ATW August 2018 : simplebooklet.com

CONTENTS Vol 17 No 2 August 2018 Editorial Jas Barley Chair of the Editorial Board ix Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting Penny Hawkins Secretary Anne McBride Giles Paiba Rita Rose Mark Prescott John Mulley Sam Izzard Deborah Ridley Luca Melotti Sarah Kappel and Robert Hubrecht 75 Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of scientific and healthcare and their pros and cons Gary Martinic 85 PAPER SUMMARY TRANSLATIONS 91 ANDREW BLAKE TRIBUTE AWARD WINNING ENTRY Refinement of tumour passage by optimisation of the tissue disaggregation protocol Melanie Galvin Alice Lallo Stewart Brown Paul Kelly Becky Bola Danielle Potter Cassandra Hodgkinson and Kristopher Frese and Caroline Dive TECH 2 TECH Blunting cutting canines of adult male monkeys a discussion by the Laboratory Animal Refinement Enrichment Forum Casey Coke Evelyn Skoumbourdis Jeannine Cason Rodgers Kelsey Finnie Louis Divincenti Marcie Donnelly Melissa Truelove Misty WIlliams Fritze Polly Schultz Vanessa Herring and Viktor Reinhardt 101 105 Phenotypes and characteristics of the Pole4 mice an exploration into what makes these animals unique Louis Kitchenham 108 Optimising mouse production good practice for efficient colony management and implementation of the 3Rs Hannah Easter 112 POSTER PRESENTATIONS A refinement to Schedule 1 Helen Read and Rhys Perry 117 Improving rabbit enrichment developing a rolling enrichment plan Katie Pearson and Jenn Gant 121 Dosing by intravenous injection in the Syrian hamster Georgia Beamish C Chapman and A Lomax 124 Refinements to animal husbandry for continuous infusion dosing in Beagle dogs Lucy Allen 126 Optimisation of workflow in a BSL 3 PET CT imaging facility for non human primate research Leo van Geest Jaco Bakker Mohammed Khayum Frank Verreck and Jan Langermans 128 Refining mouse handling do we don t we An animal technologist s perspective Alexander Swan 130 Setting up a new surgical dosing team at Envigo Huntingdon Jamie Tuck Kate Read Stuart Bradley Lydia Mills Katrina Turner Maxfield Turner Deborah Potts and Jess Hill 132 Introduction of gang caging for group housed rats David Clarke and Lena Ioannou 136 Development of a real time respiration monitoring system for in vivo optical imaging Stuart Gilchrist Ana Gomes Sheena Wallington Veerle Kersemans Marcus Westcott and Sean Smart 138 To glow or not to glow that is the question Christopher Huggins 140 Mice on the road how to move thousands of mice whilst maintaining microbiological status and high levels of welfare Rafaela Fern ndez de la Fuente 142 Going on a diet Matthew O Brien 148 ABSTRACT A survey of rabbit handling methods within the United Kingdom and the Republic of Ireland James Oxley Clare Ellis Anne McBride and Wanda McCormick 151 Instructions to Authors 152 i

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IAT REPRESENTATIVES OFFICERS President Dr Robin Lovell Badge CBE FRS Immediate Past President Professor Sir Richard Gardner MA PhD FRSB FIAT Hon FRS Vice Presidents David Anderson MRCVS Stephen Barnett BA MSc FIAT Hon CBiol FRSB RAnTech Miles Carroll PhD Brian Cass CBE Gerald Clough BSc PhD EurBiol CBiol MRSB SFZSL Paul Flecknell MA Vet MB PhD DLAS DipLECVA MRCVS FIAT Hon Penny Hawkins PhD BSc Wendy Jarrett MA Judy MacArthur Clark CBE BVMS DLAS FRSB DVMS h c DipECLAM FRAgS DipACLAM MRCVS Fiona McEwen BSc BVM S MSc MRCVS Tim Morris BVetMed PhD DipACLAM DipECLAM CBiol FRSB CertLAS MRCVS Jos Orellana BVSc MSc Clive Page OBE PhD BSc Jan Bas Prins PhD MSc Vicky Robinson CBE BSc PhD Gail Thompson RLATG Robert Weichbrod PhD RLATG Life Members Charlie Chambers MIAT RAnTech Roger Francis MSC FIAT RAnTech Pete Gerson MSc FIAT RAnTech Cathy Godfrey FIAT RAnTech John Gregory BSc Hons FIAT CBiol FRSB RAnTech Patrick Hayes FIAT DipBA RAnTech Robert Kemp FIAT Hon RAnTech Phil Ruddock MIAT RAnTech Ted Wills HonFIAT RAnTech Dorothy Woodnott FIAT Honorary Members Mark Gardiner MIAT RAnTech Andy Jackson MIAT Sarah Lane MSc FIAT Brian Lowe MSc FIAT RAnTech Sue McHugh BSc FIAT Norman Mortell BA Hons MIAT RAnTech Terry Priest MBE FIAT RAnTech Trevor Richards BEM MIAT David Spillane FIAT Wendy Steel Pete Willan DMS FInstLM MIAT Members of Council Ken Applebee OBE Matthew Bilton Kally Booth Charlie Chambers Steven Cubitt Simon Cumming Haley Daniels Glyn Fisher Nicky Gent Cathy Godfrey Alan Graham Nathan Hill Linda Horan Sam Jameson Elaine Kirkum Adele Kitching Theresa Langford Sylvie Mehigan Steve Owen Alan Palmer Allan Thornhill John Waters Lynda Westall Carole Wilson Adrian Woodhouse Council Officers Chair Ken Applebee OBE FIAT CBiol FRSB RAnTech Vice Chair Linda Horan BSc Hons MIAT RAnTech Honorary Secretary Linda Horan BSc Hons MIAT RAnTech Honorary Treasurer Charlie Chambers MIAT RAnTech Assistant Treasurer Glyn Fisher FIAT RAnTech Chair Board of Educational Policy Glyn Fisher FIAT RAnTech Chair Board of Moderators Haley Daniels MBA MSc MIAT RAnTech CIPD Chair Registration Accreditation Board Ken Applebee OBE FIAT CBiol FRSB RAnTech ATW Editor Jas Barley MSc FIAT RAnTech Bulletin Editor Carole Wilson BSc MIAT Branch Liaison Officer Lynda Westall BSc Hons FIAT DMS RAnTech EFAT Representative Charlie Chambers MIAT RAnTech Website Coordinator Allan Thornhill FIAT RAnTech Animal Welfare Officers and LABA Representatives Matthew Bilton Kally Booth Simon Cumming Nicky Gent Sylvie Mehigan John Waters ATW Bulletin Editorial Board Jas Barley Chair Matthew Bilton Nicky Gent Patrick Hayes Elaine Kirkum Carole Wilson Lynda Westall Board of Educational Policy Glyn Fisher Chair Steven Cubitt Secretary Adele Kitching Board of Moderators Haley Daniels Chair Simon Cumming Cathy Godfrey Moderators Anthony Iglesias Theresa Langford Jenny Parks Sarah Reed Communications Group Adrian Woodhouse Chair Nathan Hill Elaine Kirkum Teresa Langford Sylvie Mehigan Allan Thornhill Lynda Westall CPD Officer Charlie Chambers Registration and Accreditation Board Ken Applebee Chair Charlie Chambers John Gregory Cathy Godfrey Gerald Clough Kathy Ryder Home Office Stuart Stevenson Observers Charles Gentry Certificate Holders Forum Ngaire Dennison LAVA Congress Committee Alan Graham Chair Haley Daniels Linda Horan Adele Kitching Allan Thornhill John Waters Diversity Officer Haley Daniels MBA MSc MIAT RAnTech CIPD UK Biosciences ASG Representative Home Office Steve Owen Charlie Chambers Alan Palmer IAT OFFICERS MAY BE CONTACTED VIA IAT Administrator admin iat org uk OR VIA THE IAT WEBSITE AT www iat org uk OR VIA THE REGISTERED OFFICE 5 South Parade Summertown Oxford OX2 7JL Advertisement Managers PRC Associates Ltd Email mail prcassoc co uk 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 2018 Institute of Animal Technology All rights reserved No part of this publication may be reproduced without permission from the publisher BRANCH SECRETARIES 2018 Cambridge Edinburgh Hertfordshire Essex Huntingdon Suffolk Norfolk Ireland London Midlands North East England North West Oxford Surrey Hampshire Sussex West Middlesex West of Scotland ii Sarah Shorne Janice Young Joanna Cruden Jo Martin Lisa Watson Rebecca Towns Ian Fielding Rachel Sandy and Joanne Bland Nicky Windows April Shipton Francesca Whitmore Josefine Morsing Linda Horan cambridgebranch iat org uk edinburghbranch iat org uk hertsessexbranch iat org uk hssbranch iat org uk irelandbranch iat org uk londonbranch iat org uk midlandsbranch iat org uk northeastbranch iat org uk cheshirebranch iat org uk oxfordbranch iat org uk shsbranch iat org uk westmiddxbranch iat org uk westscotlandbranch iat org uk

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August 2018 Animal Technology and Welfare THE INSTITUTE OF ANIMAL TECHNOLOGY ETHICAL STATEMENT Our purpose is to advance knowledge and promote excellence in the care and welfare of animals in science and to enhance the standards and status of those professionally engaged in the care welfare and use of animals in science Editorial Jas Barley Chair of the Editorial Board When you think about literature it is apparent that not only have many authors studied the behaviour of humans but also that of animals Moby Dick Animal Farm Watership Down to Peter Rabbit to name a few all contain careful observation of the interaction between man and his companions on earth both wild and domesticated although I do not recall seeing many hedgehogs dressed up in pinafores Beatrice Potter may not have known that it is lactucin in lettuce that makes man and animals sleepy but she had probably observed her rabbits dozing after a feast of lettuce although nowadays many rabbit owners avoid feeding lettuce in large quantities as it can adversely affect the intestines in lagomorphs and rodents Even childrens songs and poetry involve animal behaviour and one of my earliest memories is of my mother telling me the poem by Georgia Roberts Durston There once was a rabbit developed the habit of twitching the end of its nose the poem continues with other albeit superficial comments on rabbit natural behaviour Many other songs and poems make similar comments on the animals that surround us The reason that I have been pondering on literature is that one of the trends that runs through this issue of our Journal is the need to observe particularly to study behaviour in the wild to guide us in providing the animals in our care with an environment they need rather than one that we think they require One of recommendations in the report on the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting is that we should find out more or refresh our knowledge about the wild type behaviour of the species we care for Could more be done to facilitate positive natural behaviours You might like to ask for a discussion or workshop at your establishment involving individuals and bodies such as your Home Office inspector named persons researchers AWERB and internal species specific or Three Rs groups To help you with this the Institute of Animal Technology Welfare Group is currently working on a review of the available literature which will be published as soon as possible in Animal Technology and Welfare Louis Kitchenham s presentation at the West Middlesex Technician and Trade Day back in February also dealt with the need for good observation when he described the issues and care of Pole4 mice For those of you who could not attend the meeting Louis has converted his presentation into a Tech 2 Tech article which is included in this issue If you attended IAT Congress this year I am sure you will agree that it was an enormous success I know that obviously not everyone can attend Congress so I am delighted to have access to almost all of the posters from IAT Congress 2018 and the first of these appear in this issue including a report from Rafaello de la Fuente of how 1400 strains of mice were moved from Hertfordshire to the new Francis Crick Institute in London Most of us would recognise the prospect of such a move as daunting to say the least but as Rafaello describes good planning allowed the move to happen with the minimium of problems For the few poster authors who have not supplied an electronic copy for reprinting I will be in touch I also hope that some all of the platform presentations will appear as papers in the Journal in due course rest assured if they do not it is not for the want of trying If you presented at Congress either on the platform gave a poster presentation or workshop please consider converting your work into a paper or article The Editorial Board is here to help you Refinement also features heavily in this issue and the implementation of this principle in relation to Mouse Handling gang caging for group housed rats respiration monitoring during imaging and dosing of Syrian hamsters are all dealt with along with other areas of refinement Optimising mouse production is another way of implementing the 3Rs by reducing over production of animals especially those of an incorrect genotype and Hannah Easter s Tech 2 Tech article clearly explains how this can be achieved You will notice that we have reprinted Melanie Galvin s winning entry for the Andrew Blake Tribute Award as unfortunately the original version gave an incorrect address Apologies to Melanie and her colleagues but sometimes human fraility just gets in the way mea culpae ix

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August 2018 Animal Technology and Welfare Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting PENNY HAWKINS Secretary 1 ANNE McBRIDE 2 GILES PAIBA 3 RITA ROSE 4 MARK PRESCOTT 5 JOHN MULLEY 6 SAM IZZARD 7 DEBORAH RIDLEY 7 LUCA MELOTTI 8 SARAH KAPPEL 9 and ROBERT HUBRECHT10 1 2 3 4 5 6 7 8 9 10 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS School of Psychology University of Southampton University Road Southampton SO17 1BJ Home Office Animals in Science Regulation Unit 14th Floor Lunar House 40 Wellesley Road Croydon CR9 2BY Envigo Woolley Road Alconbury Huntingdon PE28 4HS National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs Gibbs Building 215 Euston Rd London NW1 2BE School of Biological Sciences Bangor University Deiniol Road Bangor Gwynedd LL57 2UW GSK Research and Development Park Road Ware Hertfordshire SG12 0DP M nster Badestra e 13 D 48149 M nster Germany Division of Animal Health and Husbandry University of Bristol Langford House Bristol BS40 5DU UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Introduction The RSPCA UFAW Rodent and now Rabbit Welfare Group has held a one day meeting every autumn for the last 24 years so that its members can discuss current welfare research exchange views on welfare issues and share experiences of the implementation of the 3Rs of replacement reduction and refinement with respect to rodent and rabbit use A key aim of the Group is to encourage people to think about the whole lifetime experience of laboratory rodents and rabbits ensuring that every potential negative impact on their wellbeing is reviewed and minimised The 24th meeting was held at the Animal and Plant Health Inspectorate APHA Weybridge on 14 November 2017 The first session addressed meeting animals needs and aiming for a good life with the needs of female breeding rabbits as a case study Three speakers gave presentations on breeding rabbit behaviour a Home Office perspective on providing enrichment appropriate to the species and refining housing husbandry and care for these animals in practice The second session on rodent welfare comprised an update on the National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs Year of Laboratory Rodent Welfare reducing stereotypic digging in gerbils enabling group housing in rats with vascular access buttons positive emotions in rats and good practice for housing male mice The day ended with a discussion session on speaking up for rodents within Animal Welfare and Ethical Review Body AWERB meetings and more generally This report summarises the meeting and ends with a list of action points for readers to consider raising at their own establishments Behaviour and welfare needs of breeding rabbit does Anne McBride University of Southampton The Home Office Code of Practice for housing and care stresses that the physiological and behavioural needs of animals should be restricted only for the minimum necessary period of time and degree see below 1 Achieving this and providing a good quality of life for any animal requires a sound up to date understanding of the physiological and behavioural requirements of the species breed or strain and how these alter during the individual s life cycle Although most commonlyused laboratory animals have been bred and often inbred in captivity for generations wild type behaviours are still innate if animals are given the opportunity to express them A classic example is the Ratlife video 75

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting in which inbred laborator y rats released into a naturalistic enclosure rapidly began to express many wild type behaviours2 and there is evidence that naturalistic behaviour is also innate in domestic pigs3 and laboratory mice 4 Researching the behaviour of the wild type species is therefore a good starting point for refining housing and care and this presentation considered the breeding rabbit doe as a case in point Understanding the behavioural needs of the pre and post parturient doe in the laboratory or any other breeding institution means considering normal doe behaviour and that of the offspring in the wild including how interactions between the doe and young change during early development up to and beyond weaning Some basics rabbits are herbivorous grazing animals and are crepuscular and nocturnal i e they are most active at dawn and dusk and during the night In the wild they are only above ground for around 30 of a 24 hour period and when it is dark in the daytime they are in the dark below ground quite a contrast to life in the laborator y Rabbits are not especially domesticated having been selected primarily for meat and fur and they comprise some 20 of the diet of over 20 species in their native Spain and Portugal 5 The primary need to avoid being eaten underlies all rabbit behaviour including the absentee parenting style Figure 1 shows a typical rabbit nest in the wild Note the downward sloping tunnel to help the blind deaf nestling kits to re locate the nest and their siblings and the earth plug which is used to completely stop the nest so that predators cannot see hear or smell the kits You and your dog have probably unknowingly walked past these stopped up nests many times The doe returns approximately every 24 hours for a few minutes to suckle the kits The frequency of nursing can vary between days and individuals as females may suckle several times a day Other than that she displays minimal parental caring behaviour while the young are in the nest The kits can recognise their mother s scent at birth and about an hour before they are usually nursed they begin to rise to the top of the nest in preparation for suckling The rest of the time they huddle in the warm nest material and rotate between the centre and periphery of the group They begin to eat solids nest material at 8 days their eyes open fully at 10 days and they begin to explore the tunnel at around 13 days The nest stop entrance is still tightly re packed by the doe following every visit until around 18 days After this time the young begin to emerge and they are weaned about 24 days post partum They still associate very closely with adults especially their mother for several weeks Natural rabbit society comprises mixed gender and age groups to help achieve safety in numbers This is obviously ver y different indeed from the laboratory situation Some ideas are set out in Table 1 for going fur ther towards accommodating the physiological and behavioural needs of both does and kits in the laboratory setting The suggestion relating to socialising the kits is based on studies that have shown handling at pre weaning results in tamer less fearful rabbits when individuals were several weeks older 6 7 To reduce the risk of cannibalisation it is advised that early handling is best done after the kits eyes are open and scent profiles are preserved by rubbing hands in the nest material and stroking the mother before touching the kits If rabbits are then regularly handled and acclimatised to being picked up throughout their lives anxiety will be reduced with benefits for the rabbits staff and science for does for kits for both Mental and mphysical stimulation a good quality and quantity of space Ability to choose between the company of other rabbits or to be alone Ability to completely get away from the kits with no visual auditory or olfactory contact Ability to have control over timing for access to the kits Early socialisation with humans to help reduce anxiety when adult Ability to rest in the dark when not being nursed for the first 18 days of life An enclosed dark nest box accessed by a downwardsloping tunnel this could have a door for observing the kits An entrance to the nest box that the doe could seal and open herself Electronic tags for individual does to access each nest box to prevent double littering Table 1 Suggested refinements to breeding rabbit housing and husbandry Figure 1 A wild rabbit nest with earth plug Illustration by Guy Troughton in McBride A 1988 Rabbits and Hares Whittet Books Essex 76 For fur ther information on rabbit behaviour and potential refinements for breeding does see references 5 8 and 9 at the end of this report Your Named Information Officer Named Veterinary Surgeon

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting Named Animal Care and Welfare Officer and AWERB should also be able to provide further information and help you promote discussion about meeting animals needs aim to provide a good life in general for all the species in your care and fur ther encourage understanding and empathy 10 Rabbit housing and enrichment appropriate to the species a view from the Home Office Giles Paiba Home Office Animals in Science Regulation Unit The publication of the Home Office Code of Practice for the Housing and Care of Animals Bred Supplied or Used for Scientific Purposes in 2014 1 set out the minimum requirements for housing and breeding rabbits used within the confines of the Animals Scientific Procedures Act ASPA 1986 as amended As mentioned above there is a requirement that animals physiological and behavioural needs be restricted only for the minimum necessary period of time and degree but how do we interpret whether space is of sufficient complexity to allow expression of a wide range of normal behaviour and how appropriate to the species can accommodation really be in a laboratory setting The Code of Practice aims to Promote good animal welfare through the provision of consistent high quality care and accommodation Support the generation of high quality reliable scientific results through the reduction of environmental variables and Implement the principles of the 3Rs through using the minimum number of animals and causing the minimum degree of pain suffering distress or lasting harm It includes both engineering standards and performance standards An engineering standard is a defined measurable parameter e g cage dimensions temperature range or photoperiod iA per formance standard is an outcomes based requirement e g noise levels including ultrasound shall not adversely affect animal welfare Both engineering and performance standards are important Engineering standards are the welfare safety net which set clear expectations and are easy to verify with respect to compliance Per formance standards are helpful because it is not always possible to prescribe how outcomes should be achieved and they also allow for advances in science and our understanding of animal welfare needs With respect to breeding rabbits we recognise that in nature these animals live in a complex environment It is important for rabbits to have a raised area for lookout and to enable hopping exercise which will increase hind limb weight bearing strain assist in maintaining bone strength and therefore reduce the likelihood of injuries The Code includes engineering standards size and optimum height of the shelf and performance standards animals must be able to lie and sit and easily move underneath the step and be able to use the surface The Code of Practice specifically advises those breeding rabbits to ensure that the need for privacy of the nesting and parturient mother are considered In reality this would need the behavioural drivers and needs previously described by Dr McBride to be considered Thus although the provision of a nest box is not mandatory it is assumed in that the Code does provide minimum cage dimensions depending upon whether nest boxes lie inside or outside the enclosure housing the doe and her litter Refining housing husbandry and care for breeding rabbits Rita Rose Envigo Breeding rabbits it should be simple right In the wild absolutely but not so straightfor ward in a laborator y Before embarking on a breeding programme fundamental questions are needed to define the study objective or production colony needs and how you can aim to safely address both these and the needs of the animals you are responsible for Our company has establishments for animal supply as well as for Contract Research Rabbits under our care may be part of a breeding colony to supply other establishments outside our company or they may be used in a scientific study that required breeding as part of its objective e g reproductive toxicology To define good practice for housing and caring for our rabbits we use information from different sectors including guidance for pet rabbits rescue settings and farmed rabbits We also have an internal Rabbit Welfare Group which meets quarterly and reports to our AWERB Our protocol for breeding rabbits involves placing a nest box in with the doe a week before she is due to litter This stays in the cage for 10 days post littering then the whole cage is changed for a new clean one without a nest box and with tray liner for the kits to sit on We recognised that this system does not separate the doe and kits as would be the case in the wild and we wanted to identify a refinement We have therefore been trialling a nest plate which is made from stock board with a lip that is high enough to keep the kits in one area until they are strong enough to hop over into the rest of the cage We are monitoring breeding 77

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting success using the nest plates and we are hoping to be able to reduce the colony size if litter productivity increases The NC3Rs Year of Laboratory Rodent Welfare Mark Prescott NC3Rs Mice and rats account for the majority of animals used in scientific procedures in the UK and globally Throughout 2017 the NC3Rs highlighted opportunities to improve the welfare of laboratory rodents By spotlighting recent advances in animal welfare science and technology hosting workshops events and data collection projects aimed at animal technologists and focusing on the adoption of good practice we aimed to have a positive impact on the large number of rodents used in research Our Year of Laboratory Rodent Welfare included many different approaches to refine rodent use including the development of technologies for automated continuous recording of the behaviour of individual mice and rats socially housed in the home cage11 12 and a novel ultra lightweight low power device for electrophysiological recordings in unrestrained and untethered mice 13 new research awards for example to refine the capture and tracking of wild rodents for ecological studies and publication of guidelines from our data sharing working groups on refining bile duct cannulation studies in rats14 and rodent models of ischaemic stroke 15 awareness of the innate needs and behaviours of laborator y rats using the Ratlife video2 and stimulated discussion as to how more space and stimulation could be provided Ran a mouse handling workshop to suppor t trainers Named Training and Competency Officers and other named persons who wanted to move from picking up mice by the tail to using cupped hands or a tunnel instead This has been demonstrated to reduce anxiety in mice with both animal welfare and scientific benefits and some establishments have successfully stopped handling by the tail 17 20 The workshop aimed to explore potential barriers to using the refined handling methods and provide practical tips and solutions A range of resources including a video tutorial poster FAQs and downloadable video clips for in house training were also made available on our website Set up a crowdsourcing multi institute data collection project to identify prevalence patterns and triggers of aggression in laboratory mice This is commonly reported and can cause pain distress and even death so the welfare impact will clearly be significant if the causes can be better understood and used to identify practical solutions Data were supplied by over 110 participants from over 30 institutions within the UK and overseas and we will communicate preliminary findings in 2018 For information and updates on all these initiatives see nc3rs org uk rodent welfare hub and nc3rs org uk 2017 year laboratory rodent welfare In other key projects relating to the day to day welfare of rodents in the laboratory we Reducing stereotypic digging in gerbils Highlighted the work of our 2016 3Rs prize winner Joanna Makowska who demonstrated the importance of burrowing climbing and standing upright for laboratory rats 16 Despite generations of domestication the laboratory rat retains innate behaviours that cannot be expressed in standard housing such as rearing on the hind legs when exploring and socialising Height restrictions in standard caging lead to muscle stiffness which rats attempt to alleviate by lateral stretching This is not only a welfare issue it has been argued that laborator y rats are metabolically morbid and therefore poor models for human disease Joanna Makowska and her colleague Dan Weary showed that young rats in large enriched cages burrow around 30 times a day climb around 75 times a day and stand upright around 180 times Some facilities are now investing in larger caging or providing shared access to a play area for rats Promoted the concept of a play area in successful workshops at the Institute for Animal Technology Congress in 2017 in which we increased John Mulley Bangor University 78 The Mongolian gerbil Meriones unguiculatus is a diurnal inhabitant of semi desert and steppe habitats It is a social species usually living in a pair with offspring and some helpers within a central burrow system that has multiple exits Relatively few gerbils are used in the UK 236 procedures using 206 animals for the first time in 2016 in fields including research into diabetes hearing epilepsy stroke thermal regulation parasitology and desert adaptation The lack of access to a tunnel system in standard laboratory housing and the inability to construct one leads to stereotypical digging behaviour in many species of desert rodent including the Mongolian gerbil The situation is further exacerbated by the excessive chewing behaviour exhibited by gerbils which renders many traditional forms of environmental enrichment unsuitable The Home Office Code of Practice acknowledges that gerbils often develop stereotypical digging behaviour suggesting a nest box with a separation wall and a tunnel entrance including

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting a bend corner a deep layer of litter for digging and nesting and a burrow substitute at least 20 cm long 1 However in our experience this has not always been effective in ameliorating stereotypical digging and on occasion the gerbils have simply chewed the tunnels up or left the nest box because humidity levels were too high due to the angle in the entrance tunnel In a study funded by UFAW we set up a controlled behavioural study using unchewable stainless steel tubes of different lengths diameters and angles made by a company that produces roll bars for cars Ten week old gerbils were housed in single sex groups of four and the analysis of video samples of the animals behaviour was randomised and blinded We defined stereotypical digging as a bout of over 12 seconds Following an initial increase in stereotypical digging the level of digging decreased and we have also anecdotally noted that noise levels have decreased in the gerbil room with far fewer scrabbling sounds The gerbils are now provided with stainless steel rat houses with open ends for ventilation and 20 cm 50 mm diameter stainless steel tubes with a 90 mandrel bend as a low cost and low tech intervention which has reduced or entirely eliminated stereotypical digging Figure 2 Enabling group housing in rats with vascular access buttons Sam Izzard and Debbie Ridley GSK Dual cannulated rats with harnesses have previously been singly housed at our facility due to concerns that animals would interfere with one another s cannulae potentially causing distress and infection We obviously wanted to avoid this on animal welfare grounds but singly housing rats is also undesirable from a welfare perspective as they are social animals Therefore we wanted to set up a vascular access button VAB system that would enable GSK to group house surgically prepared rats We trialled placing a VAB between the scapulae under general anaesthesia with perioperative pain relief monitoring body weight daily for the first 8 days postsurger y as par t of our post operative welfare assessment protocol We found that animals gained weight more rapidly and steadily with the VABs than previous cohorts with the harnesses They can be pairor group housed before and after each study improving their welfare The system was also successfully validated for pharmacokinetic steady state infusion studies indicating that there were no issues with data quality when changing from harnesses to VABs However some issues were encountered including swelling around the button site animals scratching at the site catheters coming away from the button pin and some movement from the tether when on study These issues have been overcome by ensuring that the tethers are removed as soon as possible after the 1 hour infusion trimming the rats nails regularly and placing the cannulae on to the pins using silicone covered forceps Having successfully overcome these issues this model is now our first choice for surgically cannulated rats Figure 3 Figure 2 Tunnel configuration for reducing stereotypical digging in gerbils Photo John Mulley Figure 3 Group housed rats with vascular access buttons Photo GSK 79

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting Expression of positive emotions in rats Luca Melotti 1 Jessica Lampe 2 Kathryn Finlayson 2 Sara Hintze 3 Oliver Burman4 and Hanno W rbel2 1 2 3 4 RG Behavioural Biology and Animal Welfare Division of Behavioural Biology University of M nster M nster Germany Division of Animal Welfare University of Bern Bern Switzerland Division of Livestock Sciences Dept of Sustainable Agricultural Systems University of Natural Resources and Life Sciences Vienna BOKU Vienna Austria School of Life Sciences Joseph Banks Laboratories University of Lincoln Lincoln LN6 7DJ United Kingdom greater in PT than CT indicating that the experience of being tickled was positive in comparison with the exposure to white noise We found that Ear Colour 0 2 scale was significantly pinker in PT than in CT and Ear Angle was wider ears more relaxed in PT compared to CT Figure 4 However other quantitative and qualitative facial measures which included Eyeball height width Ratio Eyebrow height width Ratio Eyebrow Angle visibility of the Nictitating Membrane and the Rat Grimace Scale did not show significant differences between treatments Animal welfare research has traditionally focussed on preventing or reducing negative experiences but more recently attention has also been given to promoting positive emotions 21 Therefore it is crucial to develop objective methods to detect and quantify positive emotions in animals We conducted two studies with rats which aimed i to identify facial indicators of positive emotions and ii to measure positive emotional contagion between rats where one individual s emotions and related behaviours directly trigger similar emotions and behaviours in others Facial indicators of positive emotions Previous studies have identified rodent facial expressions which specifically occur in situations likely to induce negative emotional states e g pain aggression and fear This study aimed to investigate whether rats exhibit specific facial expressions of positive emotions during play with humans Fifteen adolescent male Lister Hooded rats were individually subjected to a 2 minute Positive Treatment PT and a 2 minute mildly aversive Contrast Treatment CT over two consecutive days PT consisted of playful manual tickling administered by the experimenter while CT consisted of exposure to intermittent bursts of white noise A positive emotional state is indicated by 50 kHz ultrasonic vocalisations USVs Vocalisations at this frequency were recorded to check whether rats had different emotional states in PT and CT High speed photos of the rats faces in a profile or three quarters view were taken during both treatments Novel qualitative and quantitative measures and also the established Rat Grimace Scale 22 were used to detect fine changes in facial expression Photos were scored by an experimenter who was blinded to which treatment the animals had received The number of positive vocalisations was significantly 80 Figure 4 A rat experiencing positive emotions Note the ear angle and colour Please see reference 23 for further information and picture credits This study contributes to the exploration of positive emotions and thus better welfare in rats as it identified potential indicators of positive facial expression resulting from a positive experience Pinker Ear Colour and wider Ear Angle both accompanied by the emission of positive vocalisations may reflect internal physiological changes associated with positive emotional arousal and muscle relaxation respectively 23 Positive emotional contagion This study investigated positive emotional contagion by exploring the effects of positive treatments received by one rat on the behaviour and vocalisations of the rest of their social group We are currently preparing a publication on this for another journal so will just give a summary here Adolescent male Lister Hooded rats were housed in groups of three and one rat per group received either positive or control treatments as follows The treated rat was taken to a separate room for 2 minutes where

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting they were either manually tickled by the experimenter given chocolate rewards or placed in the treatment arena without further action control condition USVs at 50 kHz and play events in the home cage attempted nape contacts to initiate play and solitary scampering were counted before and after the treatment by an experimenter who was blinded to the treatments the rats had received Our results provided some evidence of short term positive emotional contagion from one individual receiving a positive treatment to its social group Positively treated rats appeared to promote emotional contagion by becoming the target of more play initiations by untreated rats and by also inducing after tickling more play between untreated rats 24 To group or not to group Good practice for housing male mice Sarah Kappel University of Bristol Penny Hawkins RSPCA Michael T Mendl University of Bristol It is widely recommended to group house male mice because mice are social animals but male mice do not naturally share territories Laboratory mice were derived from Mus musculus a species that forms territories inhabited by a small group of individuals including one dominant male several females pups and juveniles before these disperse Territory size varies with food availability and population density e g from a few square metres close to human dwellings to several square kilometres in natural habitats Dominant males are highly intolerant of intruders and the introduction of a strange mouse provokes aggressive behaviour in the territory holder 4 25 Although laboratory mice have been bred in captivity for many generations wild type behaviours can still be innate 4 as discussed by McBride earlier in this report We can assume that a male mouse given the opportunity to choose would prefer to be housed with a group of females with enough space to drive away juvenile male offspring which is obviously impractical in the average laboratory Given that the male mouse is pre adapted to live with other mice is living with other males the best alternative to a natural group composition To try to answer this question we conducted a review of the literature on mouse husbandry behaviour and welfare with a survey of current practice for housing male mice which has been published elsewhere 26 Our key findings were Most survey respondents 120 of 147 people believe that male mice naturally prefer to live with other mice and that male mice should ideally be group housed with other males in the laboratory 123 of 147 If males were singly housed this was mostly due to aggression 122 responses or for scientific reasons 100 The literature review found that individual housing allows males to have their own territor y and eliminates injurious aggression and stressful social defeat but being housed alone causes social deprivation Group housing fulfils the need to be with other animals but injurious aggression between male mice can be a serious animal welfare issue Even without aggression not all animals within a group will be in a state of positive welfare Many male mice may be negatively affected by the stress of repeated social defeat and subordination Both single and group housing therefore raise concerns about welfare and also research validity On the basis of current Codes of Practice 1 responses to our sur vey and the economic implications of single vs group housing there is strong motivation to group house male mice and facilities will continue to do so For example of 147 survey respondents 99 reported that it was general practice post weaning to group house male mice just 4 stated that these were routinely singly housed and 44 stated that both housing conditions were applied 26 The current literature does suggest that it is generally speaking preferable from an animal welfare perspective to house male laboratory mice in groups We suggest that group housing for male mice is the less worse approach but do not positively endorse this practice because male mice would naturally prefer to live with a group of females not other males However whether group or single housing is better or less worse in any given situation is highly context dependent according to strain age social position life experiences and housing and husbandry protocols It is important to evaluate what is preferable in each case from animal welfare and ethical perspectives using the literature and current good practice for housing male mice and ensure that this is discussed within the facility e g by the AWERB with input from named persons It is possible to reduce the risk of aggression in groups of male mice via thoughtful housing husbandry and care see also the NC3Rs project on mouse aggression above in this report The literature and our sur vey repor ts measures including transferring some nesting material not litter from used to clean cages 27 providing enrichment that can be manipulated e g nesting material as opposed to solid shelters 28 housing in groups of three29 and choosing less aggressive strains if this is compatible with the research objective 81

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting To conclude it is not possible to house male mice in the laboratory in a way that is compatible with their natural behaviour nor is it possible to make sweeping statements regarding good practice for housing all male mice There is still much to learn about the behaviour of different mouse strains and how this is affected by housing husbandry and care life stage and previous experiences 26 2 The discussion session The meeting ended with a discussion session which aimed to explore how participants felt able to raise the priority given to rodent welfare within their facilities and to initiate or become involved with ethical debates around rodent use In addition to general discussion topics addressing whether the species identified as special by the ASPA dogs cats equidae and primates genuinely deserve greater consideration and which species participants would consider to be special and why We also asked participants how they had spoken up for rodents within their establishments Figure 5 answers were presented as tick boxes in a Turning Point slide and people could select as many as applied Most of the 80 participants had made colleagues think more about harms to rodents and had brought new information about rodent behaviour biology and welfare to their attention top two bars in Figure 5 Fewer had discussed these issues directly with researchers or participated in the AWERB and raising ethical issues scored lowest although around two thirds of the audience had been able to do this The discussion was helpful and it was encouraging to see that participants felt able to raise welfare and ethical issues 3 4 5 6 7 8 Could more be done to facilitate positive natural behaviours You might like to ask for a discussion or workshop at your establishment involving individuals and bodies such as your Home Office inspector named persons researchers AWERB and internal species specific or Three Rs groups Look at the NC3Rs Rodent Welfare Hub nc3rs org uk rodent welfare hub and see whether you can use it to further refine housing husbandry and care procedures and welfare assessment for rodents Do not allow stereotypical behaviour to become normalised i e an accepted aspect of standard housing Investigate the literature talk to colleagues in other institutions to see what steps are suggested to reduce such behaviours and ask for support in researching and trialling ways of reducing or eliminating any outbreaks If you care for gerbils and are concerned about stereotypical digging behaviour ask to trial the shelter and tunnel system described in this report If social animals are routinely housed individually because of concerns about externalised ports or instrumentation research the literature for ways of refining procedures approaches or husbandry to enable group housing Promote the concept of encouraging positive welfare and emotions for the animals in your care as well as reducing suffering You might like to share the link to this ar ticle with colleagues news nationalgeographic com 2016 12 happy ratsfacial expression animals emotion Read the full paper on male mouse housing and pass it on to colleagues You may also wish to use the paper to suggest to your establishment that it reviews its practice e g beginning with a discussion by the AWERB Commit to promoting knowledge of rodent biology behaviour and welfare amongst colleagues as in Figure 4 and to raising ethical issues associated with rodent use within your AWERB and more generally If your AWERB is looking for topics as part of its forum for discussion function suggest a rodent related topic like those set out in the discussion session section of this report Acknowledgements Figure 5 Responses to the question Who has challenged assumptions about rodents and spoken up for them and how The histogram shows the number of respondents who had achieved each action Thank you to all the speakers and delegates for the talks and discussions We are also grateful to APHA for providing the meeting venue References 1 List of action points based on all of the presentations and discussions 1 Find out more or refresh your knowledge about the wild type behaviour of the species you care for 82 UK Home Office 2014 UK Home Office Code of Practice for the Housing and Care of Animals Bred Supplied or Used for Scientific Purposes Rodents and Rabbits Accessed 3 January 2018

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Report of the 2017 RSPCA UFAW Rodent and Rabbit Welfare Group meeting 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Berdoy M 2002 The Laboratory Rat A Natural History Film University of Oxford UK Accessed 3 January 2018 Wischner D Kemper N and Krieter J 2009 Nestbuilding behaviour in sows and consequences for pig husbandry Livest Sci 124 1 8 Latham N and Mason G 2004 From house mouse to mouse house The behavioural biology of free living Mus musculus and its implications in the laboratory Appl Anim Behav Sci 86 261 289 McBride E A 2015 Rodents and rabbits the what and the why In Report of the 2015 RSPCA UFAW Rodent and Rabbit Welfare Group Meeting by P Hawkins et al Animal Technology and Welfare 15 1 10 11 Castadi K et al 2005 Even minimal contact linked to nursing reduces fear response towards humans in rabbits Appl Anim Behav Sci 95 123 128 Der Weduwen S and McBride E A 1999 Rabbit behaviour and the effects of early handling Proceedings of the 2nd World Meeting on Ethology and Mondial Vet Lyon France Accessed 3 January 2018 UFAW RSPCA 2008 Refining Rabbit Care A Resource for those Working with Rabbits in Research Download at Accessed 3 January 2018 Dorning J and Harris S 2017 The Welfare of Farmed Rabbits in Commercial Production Systems A Scientific Review Download at Accessed 3 January 2018 McBride E A 2017 Small prey species behaviour and welfare Implications for veterinary professionals Journal of Small Animal Practice DOI 10 1111 jsap 12681 Bains R S Wells S Sillito R R Armstrong D Cater H L Banks G and Nolan P M in press Assessing mouse behaviour throughout the light dark cycle using automated in cage analysis tools Journal of Neuroscience Methods Accessed 28 March 2018 Redfern W S Tse K Grant C Keerie A Simpson D J Pedersen J C et al 2017 Automated recording of home cage activity and temperature of individual rats housed in social groups The Rodent Big Brother project PLoS ONE 12 9 e0181068 doi org 10 1371 journal pone 0181068 Jiang Z Huxter J R Bowyer S A Blockeel A J Butler J Imtiaz S A Wafford K A Phillips K G Tricklebank M D Marston H M and RodriguezVillegas E 2017 TaiNi Maximizing research output whilst improving animals welfare in neurophysiology experiments Scientific Reports 7 8086 doi 10 1038 s41598 017 08078 8 Burden N Kendrick J Knight L McGregor V Murphy H Punler M and van Wijk H 2017 Maximizing the success of bile duct cannulation studies in rats recommendations for best practice Laboratory Animals 51 5 457 464 doi 10 1177 0023677217 698001 Percie du Sert N Alfieri A Allan S M Carswell H V O Deuchar G A et al 2017 The IMPROVE Guidelines Ischaemia Models Procedural refinements of in vivo experiments Journal of Cerebral Blood Flow and Metabolism 37 1 3488 3517 Accessed 3 January 2018 16 17 18 19 20 21 22 23 24 25 26 27 28 29 Makowska I J and Weary D M 2016 The importance of burrowing climbing and standing upright for laboratory rats Royal Society Open Science 3 160136 Download at Accessed 3 January 2018 Hurst J L and West R S 2010 Taming anxiety in laborator y mice Nature Methods 7 825 826 doi 10 1038 nmeth 1500 Ghosal S Nunley A Mahbod P Lewis A G Smith E P et al 2015 Mouse handling limits the impact of stress on metabolic endpoints Physiol Behav 150 3137 doi 10 1016 j physbeh 2015 06 021 Gouveia K and Hurst J L 2013 Reducing mouse anxiety during handling Effect of experience with handling tunnels PLoS ONE 8 6 e66401 doi 10 1371 journal pone 0066401 Gouveia K and Hurst J L 2017 Optimising reliability of mouse performance in behavioural testing the major role of non aversive handling Scientific Reports 7 44999 doi 10 1038 srep44999 Boissy A Manteuffel G Jensen M et al 2007 Assessment of positive emotions in animals to improve their welfare Physiol Behav 92 3 375 97 doi 10 1016 j physbeh 2007 02 003 PMID 17428510 Sotocinal S G Sorge R E Zaloum A et al 2011 The Rat Grimace Scale A partially automated method for quantifying pain in the laborator y rat via facial expressions Molecular Pain 7 55 doi 10 1186 17448069 7 55 Download at Accessed 3 January 2018 Finlayson K Lampe J F Hintze S W rbel H and Melotti L 2016 Facial indicators of positive emotions in rats PloS ONE 11 11 e0166446 download at Accessed 3 January 2018 Lampe J F Burman O W rbel H and Melotti L in prep Investigating positive emotional contagion in rats Crowcroft P 1966 Mice All Over G T Foulis Company London UK Kappel S Hawkins P and Mendl M T 2017 To group or not to group Good practice for housing male laborator y mice Animals 7 12 88 doi 10 3390 ani7120088 mdpi com 2076 2615 7 12 88 Accessed 3 January 2018 Van Loo P L P Van Zutphen L F M and Baumans V 2003 Male management Coping with aggression problems in male laboratory mice Lab Anim 37 300313 Van Loo P L P Kruitwagen C L J J Van Zutphen B F Koolhaas J M and Baumans V 2000 Modulation of aggression in male mice Influence of cage cleaning regime and scent marks Anim Welf 9 281 295 Van Loo P L Mol J A Koolhaas J M Van Zutphen B F and Baumans V 2001 Modulation of aggression in male mice Influence of group size and cage size Physiol Behav 72 675 683 83

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August 2018 Animal Technology and Welfare Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of science and healthcare and their pros and cons GARY MARTINIC Faculty of Science University of Western Sydney Richmond NSW Australia Introduction Perhaps better known for their ability to eliminate adversaries in war zones and to spy on people drones as they are more commonly known have also been used recreationally since at least the 1960s in the form of remote control aeroplanes and helicopters which have been flown in parks and at aeromodelling clubs across the world This recreational drone industry has grown in leaps and bounds in the last decade or so with the introduction of affordable high capacity lithium batteries and brushless electric motors making models cheaper lighter and more powerful than their nitro methanol or diesel burning predecessors This relatively new class of aircraft have emerged hand in hand with the miniaturisation of high quality cameras like the GoPro Hero and with the development of first person view FPV technology which have enabled a multitude of applications across various fields of endeavour However it should be noted that the term drone is somewhat of a misnomer as a more accurate description is to describe them either as an unmanned Figure 1 Elephants at Tarangire National Park shown during their migration being herded away from farmed areas by Forest Rangers using drones Source Dronesetc com aerial vehicle UAV an unmanned aircraft UA or Remotely Piloted Aircraft RPA Figure 2 A drone carrying medical supplies arrives at its remote destination Source Baseline of Health Foundation The technology Operating via the use of Global Positioning System Satellite GPS technology their preprogrammed altitudes and routes are controlled either by computers on board the UAV or under the remote control of a pilot on the ground or in another vehicle UAVs come in a wide variety of shapes sizes configurations and characteristics and while they have been traditionally remotely piloted they are being operated increasingly under semi autonomous control With each passing year these technologies have grown smaller faster and more sophisticated mirroring developments in the smart phone industry which essentially uses the same electronic components namely optics embedded micro processors sensors and batteries Compared to manned aircraft military UAVs can also carry out a larger range of intelligence surveillance and reconnaissance ISR and surgical strike missions they are smaller and more economical and can fly for longer without the need for re fuelling or 85

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Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of science and healthcare risk of pilot fatigue Being discreet and needing little experience to operate they also are significantly less expensive than training Air Force pilots Due to these many advantages UAVs have had a hugely successful impact on warfare in the current theatres of conflict namely Afghanistan and Iraq A myriad of civilian uses Fortunately aside from their well established military uses1 3 and even before the first successful UAV delivery made in the USA 4 non military UAVs have increasingly been used in the industrial recreational commercial and security non military sectors Some of their more popular uses are in commercial aerial sur veillance aerial photography digital mapping cinematography journalism major spor ting event telecasts law enforcement search and rescue disaster relief support operations cargo transport and even in real estate marketing They have also proven to be very effective in surveying and inspecting powerlines gas and oil pipelines distant bridges offshore oil platforms wind platforms road and rail transport infrastructure and safety checks on large commercial aircraft The reason they have been so effective is because UAVs can easily get to remote regions difficult terrain or hard to reach places and when compared to the use of manned flying aircraft UAVs can achieve this at a fraction of the cost Their ability to cover larger areas in a shorter amount of time while beaming back aerial footage in real time to base control is another major advantage Over the last few years they have also proven their utility in a number of scientific fields including wildlife research Figure 1 conservation and anti poaching operations farming and agriculture archaeology oceanography oil gas mineral exploration and forestry and remote sensing More recently and more aligned to the medical and healthcare fields UAVs have been trialled to deliver vaccines Figure 2 to remote regions of the developing world They are also being trialled as mini aerial ambulances the Dutch have developed ambulance drones capable of offering a rapid response to victims suffering a heart attack they are being used in forensics and have even had their capabilities tested in the fields of battlefield medicine 5 This article describes the application of these latter new uses of UAVs from a broader scientific and healthcare standpoint and also discusses how to minimise the impact of these technologies on wildlife in the field Uses in agriculture and the environment Precision Agriculture is a concept that observes measures and responds to field variability in crops UAVs have been very useful in this regard as they can 86 be used to identify when nitrogen levels are low Also because of their unique ability to fly precisely low over crops they can be used to check for signs of disease and for other crop anomalies in addition to their abilities to steer water supply and pesticides to needy crops As UAVs are fast becoming a great tool for crop surveys it is predicted that they will likely be one of the largest sources of big data in one of the largest businesses in the world agriculture It then follows that this may allow us to transform the way we grow our food and it is why some commentators refer to UAVs as the future of food 6 Traditionally to know the status of crops farmers had to walk their fields but most of the time they could not really see what that was in the middle of these fields Today those same farmers can fly UAVs over their fields and using the UAVs sensitive inbuilt cameras they can almost immediately receive information on growth patterns water supply and chemicals used 6 On many large consolidated farms the prophylactic spraying of pesticides is performed so as not to miss an infection that can result in crop losses not actually because an infection is present 6 Using infrared lenses on a UAV flown high above the condition of the earth can be generally assessed and on the information provided better informed farmers can then decide to use water more efficiently and to reduce or entirely eliminate chemical use These advantages are today utilised globally Aside from the area of food and UAVs have also provided real benefits in other areas of agriculture that of livestock farming where the location status and movement of farm animals can be quickly and inexpensively monitored using UAVs Uses in wildlife management and research Wildlife management and research has also significantly benefitted from the use of UAV technology This has allowed scientists to survey photograph Figure 3 Two Humpback whales can be visualised lunge feeding on large schools of Herring This image was captured by a UAV flying overhead Source dailyexpress co uk

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Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of science and healthcare Figure 3 monitor and to protect wildlife sanctuaries and marine reserves The advantage being that using this technology both thermal imaging and live video can be sent in real time to regulatory authorities tasked with protecting these environments and consequent action can be taken against those carrying out illegal activity for example poaching of wildlife or overfishing Marine mammal research has also advanced via the use of UAVs Previously samples of whale blow nasal mucous had to be collected by the researchers on small boats trying to get close enough to the whale to be able to collect these samples This practice was not only potentially dangerous for the researchers but it was also intrusive for the whales Today whale health can be monitored much more easily as researchers use small UAVs which can flow low enough over the blowhole Figure 4 of various whale species to collect these samples for data analysis The use of UAVs is also of course effective in all environments air land and sea wherever wildlife management and research is being carried out Figure 4 Image showing a small UAV hovering above the blowhole of a whale to collect blow nasal mucous for health monitoring data analysis Source Woods Hole Oceanographic Institution In many African countries elephant populations when not properly managed easily can get out of control and this results in masses literally thousands of these animals destroying and stampeding through arable acres of valuable farmland In the past farmers would react by shooting and killing these animals to protect their land For tunately today forest rangers and farmers are working together using UAVs to navigate and control elephant herds away from valuable farmed areas during times of migration One example of where this is happening is in Tarangire National Park in Tanzania Figure 1 but this practice is being used at multiple locations across Africa Uses in remote healthcare Perhaps one of the most worthy uses of UAVs in recent years has been their application as a delivery vehicle for vaccines and medicines to remote regions of the developing world 7 Originally trialled as a deployment method in mid late 2016 8 the use of small UAVs to deliver humanitarian medicines such as vaccines first aid blood products and even organs for transplantation in a few cases 9 have proven so successful that there is no doubt that this quick new cost effective technology will only increase in use in the future Mostly borne out of necessity this new supply chain delivery tool the UAV stands to essentially replace entire fleets of slower land based transport vehicles such as trucks buses cars and motorbikes which cannot easily traverse the difficult terrain10 and harsh conditions9 of the remote villages of the African sub continent where life saving vaccines and other medicines need to be delivered in an efficient and timely manner This novel deliver y method not only eliminates expensive fuel maintenance and labour costs it also delivers these medicines quickly cheaply and most importantly in a timely manner thereby reducing risk of spoilage from lack of refrigeration This amounts to not only unprecedented cost savings but also wider coverage of regions as vaccines are of no value if they do not get to the people that need them 8 These vaccines are commonly used against illnesses such as the Zika virus Dengue Fever and Malaria which many low to middle income African countries are affected by However they also include rotavirus measles tetanus and hepatitis type B vaccines normally given prophylactically in childhood Using UAVs vaccine availability improved and resulted in the immunisation of up to 96 percent of the target population wherever it was used A study by Haidari et al 8 found the use of UAVs increased vaccine and medical supply availability and reduced costs in a range of settings and circumstances when they were used frequently enough to overcome capital set up and maintenance costs Their study using computational modelling showed that the major drivers of cost savings using UAVs were the road speed of traditional land vehicles the number of people needing to be vaccinated and the distances travelled When one considers that many African countries currently cover only about 60 percent of the population using land based approaches the use of UAVs is only set to grow 8 10 Medical uses Another innovative and praiseworthy use of UAVs has been in the development of a UAV air ambulance designed by a Dutch engineering student Alec Momont 11 At the end of 2014 Delft University in the Netherlands unveiled a prototype of an ambulance drone Figure 5 purposely designed and equipped with a defibrillator and a camera microphone and speakers for the express purpose to respond to heart attack incidents in the street 12 With the ability to cover distances of 12 km in around 1 min flying at a 87

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Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of science and healthcare speed of up to 100 km h fitted with an automatic external defibrillator Source Daily Mail could be transmitted to the responding base station Other medical procedures could be administered through the drone with responder assistance before a road ambulance arrives Essentially a paramedic assesses the kind of situation he she is dealing with chooses the appropriate kit inserts in into the UAV and then it flies off to the incident scene Discussion Figure 5 An operator on scene applies transducer patches to the chest of a patient suffering a heart attack This UAV has the potential to save thousands of lives as the majority of deaths following a heart attack occur due to a slow emergency response time Therefore it promises to be literally life saving It is fitted with an automatic external defibrillator known as an AED and its purpose is to shock the heart back to life It receives emergency mobile calls and then uses the GPS signal to locate the person in distress Once it arrives at the scene the UAV is controlled by a paramedic who using the UAVs camera and speakers gives personalised instructions to a responder near the patient The defibrillator operates automatically once the transducer patches have been applied to the patient s chest The paramedic can then continue to talk to the responder and to monitor and observe the patient until they can be transported to a hospital for further treatment and follow up Due to the complex nature of systems such as these and separately due to the legal issues which yet need to be resolved innovations like this make commercial development not an easy task Delft University is currently developing the technological concept further and interestingly it plans to include additional uses such as delivering oxygen supplies to people affected by fires or for respiratory illnesses Other teams that are working on similar applications are developing UAVs to assist hazardous area response teams who deal with medical emergencies involving chemical biological and nuclear materials 13 Expanding on this concept further the author believes that UAVs could be packed and made ready to respond to certain situations For example a larger UAV such as a Hexacopter could carry larger payloads for larger scale emergencies containing a removable bay with pre made emergency kits with bandaging supplies and tourniquets to address bleeding for road accident victims the supply of auto injector type medications such as insulin for diabetics aspirin for chest pain etc afterwards and if carried out properly vital signs 88 With regards to UAV delivery of humanitarian medical aid to remote regions it needs to be remembered that regulatory issues could potentially limit the ability of UAVS to deliver these goods in countries where the law hasn t kept pace with technology Also specialised skillsets and tools are needed to operate and maintain such equipment and sourcing may be difficult to find in some developing countries Appropriate packaging and excellent coordination are also required between those dispatching and those receiving goods since no person accompanies a delivery despatched via a UAV While these technical issues remain to be sorted in some developing countries developed countries are already reaping the benefits of the use of these technologies In early 2017 it was reported that UAVs had saved at least 59 people in 18 different incidents around the world since 2013 of which 38 of those individuals were saved thanks to rescue teams and civilians using their UAVs to aid people facing life threatening emergencies 14 In effect that is one person s life saved on average every week The authors stated that there were many more unrepor ted incidents excluded from the figures because no definitive counts were available Therefore one can only assume that in fact the actual figures for that year were likely much higher In a case in point approximately 200 people were spotted and saved by UAVs during a series of devastating floods which occurred in India in Dec 2015 Again these were not included in the above figures Apart from the number of people saved it is the ingenuity of ways in which they were saved which is also fascinating For example a Canadian man who was lost in the snow fields was spotted and rescued in 2013 by an UAV and similarly in early 2017 when a group of kayakers went missing for days They were found using a heat sensing UAV as were flooding victims in an unrelated incident Back in mid 2015 two teens were prevented from drowning due to a UAV which successfully delivered life vests to them Interestingly in one third of those cases people were rescued by bystanders civilians non professionals who happened to have drones ready when the incidents occurred One can only assume that the number of life saving incidents will increase in the coming years as the technology becomes more widespread While it is always important to alert the relevant authorities at the first sign of emergencies it also stands to reason that UAVs will continue to provide a net benefit to public safety 14

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Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of science and healthcare The cons While UAVs promise everyday benefits and many would argue that they have led to a generational shift in the advancement of technology these peeping eye goanywhere devices have the serious potential in the wrong hands to be misused which is why UAV related legislation is being introduced in many countries This has been largely due to privacy and safety concerns Most law abiding citizens have a right to live in a free society without being continuously monitored by government agencies These issues represent some of the ethical and legal challenges which need to be urgently addressed but there are also regulatory issues training licencing and safety and technical issues with UAVs such as flight time endurance and range which also need to be improved in some areas No doubt with the menacing pace of technological advances these will be overcome Indeed many of these hurdles have already been resolved in the United States of America by the Federal Aviation Authority FAA with UAV related legislation having been introduced in more than 30 states while Australia s Civil Aviation Safety Authority is touted to closely follow suit Though UAVs have proven to be useful ecological survey tools for studying and protecting wildlife with their ever growing acceptance comes a need to ensure that they do not harm the animals they are trying to observe This harm may be manifested with the animals being disturbed or even stressed leading to undesirable and unintended impacts on wildlife Varying species may react differently to intrusive UAVs coming too close for example some birds may abandon their hatchlings if too stressed by the experience Researchers in the field need to find ways to minimise the impact of these technologies with the wildlife they are studying This can be by altering the type of UAV flown to a more low impact and less noisy model or by modifying the study conditions or mode of operation In an effor t to better understand manage and conserve our planet s biodiversity field biologists from the University of Adelaide have proposed a UAV Code of Best Practice 15 In this document they provide some recommendations which include adopting a precautionary principle in lieu of evidence complying with the relevant civil aviation regulations using sound institutional animal ethics processes selecting the most appropriate UAV for the study exercising minimum wildlife disturbance flight practices and publishing their methods and results in detailed reports No doubt these recommendations will be well received by the scientific community at large Summary As can be seen from some of the images in this article UAVs are very powerful ecological survey tools capable of producing high resolution quality photography and live video footage in a way that the human eye has never seen before This technology is an enabler and UAV systems as tools are all about data capture and dissemination From flying in the eye of a hurricane to flying high above forest fires or volcanoes UAVs can provide this valuable data faster cheaper and more safely and discreetly than any previous technology that has been available UAVs have profoundly transformed the military domain and are now rapidly revolutionizing many other industries Although there are some cons inherent with their use sensible regulations and policies put into place will alleviate these issues On the whole the pros far outweigh the cons as UAVs fast become an everyday technology with huge benefits to society Acknowledgements I thank Drs Edward Narayan Shane Griffin and Louise Rabbit for critically reviewing this manuscript and for offering useful suggestions Sources of images used Figure 1 Elephants at Tarangire National Park Digital Image Dronesetc com 22 Feb 2017 Web 12 Jan 2018 http kickassfacts com drones facts Figure 2 A drone delivering medical supplies Digital Image The Baseline of Health Foundation 25 Jun 2016 Web 12 Jan 2018 http jonbarron org healthcare hospitals do gooder drones Figure 3 Two Humpback whales can be visualized lunge feeding on large schools of Herring Digital Image Daily Express 17 Dec 2015 Web 12 Jan 2018 https www express co uk news nature 627521 hu mpback whales feeding drone footage herring Nor wayPatrick Dykstra Figure 4 Image showing a small UAV hovering above the blowhole of a whale to collect blow nasal mucous for health monitoring data analysis Digital Image Woods Hole Oceanographic Institution 10 Oct 2017 Web 12 Jan 2018 https www whoi edu newsrelease whale blow microbiome Figure 5 Ambulance drone Digital Image Daily Mail 29 Oct2014 Web 16 Jan 2018 https www youtube com watch v y rEI4bezWc References 1 2 3 Martinic G 2012 Drones or Smart Unmanned Aerial Vehicles Australian Defence Force Journal Issue 189 Nov Dec Martinic G 2016 The Explosion in the Use of Unmanned Weapons Systems their Increasing Utility Effectiveness and Legality in Operations and their Implications for Future Military Pilots Headmark Journal of the Aust Naval Institute Mar 06 2016 online edn Martinic G 2014 The Proliferation Diversity and Utility of Ground based Robotic Technologies Canadian Military Journal Vol 14 No 4 Oct 89

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Novel and innovative civilian uses of drones or unmanned aerial vehicles in the fields of science and healthcare 4 5 6 7 8 9 10 11 12 13 14 15 90 Gibbs S 2015 First successful drone delivery made in the US Technology The Guardian Jul 20 Martinic G 2014 Glimpses of Future Battlefield Medicine the Proliferation of Robotic Surgeons and Unmanned Vehicles Technologies Journal of Military Veterans Health Vol 22 3 Sept edn Green E 2013 Drones might be the future of food Business The Atlantic Oct 3 See also http www wsj com video cio network how drones willrevolutionize farming DEB563DD 87EF 4EE9 B6B2D8645F70EC89 html Science Daily 2016 Drones could be cheaper alternative to delivering vaccines in developing World Jun 21 issue Bloomberg School of Medicine Press Johns Hopkins University Haidari L A et al 2016 The economic and operational value of using drones to transport vaccines Vaccine 34 4062 4067 Thiels C A et al 2015 Use of unmanned aerial vehicles for medical product transport Air Medical Journal 34 104 108 Tang L and Shao G 2015 Drone remote sensing for forestry research and practices J For Res 26 4 791797 Suggested Health 2014 Grad student invents flying ambulance drone to deliver emergency shocks Advancements in Health and Science Egozi A 2014 A drone that can save lives Technology Israeli Homeland Security Nov 05 Wong S 2017 Defibrillator drones could save lives before ambulance arrives New Scientist 13 Jun issue Reisinger D 2017 Here s how many lives drones have saved since 2013 Technology Fortune Mar 14 Hodgson J and Koh L P 2016 Best practice for minimising unmanned aerial vehicle disturbance to wildlife in biological field research Correspondence Current Biology Vol 26 10 404 405

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August 2018 Animal Technology and Welfare PAPER SUMMARY TRANSLATIONS INHALTVERZEICHNIS Bericht zur tagung der RSPCA UFAWTierschutzgruppe f r Nager und Kaninchen 2017 PENNY HAWKINS Sekret rin 1 ANNE McBRIDE 2 GILES PAIBA 3 RITA ROSE 4 MARK PRESCOTT 5 JOHN MULLEY 6 SAM IZZARD 7 DEBORAH RIDLEY 7 LUCA MELOTTI 8 SARAH KAPPEL9 und ROBERT HUBRECHT10 1 2 3 4 5 6 7 8 9 10 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Gro britannien School of Psychology University of Southampton University Road Southampton SO17 1BJ Gro britannien Home Office Animals in Science Regulation Unit 14th Floor Lunar House 40 Wellesley Road Croydon CR9 2BY Gro britannien Envigo Woolley Road Alconbury Huntingdon PE28 4HS Gro britannien National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs Gibbs Building 215 Euston Rd London NW1 2BE Gro britannien School of Biological Sciences Bangor University Deiniol Road Bangor Gwynedd LL57 2UW Gro britannien GSK Research and Development Park Road Ware Hertfordshire SG12 0DP Gro britannien Bereich Verhaltensbiologie und Tierschutz Abteilung f r Verhaltensbiologie Universit t M nster Badestra e 13 D 48149 M nster Deutschland Division of Animal Health and Husbandry University of Bristol Langford House Bristol BS40 5DU Gro britannien UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Gro britannien Abstract Die RSPCA UFAW Tierschutzgruppe f r Nager und nun auch Kaninchen veranstaltet seit 24 Jahren jeden Herbst ein eint giges Treffen bei dem die Mitglieder ber Themen aktueller Tierschutz Forschung diskutieren und Erfahrungen und Meinungen ber nagerrelevante Tierschutzfragen sowie ber die Umsetzung der 3R Prinzipien Vermeidung Reduktion und Verbesserung des Einsatzes von Nagern und Kaninchen austauschen k nnen Ein Hauptziel der Gruppe ist es zum Nachdenken ber die Gesamtheit der Lebenserfahrungen von Nagern und Kaninchen als Laborversuchstiere anzuregen und sicherzustellen dass jede potenzielle Beeintr chtigung ihres Wohlergehens gepr ft und auf ein Minimum gesenkt wird Die 24 Tagung fand am 14 November 2017 im Animal and Plant Health Inspectorate APHA in Weybridge statt Die erste Sitzung befasste sich damit wie den Bed rfnissen der Tiere Rechnung getragen und ihnen ein gutes Lebens erm glicht werden kann Dabei wurden die Bed rfnisse von Zuchtkaninchen als Fallstudie betrachtet Drei Referenten hielten Vortr ge ber das Verhalten von Zuchtkaninchen und boten Informationen aus Sicht des Innenministeriums ber eine praxisnahe artgerechte Anreicherung und Verbesserung von Unterbringung Haltung und Pflege dieser 91

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August 2018 Animal Technology and Welfare Neuartige und innovative zivile Nutzung von Drohnen oder unbemannten Luftfahrzeugen in den Bereichen Wissenschaft und Gesundheitswesen sowie ihre Vor und Nachteile GARY MARTINIC Faculty of Science University of Western Sydney Richmond NSW Australien Korrespondenz g martinic westernsydney edu au Abstract Drohnen sind allgemein eher f r ihre F higkeit bekannt Gegner in Kriegsgebieten zu vernichten und Menschen auszuspionieren Sie werden aber auch schon lange zumindest seit den 1960er Jahren in Form von ferngesteuerten Flugzeugen und Hubschraubern auch in der Freizeit eingesetzt Es sei hier jedoch darauf hingewiesen dass der Begriff Drohne etwas irref hrend ist denn genau genommen handelt es sich hierbei um unbemannte Luftfahrzeuge UAV unbemannte Flugger te UA oder ferngesteuerte Flugger te RPA Erfreulicherweise werden neben den UAVs f r etablierte milit rische Zwecke und noch vor dem ersten erfolgreichen UAV Einsatz in den USA auch zunehmend UAVs in Zivilbereichen wie Industrie Freizeit Handel und nichtmilit rische Sicherheit verwendet Der Grund warum UAVs so erfolgreich sind liegt darin dass sie entlegene Regionen schwieriges Gel nde oder schwer zug ngliche Orte problemlos erreichen und dies im Vergleich zum Einsatz von bemannten Flugzeugen zu einem Bruchteil der Kosten m glich ist Ihre F higkeit gr ere Bereiche in k rzerer Zeit abzudecken und gleichzeitig Luftaufnahmen in Echtzeit an die Bodenkontrolle zu senden ist ein weiterer gro er Vorteil In den letzten Jahren haben sie sich auch in verschiedenen wissenschaftlichen Bereichen bew hrt z B in der Wildtierforschung im Naturschutz und bei der Bek mpfung von Wilderei aber auch in der Land und Agrararch ologie in der Ozeanographie in der l Gas und Mineralexploration sowie in der Forstwirtschaft und der Fernerkundung Seit einiger Zeit werden zudem mehr auf die Bereiche Medizin und Gesundheitswesen ausgerichtete UAVs erprobt um Impfstoffe in entlegene Regionen armer L nder zu liefern Sie werden auch als Mini Luftrettungswagen getestet die Niederl nder haben Ambulanzdrohnen entwickelt um schnelle Hilfe f r Herzinfarktopfer zu erm glichen finden in der Forensik Anwendung und ihre F higkeiten wurden sogar auf dem Gebiet der Schlachtfeldmedizin erprobt Dieser Ar tikel beschreibt die Nutzung von UAVs f r diese neuen Zwecke aus einem weitreichenderen wissenschaftlichen und gesundheitlichen Blickwinkel und er rtert auch wie die Auswirkungen dieser Technologien auf die Tierwelt in diesem Bereich minimiert werden k nnen 93

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Animal Technology and Welfare August 2018 CONTENU DE LA REVUE Rapport de r union entre la RSPCA et l UFAW sur le bien tre des rongeurs et des lapins PENNY HAWKINS Secr taire 1 ANNE MCBRIDE 2 GILES PAIBA 3 RITA ROSE 4 MARK PRESCOTT 5 JOHN MULLEY 6 SAM IZZARD 7 DEBORAH RIDLEY 7 LUCA MELOTTI 8 SARAH KAPPEL 9 et ROBERT HUBRECHT10 1 2 3 4 5 6 7 8 9 10 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS School of Psychology University of Southampton University Road Southampton SO17 1BJ Home Office Animals in Science Regulation Unit 14th Floor Lunar House 40 Wellesley Road Croydon CR9 2BY Envigo Woolley Road Alconbury Huntingdon PE28 4HS National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs Gibbs Building 215 Euston Rd Londres NW1 2BE School of Biological Sciences Bangor University Deiniol Road Bangor Gwynedd LL57 2UW GSK Research and Development Park Road Ware Hertfordshire SG12 0DP RG Behavioural Biology and Animal Welfare Division of Behavioural Biology University of M nster Badestra e 13 D 48149 M nster Allemagne Division of Animal Health and Husbandry University of Bristol Langford House Bristol BS40 5DU UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN R sum Le groupe de d fense du bien tre des rongeurs et depuis peu des lapins de la RSPCA et de l UFAW se rassemble chaque automne pendant une journ e depuis 24 ans afin que ses membres puissent discuter des recherches actuellement men es en mati re de bien tre animal changer leurs points de vue sur les questions touchant au bien tre animal et partager leurs exp riences concernant la mise en uvre de la m thodologie des 3R Remplacement R duction et Raffinement relative l utilisation des rongeurs et des lapins L un des principaux objectifs du groupe est d inciter r fl chir sur l exp rience que subissent les rongeurs et les lapins de laboratoire au cours de leur vie et de s assurer que chaque impact n gatif potentiel sur leur bien tre est pass en revue et r duit au maximum La 24e r union a eu lieu l Animal and Plant Health Inspectorate APHA de Weybridge le 14 novembre 2017 La premi re session portait sur la mani re de r pondre aux besoins des animaux et de leur offrir une belle vie et incluait une tude de cas li e aux lapins reproducteurs Trois intervenants ont pr sent un expos sur le comportement des lapins reproducteurs sur le point de vue du Minist re de l int rieur concernant la mani re de fournir un milieu enrichissant appropri chaque esp ce et d am liorer leur habitat et sur les techniques d levage et les soins offerts ces animaux La deuxi me session qui portait sur le bien tre des rongeurs pr sentait les derni res actualit s concernant le Centre national pour le remplacement le raffinement et la r duction de l exp rimentation animale NC3Rs les v nements c l brant l ann e du bien tre des rongeurs de laboratoire la r duction du forage st r otyp des gerbilles la possibilit de mettre en place pour les rats un habitat collectif poss dant des boutons d acc s vasculaire les motions positives observ es chez les rats et les bonnes pratiques li es l habitat des souris m les La journ e s est termin e avec une s ance de discussion au sujet de la d fense des rongeurs durant les r unions de l AWERB Animal Welfare and Ethical Review Body entre autres Ce rapport r sume la r union et pr sente en conclusion une liste de mesures qu il est sugg r aux lecteurs de prendre en compte et de mettre en uvre au sein de leurs tablissements Mots cl s Bien tre rongeurs lapins les 3R conditions de vie 94

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Paper Summary Translations Nouvelles utilisations civiles innovantes des drones ou des v hicules a riens sans pilote dans les domaines de la science et des soins de sant et avantages et inconv nients GARY MARTINIC Faculty of Science University of Western Sydney Richmond NSW Australie Correspondance g martinic westernsydney edu au R sum Peut tre mieux connus pour leur capacit espionner et liminer l adversaire dans les r gions en guerre les drones comme on les appelle plus souvent sont galement utilis s des fins r cr atives depuis les ann es 60 au moins sous la forme d avions et d h licopt res t l command s Il est toutefois important de remarquer que le mot drone est en quelque sorte un terme inappropri et qu une description mieux adapt e consisterait utiliser les expressions v hicule a rien sans pilote UAV ou Unmanned Aerial Vehicle a ronef sans pilote UA ou Unmanned Aircraft ou encore a ronef command distance RPA ou Remotely Piloted Aircraft Heureusement outre leurs nombreuses utilisations militaires et la toute premi re livraison par UAV r ussie effectu e aux tats Unis les UAV civils sont de plus en plus utilis s dans les domaines industriel r cr atif commercial et de la s curit sans aucun lien avec les activit s militaires La raison pour laquelle ils sont si efficaces est que les UAV peuvent acc der facilement aux r gions loign es aux terrains complexes ou aux endroits difficiles d acc s et en comparaison avec les a ronefs avec pilote les UAV peuvent atteindre ces objectifs moindre co t Leur capacit parcourir de vastes surfaces dans une dur e de temps r duite tout en relayant des images a riennes en temps r el la base constitue un autre avantage consid rable Depuis quelques ann es ils ont galement prouv leur utilit dans diff rents domaines scientifiques tels que la recherche sur les ressources fauniques les activit s de protection et de lutte contre le braconnage l arch ologie r alis e dans les secteurs de l exploitation agricole et de l agriculture l oc anographie l exploration p troli re gazi re et min rale la gestion foresti re et la d tection distance Plus r cemment et plus en phase avec le domaine m dical et des soins de sant les UAV ont t utilis s des fins de test consistant livrer des vaccins dans des zones loign es de pays en voie de d veloppement Ils font galement l objet de tests sous la forme de petites ambulances a riennes les hollandais ont mis au point des drones ambulanciers capables d intervenir rapidement aupr s de victimes de crise cardiaque ils sont utilis s dans la police scientifique et ils sont m me test s dans le secteur des soins dispens s sur les champs de bataille Cet article d crit l int r t de ces nouvelles utilisations des UAV d un point de vue scientifique et m dical plus g n ral et tudie comment minimiser l impact de ces technologies sur la faune et la flore pr sentes sur ces terrains 95

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Animal Technology and Welfare August 2018 INDICE DE LA REVISTA Informe sobre la reuni n del Grupo para el bienestar de roedores y conejos RSPCA UFAW 2017 PENNY HAWKINS Secretaria 1 ANNE McBRIDE 2 GILES PAIBA 3 RITA ROSE 4 MARK PRESCOTT 5 JOHN MULLEY 6 SAM IZZARD 7 DEBORAH RIDLEY 7 LUCA MELOTTI 8 SARAH KAPPEL9 y ROBERT HUBRECHT10 1 2 3 4 5 6 7 8 9 10 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS School of Psychology University of Southampton University Road Southampton SO17 1BJ Home Office Animals in Science Regulation Unit 14th Floor Lunar House 40 Wellesley Road Croydon CR9 2BY Envigo Woolley Road Alconbury Huntingdon PE28 4HS National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs Gibbs Building 215 Euston Rd Londres NW1 2BE School of Biological Sciences Bangor University Deiniol Road Bangor Gwynedd LL57 2UW GSK Research and Development Park Road Ware Hertfordshire SG12 0DP RG Behavioural Biology and Animal Welfare Division of Behavioural Biology University of M nster Badestra e 13 D 48149 M nster Alemania Division of Animal Health and Husbandry University of Bristol Langford House Bristol BS40 5DU UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Resumen El Grupo para el bienestar de roedores y ahora tambi n conejos RSPCA UFAW ha celebrado una reuni n de un d a cada oto o durante los ltimos 24 a os para que sus miembros puedan debatir sobre la investigaci n actual sobre bienestar intercambiar opiniones sobre temas relacionados con el bienestar de roedores y compartir su experiencia respecto a la implementaci n de las 3 R reemplazo reducci n y refinamiento en relaci n al uso de roedores y conejos Uno de los objetivos primordiales del Grupo es fomentar que las personas piensen sobre la experiencia vital de los roedores y conejos de laboratorio garantizando a su vez que cualquier impacto negativo posible en su bienestar sea revisado y reducido La reuni n n mero 24 se celebr en el Animal and Plant Health Inspectorate APHA Weybridge el 14 de noviembre de 2017 La primera sesi n trat sobre satisfacer las necesidades de los animales y sobre conseguir una buena vida junto con las necesidades de los conejos de apareamiento como estudio de caso Tres ponientes hicieron sus presentaciones sobre el comportamiento de los conejos de apareamiento una perspectiva del Ministerio del Interior sobre ofrecer un mayor enriquecimiento seg n la especie y refinar el enjaulamiento la cr a y el cuidado de estos animales en la pr ctica La segunda sesi n sobre el bienestar de los roedores comprendi una actualizaci n sobre el Centro Nacional para el Reemplazo Refinamiento y Reducci n de Animales en la Investigaci n NC3R el A o del Bienestar de los Roedores de Laboratorio reducci n del escarbado habitual de los jerbos permitir el enjaulamiento en grupo de ratas con botones para acceso vascular emociones positivas en ratas y buenas pr cticas en el enjaulamiento de ratones macho El d a finaliz con un debate en el que se defendi a los roedores en reuniones del Animal Welfare and Ethical Review Body AWERB y en otras sesiones m s generales Este informe resume la reuni n y finaliza con una lista de puntos de acci n para los lectores puedan considerar presentarlos en sus propias instalaciones Palabras clave Bienestar roedores conejos 3R experiencia de por vida 96

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Paper Summary Translations Usos civiles novedosos e innovadores de drones o Veh culos A reos no Tripulados en los campos de las Ciencias y la Salud y sus pros y sus contras GARY MARTINIC Faculty of Science University of Western Sydney Richmond NSW Australia Corrispondencia g martinic westernsydney edu au Resumen Quiz s conocidos m s bien por su capacidad para eliminar adversarios en zonas en guerra y para espiar a personas los drones como suelen llamarse tambi n se han utilizado de forma recreativa al menos desde los a os 60 con aviones o helic pteros teledirigidos Sin embargo cabe mencionar que el t rmino dron es de alguna manera un nombre equivocado ya que una forma m s precisa para describirlos es un veh culo a reo no tripulado UAV un avi n no tripulado UA o un avi n pilotado de forma remota RPA Por suerte adem s de su uso militar ya establecido e incluso antes de que se creara el primer UAV con xito en los EE UU los UAV no militares se han utilizado cada vez m s en los sectores industrial recreativo comercial y de seguridad no militar El motivo por el que han sido tan eficaces es porque los UAV pueden llegar f cilmente a regiones remotas terrenos complicados o lugares dif ciles de alcanzar y cuando se comparan con el uso de aviones tripulados los UAV pueden conseguir esto de una forma mucho m s econ mica Su capacidad para cubrir mayores zonas en un plazo de tiempo inferior enviando registros a reos en tiempo real a la base de control es tambi n otra de sus principales ventajas En los ltimos a os tambi n han demostrado ser tiles en varios campos cient ficos entre los que se incluyen la investigaci n de la fauna operaciones de conservaci n y en contra de la caza furtiva arqueolog a de agricultura y ganader a oceanograf a silvicultura y exploraci n mineral gas petr leo y teledetecci n M s recientemente y m s en l nea con los campos de la salud y la medicina los UAV han sido testados para el transporte de vacunas a regiones remotas en pa ses en desarrollo Tambi n est n siendo testados como miniambulancias a reas los holandeses han creado ambulancias drones capaces de ofrecer una respuesta r pida para socorrer a v ctimas de infartos tambi n se est n utilizando en actividades forenses y se ha testado su capacidad en el campo de la medicina en campos de batalla Este art culo describe la aplicaci n de estos usos recientes de los UAV desde un punto de vista cient fico y sanitario m s amplio y tambi n trata sobre c mo minimizar el impacto de estas tecnolog as en la fauna del terreno 97

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Animal Technology and Welfare August 2018 INDICE DELLA REVISTA Relazione del convegno del Gruppo per il Benessere dei Roditori e Conigli RSPCA UFAW 2017 PENNY HAWKINS Segretaria 1 ANNE McBRIDE 2 GILES PAIBA 3 RITA ROSE 4 MARK PRESCOTT 5 JOHN MULLEY 6 SAM IZZARD 7 DEBORAH RIDLEY 7 LUCA MELOTTI 8 SARAH KAPPEL9 e ROBERT HUBRECHT10 1 2 3 4 5 6 7 8 9 10 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS School of Psychology University of Southampton University Road Southampton SO17 1BJ Home Office Animals in Science Regulation Unit 14th Floor Lunar House 40 Wellesley Road Croydon CR9 2BY Envigo Woolley Road Alconbury Huntingdon PE28 4HS National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs Gibbs Building 215 Euston Rd Londra NW1 2BE School of Biological Sciences Bangor University Deiniol Road Bangor Gwynedd LL57 2UW GSK Research and Development Park Road Ware Hertfordshire SG12 0DP RG Behavioural Biology and Animal Welfare Division of Behavioural Biology University of M nster Badestra e 13 D 48149 M nster Germania Division of Animal Health and Husbandry University of Bristol Langford House Bristol BS40 5DU UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Estratto Il Gruppo per il Benessere dei Roditori e ora Conigli RSPCA UFAW organizza da 24 anni un convegno di un giorno ogni autunno per consentire aisuoi membri di discutere dell attuale ricerca in materia di benessere animale scambiarsi opinioni su queste tematiche e condividere le esperienze di implementazione delle 3 R di replacement reduction e refinement sostituzione riduzione e perfezionamento relativamente all utilizzo di roditori e conigli Una delle finalit principali del Gruppo quella di incoraggiare a pensare all esperienza a vita dei roditori e conigli da laboratorio verificando che venga analizzato e minimizzato ogni potenziale impatto negativo sul loro benessere Il 24 convegno si tenuto presso l Ispettorato per la Salute di Animali e Piante Animal and Plant Health Inspectorate APHA di Weybridge il 14 novembre 2017 La prima sessione ha trattato la soddisfazione delle esigenze degli animali puntando a una vita buona usando i conigli destinati alla riproduzione come caso di studio Tre oratori hanno illustrato rispettivamente il comportamento dei conigli da riproduzione la prospettiva del Ministero dell Interno sulla fornitura di un arricchimento appropriato alla specie e il perfezionamento di alloggiamento allevamento e cura di tali animali nella pratica La seconda sessione incentrata sul benessere dei roditori ha incluso un aggiornamento sul National Centre for the Replacement Refinement and Reduction of Animals in Research NC3Rs sull Anno del Benessere dei Roditori da Laboratorio sulla riduzione dello scavo stereotipico nei gerbilli sull alloggiamento in gruppo dei ratti dotati di pulsanti per l accesso vascolare sulle emozioni positive nei ratti e sulle buone pratiche di alloggiamento dei topi maschi La giornata si conclusa con una discussione sugli interventi a favore dei roditori durante le riunioni dell Organismo per il Benessere Animale e la Revisione Etica Animal Welfare and Ethical Review Body AWERB e in un ambito di pi ampio respiro La presente relazione riepiloga il convegno e termina con un elenco di azioni che i lettori dovrebbero mettere in atto presso le rispettive strutture Parole chiave benessere roditori conigli 3 R esperienza a vita 98

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Paper Summary Translations Nuovi e innovativi usi civili dei droni o aeromobili a pilotaggio remoto nel campo della scienza e della sanit e i loro pro e contro GARY MARTINIC Faculty of Science University of Western Sydney Richmond NSW Australia Corrispondenza g martinic westernsydney edu au Estratto Forse meglio noti per la loro capacit di eliminare gli avversari nelle zone di guerra e di spiare le persone i droni come sono comunemente chiamati sono stati usati anche a scopo ricreativo almeno fin dagli anni 60 sotto forma di aeroplani ed elicotteri telecomandati Tuttavia va sottolineato che il termine drone una sorta di denominazione impropria Una descrizione pi precisa quella di aeromobili a pilotaggio remoto APR Fortunatamente oltre ai loro consolidati usi militari e ancor prima della prima consegna di un APR negli Stati Uniti gli APR non militari sono stati usati con maggior frequenza nei settori industriale ricreativo commerciale e della sicurezza non militare Il motivo per cui sono cos efficienti risiede nel fatto che gli APR possono raggiungere facilmente regioni remote terreni difficili o luoghi impervi a confronto con l utilizzo degli aeromobili con pilota gli APR possono operare a una frazione del costo La loro capacit di coprire grandi aree in tempi pi brevi inviando al contempo immagini in tempo reale alla base di comando un altro importante vantaggio Negli ultimi anni si sono dimostrati utili anche in vari campi scientifici fra cui ricerche sulla fauna selvatica operazioni di conservazione e antibracconaggio archeoagricoltura e allevamento oceanografia esplorazione di petrolio gas minerali silvicoltura e rivelamento remoto Pi di recente e in linea con il campo medico sanitario gli APR sono stati usati per la consegna di vaccini nelle regioni remote dei Paesi in via di sviluppo Sono stati testati anche come miniambulanze aeree gli olandesi hanno sviluppato i droni ambulanza capaci di offrire un soccorso rapido a persone colpite da infarto vengono usati nelle indagini forensi e le loro capacit sono state messe alla prova anche nella medicina sul campo di battaglia Il presente articolo descrive l applicazione di quest ultimi nuovi utilizzi degli APR da un ampio punto di vista scientifico e sanitario Spiega inoltre come ridurre al minimo l impatto di tali tecnologie sulla fauna selvatica 99

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August 2018 Animal Technology and Welfare ANDREW BLAKE TRIBUTE AWARD WINNING ENTRY Refinement of tumour passage by optimisation of the tissue disaggregation protocol MELANIE GALVIN ALICE LALLO STEWART BROWN PAUL KELLY BECKY BOLA DANIELLE POTTER CASSANDRA HODGKINSON KRISTOPHER FRESE and CAROLINE DIVE Clinical and Experimental Pharmacology Group Cancer Research UK Manchester Institute University of Manchester Alderley Park SK10 4TG Correspondence melanie galvin manchester ac uk Winner of the 2018 Andrew Blake Tribute Award Abstract Surgical implantation of tumour pieces has been used regularly in the passage of tumour models This involves an invasive surgical procedure carried out under anaesthesia ensuring mice have adequate pain relief and peri operative care The procedure also requires multiple tumour bearing donor mice to generate sufficient tissue for surgical implantation particularly for large scale efficacy studies We sought to refine our tumour passage methods and subsequently reduce the number of mice required for studies by implanting viable dissociated tumour cells instead of tumour fragments Tumours are disaggregated and depleted for red blood cells and any dead residual mouse cells This allowed the enrichment for human tumour cells that were mixed with Matrigel and implanted subcutaneously in the hind flank of mice Cell viability was successfully maintained with this method and mice developed tumours Due to the high number of cells generated using this protocol we are able to implant more mice per tumour thus reducing the number of donor mice required for large scale efficacy studies Moreover subcutaneous injection is a less invasive procedure with a milder severity than surgical implant under general anaesthesia Not only does this process improve animal welfare from a 3Rs perspective it also improves other aspects of the experiment by reducing intertumour heterogeneity thus improving the science Sponsored by The Association of the British Pharmaceutical Industry Keywords Mouse Tumour passage Refinement Reduction 3Rs Animal Welfare Introduction Small cell lung cancer SCLC represents 15 20 of lung cancers with 30 of patients presenting with limited stage LS or localised disease and 70 presenting with extensive stage ES metastatic disease Median survival time with treatment for these patients is 15 20 months for LS and 8 13 months for ES High response rates including complete responses are obtained with platinum based chemotherapy radiotherapy regimens however 5 year survival remains at 5 Drug resistant relapse is common and there is little benefit from second line therapy Since 2007 more than 100 clinical trials have started in SCLC but positive results have been very limited 1 One of the reasons for the slow improvement in SCLC clinical care is that patient derived preclinical models are difficult to generate mainly because of limited access to fresh tumour tissue for research purpose To overcome this problem we recently generated patient derived mouse models from the circulating tumour cells of ES SCLC patients 2 101

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Andrew Blake Tribute Award Winning Entry In Hodgkinson et al we described the processing of SCLC patient blood samples to enrich circulating tumour cells CTCs to produce tumours in mice for experimental use 2 These tumours or Circulating Tumour Cell Derived Explant CDX models accurately recapitulate the human disease and mimic the patient response to treatment in clinic CDX models have been successfully passaged and maintained growth characteristics however passaging has involved an invasive surgical procedure implant so from one tumour of 1000mm we are able to get 20 pieces and therefore 20 implants Disaggregation of a tumour of a similar size generates 85 million cells When using disaggregated cells we implant 100 000 cells per mouse allowing us to potentially implant 850 mice from one donor tumour Figure 3 120 Method CDX tumours were grown on the right flank of 8 16 week old female NOD Cg Prkdcscid Il2rg tm1Wjl SzJ NSG mice and removed at 800mm Tumours were enzymatically digested with a Human Dissociation Kit Milteyi Biotech and mechanically dissociated using a GentleMACS dissociator Miltenyi Biotech The dissociated tumours were incubated at 37 C for two periods of 30 minutes strained through a 70 m filter and centrifuged at 250g for 5 minutes The cell pellet was run through a negative selection column using mouse and dead cell beads Miltenyi Biotech The resulting cell pellet was either mixed with 50 50 Matrigel Cornig and implanted subcutaneously on the flank of NSG mice or cryopreserved Figure 1 80 60 40 20 0 Fresh Cells Cryopreserved Cells Fresh Pieces Figure 2 Time to enrolment size 200mm post implantation of fresh frozen cells or surgically implanted tumour fragments 900 Number of Implants Per Tumour To reduce the invasiveness of this procedure we decided to trial a tumour disaggregation protocol This protocol was already being utilised for ex vivo research purposes 3 Days post Implantation 100 Mice are anaesthetised and given a small dose of analgesia before a small incision is made on the flank to enable the implantation of a 3x3 mm tumour piece The mice are then surgically clipped before being allowed to recover 800 700 600 500 400 300 200 100 0 Disaggregated Cells Tumour Pieces Figure 3 Example of total number of implants per tumour Discussion The use of the tumour disaggregation protocol has significantly improved the passaging of CDX models reduced the number of donor and recipient mice required for studies and refined tumour passage Figure 1 Dissociation of CDX tumours for implant or cryopreservation Results Tumours have been successfully generated from both fresh and frozen disaggregated cells and the average growth time to 200mm decreased when using disaggregated cells compared to tumour pieces Figure 2 Furthermore the time to reach 200mm was much less variable when using dissociated cells instead of fragments The number of possible implants per tumour increases considerably when passaging tumour pieces we cut the tumour into 3mm chunks for 102 Tumour disaggregation reduces the number of recipient mice required for a study as tumours derived from disaggregated cells reach enrolment time at a more uniform time implanting tumour fragments requires more mice to ensure a sufficient number of tumours of similar size at the same time It is possible that this is due to the more homogeneous implant of tumour cells tumour fragments are likely to contain different amounts of non viable tumour tissue such as necrotic tissue The protocol also reduces the number of donor mice required for a large scale study as one tumour produces enough cells to implant large numbers of mice

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Andrew Blake Tribute Award Winning Entry Subcutaneous implant of tumour cells eliminates the need for invasive surgery carried out under general anaesthesia reducing the severity of the procedure and improving the welfare of the mice Surgical passage requires adequate peri and post operative care of the mice providing pain relief when necessary and close monitoring of the animals to ensure full recovery from the anaesthesia Tumour disaggregation also allows depletion of dead cells and contaminating mouse stromal cells to ensure a more homogeneous tumour cell suspension for implant This subsequently reduces intertumour heterogeneity and improves the quality of the experiment References 1 2 3 4 Koinis F Kotsakis A and Georgoulias V 2016 Small cell lung cancer SCLC no treatment advances in recent years Trans Lung Cancer Res 5 1 39 50 Hodgkinson C L Morrow C J Li Y et al 2014 Tumorigenicity and genetic profiling of circulating tumor cells in small cell lung cancer Nat Med 20 8 897 903 Potter D S Galvin M Brown S et al 2016 Inhibition of PI3K BMX Cell Survival Pathway Sensitizes to BH3 mimetics in SCLC Mol Cancer Ther 15 6 Lallo A Schenk M Frese F et al 2017 Circulating Tumor Cells and CDX Models as a Tool for Preclinical Drug Development Transl Lung Cancer Res 6 4 397 408 Tumour disaggregation and subsequent ex vivo culture could also be used as a potential replacement for in vivo studies 4 Disaggregated CDX cells have been cultured in vitro and accurately recapitulate the genotypic phenotypic and pharmacological aspects of the donor CDX tumour Lallo et al submitted suggesting that ex vivo culturing of CDX cells can be used to screen several compounds and may provide an insight into the mouse CDX sensitivity Therefore we will be able to reduce the number of mice used for pharmacologic studies focussing only on those studies that looked promising in vitro More studies are being carried out and testing of direct culture of CTCs from the patients is under evaluation Taken together these data open a new window in the study of SCLC and will help reduce the number of mice used to investigate this disease Conclusion The tumour disaggregation protocol is the enzymatic digestion of CDX tumours for subcutaneous cell implant This reduces the number of mice required as donor and recipient animals and refines tumour passage by eliminating invasive surgery The removal of both dead cells and mouse cells using the negative selection column results in implantation of tumour cells only improving the success rate of tumour growth and their homogeneity Cell cultures of disaggregated CDX models recapitulate in vivo drug studies and therefore could be used as a possible replacement for in vivo studies in the future Acknowledgements I would like to thank Professor Caroline Dive and Dr Kris Frese for their continued support and the Clinical and Experimental Pharmacology in vivo team for their hard work I would also like to thank Alice Lallo for providing the cell culture data and for her constructive criticism of this manuscript 103

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August 2018 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 Blunting cutting canines of adult male monkeys a discussion by the Laboratory Animal Refinement Enrichment Forum CASEY COKE EVELYN SKOUMBOURDIS JEANNINE CASON RODGERS KELSEY FINNIE LOUIS DIVINCENTI MARCIE DONNELLY MELISSA TRUELOVE MISTY WILLIAMS FRITZE POLLY SCHULTZ VANESSA HERRING and VIKTOR REINHARDT Correspondence viktor cot net When you keep adult Rhesus macaque Macaca mulatta males adult Cynomolgus Macaca fascicularis males or adult Vervet males Chlorocebus pygerythrus in the same enclosure is it advisable to have the canines of the males blunted or cut Reinhardt Discussion I have worked with pair and group housed male macaques who have had intact blunted or pulpotomised canines My personal preference is to keep the canines fully intact or have only the tips blunted Crush injuries from amputated canines can be much more severe and dangerous than a clean cut injury inflicted by an intact or slightly blunted canine It is sometimes indicated to blunt the tips of a male s canines because of overgrowth of the teeth or in older animals who develop a slight off set of the jaw Slightly blunting will prevent the tips of the canines from injuring the interior of the lower lip or other areas of the oral cavity As for pulpotomies I ve seen too many begin to infect or develop abscess over time I reserve pulpotomies for situations when it is necessary to remove the canine due to a severe infection or break of the tooth Skoumbourdis I would not blunt a canine to reduce injury severity from aggression as the crush injuries can be just as bad or even worse due to the tissue damage If animals are 105

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Tech 2 Tech likely to fight then group compatibility should be re assessed USDA has a policy statement on canine blunting cutting Policy 3 recognises that the removal or reduction of canine teeth in non human primates can cause ongoing pain discomfort or other pathological conditions 1 In its Policy on Teeth Reduction and Removal the American Veterinar y Medical Association is opposed to removal or reduction of healthy teeth in nonhuman primates because the animals may still cause severe injury with any remaining teeth and this approach does not address the cause of the behaviour Removal or reduction of teeth for non medical reasons may also create oral pathologic conditions To minimise injury recommended alternatives to dental surgery include behavioural assessment and modification environmental enrichment changes in group composition and improved animal housing and handling techniques 2 DiVincenti We blunted and by blunting I mean slightly rounding the tip without opening the pulp the canines of male macaques before group formation Compared with males who did not have their canines blunted we saw reduced severity of bite injuries during group formation and less severe injuries requiring veterinar y intervention We did not observe more crushing injuries in the group in which the males had the tips of their canines slightly blunted and rounded These males made normal use of their canines including using them for open mouth threat displays I do not believe in canine extraction unless the tooth is seriously damaged infected We saw horrible mangled mouths in some of the adult male macaques who had their canines extracted and were later retired to us their premolars and incisors had moved and some had problems eating because over time their bite no longer lined up Coke blunted back when it was the norm Not only were they still very active self biters but many of them suffered from serious dental issues that only got worse as they aged Herring I agree we have never extracted cut or blunted canines in any macaque species but I have treated root abscesses in animals who had their canines blunted before they arrived at our facility It has been my experience that crushing bite injuries inflicted by blunted canines are more complicated to heal than puncture bite injuries inflicted by sharp intact canines Donnelly Two incidents have been documented of male rhesus macaques killing other social partners as a result from fighting even after their canines had been cut at the level of adjacent teeth 3 Reinhardt Blunting canines on male macaques in an attempt to minimise serious injury in the event of a fight is well intentioned Nobody wants to walk into a room in the morning and find a dead macaque from an unexpected overnight fight but is it really the best thing to do We ve put these animals in an unnatural challenging position by creating tight boundaries in which they can t escape or flee from a threat be it an approaching human or a dominant conspecific We thought years ago that minimising the risk of bite injuries by simply removing or at least blunting the males long canines was a humane option I questioned this assumption after talking with a researcher who was conducting a In my experience if adult males are compatible with one another blunting their canines is not necessary In fact I have seen tooth root abscesses resulting from improperly blunted canines Although sharp canines can be very damaging blunted ones can too WilliamsFritze Agreed if adult male companions are compatible blunting their canines is not necessary Truelove I am very much against blunting canines Yes large canines can inflict deep puncture injuries but they also tend to be relatively uncomplicated to heal and repair In my opinion crush injuries inflicted by blunted canines are usually more damaging and take considerable time to heal properly I managed a severe self injurious biting SIB colony of male rhesus macaques for many years The males came from other facilities where they were listed for euthanasia but donated to us instead Many of them had their teeth 106 Figure 1 Display of canines to intimidate opponent Photo Mark Dumont

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Tech 2 Tech study on the impact of blunted canines vs natural canines on fighting and injuries in their breeding colony of rhesus macaques It turned out that males with blunted canines fought more frequently and for longer durations than those with intact canines Males with intact canines were able to avoid fighting in most cases because by flashing their big intimidating weapons it was easier for them to make opponents submit or flee rather than pick a fight In light of these findings we felt it was better to leave the canines of our males intact and implement every precaution possible to prevent or minimise the risk of bite lacerations It is a lot more work for us but in the end much better for our animals We searched for and discovered ways to reduce or eliminate stressors and other environmental contributors that might lead to fighting We are always on our toes because every time we think we have every possible issue covered another macaque comes up with a completely unique and new idea of how and why he is going to bite his cousins For us learning and refining is likely to never stop Schultz All comments to our present discussion seem to be related to macaques Can anybody who is working with vervets African Green Monkeys share observations to this discussion Reinhardt Oh boy the first time I worked with vervets I was like MAN those are big canines Sabre tooth monkeys I am in the middle of trying to pair male AGMs in a colony of 20 It is pretty much a nightmare Neither one of our vets has even suggested blunting Those canines are so long and so sharp I guess blunting them might help to minimise damage Cason Rodgers Historically we have blunted the canines of male vervets and some females as their canines are just as large as the males to reduce severity of injury during altercations I cannot objectively say if it helped It certainly does not reduce aggression nor the number of injury incidences but it may reduce severity of sustained wounds In general if done correctly without opening the pulp blunting the teeth does no harm We have plenty of old vervets

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Animal Technology and Welfare August 2018 Phenotypes and characteristics of the Pole4 mice an exploration into what makes these animals unique LOUIS KITCHENHAM Sainsbury Wellcome Centre 25 Howland Street Fitzrovia London W1T 4JG Correspondence louis kitchenham ucl ac uk Adapted from a 2018 IAT West Middlesex Branch Technicians and Trade Day Young Presenters paper Pole4 mice How do they occur Why are they important How do we care for them Pole4 were an integral component of research at the Cancer Research UK laboratories Pole4 DNA Polymerase Epsilon 4 Accessory Subunit is a Protein Coding gene Among its related pathways are Pyrimidine metabolism KEGG and CDK mediated phosphorylation and removal of Cdc6 Gene Ontology GO annotations related to this gene include sequencespecific DNA binding and DNA directed DNA polymerase activity Their origins arise from a hybrid C57BL 6 background and the majority of animals looked like a typical laboratory mouse They were commonly crossed with various other strains within the colony with no issues However occasionally they did develop rather strange physical characteristics These changes affected various parts of the animal s body White patch on the belly Figure 1 Pole4 mouse showing white patch on abdomen Abnormal face and nose This is usually detectable from an earlier age typically 6 7 days The facial structure of the animal seems distorted and pushed in This can usually be spotted from an early age typically 8 9 days old when fur begins to develop A small white patch becomes visible on the underside of the animal which strangely does not seem only to be part of the coat or fur colour but actually the skin of the animal itself It can be a large or small patch or sometimes adjoining dots or lines Since starting work at my current facility my line manager tells me that she had encountered these animals before She also said that this phenotype the white patch was used as an alternative method of genotyping as the white patch would indicate that the animal had the desired genetic make up This helps to adhere to the 3Rs goal of refinement less stress for the animal and no need for a biopsy to be taken 108 R L Figure 2 Shows abnormal facial features of Pole4 mouse right compared to those of normal mouse left

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Tech 2 Tech The nose is slightly flattened while the eyes are more closely pushed together While it looks quite strange there are no signs that it impairs the animal s health or quality of life Bent kinky tail This is typically visible from 9 10 days and is expressed as a dramatic bend or kink in the tail It can range from a small kink in the centre or the end of tail to a W shape completely distorting the appearance To touch it feels broken but it appears not to cause the animal any pain or discomfort However there have been instances when animals have been found dangling from the food hopper as the tail was bent in such a way that it was like a hook While this at first can seem quite comical it could have caused a real welfare issue if staff were less than vigilant and the animal had been left overnight or over a weekend in that situation This defect can affect the animal s early life and they must be provided with a wet mash diet of small pellets of high protein extruded rodent diet which have been soaked overnight in water This enables the animals to eat more easily as the odd shape of their heads appears to restrict their ability to access the overhead food hoppers It seems to help these animals mainly due to the fact they were quite small and runty anyway and the higher protein ratio gives them a physiological boost Figure 4 X ray of Pole4 mouse skull showing domed characteristic Various examples of skull defects Figure 3 Mouse exhibiting kinked tail Trembling and walking This is typically visible when the animals begin to walk freely from 10 days onwards They seem very unsteady and tremble slightly as if trying to regain balance If you can imagine a little drunk mouse Caused by mild scoliosis of the spine the result of a genetic mutation Figure 5 Shows mice carrying the Pole4 gene compared to mouse of homozygous genotype Domed head This is usually not apparent until animals are between 14 21 days old The head of the animal has a domed appearance and it looks as if the cause may be hydrocephalus fluid on the brain 109

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Tech 2 Tech Abnormal Kink Curly Abnormal White face in tail tail walking patches 35 29 83 20 3 28 80 8 23 57 8 5 It is characterised by intrauterine growth retardation and postnatal dwarfism with a small head narrow birdlike face with a beak like nose large eyes with downslanting palpebral fissures receding mandible and intellectual disability All 3 phenotypes 2 phenotypes 1 phenotype only 15 43 15 43 2 6 More closely comparable with Facial Dysmorphism Immunodeficiency Livedo reticularis and Short Stature FILS Syndrome which is a syndrome caused by a Pole1 mutation The reason for these phenotypes seems to arise during the embryonic stage Total Table 1 Incidence of phenotypes What next The tables illustrate the rate at which these physical characteristics occur within a specific cohort of the mice The data helps to show how these animals can be affected by one or a combination of all phenotypes It is clear to see that on many occasions the mice exhibit 2 or 3 different phenotypes and quite rarely are they only affected by one Why does this happen When creating the Pole4 strain the mice unexpectedly began to exhibit many features suggestive of Seckel Syndrome When an animal within the colony displaying these phenotypes was identified the user of the phenotype was notified The animal would then be transferred to the researchers whereby an aging study would be conducted Preliminary PM analysis showed Pole4 mutants have a higher incidence of lymphomas over the course of their life Due to the tumour susceptibility of the Pole4 animals tumour growth and how this may be affected by the phenotypes and mutations would also be monitored How does this help us A mouse model of Seckel syndrome has been developed This mouse model is characterised by a severe deficiency of ataxia telangiectasia and Rad3related protein ATR These mice suffer high levels of replicative stress and DNA damage Adult Seckel mice display accelerated aging These findings are consistent with the DNA damage theory of aging The p53 transcription factor is commonly known as a tumour suppressor but also plays a fundamental role in regulating mammalian ageing which would help to explain the interest for this project As a result of this information and the various other similarities the researchers anticipated that the Pole 4 animals may exhibit the same adverse phenotypes as the Seckel mice In basic terms these phenotypes occur when the mouse is growing as an embryo the replicative stress means the foetus does not grow correctly causing these mutations It was deduced this must be happening at the same rate in the Pole4 mice Editor s note Seckel Syndrome or microcephalic primordial dwarfism also known as bird headed dwar fism Harper s syndrome Virchow Seckel dwarfism and Bird headed dwarf of Seckel is an extremely rare congenital nanosomic disorder Inheritance is by an autosomal recessive gene Beneath the surface Helped me to learn more about the reasons behind some of the physical characteristics which are present due to genetic manipulation It is all very well seeing phenotypes it s another task to fully understand why they occur Different strokes These phenotypes affect the mouse s anatomy in various different ways as such our care for them must be tailored to suit this It is fine to refine Adhering to the principles of the 3R s is of paramount importance to any scientific research carried out in the UK Technologists and researchers should at all times be looking for ways to do so Alternative handling methods for mice with kinky tails using the white patch as an indicator of genotype are examples of ways in which we as technologists can refine our techniques to suit the welfare needs of these animals Welfare above all While severity limits and protocols govern how far an experiment can go we must never lose sight of the fact that these are living animals and we have the responsibility of ensuring their care is of the highest standard possible We must understand the science but also be conscientious of the lives that are at stake and the lives that allow us to push our knowledge of the world forward 111

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Animal Technology and Welfare August 2018 Optimising mouse production good practice for efficient colony management and implementation of the 3Rs HANNAH EASTER The Francis Crick Institute 1 Brill Place London NW1 1BF Correspondence hannah easter crick ac uk Based on a Poster displayed at IAT Congress 2018 Introduction This poster provides an overview of the methods used to minimise mouse production through various techniques It will look at ways for scientists and technicians to produce the required numbers of genetic models of interest from fewer litters and reduce the overall numbers of mice produced The main techniques utilised and described in this poster include the use of a database to see live breeding data calculating number of mice required for experiments using punnet squares to decide on best breeding pairs and pedigree charts to record lines the efficient use of cryo preservation and tick over protocols for minimally used lines and using economic breeding life to maximise productivity of pairs Optimising breeding performance at the Francis Crick Institute The Francis Crick Institute is a newly opened institute with scientists and staff coming from multiple sites into one building Being a central London location effective use of space allocation is critical One of the measures that the Biological Research Facility BRF is implementing is to have clear information and guidelines on effective colony management We are providing various tools and services that can assist the people responsible for mouse colonies to plan their experiments and optimise breeding performance Providing information of basic colony management and how to set up breeding schemes to fit in with the economic breeding life of the mice Cryopreservation of lines which are not needed live on the shelf or as a safeguard in case anything happens to a mouse colony e g disease or failed experiment Breeding of communal lines which any user group can use to minimise repeat colonies and reduce genetic variation within the Institute 112 Workshop training is provided for user groups and colony managers which provides practical advice on how to estimate animal cohort size carry out statistical tests experimental analysis breeding calculations and randomisation Placing strains on tick over when suitable and providing assistance to help the colony managers do this effectively Providing an effective database so colony managers can look at live breeding data and also call up reports to allow them to check colony breeding data and then request alterations to current breeding pairs if needed Using methods to optimise breeding such as checking a female s oestrus phase before placing in a timed mating to increase the chances of successful pairings This has seen around an 80 increase in successful timed matings within the Francis Crick Institute breeding units These measures together with good communication between the technician and researcher is helping to optimise breeding within The Francis Crick Institute and reduce cage and mouse numbers Considerations for breeding animals It is important to consider various factors when choosing which mice to use for breeding and the appropriate breeding strategy Relevant to all strains Consider the economic breeding life of the mouse which includes looking at the size health survival rates sex ratio of the litters produced and the ability to nurture the litters in the female

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Tech 2 Tech Is the strain free from unexpected abnormalities and phenotypes Correct genotype to give the best ratio of useful litters Correct temperament for breeding were the previous generations good breeders Do you want to have pairs or trios Trios will produce more mice but you may not know which female is the mother of the litter Strain dependent What background strain is the mouse Some background strains such as CD1 have larger litters compared to C57Bl 6J mice Is there an issue with the breeding of this strain Some Genetically Altered GA lines are known to be sub fertile when homozygous Are there any phenotypes in this strain Phenotypes such as APC Min are health status dependent and mice could get sick from 3 6 months of age Do you need to set up more breeders to compensate Does the strain have Green Fluorescent Protein GFP or similar incorporated This will allow for easier identification of the correct allele without biopsies Can your line become homozygous increasing the number of suitable mice per litter Or is there a lethal phenotype pertaining to the genotype Do you have a low requirement of mice for your experiment or will there be periods when your usage is lower than at other times Can it be put into a tick over breeding strategy Depending on what is required this can be based around number of mice age or the genotype of the animals As you can see below Figure 2 you only achieve a 1 64 chance of obtaining a triple KO but there are also many potential genotypes which can be used for a fur ther cross to increase triple knockout KO generation Figure 2 Table showing incidence of genotypes in triple KO production In the second mating Figure 3 if you cross any combination of single KO het het with each other a 1 16 chance of the triple KO required is achieved Figure 3 Table showing increased incidence of triple KO If you crossed a double KO het you would increase your chance of getting the triple KO to 1 4 with all offspring being useful for further matings Figure 4 Use of Mendelian genetics to set up pairs Observing the rules of Mendelian Genetics when choosing mice for pairs can help to generate more of the required genotype mice in fewer litters This can be done with simple single allele strains or can be used for multiple allele Genetically Altered GA lines Het x Het x Het x Het x Het x Het AaBbCc x AaBbCc Figure 4 Table showing higher frequency of triple KO Breeding Having a good database where you can easily see all the live breeding data makes colony management a lot easier The first step is to ensure that all users of the database are correctly trained in its use At The Francis Crick Institute we provide training to all new starters both researchers and technicians Being able to see the following data will help plan experiments as well as assess the current breeding levels Here are some data points which can help G G G G Figure 1 Table showing incidence of inheritance WT Wild Type HET heterozygous HOM Homozygous G G number of pairs in each strain date of birth of offspring number of offspring each pair has had number of males and females in each litter number of days since pairing number of failed litters if any 113

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Tech 2 Tech Good databases will have the ability to generate reports which give you breeding statistics so you can see how successful a strain is in regards to G G G G G litter size sex ratio time between litters pre weaning loss this information will allow you to adjust the breeding scheme accordingly It is also a good way for you track individual mice back to the founding pair so the ability to pull up a pedigree chart of breeders for that strain is an invaluable tool to have on a database Figure 7 shows an example of what a colony dashboard looks like on a database the main features are G G G G genotype variations and how many mice of each there are how many breeding pairs the strain has any homozygous lethality related to specific alleles also any other notes or flags the strain may have Calculating the number of mice matings needed By calculating the number of mice needed before starting any experiments you can reduce the chance of over under breeding Some experiments will have special requirements such as only needing male mice etc This will need to be taken into consideration when designing your experiment There are many different tools online which can help with planning cohort size Two examples are Figure 5 Example of pedigree chart It shows a founding breeder and all the breedings that have come from this original mating Experimental Design Assistant Found on the NC3Rs website This is a free online tool which is designed to help researchers ensure they use the minimum number of animals for their specific objectives https www nc3rs or g uk experimental designassistant eda Breeding Colony Size Planning Worksheet The Jackson Laborator y have a worksheet you can download which uses a calculation to help you decide how many pairs you need to set up and how many mice you will need for it https www jax org jax mice andser vices customer suppor t technicalsuppor t breeding and husbandr y suppor t colonyplanning Cryopreservation Figure 6 Example of a pair and litter housed at The Francis Crick Institute The mice are displaying the variant of the Tomato allele which gives them the distinctive pink colouring Cryopreservation in regards to laboratory animals is the process of freezing genetic information for a particular strain which can then be used in the future to re derive when needed This is useful if a strain is not going to be used for a long time it can be cryopreserved and then the live animals can be removed from the shelf The strain can then be brought back when required It is also good to freeze strains even if they are still needed live on the shelf so there is always a back up if something happens to the colony Why is it important At The Francis Crick Institute we are trying to reduce the number of mice produced using the methods featured on this poster It is not only ethical to try and reduce overall mouse numbers but also helps when working to set total cage allowances within the BRF Figure 7 Colony dashboard 114 This is an industry wide issue in 2016 1 65 million GA mice were created but were not used in further experiments

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August 2018 Animal Technology and Welfare POSTER PRESENTATIONS Originally presented at IAT Congress 2018 A refinement to Schedule 1 HELEN READ and RHYS PERRY Department of JBIOS Cardiff University College of Biomedical and Life Sciences College House King George V Drive East Heath Park Cardiff CF14 4EP Correspondence perryr2 cardiff ac uk Winner of the LBS Biotech Best Poster Award Introduction For the last decade Joint Biological Services JBIOS at Cardiff University have inver ted animals for intraperitoneal IP injection It was thought that tipping the head and body downward would move viscera away from the surface of the ventral abdomen creating space to inject 1 2 Due to the risk of injecting into the caecum on the left hand side 3 4 intraperitoneal injections are carried out by injecting into the lower right quadrant of the animal s abdomen However within the context of animal research there are varying methods used for animal restraint and angle of injection this appears to be due to personal preference and experience It remains unclear whether one method is more accurate than the others the time of death and animal stress administering an anaesthetic overdose when Aims and objectives The overall aim was to investigate if the Schedule 1 method of Overdose of Anaesthetic via IP injection could be refined The primary objectives were to examine whether inverted restraint resulted in more effective outcomes and whether this depended on the angle of injection In addition to assessing the effectiveness of injections through the time taken for the overdose to be effective we also examined the stress effects of the manipulation Method Under the Animals Scientific Procedures Act 1986 5 an Overdose of Anaesthetic is a Schedule 1 method commonly carried out via IP injection Most animals succumb to the overdose of anaesthetic within ten minutes however on numerous occasions we have observed animals that are still breathing after thirty minutes This is an animal welfare issue Schedule 1 should be as quick and humane as possible To address these issues we carried out a study to examine the impact of restraint and injection angle on It would have been unethical and unnecessary to breed or order animals for this study so we utilised 126 animals that had reached the end of their experiment and were marked up to be culled This provided us with accurate data of the effect as information was obtained from rats of varying strains ages weights and experimental backgrounds Within each cohort every animal was weighed and numbered 1 6 prior to random selection for each injection 117

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Poster Presentations The following information was recorded for each animal G G G stress level time conscious time of death Stress level was scored on a scale of 1 3 assessing animal resistance and vocalisation levels while carrying out the technique To ensure consistency the technologist restraining the animals was solely responsible for stress scoring Stress Score 1 Mild 2 Moderate 3 High Figure 5 Stress scoring chart In terms of correlations the single best predictor of time of death was animal weight R 0 27 BF10 92 0 which is consistent with the fact that the same dose was administered regardless of weight There was no evidence that stress was related to any other measure largest R 0 18 BF10 0 74 for the stress and time conscious correlation Conclusions Even though we used a wide variety of animals there was little difference in the timings of loss of consciousness and death for all of the IP injection methods used Unexpectedly changing the angle of animal restraint and injection did not appear to have an effect on Euthatal uptake However this needs to be explored further to investigate needle placement in the hope of not only refining this method for other species but also for procedural IP injections We observed that restraining rats upright reduces the stress of an IP injection Therefore we strongly recommend using this technique to refine this Schedule 1 method As soon the as the injection was completed a timer was started the animal was monitored until death The time for loss of consciousness was recorded once the animal had lost its pedal reflex while time of death was recorded once permanent cessation of circulation was confirmed This was assessed by the Animal Technologist applying gentle pressure to the animal s thorax with their first two fingers Results Time Conscious Bayes ANOVA analysis demonstrated that restraint angle did not influence time conscious BF10 0 09 and although perpendicular injections resulted in numerically longer times this Bayes analysis suggests the evidence was more consistent with no effect BF10 0 46 There was no interaction between these factors BF10 0 03 Time of death Bayes ANOVA analysis demonstrated that restraint angle did not influence time of death BF10 0 14 Perpendicular injections did result in longer times BF10 4 43 Although there was no interaction between these factors BF10 0 12 the main effect of restraint angle was predominantly driven by the animals injected while inverted as this was the only case in which there was evidence that perpendicular and shallow injections differed BF10 3 70 Stress Bayes ANOVA analysis demonstrated that restraint angle had a large effect on stress BF10 1000 There was no evidence of an effect of injection angle BF10 0 26 or interaction between the two factors BF10 0 60 Acknowledgements A huge thank you to Dominic Dwyer who has been a great help throughout this study and has coached us through the statistics 119

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August 2018 Animal Technology and Welfare Improving rabbit enrichment developing a rolling enrichment plan KATIE PEARSON and JENN GANT Fera Science Ltd Sand Hutton York YO41 1LZ Correspondence katie pearson fera co uk jenn gant fera co uk Introduction Enrichment should provide animals with an environment that meets their needs be specific to the physiological and ecological traits of the species and increase the frequency and diversity of positive natural behaviours 1 2 An ethogram was developed to aid in consistent identification and description of behaviours 3 A time budget was formulated listing common rabbit behaviours from the ethogram 4 A CCTV camera was placed in the corner of the pen to give a wide field of view to observe behaviour Enrichment may have beneficial neutral or harmful effects Therefore it is important to evaluate such effects before implementation The study 1 Data was recorded initially for 3 days with no enrichment as a control 2 The pen was recorded for 18 hrs containing each item individually 3 Behaviour from 5 sets of one minute samples of CCTV footage for each item was observed and recorded on the time budget Figure 1 Female New Zealand White Rabbit A three week enrichment preference study trialling different substrates food and pen structures was carried out in three group housed female New Zealand White Rabbits NZW Oryctolagus cuniculus Figure 2 A circular table which is a repurposed empty wooden cable drum Objectives 1 Determine preference of enrichment items 2 Evaluate items resulting in an increased frequency of positive behaviours 3 Develop a rolling enrichment plan that provides a varied programme of stimulation Method Observation and data recording 1 A checklist was placed in the animal room to track introduction of enrichment items Figure 3 Organic cabbage hung by wooden pegs from a natural fibre washing line 121

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Poster Presentations Figure 4 A bed of paper wool nesting material Figure 7 The total proportion of time within 5 1 minute samples that three female rabbits were observed interacting with 14 distinct enrichment items individually added to the pen The dashed line highlights incidences when 15 of observed time was spent interacting with enrichment items Food The highest proportion of time was spent with the washing line Figure 2 28 1 Substrate Straw Figure 5 ball pit and paper wool Figure 3 substrates were interacted with the most at 38 8 35 2 and 32 9 respectively Figure 6 Figure 5 A variety of cardboard items including egg boxes tubes and urinal bottles Behavioural Results Figure 6 A large bed of straw Results Preference results Structural The largest proportion of observed time was spent with the circular table Figure 1 45 9 122 Figure 8 The total proportion of time within 5 oneminute samples that three female rabbits were observed stationary when 14 distinct enrichment items were added to the pen and when no enrichment items were added The dashed line highlights incidences when the same proportion of time was spent stationary compared with a non enriched pen 45 45

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Poster Presentations Structural The highest percentage reduction in stationar y behaviour compared to the non enriched pen was seen in the hessian at 19 3 Food The highest percentage reduction was seen in the washing line 33 9 Substrate The addition of paper wool 32 7 caused the highest stationary reduction Figure 7 pit was replaced with small cardboard items Figure 4 Fera Science Ltd Form Acknowledgements Our thanks goes to Heather Warrender for creating the initial rolling enrichment template and ethogram design Kelly Hunter and Nikki Cook for their continued assistance implementing the enrichment Matt Brash NVS and Liz Brown NACWO for guidance on enrichment item suitability and animal welfare advice Patricia Pimlott for creation of the rolling enrichment form and poster editing and lastly to the Animal Technicians at Fera Conclusion Preference Preference was defined by interaction for 15 or more of the rabbits total observed time This was noted for the circular table hessian and platform structures the washing line and the hay rack food items and the straw ball pit paper wool and bark chipping substrate References Wolfensohn S and Lloyd M 2013 Handbook of Laborator y Animal Management and Welfare Oxford University Press Oxford Behaviour A correlation between the addition of enrichment items and reduction in stationary behaviour was suggested in almost all items This indicates that the pen environment was more stimulating for the animals when enrichment is added Figure 9 A rolling enrichment form for use in animal rooms This example includes rabbit enrichment items tested in this preference study Rolling enrichment plan A rolling enrichment plan was created using the preferred items A form was created to enable technicians to plan and record the use of enrichment items and can be used for other species Figure 8 Post study welfare advice The ball pit and hessian were eliminated due to chewing and potential ingestion of the items The ball 123

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Animal Technology and Welfare August 2018 Design by intravenous injection in the Syrian hamster GEORGIA BEAMISH C CHAPMAN and A LOMAX Envigo Woolley Road Alconbury Huntingdon Cambridgeshire PE28 4HS Correspondence val summers envigo com Introduction When testing the safety of novel pharmaceuticals at Envigo we often need to administer materials intravenously to replicate the intended therapeutic route This is routine in rats and mice using the tail vein Recently a customer asked us to use this route to administer their test material to hamsters as other rodent models were not suitable for this compound Clearly the tail was not an option In this poster we explore the development and refinement of intravenous dosing in the Syrian hamster via the jugular vein and explain our current method and equipment Method development time and resources would be outweighed by the benefit of having non obstructed and clear access There was also the potential for the anaesthetics analgesics and antibiotics if used to interfere with our ability to determine any treatment related effects of the test material Additionally we needed to administer the test material on multiple occasions the welfare cost of repeated surgical access was considered unacceptably high We stepped back and looked again at the method for jugular bleeding in the rat By placing a hamster under light anaesthesia approx 1 to 2 minutes it was possible to perform a secure jugular sampling hold with access to the vein on both sides Where did we start There was ver y limited information available for intravenously dosing hamsters but during our initial investigations we learned that technologists in Spain had been administering intravenously to hamsters via the cephalic vein Our technicians went to Spain to observe the procedure and although we discovered the cephalic vein was an option the success rate was low at only 50 with 0 1 mL we were required to administer 0 27 mL What other route could we use As we were unable to achieve a high success rate using the cephalic vein we needed to investigate a reliable alternative We already routinely collect blood samples from rats and mice via the jugular route so this seemed to be a sensible option to investigate The hold for jugular bleeding is a secure way of keeping the animal still and giving access to the jugular vein To achieve the hold the handler must tuck both left and right forelimbs of the animal between their middle finger and thumb behind the animal s back and pushing the clavicles and sternum slightly forwards The index finger is then placed on top of the back of the animal s head and brought downwards along its back causing its head to raise This gives full access to the neck area see illustrations Each step of hold The jugular vein on a rat is visible under the clipped area of skin on the ventral neck and rats can be successfully restrained and sampled without anaesthesia however on a hamster the visibility of the jugular vein is more limited and the loose skin and movement of the animal would make restraint and sampling very challenging Due to these factors we discussed surgical administration surgical anaesthesia and excision of the skin to enable full visibility and access to the exposed jugular vein Although having full access to the jugular vein would improve the success rate hugely we had to decide whether the welfare costs for the animal as well as 124 Figure 1 Illustration of holding a hamster for intravenous administration

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Poster Presentations Method Once the hamster was anaesthetised and in a secure hold the Animal Technologist performing the procedure would remove the fur from the area over the vein with small animal clippers to provide a clear view As can be seen from Figure 2 two small darker areas are visible this is where the jugular vein is located If the vein is still not clearly visible a small piece of dampened cotton wool wiped over the skin can be used to encourage the vein to show Once the Animal Technologist is confident that the vein has been accessed the test substance can be administered The area in which the needle has entered the skin is observed during administration to ensure the test substance is continuing to enter the vein If the needle is no longer in the vein a visible swelling under the skin would arise If this occurs administration is paused and a fresh needle attached to the syringe before attempting to relocate the vein When the test substance has been fully administered the needle is carefully removed and discarded and a swab is pressed on the area to stop any bleeding from needle insertion The animal is then placed into its home cage and observed until fully recovered from being anaesthetised Figure 2 Dark colouration of skin indicates location of the jugular vein Results Three studies have been performed with a total of 452 hamsters dosed 120 of these being repeat dose administration once weekly for 4 weeks alternating between left and right jugular veins with 100 dosing success rate Bruising post dosing at the injection site has been negligible Conclusion Repeated weekly intravenous administration in hamsters can be reliably achieved using the method described Figure 3 By using a diagram from an anatomy textbook 1 the needle can then be lined up accordingly and inserted under the skin at the point of the dark area Acknowledgements Building 5 Animal Unit Staff Project Licence Holder Building 5 NACWO Laboratory Animal Technologies Management Reference 1 Popesko P Rajtova V and Horak J 2003 A Colour Atlas of Anatomy of Small Laboratory Animals Volume 2 Rats Mouse Hamster Pub Elsevier Health Sciences ISBN 10 0702027030 ISBN 13 9780702027031 currently unavailable Figure 4 As with standard intravenous dosing procedure the syringe plunger is pulled back to confirm whether the vein had been successfully located This would be shown with a small amount of blood flashing into the needle hub and syringe 125

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Animal Technology and Welfare August 2018 Refinements to animal husbandry for continuous infusion dosing in Beagle dogs LUCY ALLEN Envigo Woolley Road Alconbury Huntingdon Cambridgeshire PE28 4HS Correspondence lucy allen envigo com Abstract At Envigo we are contracted by our customers to conduct studies usually to evaluate the safety of new drugs and chemicals A rodent and non rodent species are usually involved in this testing with the method of administration designed to replicate the intended clinical administration A frequently used route of administration is continuous intravenous infusion In dogs this method requires animals to wear a specially designed jacket to carry and protect the test material and infusion apparatus combined with an anti inter ference collar unlike smaller species where tethers may be used this method allows our dogs to be infused continuously or intermittently via a previously implanted access port and catheter while still having full ambulator y movement training is started with short sessions of one hour and gradually extended to 24 hours to ensure they are well acclimatised to wearing the jackets and padded collars continuously For this training the dogs are encouraged to walk out of the animal room and into an adjoining annexe where they are placed onto a trolley and offered beef biscuit rewards as the jacket is placed on they are then walked back to the home pen with a further treat and positive praise This means that if an animal needs to be brought out for start of dosing or for repairs to equipment they are happy to come out and used to surroundings During this training phase animals are singly housed to avoid interference with one another s jackets however they are still exercised daily in groups with supervision to allow socialisation However this route of administration in Beagles presents many challenges At Envigo we are passionate about the 3Rs and this poster describes the refinements we have implemented to overcome some of these challenges in Beagle dogs Introduction Because of the potential for animal interference infusion dosing using an ambulatory pump in Beagle dogs can require the animals to be housed individually after surgery while being trained and during dose administration Enrichment and husbandry procedures can be refined during this period to ensure the social and welfare needs of Beagles are improved that they are well acclimatised to the infusion equipment and housing prior to the dosing procedure and to avoid interference to the infusion system from animals Procedures refinements Figure 1 Beagle on the trolley wearing infusion jacket Training pre surgery After a two week general acclimatisation and socialising period Beagles begin training for wearing collars and jackets daily the collars and jackets are designed to carry the pump and dose as well as protect infusion lines and equipment from interference This 126 Routine husbandry procedures such as daily feeding are also performed with animals wearing jackets and collars once the training time increases to ensure no drop in food consumption occurs from wearing their infusion equipment

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Poster Presentations Care post surgery For intravenous infusion over prolonged periods dogs are surgically implanted with an intravenous catheter connected to a subcutaneous Vascular Access Port VAP Following surgery the dogs are returned to a recovery animal unit where they can recover in the quiet with only other surgery dogs present A large plastic dog bed is placed in each animal s pen containing soft vet bedding to provide a comfortable area to sleep A selection of toys are placed in each animal s home pen and these are rotated daily to provide new enrichment and interest A warm heat pad is provided in a dog bed for extra comfort and a sensitivity meat dog diet is offered twice daily which is easily digested and aids recovery At the end of the day following surgery provided the Beagles are displaying normal behaviour and are bright and alert they are returned to their permanent animal unit as we put them back on the same day on the last few studies and can be group housed and exercised we have not seen any wound interference as a result of group housing our dogs post surgery Animals are reintroduced to jacket training on the 7 days running up to dosing and at this point a soft undershirt is placed under the jacket to provide a snug comfortable fit and to form a protective layer over surgery sites During dosing During this period the Beagles remain singly housed to avoid them chewing other animals jackets Dental chews are offered once daily to provide a longlasting chew for enrichment and which also promotes good teeth and gum health Rubber puzzle ball toys containing meaty treats inside are offered twice daily which provide the dogs lasting entertainment and interest to remove the treats from the balls Figure 3 Beagle investigating a toy whilst wearing infusion jacket Discussion By implementing these refinements the Beagles become well acclimatised to the jackets being singly housed and to their environment before the study commences meaning they are comfortable and normal behaviours and food consumption are not affected from the procedures involved in dosing On previous infusion studies an Animal Technologist would return during the night to perform a check to ensure no dose equipment had been chewed or removed however it was found this could disturb the Beagles sleep and possibly encourage chewing A trial was conducted with these extra enrichment provisions during the day to see if the night check could be obviated and it was found that the Beagles slept well throughout the night when occupied through the day and less interference to the dose equipment was seen without the need for a night check Our dogs are still well socialised with other dogs daily and have a variety of toys and treats which provide stimulation and promote natural behaviour throughout the day along with extra time with technologists Interference with dose equipment and chewing has been greatly reduced and such studies can be run reliably and accurately with minimal disruptions in infusion Figure 2 Beagle pen showing bed and enrichment toys References Extra one to one time with technicians is given each day in the dogs home pens where lots of praise is given as well as playtime The Beagles are exercised in pairs or trios daily where they can socialise and interact with each other whilst being supervised to ensure no chewing takes place 1 2 Scullion Hall Laura E M Robinson Sally Finch John and Buchanan Smith Hannah M 2017 The influence of facility and home pen design on the welfare of the laboratory housed dog Journal of Pharmacological and Toxicological Methods Vol 83 Jan Feb 2017 Green Owen P and Healing Guy 2013 Non clinical Vascular Infusion Technology Volume II The Techniques CRC Press 2013 127

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Animal Technology and Welfare August 2018 Optimisation of workflow in a BSL 3 PET CT imaging facility for non human primate research LEO VAN GEEST JACO BAKKER MOHAMMED KHAYUM FRANK VERRECK and JAN LANGERMANS Biomedical Primate Research Centre Lange Kleiweg 161 2288 GJ Rijswijk The Netherlands Correspondence geest bprc nl Introduction Sandwiched Positron emission tomography computed tomography PET CT has become an influential imaging modality in diagnosis and research of infectious diseases e g tuberculosis in non human primates The Biomedical Primate Research Centre a leading institute in tuberculosis TB research has recently implemented a PET CT in their experimental animal facility All procedures performed at the BPRC are in agreement with the regulations for animal handling and welfare as described in the EU Directive 63 2010 Tuberculosis research is performed in a high containment BSL 3 facility and use of PET CT is a challenge with respect to biohazard and radiation safety To limit hazards not only laboratory design but also well written procedures and an optimal workflow must be created implemented and maintained We have developed a PET CT facility with optimal shielding along with an efficient workflow to perform high volume procedures and minimise the radiation exposure and biohazard risk to the staff and environment by the implementation of As Low As Reasonably Achievable ALARA principles Figure 1 Floor Plan of BP PET CT imaging facility 128 Figure 2 Example of Pet CT images

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Poster Presentations G G G 2 pairs of gloves safety footwear disposable overall Personnel training items A Radiation safety radioactive substance and animal handling G G Figure 3 Pet CT and anaesthesia set up Safety operational aspect Practical training to understand the nature of the hazards and remedies Plan to minimise time spent handling radioactivity Distance yourself appropriately and use shielding from sources of radiation Contain radioactive materials in defined work areas G wear appropriate protective clothing and dosimeters G monitor the work area frequently swipe test for contamination control G follow the institutional code of conduct for work safety G minimise accumulation of waste and dispose of it by appropriate routes e g autoclave due to presence of TB G after completion of work monitor yourself and shower out disinfection to prevent TB spreading and regulatory specific radiation training for dedicated personnel super visors holding additional certification for radiation BY research G biological radioactive waste handling G disinfection of radioactive syringe containers by Hydrogen peroxide H2O2 gassing B Biohazard safety and handling G G G G biosafety training for dedicated personnel respiratory protective equipment RPE obser vation and scoring of animal s wellbeing behaviour clothing and shower procedure Conclusion High throughput PET CT lab was established in line with European standards Workplace safety training gave our facility workers the knowledge and skills they need to protect equipment facilities animals and most importantly each other against unintentional incidents They were trained to recognise safety hazards and correct and report them The implementation of the PET CT results in refinement and reduction as good quality imaging can be done in the same animal during the whole infection period leading to a decrease in animal numbers needed Animal health monitoring during PET CT acquisition Observed by experienced veterinarians during 2hr long procedure induction of anaesthesia radioactive substance uptake and PET CT acquisitions Anaesthesia is induced with ketaminemedetomidine and maintained with isoflurane gas anaesthesia After wards atipamezole is administered for recovery G body temperature of the experimental animal is maintained by warm air flow G strict monitoring of physiological parameters by using capnograph and pulse oximeter from induction of anaesthesia until recovery from anaesthesia Personal protective equipment PPE G G respiratory protective equipment RPE hood 129

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Animal Technology and Welfare August 2018 Refining mouse handling do we or don t we An animal technologist s perspective ALEXANDER SWAN BSF Department of Biology University of York York YO10 5DD Correspondence alex swan york ac uk Introduction 1 Due to recent research in the industry identifying that the way we handle mice significantly influences their stress levels many research facilities have been moving towards new methods of handling These new methods include picking mice up in a tunnel or cupping them in the hand As Animal Technologists responsible for the daily husbandry of rodents we could see advantages and disadvantages with this change for the animals staff and researchers We decided to do a pilot study within our facility at York prior to making the decision as to whether to implement the new methods across the entire facility This poster presents the methods we used for the pilot study findings of the study and what the study concluded from an Animal Technologist s perspective this very gently so that we would not cause any stress to the animal If the mouse resisted slightly we would move onto another mouse and return to the other animal later Methods We started off by leaving the clear plastic tubes in the cages for an extended period to habituate the animals to the tubes This ensured the tubes had the mice scent on and that the tubes were not considered foreign objects to the mice Figure 2 Mouse in the tube with hands blocking the ends Once the mouse was in the tube we would place the other hand flat against the opposite side While transferring the mouse into the next cage we would examine the mouse for any health issues if this was a breeding mouse we would also check for pregnancy If any health issues were identified the animal would be coaxed out of the tube for further inspection Figure 1 Mouse being exposed to clear tube When it came to moving mice into clean cages we would place a flat palm of a hand against one side of the tube this ensured the mouse would not escape We then gently guided the mouse into the tube we did 130 Figure 3 Examination of mouse whilst in tube

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Poster Presentations Pilot study This pilot study is purely based on observational analysis With the pilot study we decided to minimise variables as much as possible We made sure the same member of staff conducted the husbandry in the room when required We also decided to take mouse stress levels into account when judging the time it took us to clean out the room The more stressed out the mouse the longer it would take us to clean out an individual cage as we would spend a lot of time chasing the mice round with the clear plastic tube 2 3 The week before starting this study we decided to time ourselves and see how long it would take us to clean out the room without using the clear plastic tubes We chose to do this because a benchmark was required This allowed us to compare our times when using the tubes and not using the tubes 4 When timing each week we would stop the clock whenever we had to leave the room to get more food or assist other people We did this because we knew if we had been asked to help a researcher it would not affect our time 5 We chose to use an average time per clean out of a single cage for the graph as each week there would be a different number of cages in the room we were conducting the pilot study in This average time was reached by working out how long it took us to clean out all the cages We then divided the time by how many cages were in the room this resulted in an average time per cage Data the BSF When we first started this study we believed it would have taken us twice as long to move mice over with a clear plastic tube but as you can see by the graph we dramatically cut the time it took us per cage to clean out and almost got to the base time of cleaning without a tube After conducting the pilot for 3 months we personally felt that by the end of the study these mice seem much more relaxed when the cage lid was open and we were moving their environment around Another worry was would the breeding mice be disturbed so much that they would harm their pups or experience birthing complications However due to reduction in stress by using these tubes we saw no negative impact at all on mouse behaviour or breeding When we originally started this study we believed the mice would be afraid of this enrichment and they would not use it once it was in their cages However the majority of breeders used this as their main nesting area showing that they felt safe and protected We believe that the positives outweigh the negatives on this study but we did find that some mice even though only a small number did completely refuse to comply with getting into the tube We believe that chasing them around the cage stressed the mice out even more We believe that in some cases it is still necessary to pick mice up by the base of the tail or cup them when transferring to a new cage at the Animal Technologist s discretion Acknowledgements John Waters Kelly Gouveia and Jane L Hurst Mammalian Behaviour and Evolution Group University of Liverpool Reference 1 Hurst J L and West R S 2010 Taming anxiety in laboratory mice Nature Methods 7 825 826 Figure 4 Graph showing how cleaning time reduced with familiarisation with the technique Conclusion After reviewing the data produced from this study we arrived at the following conclusions 1 One of our biggest concerns was the time impact on 131

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Animal Technology and Welfare August 2018 Setting up a new surgical dosing team at Envigo Huntingdon JAMIE TUCK KATE READ STUART BRADLEY LYDIA MILLS KATRINA TURNER MAXFIELD TURNER DEBORAH POTTS and JESS HILL Envigo Woolley Road Alconbury Huntingdon Cambridgeshire PE28 4HS Correspondence jamie tuck envigo com Summary This poster describes the development of a dedicated surgical team to undertake a new project requiring intra striatal dosing in the rat Introduction At Envigo our Contract Research Services receive requests for a diverse range of study types in a range of species A recent enquiry required us to develop the competencies for intra striatal dosing in rats This was a challenging study that required us to expand our surgical resources as our existing resource was being fully utilised on other programmes of work This poster describes this expansion and the development of the competencies to successfully undertake this study Challenges 1 Development of the surgical dosing method 2 Development of a team of seven technicians with the required legal authorities and the competence to perform their designated roles 3 Identifying and securing the necessary equipment required for the preparation dosing and recovery of the animals involved in the study 4 Developing suitable areas to house prepare and perform surgical dosing and allow the rats to recover inside a conventional animal facility that met the needs of the study design Development of the surgical dosing method Our veterinary team develop and perform a wide range of surgical procedures on rodents rabbits dogs monkeys pigs and sheep Our veterinary surgeons researched developed and refined the method for intra striatal dosing in rats by looking at all the parts of the study plan describing the method required for the dose administration The stereotaxic procedure is well recognised however refinements were possible by ensuring high quality asepsis ensuring a very low risk of infection Consequently prophylactic antibiotics were 132 not necessary The analgesic regime included local anaesthesia non steroidal anti inflammatories and opioids to ensure it met Envigo s standard for animal welfare in preventing any pain or suffering that could have been caused to the rat through the procedure Dosing method The study design required single intra striatal administration of a novel formulation to 264 mature rats The rats were positioned in a stereotaxic apparatus such that the ear bars and incisor bar maintained a flat skull position A skin incision was made over the skull and the subcutaneous tissue was blunt dissected from the skull with dry sterile cotton tipped applicators A drop of local anaesthesia mix was applied to the exposed tissue The tip of the drill bit was moved to the appropriate anterior posterior and medial lateral coordinates The drill was activated and advanced through the bone of the skull with the rotational speed kept slow so thermal damage to the tissues was reduced Saline was dripped onto the skull while drilling to prevent thermal damage The drill bit was removed and any bone shavings wiped away This was then repeated for the second hole The micromanipulator holding the drill was replaced with the syringe pump holding the syringe The needle was positioned over the skull with the bevel facing caudally The needle was moved to the appropriate co ordinates in the same manner positioning the needle directly over the drilled holes The needle tip was advanced through the skull hole until the bevel of the needle passed the skull sur face this coordinate was considered the dorsal ventral zero point The dose was injected using the syringe pump at an injection rate of 1 ul minute The needle was removed two minutes after infusion had completed while putting downward pressure on the animal s skull the needle was backed carefully out of the skull monitoring for signs of any dose leakage Dosing was repeated through the second cranial hole After completing the second dose the incision was sutured closed and the animal removed from the stereotaxic frame and placed in the recovery cage

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Poster Presentations Development of the team Once the method had been developed we had to plan how we would perform the dosing effectively and efficiently We identified the following roles Figure 1 Drilling the 2 holes through the bone of the skull Figure 2 Prior to manoeuvring the syringe holder into position to administer the test substance One member of the team to look after the preparation of the animals prior to surgery the filling of the syringes and attachment to the pump and the recovery of the animals before being returned to their home cages back in the animal room Two members of the team to be scrubbed up and sterile to perform the opening and closing of the dose site and all the sterile parts of the procedure required One member of the team to oversee monitoring the animal s condition while under the anaesthetic and adjusting the required depth accordingly One member of the team oversaw operating the stereotaxic frame and monitoring the syringe pump Although five technicians would be involved every time we went into surgery we developed a further two technicians to allow for breaks holidays and other absences This created a team of seven technicians Training and supervision for all team members to be competent in the different roles had to be carefully planned Five members of the team were new to surgical procedures and required Home Office personal licensee training to pass module 4 PILC and obtain the required legal authority Training in best practice for working aseptically as this was something the team had not had to do in any previous non surgical work Instruction on how to sterilise equipment and prepare surgical rooms ready for the work to be performed Learning about the pre and post dose pain relief and antibiotics and planning monitoring and administration schedules Training in and then practicing the different stages of the dose administration and surgical procedures on cadavers ensuring we were comfortable and confident at performing the different tasks Our veterinary surgeons supervised our training and development Rooms and equipment We needed to modify two of our rooms in our conventional animal facility as surgery suites This required an amendment to the establishment license and for the rooms to be inspected and approved by our Home Office inspector Figure 3 Closing the incision site with suture All the equipment needed to perform the procedures had to be obtained and prepared everything from 133

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Poster Presentations anaesthetic equipment and stereotaxic frames to the cotton bud tips We had to purchase syringe pumps to be able to administer less that 5 l over four minutes To do this we bought four KD Scientific Legato 100 syringe pumps that were capable of fulfilling this requirement Syringes that where compatible with this pump and the accuracy requirements were sourced and purchased we used Hamilton Microliter 10 l glass syringes with 26 gauge 51mm removable needles Once purchased the equipment was calibrated as per our Standard Operating Procedure SOP for the calibration of infusion pumps in order to verify their accuracy before use Figure 7 Sterile surgical equipment table Housing post surgery Figure 4 KD Scientific Legato 100 syringe pump capable of administering 1 l a minute We decided to group house the animals post surgery We used a Tecniplast 2000p cage with a layer of vet bed on top of a heat mat with the cage also on top of a water bed set to maintain a temperature of 37 C to recover individual animals immediately post surgery Once each animal was able to walk around freely it was transferred to another Tecniplast 2000p cage with vet bed on top of a water bed set to 37 C again with access to food and water where it was joined by each recovered cage mate and monitored until all four animals were fully recovered All four cage mates were then returned to the animal room and their home cage Post operative care After each animal had received its dose and recovered from their procedure they were monitored throughout the day of the procedure then each day after dosing the animals where handled and the surgery site examined and monitored Each animal received post dose analgesia of Buprenorphine 0 05 mg kg approximately ever y 12 hours for 3 doses and Meloxicam 1 mg kg approximately every 24 hours for 4 doses If any extra analgesia was felt to be required a discussion with the veterinary surgeons would take place and any decisions documented in the study plan Figure 5 Hamilton Microliter 10 l glass syringe Figure 6 Animal preparation area 134 Figure 8 24 hrs post dose

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Poster Presentations Conclusion With the support guidance and collaboration with our veterinary team we successfully prepared a large surgical team secured and prepared the equipment and rooms to undertake this demanding project and substantially increase surgical capacity and capabilities of Envigo Figure 9 7 days post dose Figure 10 28 days post dose Discussion After all the planning and equipment was set up and ready to use we were able to start this study confident with what was required from us We worked closely with our veterinary surgeons who continued offering advice and assistance to us over the first couple of weeks of surgery They were happy to sign our training records that we were competent to perform the procedure allowing us to carry out the procedure without the direct supervision of the veterinary surgeons although they remained in regular contact with us ensuring we were happy and were always available if we needed any assistance from them After starting to successfully perform the procedures and following discussion with the vets we felt we could utilise the members of the team in a different way to make the procedure more efficient We did this by merging the two members of the team that where monitoring the anaesthetic and operating the stereotaxic frame so there would be one sterile person and one non sterile person per animal This meant the non sterile person would be required to monitor the animal under anaesthesia as well We found this worked well as the sterile person was also able to keep their eyes on the rat and the member of the team performing the preparation of the animal was able to assist as required 135

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Animal Technology and Welfare August 2018 Introduction of gang caging for group housed rats DAVID CLARKE and LENA IOANNOU Life and Medical Sciences Biological Support Unit University of Hertfordshire College Lane Hatfield Hertfordshire AL10 9AB Correspondence l m ionnou herts ac uk Introduction Enrichment includes Housing conditions can impact directly on laboratory animals welfare Confined spaces severely restrict strongly motivated natural behaviours such as foraging nesting hiding climbing exploring and choosing social partners G G G G 6 2 2 1 suspended PVC tunnels floor based PVC tunnels Aspen wood chew blocks cardboard tube Caging imposes restrictions on normal social dynamics in rodents their quality of life can be severely compromised by the effects of loss of freedom and the ability to exercise control over their environments At the University of Hertfordshire UH we endeavoured to develop a new system of gang caging which allowed our stock rats to be housed in larger social groups establish complex social hierarchies and to enable foraging hiding nesting and rearing Figure 2 Interconnected rat gang cages Figure 1 Rat gang cage Method Using an existing gang cage designed for ferrets we developed a version that would be suitable for housing rats Caging consists of 3 floors with a combined floor area of 11620m connected by 2 ladders Each individual unit can be interconnected by a PU Flex Tube 136 Figure 3 Bedding and nesting options

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Poster Presentations Diet G G G G 5LF2 ad lib sunflower Seeds every other day wheat Grain every other day forage Mix once weekly x10 Rats observed for this poster were CRL WI males 326 350g Observations include G G G G G G G G G increased social interactions hopping gait co operative hoarding nesting calmer animals improved handling exercising of choice over sleeping areas climbing jumping extensive foraging Figure 5 Interchangeable enrichment Acknowledgements IPS NIBSC Animal Techncians RVC Camden Animal Technicians Tecniplast UK Figure 4 Foraging Discussion The general demeanour and behaviour of the stock rats housed in the gang cages is markedly more positive and calm compared with that of rats housed in traditional P2000 caging Gang caged rats do not startle when a technician or visitor enters the room nor do they make any effor t to escape their cage environment when the doors are opened or left open for a short period They interact more positively with technicians although regular handling is needed to retain familiarity with the technician and this is facilitated during the weekly health check and during cage cleaning Co operative behaviours have been seen including feeding and nesting The hopping gait is adopted fairly quickly after housing and extensive rearing and jumping can be seen Little conflict arises although the likelihood of fighting is observed to increase with weight ie the heavier older the rats are when initially housed increases the chance of conflict in the cage Optimum weight for harmonious housing appears to be up to 350g Figure 6 Rat gang cage tower interconnected References Balcombe J P 2010 Laboratory rodent welfare Thinking outside the cage Journal of Applied Animal Welfare Science 13 1 77 88 Found at http animalstudiesrepository org cgi viewcontent cgi artic le 1004 context acwp_lab Hurst J L Barnard C J Tolladay U Nevison C M and West C D 1999 Housing and Welfare In Laboratory Rats Effects Of Cage Stocking Density Behavioural Predictors Of Welfare Animal Behaviour 58 p 563 586 Found at http www liv ac uk mbe pdf 99_HurstAB pdf Berdoy M 2003 The Laborator y Rat A Natural Histor y Available http www ratlife org Home 0Mainframeset Mainframeset htm Last accessed 10th Nov 2015 137

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Animal Technology and Welfare August 2018 Development of a real time respiration monitoring system for in vivo optical imaging STUART GILCHRIST 1 ANA GOMES 1 SHEENA WALLINGTON 1 VEERLE KERSEMANS 1 MARCUS WESTCOTT2 and SEAN SMART1 1 2 CRUK MRC Institute for Radiation Oncology Department of Oncology University of Oxford Old Road Campus Research Building Roosevelt Drive Oxford OX3 7DQ PerkinElmer UK LTD Chalfont Road Seer Green Beaconsfield Buckinghamshire HP9 2FX Correspondence sheena wallington oncology ox ac uk Introduction Anaesthesia induces autonomic ner vous system depression that leads to cardiovascular and respiratory depression The most reliable method to monitor the depth of anaesthesia in gas anaesthetised small animals is by monitoring the respiration rate If the rate is high the animal is too lightly anaesthetised and at risk of waking up If the rate is too low basic physiology including blood oxygenation can be adversely affected and can lead to death Commercial bioluminescence scanners provide an anaesthetic gas delivery system for use in mice but do not provide a means for monitoring the depth of anaesthesia When starting the imaging session the door of the dark chamber is closed Opening the door is either not possible or will abort the imaging session As such once the door is closed no monitoring of the animal is achievable This in turn means that the user is unable to confirm the depth of anaesthesia throughout the imaging procedure Therefore animals are at risk of being under or over anaesthetised This is particularly important when performing kinetic assays which can take more than 1hr to complete A simple retrofit to a commercial optical imaging scanner allowed real time Figure 1 Respiration monitoring schematic 138 respiration procedure monitoring throughout the imaging Method Imaging was performed on an IVIS Spectrum scanner Perkin Elmer Five piezo electrical sensors LDT0028K TE Connectivity were arranged such that mice need only to be placed upon them to enable respiration monitoring These sensors were set into corrugated channels which allow for any length of mouse to be placed in either prone or supine position and still generate a good signal Signals were routed to a commercial physiological amplification and recording system DA100C amplifiers and MP150 recording system Biopac Inc which displayed and recorded the signals and respiration rates in real time For system validation 10 female CBA mice 2 sets of 5 with weight range 1 being 20g and weight range 2 being 30g were anaesthetised and placed on the respiration Figure 2 A 5 channel corrugated base was 3D printed in house

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Poster Presentations sensors for a 5 minute imaging session in both prone and supine positions before being removed from the scanner Figure 1A C The procedure of mouse positioning imaging and removal was repeated 5 times before the animals were recovered Results Respiration rate was monitored successfully and respiration traces responded to changes in respiration rate due to different concentrations of isoflurane in all test cases Conclusions Flexible control of the respiration rate enabled adaptive control of the depth of anaesthesia The operator was able to ensure that animal welfare was paramount by confirming that the correct level of anaesthesia was given and maintained which in turn avoided anaesthesia either too deep or too light The former presents a welfare issue the latter affects welfare and safety Should any animal wake up on the imaging platform it could move and get caught in the scanner mechanism and an animal dosed with radiotracer or any other toxic tracer could wake up and contaminate the scanner These events must be avoided and the use of respiration monitoring reduces the chance of either happening Figure 3 5 piezo sensors running the length of the channels were affixed to the base and their wiring was fitted into a sealed compartment Acknowledgements Funding Cancer Research UK CRUK Medical Research Council MRC Engineering and Physical Sciences Research Council EPSRC and the Oxford Centre for Drug Delivery Devices OxCD3 Figure 4 Real time changes in respiration rate due to different concentrations of isoflurane were recorded in real time for all mice Figure 5 Shows 20g mice fitting with plenty of room to spare The base can be moved freely to allow for animals of varying lengths 139

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Animal Technology and Welfare August 2018 To glow or not to glow that is the question CHRISTOPHER HUGGINS Comparative Biology Centre Newcastle Medical School Newcastle upon Tyne NE2 4HH Correspondence chris huggins ncl ac uk Introduction Over the past few years different methods of in vivo imaging have become common place and are often used to look at tumour growth or brain function When carrying out bioluminescence scans our researchers reported seeing some inconsistencies in their control animals Was this a fault with the scanner or could it be something else Previous research had shown that standard animal feed and forage items like sunflower seeds had produced strong bioluminescence signals and this may account for false signals The animals in question had been given a special feed devoid of chlorophyll so diet was ruled out On completion of the studies the mice were humanely culled and were sampled Necropsy results showed they had undigested wooden particles from the cage substrate in their digestive tract This posed the question was this the cause of the abnormalities all along For example had the animals been eating the bedding This poster looks at the bioluminescence properties of common cage substrates and forage foods Figure 2 Shows bioluminescence proper ties of common forage foodstuffs Figure 3 IVIS scans of mice that had been housed on corn cob bedding showing high fluorescence from ingested particles Method Figure 1 Showing bioluminescence properties of bedding 140 To determine the cause of the rogue bioluminescence signals a two fold non invasive study was devised to

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Poster Presentations image the various materials the animals came into contact with These common substrates aspen chips Celudry and two grades of corn cob were first placed in Petri dishes The dishes were then imaged side by side at the wavelengths used commonly used to image animals namely 745 800 820 Ex Em NIR use Celudry when animals enter an imaging study This way they can be confident that their results are accurate enabling a reduction in the number of animals used The second part of the study used control animals due to be imaged for an on going study These animals were placed on each of the four test substrates for 48 hours prior to being imaged As per the study they were undergoing the animals were fed an imaging diet but not given any forage foods before being imaged After being imaged the animals were culled using a Schedule 1 method and gut samples collected to investigate its contents Dr Jill Hunter ICM Newcastle University Imaging Welcome Trust IVIS Machine Grant funding Body CBC Rodent area Technical Staff and Chief Technician Study assistance Acknowledgements Results Looking at the IVIS images from the bedding it is apparent that that they do give off bioluminescence signals of various strengths The intensity of the signal increases with the increased amount of organic material contained within the substrate Celudry gave the lowest signal followed by aspen chip which gave a medium signal The highest signals were seen from corn cob with the larger of the two samples giving the highest signal of all The imaging results from the control animals were interesting These animals should not have given any signals but that was not always the case Animals housed on Celudr y showed no rogue signals However animals housed on aspen showed weak unexpected signal and the animals housed on corn cob gave very strong signals Looking at the contents of the gut there was no evidence of the animals consuming the Celudry A few 6 small chewed pieces of aspen were found in the animals that showed a weak signal when imaged The animals housed on corn cob had between 2 and 3 pieces in their gut but these animals gave the strongest signals of all despite having far fewer pieces in the gut than other groups Discussion After reviewing the results it is apparent there is evidence of animals consuming their bedding and that ingestion of the bedding material has the potential to influence experimental results when bioluminescence imaging is used This influence could be detrimental if for example the target organs requiring imaging are located around the digestive tract With the increased use of bio markers it will be important to not only consider a diet s impact on bioluminescence signalling but also that of the bedding substrate used At Newcastle University we now advise researchers to 141

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Animal Technology and Welfare August 2018 Mice on the road how to move thousands of mice whilst maintaining microbiological status and high levels of welfare RAFAELA FERN NDEZ DE LA FUENTE The Francis Crick Institute 1 Brill Place London NW1 1BF Correspondence rafaela fernandezdelafuente crick ac uk Abstract Materials needed This poster provides an overview of the processes involved in successfully moving over 1400 lines of mice from Hertfordshire to Central London as a part of a large scale migration process following the opening of the new Francis Crick Institute building To maintain and ensure a clean microbiological status IVC cages The animals were transported in their home Transpor t is a significant stressor that has the potential to impact on both animal welfare and research outcomes Staff across the BRF have worked very hard to ensure the safe arrival of thousands of cages with minimal distress while maintaining a clean microbiological status for every single mouse adhering to tight deadlines for vacating the old facility and ensuring minimal disruption to research Why the move was needed In 2015 The Francis Crick Institute was formed and in 2016 it moved into a brand new state of the art building in central London making it the biggest biomedical research facility under a single roof in Europe Figure 1 IVC cage The MRC National Institute of Medical Research NIMR and the London Research Institute LRI of Cancer Research UK became part of The Francis Crick Institute in 2015 Prior to the migration in 2014 BRF teams together with Genetic Manipulation Services GeMS started rederiving over 1400 different mouse lines from both founding Institutes into P block at Clare Hall Hertfordshire This ensured that the new Institute held animals at the highest levels of microbiological status to allow world class research be performed and to maintain excellent standards of welfare After 2 years of hard work in September 2016 the first animals started to be moved to The Francis Crick Institute This process was finally completed in December 2017 142 Figure 2 Schematic of isolator showing features

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Poster Presentations cage to ensure they were fully protected from all microorganisms Every cage had bedding material a house enough food for the journey and a pot of hydrogel as the bottles were removed to avoid leaking during transport Contact points and lock down mechanism fitted with rubber dampers to minimise vibration Isolator This is made of a PVC canopy and all seals are maintained under positive pressure ensuring that air cannot leak into the isolator The user carries out manipulations by working through the sleeves The isolator is fumigated every day after every use Van A specialist vehicle designed to securely lock and provide power to 4 transfer isolators No more than 3 isolators were transferred at the same time due to load weight restrictions Figure 5 Schematic of contact points and lock down mechanism Figure 3 Schematic of van Thermally insulated and fully antibacterial hygiene coated Fresh air ventilation and LED interior roof lighting Load area climate controlled at 21 C 2 C and relative humidity 55 10 to comply with the Home Office Code of Practice Fitted with a power supply and dash mounted audiovisual alarm to indicate system failure to the driver Fitted with a tail lift Figure 4 Lorry Tail Lift side and tail flaps drop down to allow easy movement onto lift Figure 6 Rapid Transport Port Figure 7 Arrangement of Rapid Transpor t Por t doors locking mechanism to prevent accidental contamination 143

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Poster Presentations Rapid Transport Port Designed to avoid mishandling which could interfere with the chain of containment during the transfer phases Isolator is connected to the port Door cannot be opened unless the isolator is properly connected and it cannot be removed if the double door is not locked shut Double door cannot be opened inside the unit when it is not equipped with a container door Process At full capacity the IVC area of P block held almost 15 000 cages plus 2 Isolator Halls holding 2 500 cages 2 or 3 isolators each with a 25 cage capacity were transported daily P block stopped breeding in October 2017 and weaned the last litters by Mid November Last animals arrived at The Francis Crick Institute in December 2017 Before sending the animals 1 A list was sent identifying the animals to be issued 2 Cages were checked by responsible technicians to ensure details matched the cage labels and the list 3 No females over 14 days pregnant were sent 4 No pups younger than 5 days were sent 5 Cages with males were double checked to avoid fights between them Figure 9 Isolator is connected to the Rapid Transport Port Figure 10 Animals are transferred through Rapid Transport Port to the Isolator Figure 8 Clare Hall Hertfordshire First animals start to be moved 144

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Poster Presentations Animals were closely monitored for a few days to ensure they had adapted to the new watering system and recovered from the journey Sentinels were used to check the microbiological status Faeces were collected from different cages and added to a sentinel cage which is sent to the microbiologist every 3 months for analysing Conclusions There was minimal effect on breeding a good sign that the mice had adapted to their new environment There was no observed detriment to animal welfare From the latest screening the sentinels show a clean microbiological status Research and science continued during the whole process 1400 lines were moved in total The process took 3 years 4684 embryo transfers and the transport of over 14 000 cages covering over 14 000 road miles Special thanks Special thanks to Barbara Rudling Hannah Sexton and Cheryl Young for all the information and pictures from Clare Hall they have given me as well as Helen Bailey and Alan Palmer for their guidance throughout the creation of this poster References PFI Systems Ltd Getinge La Calh ne group Vehicle Overview pdf document www crick ac uk www tecniplast it 147

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Animal Technology and Welfare August 2018 Going on a diet MATTHEW O BRIEN BMS University of Oxford John Radcliffe Hospital Headley Way Headington Oxford OX3 9DU Correspondence matthew obrien bms ox ac uk Abstract Time for change Going on a Diet Changing our practices when it comes to feeding Cage design 1 Currently can hold 415 1g in a full hopper The new size holds up to 302 9g giving us a difference of 112 2g After a recent review into the costs of consumables at our facility the topic of wasted diet was raised We use approximately 1000kg of diet per month in the facility The diet is irradiated adding to our costs Our aim was to review what the human perception was in regards of how much diet should be in the actual food hopper The current practice is to top up the food hopper when performing the weekly service of the cage even if the amount of food left and the number of animals did not warrant it Given this practice we discussed what the effect would be of reducing the size of the hopper so that it contains less weight but still enough for the number of animals in the cage It should be noted that there was no intention to food restrict any animal and that all animals and cages are checked daily to ensure enough adlib food and water are available Cage design 2 Can hold 800g in a full hopper The new size can hold 500g giving us a difference of 300g The graph below shows the difference in diet consumption using the altered diet hopper Our facility houses on average 2500 per week Old vs New by week From a welfare prospective it does mean that the animal is not constantly consuming diet that is older than necessary In this presentation we are able to demonstrate our first findings and what impact it had on the amount of wasted diet seen It is not meant to be the final chapter in this discussion but will hopefully encourage further debate from all interested parties The idea Using the Handbook of Laborator y Animal Management and Welfare Sarah Wolfensohn and Maggie Lloyd 1 our study was based on mice eating 5g a day We weighed out how much diet each hopper can hold then divided it by how much mice eat Figure 1 Graph showing the difference between the amounts of diet used in Kg each week in the month of Nov 2017 Results The amount of diet we add to a cage is still rather high An old deep seated idea was that we had to take into account the weight of the diet in the hopper to help the animals feed We now know that is not the case we have seen mice eat from the middle of the hopper not the bottom as this is often soiled We have also always had this idea that we fill the hopper because it looks better Better for who Us or the animals 148 NEW OLD

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Poster Presentations CAGE DESIGN 1 Mouse number Days of food 5g per mouse Full hopper Full changed Normal hopper 1 83 60 2 41 30 3 27 20 4 20 15 5 16 12 6 13 10 7 11 8 5 8 10 7 5 OLD OLD NEW RAT CAGE 1 Days of food 20g per mouse Mouse number Full normal Full smaller 1 63 21 2 31 5 10 5 3 21 7 NEW CAGE DESIGN 2 Days of food 5g per mouse Mouse number Full normal Full smaller 1 160 100 2 80 50 3 53 33 4 40 25 5 32 20 6 26 16 7 22 14 8 20 12 5 9 17 11 10 16 10 11 14 9 12 13 8 Rat cages Redundant cages We on average due to experimental and surplus reasons retire 150 175 cages a week Assuming 175 cages being used have full hoppers 175 x 415g 72625g or 72 6kg which is equal to 5 8 of 12 5kg bags of diet Compared to the smaller hopper 175 x 302g 52850g or 52 8kg which is equal to 4 2 bags of 12 5kg diet Multi species This refinement can be applied to many different diet hoppers We have decreased the rat hopper size by 60 and the mice hopper by 27 We are in the process of changing the hoppers for other species 149

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August 2018 Animal Technology and Welfare ABSTRACT A survey of rabbit handling methods within the United Kingdom and the Republic of Ireland JAMES OXLEY 1 CLARE ELLIS 2 ANNE McBRIDE3 and WANDA MCCORMICK4 1 2 3 4 Independent Researcher Measham Swadlincote Derbyshire Arts Science and Technology University of Northampton Avenue Campus St Georges Avenue Northampton Northamptonshire NN2 6JD Psychology University of Southampton Southampton SO17 1BJ UK Animal Welfare and Equine Moulton College West Street Moulton Northamptonshire NN2 7RR Correspondence james oxley1 hotmail com Reprinted with permission of the authors and the Journal of Applied Animal Welfare Science Reprinted from the Journal of Animal Welfare Science April 2018 E publication https www tandfonline com doi full 10 1080 10888705 2018 1459192 Abstract Rabbits are commonly kept in a variety of settings including homes laboratories and veterinary clinics Despite the popularity of keeping this prey species little research has investigated current methods of handling The aim of this study was to examine the experience of caregivers owners and keepers in using five handling methods commonly referred to in books written for companion animal pet owners and veterinary and or laboratory personnel An online sur vey was completed by 2644 respondents representing all three of these groups and breeders Data were acquired to determine sources that participants used to gain knowledge of different handling methods the methods they used and for what purposes they used them and their perceptions of any associated difficulties or welfare concerns Results indicated that participants most frequently used the method of supporting a rabbit s body against a person s chest which was considered the easiest and most welfare friendly method of the handling methods explored Scruffing with rear support was the least used method and was considered to be distressing and painful for the rabbit As rabbits are a terrestrial prey species being picked up is likely an innately stressful experience Additional research is encouraged to explore the experience of rabbits during handling to identify methods that can be easily used with the fewest welfare compromises Open access link https www tandfonline com doi full 10 1080 1088 8705 2018 1459192 151

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Animal Technology and Welfare August 2018 Instructions to Authors Subjects considered for publication may include original articles technical notes and reviews pertaining to all aspects of animal science and technology management and education The Editorial Board wishes to offer particular encouragement to papers leading to improvements in environmental enrichment the general care and welfare of the animals used in particular those species and strains exhibiting harmful genetic defects and papers describing refinements in techniques a reduction in the number of animals that need to be used or alternatives to animal use Papers describing experimental procedures will only be accepted for publication if authors clearly state that the procedures conform to the prevailing principles and Codes of Practice of the Animals Scientific Procedures Act 1986 Papers submitted from outside the U K should state what legislation and or ethical approval the work has been carried out under In addition authors who describe surgical techniques with recovery should include details of post operative care and any analgesic therapy provided All submissions should follow the ARRIVE Animal Research Reporting of In Vivo Experiments guidelines Kilkenny C Browne WJ Cuthill IC Emerson M Altman DG 2010 Improving Bioscience Research Reporting The ARRIVE Guidelines for Repor ting Animal Research PLOS Biol 8 6 e1000412 doi 10 1371 journal pbio 1000412 The Editorial Board reser ves the right to seek independent advice on any aspect of the content of an article but the final decision on acceptance or rejection remains with the Board Hard copy The original manuscript plus two copies should be sent to the address below together with a copy on disk CD or DVD All sheets should be typewritten on one side in double spacing and serially numbered Any photographs or graphs should be supplied as originals and conform to the format in 4 below Address for submission Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL No responsibility will be accepted for loss or damage to such articles Electronic files of submissions are required together with separate files of photographs and any graphics that appear in the manuscript Electronic submissions should be sent via email via atw iat org uk alternatively manuscript plus two copies may be sent as hard copy to the address below All sheets should be typewritten on one side in double spacing with 4 cm margins and serially numbered Additionally a copy on disk should be provided or sent by email via atw iat org uk Articles for submission should be sent to Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL No responsibility will be accepted for loss or damage to such articles Submission Material submitted for publication will be considered provided that it is contributed exclusively to Animal Technology and becomes the property of the Institute of Animal Technology 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 152 Format 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 Summary Introduction Methods Results

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Instructions to Authors v Discussion vi Acknowledgements vii References 3 Measurements should be given in metric units see The use of S I Units 1969 British Standards Institution publication and spelling should follow that of the Oxford English Dictionary Abbreviations must be defined in full at their first appearance in the text The 24 hour clock should be used for times Words to appear in italic type should be underlined Designation of inbred strains should be in accordance with the International Index of Laboratory Animals 6th edition compiled edited and published by M W Festing 1993 4 Photographs should have clear and well contrasted tone values and be in colour All illustrations charts e g histograms and graphs and photographs should be submitted separately and bear on the reverse side the author s name a number corresponding to the order in which it appears in the text e g Figure 1 and an arrow pointing to the top 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 Only essential references should be included Authors are responsible for verifying them against the original source material ATW uses the Vancouver referencing system references should be identified in the text by superscript Arabic numbers e g 12 after any punctuation and numbered and listed at the end of the paper in the order of when they are first cited in the text Automatic numbering should be avoided References should include the names and initials of up to six authors If there are more than six authors only the first three should be named followed by et al Publications for which no author is apparent may be attributed to the organisation from which they originate Simply omit the name of the author for anonymous journal ar ticles avoid using Anonymous 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 e g Saigeman S 1998 Environmental enhancement of cats what why how Animal Technology Vol 49 No 3 145 154 Books Surname and initials of author s date title of book Name of publisher Town of publisher e g Flecknell P A 1987 Laborator y Animal Anaesthesia Academic Press London Chapter from a multi author book Surname and initials of chapter author s date title of chapter In title of book surname and initials of book editors Name of publisher Town of publisher first and last page numbers of chapter e g Gregory J A 1985 Principles of Animal Husbandry In Laboratory Animals An Introduction for Experimenters Second Edition Tuffrey A A John Wiley Sons Ltd Chichester 87 105 Papers accepted for publication but not yet published should be included in the list of references followed by in press Papers in preparation personal communications and unpublished observations should be referred to as such in the text only 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 Number of animals and any pretreatment 153