environmental enrichment can modify disease progression12 but this is addressed by carefully ensuring that equal enrichment is provided to all mice In addition to the kinds of refinement outlined in the table it is also ver y impor tant to ensure that experiments are properly designed so as to use the optimum number of animals minimise wastage and ensure that all variables are recognised and taken into account Regrettably however there is evidence of Housing G G Adaptations for disabled animals G G G G Health checks and veterinary care G G G Experimental protocol Interventions and humane endpoints G G G All cages contain ALPHA dri nesting material 2 nestlets and 2 fun tunnels Males are housed in groups of three as this has been found to be the optimal group size to minimise fighting Fun tunnels are removed at 100 days as mice may begin to show signs of hind limb weakness and paralysis and could become trapped or injured Non particulate litter and nesting material are provided as animals may have difficulty grooming themselves Cages are fitted with long sipper tubes and mash is provided daily on a Petri dish lid in the corner of the cage from 100 days The length of the sipper tubes is carefully controlled to prevent contact with litter and nesting material which can lead to leaks General health and the righting reflex are checked twice daily from 100 days or earlier if motor problems are apparent If mice have eye problems the eyes are cleaned twice daily with sterile saline and lubricating drops applied Tears Naturale Alcon If mice are dehydrated they are given an i p injection of sterile saline 100 ml kg 24 hours which works out as 150 200 microlitres every 2 hours and closely monitored Early screening of motor function and muscle characteristics should be used to select which drugs to take through to survival studies to minimise the number of mice suffering extreme motor deficits11 Mice are separated from their cage mates as soon as they show signs of hind limb paralysis or if healthier cage mates show them undue attention Animals are humanely killed if they are unable to right themselves within 20 seconds of being placed on their sides Any mouse taking longer than 15 seconds to right in the evening check is euthanased and one day added to their survival time Table 2 Standard Operating Procedure for caring for SOD1 mice 57

Tech 2 Tech suboptimal study design in most of the studies performed in the SOD1G93A mouse13 This includes poor recognition of confounding variables such as non ALS mortality clustering of littermates and imbalance of the sexes Scott et al 200813 proposed clear guidelines for improving ALS study design but the majority of post 2008 publications have failed to adopt this best practice A clear and immediate improvement would be to adhere to these guidelines thus avoiding animal wastage on misleading experiments5 Action points G If using or caring for SOD1 mice review housing husbandry and care and the experimental protocol using table 2 G Use earlier timepoints when screening possible treatments wherever possible to avoid progressing to survival studies unless there has been a positive finding G Adhere to the guidelines on study design in Scott et al 200813 and mention these in publications and presentations G If not working with SOD1 mice but involved with other severe procedures consider whether the approach to reducing and refining ALS studies could be applied in your own protocols References 21 22 23 24 25 26 Acknowledgements Many thanks to GlaxoSmithKline for generously providing the venue free of charge and for administrative and IT support with special mention to Jo Cruden for all her help Thanks also to all of the delegates who attended and par ticipated in the meeting 27 28 5 See also the discussion of experimental design and reporting in Rice earlier in this report 29 10 11 12 13 58 Rice A S C Cimino Brown D Eisenach J C Kontinen V K Lacroix Fralish M L Machin I Preclinical Pain Consortium Mogil J S and St hr T 2008 Animal models and the prediction of efficacy in clinical trials of analgesic drugs A critical appraisal and call for uniform reporting standards Pain 139 243 247 Rice A S C 2010 Predicting analgesic efficacy from animal models of peripheral neuropathy and nerve injury A critical view from the clinic In Pain An Updated Review Refresher Course Syllabus Mogil J S IASP Press Seattle 415 426 Sena E van der Worp H B Howells D and Macleod M 2007 How can we improve the pre clinical development of drugs for stroke Trends Neurosci 30 9 433 439 Macleod M M Fisher M O Collins V Sena E S Dirnagl U Bath P M W Buchan A van der Worp H B Traystman R J Minematsu K Donnan G A and Howells D W 2009 Good Laboratory Practice Preventing introduction of bias at the bench Stroke 40 3 e50 e52 Kilkenny C Browne W J Cuthill I C Emerson M and Altman D G 2010 ARRIVE guidelines Animal Research Reporting In Vivo Experiments PLoS Biol 8 6 e1000412 download at www nc3rs org uk arrive Andrews N Harper S Issop Y and Rice A S C 2011 Novel nonreflex tests detect analgesic action in rodents at clinically relevant concentrations Ann N Y Acad Sci 1245 11 13 doi 10 1111 j 17496632 2011 06342 x 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 Dev Psychobiol 50 3 266 77 Cinelli P Rettich A Seifert B B rki K and Arras M 2007 Comparative analysis and physiological impact of different tissue biopsy methodologies used for the genotyping of laboratory mice Laboratory Animals 41 174 184 Hurst J L and West R S 2010 Taming anxiety in laboratory mice Nature Methods 7 825 826 Gardiner M Chrobot N Lizor L Hutchison M and Wells S 2012 Care of the aged meeting the welfare needs of ageing mouse colonies Animal Technology and Welfare 11 3 206 208 Mead R J Bennett E J Kennerley A J Sharp P Sunyach C Kasher P Berwisk J Pettmann B Battaglia G Azzouz M Grierson A and Shaw P J 2011 Optimised and rapid pre clinical screening in the SOD1G93A transgenic mouse model of Amyotrophic Lateral Sclerosis ALS PLoS ONE 6 8 e23244 doi 10 1371 journal pone 0023244 Stam N C Nithianantharajah J Howard M L Atkin J D Cheema S S and Hannan A 2008 Sex specific behavioural effects of environmental enrichment in a transgenic mouse model of amyotrophic lateral sclerosis Eur J Neurosci 28 717 723 Scott S Kranz J E Cole J Lincecum J M Thompson K Kelly N Bostrom A Theodoss J AlNakhala B M Vieira F G Ramasubbu J and Heywood J A 2008 Design power and interpretation of studies in the standard murine model of ALS Amyotrophic Lateral Sclerosis 9 4 15

April 2013 Animal Technology and Welfare POSTER PRESENTATIONS Originally presented at LASA Winter Meeting 2012 The use of contrast agents to enhance soft tissue imaging using micro CT in laboratory rodents JORDI LOPEZ TREMOLEDA and WILLY GSELL Biomedical Imaging Centre BIC MRC Clinical Sciences Centre Imperial College London Corresponding author jordi lopez tremoleda csc mrc ac uk Introduction G Micro CT has proven ver y efficacious imaging technique for hig density structures such as bone providing excellent spatial resolution through rapid acquisition times allowing for high throughput preclinical screenings G However its application for imaging low density organs and body fluids is limited by their poor contrast G To improve their visualisation exogenous contrast agents have been successfully applied in clinical practice but their implementation in preclinical models has proven challenging due to the rapid clearance of the control agents along with the limitations of preclinical CT systems for carrying ultrafast acquistion Contrats Enhances CE Micro CT Preclinical Methodology Limitations for contrast enhanced preclinical CT High heart rates of rodents 10xfaster than humans with short circulation times around 30s in humans v 12s in mice Limited speed of acquistions in Micro CT systems compared to clinical systems scan times 2 4 minutes for micro CT vs 330mins for clinical sets Development of new generation of CT contrast agents G Blood pool selective contrast agents with longer retention in the vascular circulation G Tissue organs specific contrast agents G Gated acquistions to control for cardiac and respiratory motion minimising motion artefacts Applications of CE Micro CT Cardiovascular imaging G Visualisation of heart anatomy chambers and major vessels G Characterisation and monitoring cardiovascular models atherosclerosis stroke MI G Co registration of anatomical with functional data from PET SPECT 59

Poster Presentations Transaxial and coronal images of mouse heart before and after injection of Exitron nano 12000 Spatial resolution 35 m with cardia and respiratory gating 80KVp 500 Amp 330 projections of 5 frames from 40ms exposure total exposure time of 200 ms Soft tissue imaging G Visualisation of organ anatomy and function G Co registration anatomical with function data PET SPECT Optical Liver imaging Coronal images of a WT mouse injected with Exitron nano 600 i v injection Gastro intestinal imaging 3D MIP images of a surgically induced bypass rodent model after oral administration of Gastrografin Bayer plc UK CE CT scan Angiography CTA with Exitron nano 12000 3D maximum intensity projections views of a mouse body and heart Spatial resolution of 35 m a also bone segmentation and 103 m b pelvic vascular network c whole body with cardiac and respiartory gating 80KVp 500 Amp 330 projections with total exposure time 200 ms 60 CT images acquired with respiration gating 80KVp 500 Amp at different times after doing of Gastrografin showing the oesophagus a and gut pouches b Seyfried et al 2012

Poster Presentations Kidney imaging Coronal images of a WT mouse addomen before and after i v injection with eXIA 160KL Multimodality approach with CE Ct and PET for imaging a flow disturbing cuff in a mouse carotid A Contrast Enhanced CT Angiography with Ultravist370 B CT C PET for 18F FDG uptake D co registration CT Angiography PET E F CTA PET impantation of a flowdisturbing constrictive cuff induced as significant elevation of FDG uptake at the ergion of low oscillatory shear compared to the response detected for controlnon flow disturbing cuff Cuhhlmann et al Circ Res 2011 Co registration and optical imaging 3D Fluorescence images FLIT IVIS Spectrum Caliper LifeSciences to localise tumour vells co registered with CT images and organ maps 100 m CE CTA 3D MIP of the kidney vasculature 40 m Multimodality imaging Co registration CE CT and PET 61

Poster Presentations Discussion G CE Micro CT provides an enhanced contrast in soft G G G G G 62 tissues and blood allowing visualisation of organs anatomy and function Improvement on the functional versatility of the Micro CT technology for imaging cardiovascular preclinical models Provide reasonable quick 3D acquisitions 10 15 compare to other modalities i e MRI 30 1hr Minimally invasive feasible and repeatable BUT it is important to account for the radiation exposure Important tool for co registering anatomical imaging with functional data from PET SPECT modalities and can be used for attenuation correction for these radionuclide based techniques Future challenges development of ultrafast acquistions and detector senstivity minimise radiation dose and optimising post processing software

April 2013 Animal Technology and Welfare Tumour models core Biological Resource Unit PAUL MACKIN1 LISA YOUNG and STEVEN KUPCZAK Cancer Research UK Cambridge Research Institute Robinson Way Cambridge Cambridgeshire CB2 0RE 1 Correspondence paul mackin cancer org uk Mouse Hospital our past Dr David Tuveson joined Cambridge Research Institute CRI in August 2006 continuing his research into pancreatic cancer saying despite research efforts treatments for pancreatic cancer have not improved survival over the last 30 years KPC production was slow waiting times for cohorts were long multi arm studies taking months to complete Dave eventually employed his own animal technician to manage the Mouse Hospital KPC production increased waiting times for cohorts fell studies completed in weeks Due to its growing success the Mouse Hospital struggled to meet demand 2010 with two dedicated staff focusing on the Mouse Hospital production increased to 8 9 enrolable KPC mice week Multi arm studies being completed in a matter of weeks other CRI research groups and collaborators were requesting the KPC requests needed to be prioritised December 2011 Dave announced he was moving back to the USA Tumour Models Core The Tumour Models Core TMC was set up as a result of Dr David Tuveson s departure in September 2012 The Mouse Hospital was established to manage and maintain his KPC mouse model of pancreatic ductal adenocarcinoma PDA Supplying year round 3 5 enrollable tumour mice weekly for therapeutic studies and translating to the patient in the clinic Originally researchers managed their own colonies Senior Scientific Officer Production Manager Senior Scientific Officer Study Manager Senior Technician NACWO As two former members of the Tuveson Laboratory we have a wealth of experience working with the KPC pancreatic cancer model 63

Poster Presentations We can offer Supply of KPC mice suitable for therapeutic study Palpation of tumour bearing mice High resolution ultrasound imaging Therapeutic study management Short term and long term survival study management Dedicated drug preparation dosing surgical necropsy areas Drug dosing scheduling Surgical intervention Necropsy Tissue fixation Study database management Training guidance and assistance Tumour models core our approach This new initiative established itself within the Biological Resources Unit at the Cambridge Research Institute with the full support of CRUK We also supply experimental control and or parental mice to recognised scientific establishments outside of the UK Production and maintenance The TMC streamlines the production of experimental mice suitable for enrolment onto therapeutic studies Weekly palpation of the triple mutants HE HE HE confirms tumour presence Tumours are measured by non invasive ultrasound imaging to confirm and follow the growth of the PDA Mice with a defined tumour burden are enrolled onto study Parental lines KrasLSLG12D p53LSL R127H R172H 20 30 breeding cages 40 70 stock cages Pdx1 cre Pdx1 cre 15 20 breeding cages 30 50 stock cages Experimental production Our primary aim is the continued central supply of experimental KPC and control mice suitable for therapeutic studies and others such as PK PD or allograft studies Mice are generated maintained under the BRU service PPL KPC mice once identified as suitable for enrolment will be transferred to other project licenses with availability at CRI KrasLSLG12D p53LSL R127H R172H x Pdx1 cre Pdxcre 40 55 breed pairs 200 350 stock cages to wean offspring genotype etc Producing approximately 60 150 KPC mice month KPC KrasLSLG12D p53LSL R127H Pdx1 cre Mice generated and not suitable for a study e g health reasons or tumour not appropriate may be killed by a Schedule 1 method Wherever possible tissues from these mice will be made available and used either for the TMC tissue bank or for other in vitro experiments Periodically as part of quality control for the KPC colony the response to gemcitabine will be monitored for any significant changes in efficacy Tumour Models Core Committee meet bi monthly discussing prioritising and approving researcher mice requests at the CRI including their affiliates and collaborators 64 We can manage and maintain your parental and experimental mouse colony

Poster Presentations Management and tumour watch Palpation KPC mice typically develop pancreatic tumours pancreatic ductal adenocarcinoma between 2 10 months of age Presence of tumours identified by palpation Typically 300 400 mice are palpated every week Suitable candidates are then scheduled for high resolution ultrasound imaging High resolution ultrasound imaging Monitoring tumour development Quantification of tumour volumes is possible using 3D ultrasound images Ultrasound Imaging 25 45 scans are performed every week and 3D images of tumours obtained Identify 10 50 tumour bearing KPC mice weekly Typically 8 9 mice are produced each week suitable for therapeutic study with an average tumour size of 69mm in diameter Total colony size is approximately 600 cages for this program 65

Poster Presentations Pancreatic Tumour Allograft Tumour Split Screen Power Doppler liver Pancreatic Tumour with enlarged kidney 3D volume Mouse Embryo Kidney Nephritis 66

Poster Presentations Preclinical studies Refining the assessment of pre enrollable tumours by precise palpations Reducing the stress of mice being repeatedly ultrasound scanned to follow the growth pattern of the tumours as well as the associated implications of repeated exposure to anaesthetic Our dedicated team provide comprehensive technical assistance with your study including skilled obser vations palpations imaging pre post enrolment necropsy tissue and blood collection Necropsy records study data and samples are delivered direct to the researcher Do you have therapeutics for evaluation Could we do similar with your model We offer new and existing research groups within CRI the opportunity to perform therapeutic studies in a well established mouse model of pancreatic cancer 67

Poster Presentations The future The Tumour Models Core Facility will continue the efforts of pancreatic cancer research by providing a central supply of experimental and control mice KPC mice Researchers will be able to continue on going research and we will explore the possibilities of refining the model We have the experience and flexibility to adopt alternative tumour models within the CRI and beyond Applying the same principles used to facilitate and expand the KPC model to what it is today We have in the pipeline variations to the KPC model as well as new genetically engineered mouse models of Breast Prostate Lung and Kidney cancers 68

April 2013 Animal Technology and Welfare Let the fish decide How group size gender and dominance affect preference for environmental enrichment in zebrafish PAUL SCHROEDER1 SOFFIA JONES IAIN YOUNG and LYNNE U SNEDDON Former address Department of Biomedical Services University of Oxford The Old Observatory South Parks Road Oxford 1 Address for Correspondence Paul Schroeder Fish Welfare Project Systems Biology Laboratory UK cic 5 7 West Central 127 Olympic Avenue Milton Park Abingdon Oxfordshire OX14 4SA Objectives Environmental enrichment often improves the welfare of captive animals yet is rarely applied to laboratory fish Fig 1 Apopka Florida USA were divided lengthwise using an opaque grey plastic partition creating two equal 5 litre compartments Fig 2a Each partition had a 5 x 5cm opening to allow transfer of the fish between the compartments Enrichment cues included alongside a barren compar tment ar tificial submerged and floating plants Table 1 Fig 2b c as well as substrates Fig 2d e and combinations of plants and substrates Fig 4 airstones and substrate images Cue One Cue Two Figure 1 In European facilities a large but yet unspecified proportion of zebrafish are housed in barren tanks with no added complexity such as refuge plants or substrate Wild zebrafish habitat features range of substrates such as silt sand and gravel and both overhanging nonaquatic and rooted aquatic vegetation1 2 Studies investigating enrichments compatible with toxicology requirements such as glass or plastic rods found little or no welfare benefits3 Our study uses preference testing letting the animals choose from a range of largely naturalistic designs rather than imposing objects selected for their hygienic qualities By applying the enrichment preference assay to two distinct social contexts this study also aims to evaluate how group sizes affects the fishes capacity to exhibit preferences Design of assay PVC tanks 26 x 22 x 14 5cm Aquatic Habitats Gravel Sand Gravel Submerged plant Floating plant Floating plant Gravel and floating plant Gravel and submerged plant Gravel image Sand image Air stone Barren Barren Sand Barren Barren Submerged plant Sand and submerged plant Sand and floating plant Barren Barren Barren Table 1 List of enrichment cues used for the preference assay Group study only Data was collected over 5 days per tank design on occupancy in the left or right chamber for pairs n 6 1 male 1 female and groups of 8 n 5 4 males 4 females In addition the animals were categorised into dominant and subordinate for each observation session pair study only 69

Poster Presentations Preferences of zebrafish in pairs Preference for substrate over a barren side was largely associated with dominance while subordinate individuals were excluded from substrate enriched compartments Fig 3a d No preferences were displayed by either sex or dominance categor y for plants plant substrate combinations or airstones Preferences of zebrafish in groups a d b c Occupancy rates in substrate enriched compartments were significantly higher than in barren compartments Fig 5a b Both sexes preferred gravel over sand substrate Fig 5c and a tank compartment with floating plants over a barren one Fig 5d Gravel was preferred over sand substrate also in combinations Fig 4 and floating plants over submerged ones Fig 5c f j Zebrafish also showed a strong preference for gravel images over a barren compartment Fig 5k m Both males and females spent more time in the barren compartment when airstones were installed on the other side Fig 5h e Figure 2 Tank design with gravel vs a barren cue a Structural enrichment cues submerged plant b floating plant c gravel d sand e Figure 4 In this study zebrafish preferred enrichment combinations featuring gravel substrate Figure 3 Pair study occupancy time in tanks with sand vs barren a b and gravel vs barren c d p

Poster Presentations Figure 5 Group study occupancy in tanks with gravel a and sand b vs barren gravel vs sand c floating d and submerged plants e vs barren floating vs submerged plants f airstone vs barren g combinations of substrates and plants h j substrate images vs barren k m p

Poster Presentations Discussion In a group context zebrafish show clear preferences for substrates and plants over barren conditions This is consistent with a recent study reporting preference for structural enrichment4 Similar results all reporting preference for gravel over other substrates have also been published for other fish species including brown trout5 three spined stickleback6 and red snapper7 In contrast when zebrafish are held in pairs habitat choices are confounded by dominance relationships Rather than simply imposing enrichment without really understanding what zebrafish actually want in their environment it is vital that we provide these fish with choices to understand their subjective preferences Acknowledgements We are grateful to Gregor Govan and Abigail Sutherland for technical support at Liverpool PS is funded by the Wellcome Trust LS and IY thank NC3Rs for grant funding References 1 2 3 4 5 6 7 72 McClure M McIntyre P and McCune A 2006 Journal of Fish Biology 69 553 570 Engeszer R E Patterson L B Rao A A and Parichy D M 2007 Zebrafish 4 21 40 Wilkes L Owen S F Readman G D Sloman K A and Wilson R W 2012 Applied Animal Behaviour Science Kistler C Hegglin D W rbel H and K nig B 2011 Applied Animal Behaviour Science Johnsson J I Carlsson M and Sundstr m L F 2000 Behavioral Ecology and Sociobiology 48 373 377 Webster M M and Hart P J B 2004 Animal Behaviour 68 1071 1077 2004 Szedlmayer S T and Howe J C 1997 Environmental Biology of Fishes 50 203 207

April 2013 Animal Technology and Welfare A review of German animal research applications from 2010 to identify which anaesthetic and analgesic regimens are used in experiments involving rodents KATHRIN HERRMANN1 and PAUL FLECKNELL2 1 2 Institute of Pharmacology and Toxicology Veterinary School Free University Berlin Comparative Biology Centre The Medical School Newcastle University Corresponding author Kathrin Herrmann fu berlin de Background Directive 2010 63 EU is to become part of national law on 1st January 2013 requiring all EU member states to implement the Principles of the 3Rs This PhD project was set aims to appraise the current situation in Germany with regard to one of the 3Rs Refinement Animal research applications submitted in 2010 are being used to make a comprehensive assessment of the refinement methods currently used in laboratories The study is limited to rats and mice as they are the two most frequently used species The primary aim of the project is to determine the efficacy of proposed anaesthetic and analgesic regimens by examining applications in which rodents undergo surgery The aim is to pinpoint areas where further refinements can still be made since the former is by far the most commonly used species in Germany Anaesthetic intraoperative analgesic regimes Isoflurane was the most widely used inhalant anaesthetic in both rats and mice Isoflurane was either combined with an analgesic or used as the sole agent Ketamine xylazine and midazolam fentanyl medetomidine were the most frequently used injectable anaesthetic combinations in mice Rats were most commonly anaesthetised with the injectables ketamine xylazine followed by pentobarbital Pentobarbital not having analgesic properties was often used in combination with a local or systemic analgesic but there were also cases where no analgesic was administered Methods Research applications from 14 of the 16 German federal states for the year 2010 which required surgical procedures in rats or mice were collected and examined For this initial survey data were analysed to assess the types of surgical inter vention the anaesthetic regimens and the intraoperative and postoperative analgesic regimens In addition the degree of severity according to the provisions laid down in Annex VIII of Directive 2010 63 EU and or according to the Berlin Pain Catalogue and the severity classification given by the researcher were determined Preliminary results Analysis of 285 out of a total of 525 research applications has been completed and certain trends have become apparent The surgical procedures most commonly per formed with rats and mice were laparotomies followed by craniotomies in rats and by skin incisions in mice More studies were done involving surgeries on mice 188 than on rats 97 Figure 1 Distribution of Anaesthetic regimens used in mice and rats with most regimens containing an analgesic component Ketamine fentanyl alpha 2 agonists Preliminary results The common agents administered intraoperatively to 73

Poster Presentations assure analgesia when using isoflurane were buprenorphine carprofen fentanyl metamizole meloxicam or nitrous oxide The time of the administration of the analgesic varied from 45 minutes prior to induction of anaesthesia until immediately before the start of surgery A noticeable amount of the surgical interventions performed under isoflurane did not use an analgesic agent and thus were done without any preventative analgesia Figure 2 Use of isoflurane with various or with no analgesic Postoperative analgesics used Depending on the severity of the inter vention buprenorphine was the most common agent used after surgical interventions rated moderate or severe 45 in mice 43 in rats followed by metamizole approx 25 The administration of an analgesic in water and sometimes in feed after a single injection of the same agent was common practice Metamizole midazolam tramadol and paracetamol were the agents most frequently administered orally However there was no indication in the applications as to whether the daily water intake was measured in order to monitor the intake of the analgesic Multimodal analgesic regimens were rare approx 9 and were considered by experimenters only in exceptional cases following severe surgeries The most common multimodal regimen identified consisted of carprofen and metamizole Severity classification Of the 285 applications analysed 57 were classified as mild 175 as moderate and 53 as severe This classification was compared to the anticipated classification given by the experimenters In 46 of the applications the researcher s severity classification differed from the classification in the above mentioned documents In the majority of cases the researcher underestimated the severity Discussion The current trend in the use of anaesthetic agents can 74 be regarded as an improvement However it was discovered that injectable agents are still in common use without antagonizing the agents at the end of the surgery even though this would be possible The prolonged post surgery sleeping period is considered the most common factor causing mortality in rodents The intraoperative administration of analgesics after induction of anaesthesia was more common than the preoperative one In 10 of the cases there was no analgesic administered postoperatively The most frequent justifications given were that no pain was anticipated that the analgesia would interfere with the study or that pain relief would be administered if pain was observed The problem pinpointed here is that in the applications assessed so far there was little indication of frequent comprehensive pain assessment being carried out The administration of analgesics should correlate with the severity of the experimental procedure and the perioperative preventative use of analgesics should become the standard

April 2013 Animal Technology and Welfare Non surgical embryo transfer with the NSETTM device is a 3Rs refinement technique that reduces stress in CD 1 mice KENDRA STEELE1 BARBARA STONE1 JAMES HESTER1 BRETT SPEAR2 and ANGELIKA FAITH GOODIN1 1 2 ParaTechs Corporation 1122 Oak Hill Drive Lexington KY 40505 USA The Markey Cancer Centre University of Kentucky 800 Rose Street Lexington Kentucky 40537 0293 USA Exhibited at the 2012 LASA Winter Meeting Introduction Non surgical embryo transfer NSET can be used to transfer blastocysts to recipient mice during the generation of transgenic mice after cryopreservation or embr yos after in vitro fertilisation and during rederivation of mouse strains Non surgical embryo transfer is a refinement one of the 3Rs of the surgical embryo transfer procedure The NSET device is a small tapered catheter that attaches to a P2 pipette and deposits embryos directly into the mouse s uterine horn Use of an NSET device in mice should substantially reduce pain suffering and recovery time of mice as the surgical procedure is replaced with an alternative method No anaesthesia is required for nonsurgical embryo transfer and the mice require no postprocedure recovery Because the NSET procedure is non surgical and does not need anaesthesia we hypothesised that use of the NSET device is less stressful than surgery for the mouse We therefore compared the stress responses of pseudopregnant CD1 mice 2 5 days post coitum following uterine embryo transfer surgery and nonsurgical embryo transfer procedures using the NSET device ELISA analysis was used to quantify levels of faecal cortisone a biomarker of stress Electrocardiograms were performed using a non invasive device the ECGenie by Mouse Specifics Inc to measure cardiac indicators of stress 75

Poster Presentations Conclusions G G G G G The NSET procedure is effective for uterine transfer of blastocyst stage embryos Surgery causes a significant increase in faecel corticosterone levels whereas the NSET procedure does not Anaesthesia causes a 38 50 decrease in the number of heart beats per minute whereas the NSET procedure does not alter heart beats per minute Surgery increases cardiac variability by 8 whereas anaesthesia alone and the NSET procedure does not significantly increase cardiac variability Use of the NSET device is less stressful to mice than surgical embryo transfer Acknowledgements We thank Kim Carrico for her contribution towards developing the ELISA protocol and Mouse Specifics Inc for the generous support regarding the EC Genie device and analysis of the data Research reported in this publication was supported in part by the Office of the Director National Institutes of Health under Award Numbers 2R44RR025737 02 8RR440D010958 and the Kentucky Cabinet for Economic Development Office of Commercialisation and Innovation under the grant agreement KSTC 184 512 11 115 with the Kentucky Science and Technology Corporation 76

April 2013 Animal Technology and Welfare Instructions to Authors Subjects considered for publication may include original articles technical notes and reviews pertaining to all aspects of animal science and technology management and education The Editorial Board wishes to offer par ticular 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 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 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 Articles for submission should be sent to Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL Submission No responsibility will be accepted for loss or damage to such articles 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 Format Articles may be submitted either electronically or by hard copy as follows Electronic 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 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 1 The first sheet of the article should contain the following 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 2 For the remainder of the paper the text should be 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 77

Instructions to Authors 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 Illustrations charts and photographs supplied on disk should be in JPEG TIFF or EPS formats and have a resolution of no less than 300dpi The captions for illustrations charts and photographs should be typed in double spacing in numerical order on a separate sheet of paper 5 References to other literature should be indicated in the text by superior figures12 and should be typed in numerical order in the order in which they appear in the text on a separate sheet of paper References should be set out as follows Journals Surname and initials of author s date title of article Name of journal in full volume number first and last page numbers Content 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 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 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 e g Saigeman S 1998 Environmental enhancement of cats what why how Animal Technology Vol 49 No 3 145 154 Number of animals and any pretreatment Time of day of procedure s Quantity and frequency of any samples Books Surname and initials of author s date title of book Name of publisher Town of publisher Statistics e g Flecknell P A 1987 Laborator y Animal Anaesthesia Academic Press London Reprints 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 78 Tests used should be named 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

Call for nominations now open for the 2013 AAALAC International Fellowship Award If you are an IAT Registered Technician RAnTech apply today Deadline is 1 June 2013 The AAALAC International Fellowship recognises two outstanding technicians The AAALAC International Fellowship recognises two outstanding technicians one Registered RAnTech and one AALAS Registered RALAT RLAT one IATIAT Registered RAnTech and one AALAS Registered RALAT RLAT RLATG CMAR who have made or or have the potential to make signi cant RLATG CMAR who have made have the potential to make signi cant contributions the eld of laboratory animal care and use This fellowship contributions to to the eld of laboratory animal care and use This fellowship is valued at more than 3000 The winner will receive a a week long guest is valued at more than 3000 The UKUK winner will receiveweek long guest visit a a prestigious biomedical research facility in USA this Autumn plus visit to toprestigious biomedical research facility in thethe USA this Autumn plus Pete Willan receiving his his Pete Willan receiving complimentary attendance at AALAS National meeting the USA s largest complimentary attendance at thethe AALAS National meeting the USA s largest award from Dr Christian award from Dr Christian laboratory animal science and technology meeting registration travel and E Newcomer Executive laboratory animal science and technology meeting all all registration travel and E Newcomer Executive Director of AAALAC accommodation expenses included along with a a daily allowance meals accommodation expenses areare included along withdaily allowance forfor meals Director of AAALAC in in 2008 2008 and out of pocket expenses and out of pocket expenses TO TO APPLY APPLY 1 Write a brief letter nominating yourself have someone write this 1 Write a brief letter nominating yourself or or have someone write this letter your behalf letter on on your behalf 2 Include 500 word written statement describing why you should be be 2 Include a a 500 word written statement describing why you should selected selected 3 Include a copy of your 3 Include a copy of your CV CV 4 Include more than three letters of of support 4 Include no no more than three letters support Carol FIAT RAnTech Carol FoxFox FIAT RAnTech TIPS TIPS waswas the 2009 recipient the 2009 recipient Be Be sure address these points in your application sure to to address these points in your application What have you done to promote animal welfare and re nement in your What have you done to promote animal welfare and re nement in your workplace workplace What have you done to promote animal welfare and re nement outside What have you done to promote animal welfare and re nement outside of your workplace of your workplace How have you shared your knowledge laboratory animal care with How have you shared your knowledge of of laboratory animal care with others others If you selected how will this award bene t your career and your If you areare selected how will this award bene t your career and your Laura Foster MIAT associates Laura Foster MIAT associates How will you translate this experience back to your workplace and your RAnTech Senior How will you translate this experience back to your workplace and your RAnTech Senior Animal Technologist Animal Technologist at at local association local association AstraZeneca 2010 winner AstraZeneca 2010 winner Does your application include evidence leadership or leadership Does your application include evidence of of leadership or leadership potential and participation with posters presentations or publications potential and participation with posters presentations or publications Have you participated in overseas meetings in past Have you participated in overseas meetings in thethe past Winners expected to to share their experiences brie y preparing a written Winners areare expected share their experiences brie y by by preparing a written report AAALAC International and publish a paper in in Animal Technology and report forfor AAALAC International and publish a paperAnimal Technology and Welfare The of cial Journal of IAT and EFAT They will also be expected to Welfare The of cial Journal of thethe IAT and EFAT They will also be expected to present their experiences at a national or local conference present their experiences at a national or local conference left left to right Mark Gardiner to right Mark Gardiner Mail or or e mail your application materials 1 June 2013 to Mail faxfax e mail your application materials by by 1 June 2013 to JoseJose Pepe Figueroa President Pepe Figueroa President AAALAC International AAALAC International of Priority Services Inc of Priority OneOne Services Inc sponsors Dr Dr Christian sponsors and andChristian E E c o c o The International Fellowship Selection Committee The International Fellowship Selection Committee Newcomer Executive Director Newcomer Executive Director 5283 Corporate Drive Suite 203 5283 Corporate Drive Suite 203 of AAALAC right of AAALAC right Frederick MD 21703 Frederick MD 21703 USA USA Fax 301 696 9627 E mail accredit aaalac org Fax 301 696 9627 E mail accredit aaalac org ForFor complete details and a list selection criteria visit http www aaalac org complete details and a list of of selection criteria visit http www aaalac org about fellowship cfm about fellowship cfm The AAALAC International Fellowship Award is presented by Association for The AAALAC International Fellowship Award is presented by thethe Association for Assessment and AccreditationLaboratory Animal Care AAALAC International Assessment and Accreditation of of Laboratory Animal Care AAALAC International through a grant Priority One Services Inc and in in co operation with AALAS through a grant by by Priority One Services Inc and co operation with AALAS Martin Heath the 2012 IAT NC3Rs Medical Research Council and the National Institutes of of Health Martin Heath the 2012 IAT NC3Rs thethe Medical Research Council and the National Institutes Health Don t delay get your application now Don t delay get your application in in now award winner holding award winner holding thethe attractive inscribed attractive inscribed crystal globe crystal globe

INDEX TO ADVERTISERS April 2013 AAALAC International xvii Allentown Inc iii Aston Pharma xv a tune software AG v Bell iv Charles River Laboratories IFC Datesand Ltd xi Harlan Laboratories viii Institute of Animal Technology xiii xviii IPS Product Supplies Ltd IBC Lillico vi PFI Systems xvi Shepherd Speciality Papers Lillico xv Special Diets Services xiv Surrey Diagnostics xii Sychem Ltd x Tecniplast UK OBC VetTech Solutions vii Institute of Animal Technology Continuing Professional Development CPD Introduction to Zebra Danio Workshop Wednesday 10th April 2013 Venue Midlands This is a half day workshop designed for those animal technologists who care for Zebra Danio and who wish to learn new skills and refresh existing skills The workshop combines formal presentations and interactive practicals Topics include An overview of zebrafish in research Basic husbandry Housing Practical aspects of colony maintenance Ill health and disease prevention Advancing and promoting excellence in the care and welfare of animals in science THIS COURSE CARRIES A 4 HOURS CPD CREDIT FOR SUCCESSFUL ATTENDANCE Registration details can be found on the Members Section of the IAT website www iat org uk or by emailing cpd iat org uk