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IAT Journal Animal Technology and Welfare G Andrew Blake Tribute Award 2015 winning article G Enhanced housing for male rabbits G Final 2014 Congress posters G AS ET 2014 part two Official Journal of the Institute of Animal Technology and European Federation of Animal Technologists ISSN 1742 0385 Vol 14 No 1 April 2015

CONTENTS Vol 14 No 1 April 2015 Editorial Jas Barley Chair of the Editorial Board ix Refinements in action Moving from head cap to harness for cannulating laboratory rats Clare Sims 1 Enhanced housing for male laboratory rabbits Joanna Cruden Jonathan Cooper and Oliver Burman 7 Report of the 2014 RSPCA UFAW rodent welfare group meeting Penny Hawkins Secretary Manuel Berdoy Charlotte Bur Debbie Bursnall Joanna Cruden Wanda McCormack Amy Miller Helen Proctor David Whittaker Maggy Jennings and Robert Hubrecht 19 PAPER SUMMARY TRANSLATIONS 29 TECH 2 TECH Improving animal welfare reducing energy cost and increasing flexibility Preben Leonhardt and Andy Domone 41 AS ET SPECIAL TRAVEL BURSARY 2014 ESSAYS Advantages and disadvantages of animal technologists performing regulated procedures in research Angelika Kosciolek 44 Advantages and disadvantages of animal technologists performing regulated procedures in research Adam Johnson 47 Advantages and disadvantages of animal technologists performing regulated procedures in research Melanie Dunscombe 49 POSTER PRESENTATIONS Introducing an objective animal health and welfare assessment system into the biological resources unit of Cancer Research UK London Research Institute Danielle Cox Valerie Borel Vannier Claire Harper Gary Martin Mercedes Sanchez and Gary Childs 51 The development of hormone loaded diets to promote xenograft growth Alison Ritchie Pam Collier Niovi Nicolaou and Anna Grabowska 55 Managing emotions in the workplace for those working with research animals Alison Hopkins 57 A simple solution to prevent the abdominal migration of temperature loggers and to facilitate their smooth retrieval post study in macaques Leo van Geest Merei Keehnen Rianne Klomp Jacco Bakker Robin van der Schilt and Jan Langermans 59 Welfare changes in high disease containment research using farm animals Hugh Simmons 62 Choosing a system for managing training records Angela Kerton Alison Hopkins and Andreas Staubi 64 Experimental and husbandry procedures impact results and animal well being Laura Anne Roberson Anna Karin Gerdin Natalia Igosheva Ozama Ismail Natasha Karp Mark Sanderson Emma Cambridge Carl Shannon David Sunter Ramiro Ramirez Solis James Bussell and Jacqueline K White 67 An evidence based approach to determine the environmental requirements for animal holding rooms Steven Cubitt Fintan Lyons Peter Barrington and Gene Murray 69 The Siberian hamster breeding and husbandry techniques Melissa Bean 71 Instructions to Authors 74 Index to Advertisers xx ATW PROFILE Animal Technology and Welfare aims to be the medium for animal technologists and all those concerned with the care and welfare of animals used for research purposes to communicate best practice ATW especially aims to promote and develop the 3Rs particularly in respect of Refinement More importantly ATW promotes the generally accepted 4th R Responsibility The responsibility that all animal technologists have in ensuring dissemination of best practice to every institution using animals in research ATW enjoys a unique position as the scientific publication for the leading organisations IAT and EFAT for the welfare of animals in research Editor Jas Barley atweditor iat org uk i

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April 2015 Animal Technology and Welfare THE INSTITUTE OF ANIMAL TECHNOLOGY ETHICAL STATEMENT IN THE CONDUCT OF THEIR PROFESSIONAL DUTIES ANIMAL TECHNOLOGISTS HAVE A MORAL AND LEGAL OBLIGATION AT ALL TIMES TO PROMOTE AND SAFEGUARD THE WELFARE OF ANIMALS IN THEIR CARE AND TO RECOGNISE THAT GOOD LABORATORY ANIMAL WELFARE IS AN ESSENTIAL COMPONENT OF GOOD LABORATORY ANIMAL TECHNOLOGY AND SCIENCE Editorial Jas Barley Chair of the Editorial Board It seems rather odd to be writing this in January and still be thinking in terms of a New Year as well as a new volume of Animal Technology and Welfare ATW when I know that you will not be reading this until mid March However as with all aspects of Animal Technology forward planning is essential Preparation for each issue takes approximately three months so the earlier you provide copy the stronger the possibility of getting your article published in a timely manner Having said that sometimes the volume of material is such that we cannot publish everything at once and this has been the situation in both the case of the Congress 2014 posters and the AS ET Special Travel Bursary 2014 entries The fact that some of this material is only just being published is in no way an indication of its value but is simply due to pressure of space Although the AS ET entries in this issue did not win the bursary they all offer different views of the advantages and disadvantages of animal technologists performing procedures and there is a strong sense of how much they care about the animals in their charge Whilst I am on the subject of forward planning the December issue for 2015 will once again be themed and will take as its subject Animal Behaviour and the natural history of the species used in biomedical science Therefore if you have any projects which will be of interest to our readers please start preparing your manuscripts as soon as possible I would be pleased to hear from you in advance if you would like to discuss your offering Returning to a more immediate timescale as well as including the final Congress 2014 poster this issue includes the winning paper of the 2015 Andrew Blake Tribute Award The award is given to the Animal Technologist judged to have made the most significant contribution to improving animal welfare over the previous twelve months This year s winner is Clare Sims from the University of Bristol with her entry Refinements in action Moving from head cap to harness for cannulating laboratory rats The paper describes how technologists and researchers worked together to refine cannulation surgery for the benefit of both the animals and the research As well as a cash award of 250 and an engraved plaque the author of the winning entry is invited to Congress as a guest of the Institute of Animal Technology to receive their award Considerable preparation and observations have also been necessary to produce the paper from Jo Cruden and colleagues which reports on a pilot study to enhance the housing for male rabbits Well having put 2014 finally to bed Congress 2015 is very much on the horizon and my thoughts are on the August issue which means I would like to make my annual plea to Congress platform presenters and poster authors to make their contribution to the meeting available to ATW so that those members readers who cannot get to Congress can benefit from the information you are making public Remember that the Journal goes to over 2500 members and subscribers in 28 countries worldwide Many of our subscribers are libraries of large Universities and global companies so the potential audience for your results is enormous I will not promise that material will definitely appear in August but if you do not write it I am unable to publish it ix

April 2015 Animal Technology and Welfare Refinements in action Moving from head cap to harness for cannulating laboratory rats CLARE SIMS Dorothy Hodgkin Building University of Bristol Whitson Street Bristol BS1 3NY Corresponding author svces bristol ac uk Winner of the 2015 Andrew Blake Tribute Award Abstract Rats are jugular vein cannulated to enable blood samples to be taken using an automated blood sampling ABS system These blood samples are used to measure hormone levels present in the blood over time In the original protocol the jugular vein cannula is exteriorised from the body of the rat via an incision on the top of the head It is then fed through a protective metal spring and attached to the ABS system The metal spring is anchored in place by two metal screws implanted into the skull of the rat The refinement to this protocol is that the cannula exteriorises on the back of the rat and a vascular access harness anchors the protective spring in place The results show that by refining this protocol animal welfare and husbandry has been improved and the number of animals used overall in the study has been reduced It is also shown that the amount of stress response hormone corticosterone in the blood does not increase with the use of the vascular access harness Key words Rats jugular cannulation access harness corticosterone refinement stress response hormone corticosterone cortisol in man The HPA axis is the main hormonal system that provides a rapid response to stress Corticosterone is released from the adrenal gland For normal metabolism corticosterone is released in hourly pulses over the 24 hour period and exhibits a circadian and ultradian rhythm which is controlled by the hypothalamus In response to environmental stressors corticosterone levels in the circulating blood will increase in an attempt to counteract the stress on the body Levels of corticosterone in the circulating blood of the rat can be measured using an automated blood sampling ABS system This system collects small blood samples from the rat via a flexible cannula implanted into the jugular vein over time while the animal is conscious giving an accurate account of how corticosterone levels in the blood change during the 24 hour cycle The ABS system is controlled by a computer and takes up to 172 samples of no more than 40 l at a frequency of one sample every 5 10 minutes over 12 to 24 hours It does this using a peristaltic pump which replaces the blood volume taken from the animal with an equivalent volume of sterile heparinised saline so that the blood volume of the animal remains unchanged during the experiment ABS samples are taken in a soundproof room so there are no outside influences on the circulation corticosterone levels Introduction The main focus for the research that this specific surgery facilitates is into the hypothalamic pituitaryadrenal HPA axis and rats are used as the HPA axis in the male rat is very similar to that of man The rat is also the species most widely used in research into the function and control of the HPA axis The HPA axis activity is characterised by a negative feedback loop within the body that regulates the levels of circulating glucocor ticoid hormones 1 Glucocor ticoids regulate metabolism and resistance to stress2 and include the The surgery to implant the jugular vein cannula is carried out under general isoflurane anaesthesia maintaining asepsis at all times In the original surgical protocol the cannula is exteriorised from the rat through a scalp incision and is fed through a protective metal spring The spring is anchored in place via two stainless steel screws that are embedded into the skull of the rat In order to do this two very shallow holes are drilled into the bone Dental acrylic is then applied over the screws and incision to form the head cap Figure 1 1

Refinements in action Moving from head cap to harness for cannulating laboratory rats Figure 1 WKY rat with head cap Figure 4 Lister Hooded rat wearing the harness The spring and cannula are attached to a swivel mounted above the cage on scaffolding to allow full free movement around the cage Figure 2 This protocol has been refined to include the use of a vascular access harness Instech Laboratories USA 3 to replace the head capping technique The harness Figure 3 fits around the rat s body and consists of soft adjustable belly bands and a hard plastic dome on the back The cannula now exits the body further back from the head and is fed through the plastic dome which adds protection to the incision site and into the protective spring which fits into a hole in the plastic Figure 4 One of the adverse effects outlined in the project licence is intra cranial haemorrhage from tissue damage when drilling and securing the screws into the skull expected incidence

Refinements in action Moving from head cap to harness for cannulating laboratory rats cardboard tubes within the cage for enrichment to avoid entanglement Therefore they are provided with Aspen wood chew blocks Nestlets and paper bedding for enrichment Figure 7 6 The animals are allowed to recover from surgery for 5 days to allow the circulating Cort levels to return to normal basal value During this time the cannula are flushed daily with sterile heparinised saline 7 Following recover y in the last 24 hours the experiment starts when the animals are connected to the ABS system Personnel do not enter the room during this time and the animals are checked by CCTV cameras above each cage viewed on a monitor outside the room Figure 5 Close up of swivel and cannula All procedures were performed under the prevailing principles and authority of the Animals Scientific Procedures Act 1986 Results Figure 6 Soundproof room set up Figure 8 shows the amount of circulating corticosterone in the blood stream of the rats over a full 24 hour period and clearly compares the base line data for those fitted with a head cap and those fitted with a harness The chart illustrates the pulsatile nature of the release of corticosterone which will naturally be at higher levels during the night or the dark phase as this is when the animals are most active due to their nocturnal behaviour It can be seen that the circulating cor ticosterone at several time points notably 8 30am 11am and 9pm is higher for the head cap method but overall the two methods have roughly the same pattern on the chart In addition to no changes in their circulating Cort levels it has also been observed that no animals have suffered intra cranial haemorrhage or other surgical complication in the last 12 months since using the harness method Figure 7 Cage set up Figure 8 Pulsatile baseline circulating corticosterone levels over 24 hours 3

Refinements in action Moving from head cap to harness for cannulating laboratory rats Discussion The results illustrate a very similar 24 hour pattern of circulating corticosterone for both the head cap and the harness method This base line data shows no exaggerated corticosterone response in the animals wearing the harness As corticosterone is a stress response hormone it indicates that wearing the harness is not more stressful for the animals than the head cap method This means the refinement has been successful and good results can be obtained It also means that data obtained before and after the refinement are still comparable The movement from one method to the other has not been without its problems however Initially when the experiments were carried out using the new harness method the animals were not habitualised to the harness for 24 hours prior to the surgery The harness was first fitted to the rat when they were under general anaesthesia for the jugular vein cannulation This meant that the animal was coming round from anaesthesia after a major surgery housed alone with a strange harness fitted around their bodies with the cannula and protective spring fixed to the swivel It was observed that all these things combined were quite stressful for the animals indicated by increased porphyrin staining around the eyes and mouth upon recovery from surgery which had not previously been observed in the head cap method Although the animals had become acclimatised by the last experimental day and this did not affect the results it was felt that in terms of animal welfare that this was a problem that needed to be addressed It was therefore decided that the animals would be introduced to the harness for 24 hours prior to surgery to allow them to get used to the feeling of the soft adjustable belly bands of the harness around their body so eliminating one stressful factor from the recovery period Acclimatising the rats to the harness for 24 hours did however raise other points The rats would now have to be singly housed for an extra day Singly housing social species such as rats is well known to be a stressful experience for them An attempt was made to see if two rats could be acclimatised for 24 hours to the harness while being housed together This however just resulted in two harnesses being completely chewed to pieces overnight So this was not an option After discussions with the Named Veterinary Surgeon NVS it was decided that the potential refinement to the protocol outweighed the stress that might be incurred from singly housing for an extra day This decision however brought on the next problem Having to dress the rats in their harness while they were awake proved to be rather difficult As the harness had initially been put on under general 4 anaesthesia this had not yet been an issue Due to the fact the rats had not been handled other than when they arrived and when cleaned out they were not used to this higher level of human interaction and resisted the harness For some individuals it was impossible to dress them without the use of general isoflurane anaesthesia It was discussed and decided that this process in itself was quite stressful and also the project licence in question only allows anaesthesia for a maximum of three times per animal for this protocol Using one for this when it may be needed after the surgery for repair of stitches etc was thought to be too risky So a handling regime was devised for the animals to allow them to get used to humans and to facilitate the dressing with the harness and so reduce stress The harnesses themselves have taken a bit of getting used to including getting the tightness of the bands right around the rat so they are not restricting but neither so loose that the rat slips out They are easily adjustable and soft but it was found that the free ends of the belly bands were irritating the rat s ears causing them to try and shake it off This increased shaking was sometimes opening the cage clasps and for a while it was thought that we had been sent rather intelligent rats Extra cage clips have been added and the free ends of the harness are now always tucked in Also the protective spring that was sent with the harness from the company was too stiff and did not provide full flexibility and restricted movement around the cage It was decided to go back to using the old springs which were custom made for purpose With all of these refinements finally in place it was evident that the protocol had been vastly improved and the move from the head cap to harness facilitated well The handling has greatly reduced stress as the animals now trust their handlers and they can be easily dressed in the harness and are much easier to work with throughout the whole process There is now little to no porphyrin staining after the surgery and any effect from singly housing the animals has been reduced because they now interact with the staff instead of an animal companion and seem bright and happy This new technique has also refined the surgery time as the animals are under general anaesthesia for a much shorter period which is better for their recovery This is because the head cap part of the surgery is the part which took the most time The results also show there have been no surgical complications since using the harness method which has reduced the number of animals used for these experimental procedures In addition to implantation of the jugular cannula for blood sample collection this new technique has also been found to be an excellent refinement for other types of cannulation such as sub cutaneous cannulation for drug administration

Refinements in action Moving from head cap to harness for cannulating laboratory rats Conclusion The whole process of facilitating this change has shown many refinements and improved animal husbandry Simplifying the surgical procedure has eliminated the surgical complications associated with the head cap method and has improved recovery for the animals All of which are positives and are prime examples of the animal care staff and the academic staff working closely together and listening to each other s ideas which is of great importance for good scientific practice and significant results from animal experiments It also shows that a major change to facilitate a refinement to an existing protocol can be introduced in the middle of long term work with little consequence if everyone works together There is often reluctance to change things in the middle of a study even if it is a refinement or improvement to animal welfare for fear of altering results but it can be done Throughout this process there has been involvement and full support from the Named Veterinary Surgeon NVS Named Animal Care and Welfare Officer NACWO and the Home Office Inspector here at the University of Bristol and efforts are being made to encourage other groups to star t using this method and also educating individuals on the benefits of handling animals for experimentation Acknowledgements Francesca Spiga John Roe Gareth Gough and Peter Gardiner References 1 2 3 Spiga F Waite E J Liu Y Kershaw Y M Aguilera G and Lightman S L 2011 ACth Dependant Ultradian Rhythm of Cor ticosterone Secretion Endocrinology 152 4 pp 1448 1457 Tortora G J and Derrickson B 2011 Principles of Anatomy Physiology Maintenance and Continuity of the Human Body 13th ed John Wiley and Sons Ltd Instech Laboratories 2014 Tethers Online Available at http www instechlabs com Infusion tethers 5

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April 2015 Animal Technology and Welfare Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens JOANNA CRUDEN1 JONATHAN COOPER2 and OLIVER BURMAN2 1 2 GlaxoSmithKline Park Road Ware Hertfordshire SG12 0DP Animal Behaviour Cognition and Welfare Research Group School of Life Sciences University of Lincoln Joseph Banks Laboratories Lincoln LN7 6TS Corresponding author joanna l cruden gsk com Abstract At the current facility we usually individually house male rabbits in cages with twice weekly access to floor pens measuring 2 5m2 After a review of rabbit housing the decision was made to transfer a population of male rabbits to floor pens on a permanent basis This offered an opportunity to view behavioural changes in rabbits when they are moved from cages to floor pens Before transfer to floor pens a cohort of 4 individually housed male rabbits were videoed for one 24 hour period in their home cages this was repeated after they had been housed in floor pens for two weeks Video records were analysed using fixed inter val instantaneous focal sampling with a record of activity posture and position in the cage or floor pen made every 15 minutes Rabbits spent significantly less time in the central area of the cage compared to central area of the floor pen p 0 012 ate less p 0 002 and there was a significant reduction in grooming p 0 002 and sitting p 0 019 in the floor pens Rabbits also appeared to spend less time at the back of the floor area and less time engaged in interaction with enrichment in the floor pen though there appeared to be an increase in exploratory behaviour rearing and time spent fully stretched out However none of these differences were statistically significant at the sample sizes used in this study 0 1 p 0 05 This pilot study suggests there are behavioural benefits to floor housing male rabbits due to increased space and behavioural oppor tunities though fur ther studies would be beneficial to assess if these findings are representative of the general laboratory rabbit population Key words laboratory rabbits floor pens behaviour welfare Introduction Rabbits are routinely housed in cages that do not offer the opportunity to perform a wide range of natural behaviours 1 for example they may be restricted in opportunities to dig rear or lie fully stretched In biomedical science it is generally accepted that robust scientific results rely on animals which are healthy and do not display signs of physiological or psychological abnormalities A fundamental part of ensuring the wellbeing of captive animals is meeting their behavioural requirements through increasing the variety and range of species appropriate opportunities 2 Species appropriate accommodation and consistency throughout studies as well as husbandry procedures such as gentle handling also help to reduce variables in experimental design 3 The rabbit is a well established animal research model In 2010 the total number of rabbits used in the EU was 335 910 representing 2 8 of all laboratory animals 4 The most common form of housing for laboratory rabbits is individual caging an approach that has remained largely unchanged over 30 years At the time of the study in 2013 the size of cages for singly housed adult rabbits in the UK Code of Practice was 4000 cm2 floor area and 45 cm height 5 Housing rabbits in large floor pens may encourage a more natural behavioural repertoire but space alone is unlikely to address all of a captive animals behavioural needs A number of studies have suggested that when provided with appropriate environments captive rabbits display few behavioural differences to their wild counterparts6 7 8 which could suggest that they would value access to a larger space with appropriate furnishings For example rabbits have been observed to forage when given the opportunity 8 including sifting or scratching at foraging substrates such as hay thrown on the floor of the pen or cage Currently the majority of laborator y rabbits are housed in cages where opportunity to forage is limited as pelleted food tends to be offered in hopper whilst hay or other forages if 7

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens provided are placed in metal hay racks Furthermore existing laboratory housing with limited environmental resources offer the rabbit limited control over their micro environment Research with other species is often aimed at improving their wellbeing through enhancing the control that they have over their microenvironment for example by offering them motivationally significant or other wise valuable resources such as nesting materials and nest boxes10 11 12 and or more complex environments Previous studies investigating the behaviour of rabbits housed in cages against those in floor pens involved two independent sets of rabbits12 13 with comparisons between the two populations rather than using a within rabbit design whereby the same rabbit is observed in both a cage and a floor pen This pilot study used a within rabbit design to control for individual variation in behaviour as well as potentially reduce the number of animals required for valid comparison All animal studies were ethically reviewed and carried out in accordance with Lincoln University Ethical Review Procedure and the GlaxoSmithKline GSK Policy on the Care Welfare and Treatment of Animals which determined the work did not constitute a procedure under Animals Scientific Procedures Act 1986 ASPA the minimum allowed by law in UK 4000cm2 floor area and minimum height 45cm and contained more cage furniture than many laboratory rabbits may experience The floor pens were 308cm x 160cm giving a floor area of 42980cm2 or about 7x the floor area of the cages with side walls 1m tall and no roof Environmental resources in the floor pen included a ramp the fun tunnel chew block sawdust on the epoxy resin flooring and an upside down guinea pig cage The home cages were adjacent cages within a single room the floor pens were adjacent to each other in a separate room to the cages Rooms were maintained at 20 C during study period on a 12L 12D light cycle Time lapse low light CCTV cameras and web video CCTV video manager software equipment were supplied by Tracksys Ltd The night vision camera had an infra red light source this allowed recording of rabbits behaviour during night hours without undue disturbance The PC screen was only switched on when the observer was in the room normally at 10am on test days for no more than 10 minutes at all other times the screen was turned off There was no discernible noise to humans although this was not tested for rabbits there was no perceptible change in rabbit behaviour when the recording equipment was switched off and on The aim of the study was to investigate the effects on behaviour of individually housed male Chinchilla rabbits when housed in cages and when they were moved to a floor pen Materials and methods Four adult male Chinchilla rabbits from Charles River Laboratories in Germany were assigned to the study and initially housed individually in 4 large rabbit cages Arrowmight with clear Perspex panels and door floor space 70cm x 85cm floor area of 5950 cm2 floor area and 50 cm height a shelf cardboard fun tunnel and chew block The cages were therefore larger than Figure 1 Rabbit lying on an upside down guinea pig cage 8 Figure 2 Rabbit cage

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens The rabbit cages were cleaned out once a week and the tray liners were replaced daily or as required Rabbits received water via bottles and were fed a high fibre maintenance diet 5325 from PMI with a handful of irradiated hay each day Rabbits had been acclimatised to the cages for 14 days prior to being videoed and videoed for a 24 hour period on the 15th day in the home cages floor pen made every 15 minutes over a 24 hour period see working ethogram below The rabbits were then moved to the individual floor pens by technologists who were not aware of which rabbits were being filmed to negate any pen location selection bias being made for the 4 selected rabbits Videoing did not commence until the rabbits had been acclimatised to the pens for 14 days The observer was blind to rabbits identities when they were moved to the floor pens to avoid familiarity bias Figure 5 Camera overlooking floor pen Figure 3 Rabbit floor pen Rabbits were a time lapse gave a high requirements for analysis videoed continuously for 24 hours using schedule of 3 frames per second which quality image whilst reducing memory thus enabling more footage to be filmed Figure 6 View into floor pen Figure 4 Floor pen room video camera set up The video was analysed by a trained observer using fixed interval instantaneous focal sampling with a record of activity posture and position in the cage or Figure 7 Rabbit in cage 9

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens Posture of the rabbit Ambulate Movement around the cage16 or pen but normal locomotor activity is slow usually observed while rabbits are foraging or exploring Body Resting Rabbit is in sternal recumbent position with legs tucked under the body Eyes open but rabbit is relatively inactive 16 or the back is relaxed at the shoulders with a slightly hunch appearance and ears may or may not be erect or the rabbit has a squat appearance and looks to be tucked up with its back arched 14 Figure 8 Rabbit in floor pen Working ethogram The working ethogram or list of behavioural categories was developed from published ethograms14 15 16 with alterations derived from pilot observations of rabbits in similar cages and floor pens This enabled rabbit activities to be defined for recording using the following working ethogram of behaviours For the purpose of instantaneous sampling an activity was defined as what an animal was doing whilst it was mobile or stationar y posture was defined as the exact deportment of the body and position was defined as the location of the rabbit in the cage or pen Activity Alert Rabbit s ears are forward during an activity such as ambulate hop and sit Doze Lying or sitting down with eyes partially open The rabbit appears to have an awareness of the environment which includes small movements in response to the activity and noise and attending technologists One or both ears of the rabbit is erect and may rotate in response to auditory stimulation or both ears will be dorsal with eyes half open 16 Eat Feeding from the hopper and chewing rabbit pellets or eating hay 1 16 14 Body Relaxed Rabbit in a left or right lateral recumbent position with forelimbs and hind limbs partly extended beyond the body and the backbone in a convex position or the forelimbs and hind limbs are extended fully extended Hop Rabbit has an arch of the back as it moves forward a faster pace than ambulate Lay fully recumbent The rabbit is lying in a fully recumbent position with the trunk of the body and all limbs fully stretched out usually in a straight line across the floor Sit alert The forelimbs are not folded beneath the body but are straight so the thorax and abdomen are clear of the floor and visible with ears erect All paws are still on the floor distinguishing this behaviour from rear 16 Stand alert Standing with ears upright or forward space between abdomen and floor rabbit is prepared for flight Rear Sitting on haunches or hind feet both forepaws clear of the floor rabbit is partially or fully stretched and often uses this stance to sniff the roof of the cage ears back or partially back 7 Rabbits may rest forepaws on the sides of the cage or be freestanding Both front feet sometimes leave the floor during washing or grooming but this was not recorded as rearing 16 7 Position Exploratory Rabbit ambulates across the area sniffing nibbling and nosing through sawdust hay sniffs at sides of the cage or pen and is alert when mobile at a slow pace Groom The teeth of the rabbit are used on the coat Fur is passed through the mouth unlike washing where only the tongue is used The rabbit appears to nibble at its coat using quick movements of the jaw 16 Tunnel interaction Rabbit enters the tunnel and either strips the inner part of the tunnel or hops through 10 Cage or pen location of rabbit see Figure 9 Back of the cage or pen 1 rabbit is at the back of the cage pen either under the shelf or in the general cage pen area Middle of cage or pen 2 rabbit is in the mid area of the cage pen Front of cage or pen 3 rabbit is in the front of the cage or pen either under the shelf or at the front of the cage outside of the shelf

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens Under the shelf or hide 4 rabbit is under the cage shelf or fully within the hide of the pen hidden from view On top of the shelf or hide 5 rabbit is on top of the shelf or hide Hidden from view not in vision rabbit has entered the hide found in the exercise pen but cannot be seen or heard only available in the pen 1 Shelf 5 4 Figure 10 There was a significant decrease in the number of times rabbits were observed eating in the floor pen t 9 67 p 0 002 The rabbits showed a tendency to carry out exploratory behaviour more often in the floor pens than cages though this was not significant t 2 90 p 0 062 including moving around the available floor space sniffing part of the environment or moving sawdust There was a significant reduction in self grooming after the rabbits were moved to the floor pens t 9 92 p 0 002 The pens and cages had enrichment items included however the only interaction with an enrichment item recorded during the observation period was with the cardboard tunnel and whilst this interaction diminished when the rabbits were moved to floor pens it was a not significant t 2 63 p 0 077 2 3 Front of cage Figure 9 The rabbit in the diagram would be counted as middle of the cage Statistical analysis The data was prepared for analysis using Microsoft Excel to count the number of time points each rabbit was observed engaged in a behavioural category listed in the working ethogram Because each rabbit was observed in both environments a paired two tailed Ttest Statistica was used to investigate differences in behaviour between cage and floor pen As this was a pilot study the differences were considered significant at statistical probability p 0 05 though it is important to note firstly that there was potential for type 1 errors due to multiple comparisons and secondly type 2 errors due to the small sample size As a consequence of the small sample size there was only a significant difference for the whole group if there was large difference in behaviour across all rabbits If one of the rabbits showed no difference in behaviour between the floor pen and cage or these differences were small for some rabbits then the difference only approached significance For these reasons behaviours where statistical probability p was below 0 1 but above 0 05 are also presented in results Results During the observation period there was no significant difference in time spent alert Paired t test t 2 14 p 0 121 or dozing t 0 61 p 0 583 when rabbits were moved from cages to the floor pens Table 1 Table 1 Total observation counts of the activity of each rabbit in the cage or floor pen Figure 10 Table showing mean SE of each activity for all rabbits when housed in cage or floor pen There was no significant difference between the number of times rabbits were obser ved to be 11

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens ambulating t 2 06 p 0 130 Table 2 Figure 11 There was no difference between rabbits in the resting position t 0 46 p 0 675 the relaxed position t 1 70 p 0 185 hopping t 1 05 p 0 369 when they were moved to floor pens The data suggested an increase in the number of occasions rabbits lay fully extended in the floor pens this was statistically not n significant t 2 50 p 0 087 There was a significant reduction in sitting after the rabbits were moved to the floor pens t 4 56 p 0 019 When the rabbit was recorded as in the sit position it was always alert When the rabbit was recorded in a stand position it was always alert and about to move there was no statistical difference time spent standing when the rabbits were moved to the floor pen t 0 77 p 0 495 The rabbits were not often observed to rear however there was an increase in how many times rearing was observed when they were moved from the cage to the floor pen which approached significance t 3 00 p 0 057 e lat bu am p ho l t ful no lie re stu po r rea d ng axe resti rel sit nd sta Figure 11 Table showing mean SE of each posture for all rabbits when housed in cage or floor pen To simplify the comparisons of position the ramp and Guinea pig cage were counted as a shelf in the floor pen Table 3 Figure 12 There appeared to be a decrease in the number of times rabbits were seen at the back of the floor area when they were moved from cages to floor pens but this was not significant t 2 66 p 0 076 There was a significant increase in the number of times rabbits were observed to be in the middle of the floor area when they were moved to the floor pen t 5 46 p 0 012 There was no significant difference shown in time at front of the cage t 2 20 p 0 115 when the rabbits were in the cages compared to when they were moved to the floor pen Table 3 Total observation counts of the position of individual rabbits when housed in either cage or floor pen There were no significant differences shown between when the rabbits were in the cages and when they were moved to the floor pens for either the number of counts observed on the shelf t 0 16 p 0 882 or under the shelf t 0 22 p 0 843 Table 2 Total observation counts of the posture of each rabbit in the cage or floor pen 12 Rabbits were awake more often during the dark period

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens Figure 16 and spent more time dozing in the light period Figure 17 regardless of whether they were in a cage or floor pen Figure 12 Table showing mean SE of each position for all rabbits when housed in cage or floor pen Figure 15 Rabbit lying fully recumbent Figure 13 Rabbit resting in the floor pen Figure 14 Rabbit relaxing in the floor pen Figure 16 Number of counts for awake 13

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens rabbits housed in cages when they were offered a fixed amount of food 8 Our rabbits were fed ad lib chow and offered hay during the afternoons This may explain the difference in the eating pattern which could be further explored as eating less while exercising more may help with obesity increase bone density and muscle tone Water intake was not recorded in this study as the bottles in the floor pens were in a blind spot for the camera and rabbits pull at the bottles which may skew the results if bottles were weighed Exploratory behaviour Figure 17 Number of counts for doze regardless of pen or cage housing Studies have shown that rabbits have a positive reaction to more stimuli in the environment 18 19 In the pilot study there was a non significant increase in explorator y behaviours including environmental interactions when the rabbits were moved to a floor pen This was maybe due to either greater floor space or increase in furniture or the combination of these two factors A number of environmental interactions were rare or never scanned in the home cage For example a new behaviour that they carried out in the floor pen was pushing their nose through the sawdust in a similar way to wild rabbits nosing through natural substrates such as vegetation Discussion Despite the relatively small sample size changes in rabbit behaviour between the cage and floor pen were sufficiently large to find significant differences in location posture and activity including less time eating grooming and sitting in the floor pen compared to the home cage Furthermore a number of behaviours of welfare importance17 were potentially more common in the floor pen than the cage including exploratory behaviour lying fully stretched and rearing Whilst these differences were not significant at the sample size used in this pilot study they would also be of interest if replicated in larger samples For these reasons and to provide insight into the value of these activities this discussion will talk about the impact on each behaviour in turn Activity Feeding behaviour When moved to the floor pens the rabbits spent less time eating Increased food consumption as well as other activities such as increased grooming has been related to lack of behavioural opportunities or low environmental stimulation in captive animals Furthermore elevated feed intake coupled with lower exercise can lead to obesity and poor body condition particularly in rabbits In this study there was no evidence of obesity and lower feed intake has not been noted in other studies of floor pen use For example no dif ferences were found in feeding behaviour between rabbits housed in floor pens vs 14 Figure 18 Picture sequence 1 4 rabbit pushing the tunnel around a floor pen In contrast there were a few occasions that rabbits in the cage used escape behaviours such as scratching at the bars or cage corners and these behaviours were not observed in the floor pens Indeed no potentially abnormal behaviours were observed in this study with the exception of one circling incident i e one record in the cage The absence of these activities in the floor pen is another potential benefit of floor housing The relatively low frequency of escape attempts or redirected activities in the cage is also noteworthy which may be a consequence of its relatively large size almost 6000 cm2 floor area height 50 cm and provision of enrichment resources

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens Grooming There was a significant reduction in self grooming that correlates with previous studies of floor housing8 which found that whether rabbits were housed in cages or in pens more enrichment in their environment appeared to reduce abnormal or excessive grooming Whilst the level of grooming observed in our rabbits was not high and there was no evidence of injury due to excessive grooming over grooming can be a behavioural problem for captive animals leading to fur loss and skin lesions in extreme cases Although some cases of overgrooming have been associated with specific illnesses in the general population low environmental complexity leading to more free time has been implicated in the development of over grooming The increased space as well as providing opportunity for more behavioural expression can indirectly improve health and welfare by reducing incidence of undesirable or harmful activities 16 17 Posture Ambulating and hopping Rabbits were recorded as ambulating when they moved slowing through the cage or floor pen this included when they nosed through hay and moved forward as they ate There was no difference in the number of times rabbits were seen ambulating or hopping There was however a difference in the form of hopping behaviour between the floor pen and the cage noted during observation In the cage the rabbit could complete one large hop in any direction whereas in the floor pens it could complete up to six large hops or many small hops across the floor area The rabbits were rarely seen only performing having one hop in the floor pens which suggests that they fully utilise the space that they are offered It is interesting that pet rabbit legislation such as that adopted by the Welsh Assembly24 requires hutches to allow room for at least 3 hops full height in the floor pens without their ears touching the top Related work in pet rabbits17 has found that rabbits with 45 cm roof height reared less than those with 75 cm roof height but that removing the roof altogether further increased rearing even in smaller rabbits able to fully rear in 75 cm high enclosures This suggests that low cage height may place both physical and psychological restrictions on the rabbit s ability and willingness to rear Position of rabbit in the cage Each floor pen had a ramp Figure 19 under which rabbits were observed to rest underneath during the day and spend time on top at night The reason for this may need further investigation although it may be as it is the highest point in the pen the rabbits could reach to look out for the other rabbits Studies carried out by Seaman et al 2008 1 showed that rabbits worked hard to gain access to a shelf but did not always use it 1 This finding concurs with the current study where the rabbits in the floor pen did not spend time on top of the higher areas offered They were usually observed under the ramp with their noses pointing towards the lowest angle This may help them feel more secure while allowing them to have a good look out position when staff entered and exited the room Similarly in the cages the rabbits were usually observed under the shelf with their noses at the front which would give them maximum observation of activity in the holding room The cage and pens were divided into thirds for the observation period While every effort was made to ensure the sections were an equal size there was a possibility that the middle of the pen incorporated a higher proportion of the floor area than middle of the cage and thus increased the amount of times rabbits were clearly observed in this area However the difference between the cage and the pen was highly significant for the middle of the pen Lay fully recumbent Rabbits were observed to be fully recumbent with hind legs fully extended in the cages on a number of occasions this behaviour was frequently observed in the floor pens but rare in the home cages The increase in the number of times they were observed to be fully recumbent in the floor pen may be an indication that when given more space rabbits will have a preference to lay fully recumbent 17 Further research is needed to investigate this resting or stretching behaviour as it may be an indication that current cage sizes are too small Rear The rabbits were noted to rear significantly more in the floor pens than the cages this may be an indication the rabbits were more aware of each other in the floor pens and may be because they had the ability to rear to their Figure 19 Top left ramp top right rabbit curled round feed bowl 15

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens When housed in floor pens the rabbits were offered food in a large round food bowl in the centre of the pen and were often observed to have their body wrapped around the food bowl Figure 19 suggesting that if housing more than one rabbit together providing a food bowl per rabbit may reduce fighting and or any potential for rabbits to avoid feeding due to the close proximity of another individual at the food bowl It would be interesting to determine whether they would continue to spend time resting by the food hopper if it was in a different position within the pen The position of the food resource may have been the reason the rabbits spent significantly more time in the middle of the floor pen although resting close to the food hopper was not observed often in the cage 22 23 24 25 Light and dark phase During the light phase the rabbits appeared to be generally inactive Figure 16 except when they were disturbed by technical staff which was then followed by a period of activity by the rabbits whether in the cages or floor pens When the dark phase began rabbits had periods of inactivity followed by long periods of activity Figure 17 This diurnal pattern of behaviour needs further investigation over a longer period of time and with more animals For example observation during day time only may overestimate resting time and underestimate activity periods and consequently the effects of providing additional space and environmental enrichment 26 27 28 29 10 Conclusion In conclusion moving to floor pens had a number of positive benefits for rabbit behaviour and welfare increasing time in key activities such as exploration rearing and stretching whilst reducing grooming and eating Although there were few signs of frustration in the large enriched cage the additional space in the floor pen increased behavioural opportunities In the current study the significant reduction in grooming and feeding suggested the rabbits were otherwise occupied with different activities when moved to a floor pen Having more useable space encouraged the rabbits to spend more time in the open area of the pen and increased the interactions with their environment 11 12 13 14 15 Acknowledgements My manager at GSK Dr Greg Whelan for his help and advice and Dr Mark Lennon for his help with the data analysis For their diligent study support all of the team at Laboratory Animal Science Stevenage especially Terry Ballard Mervin Saville and Will Traylen 16 17 18 References 21 16 Seaman S C Waran N K Mason G and D Eath R B 2008 Animal economics assessing the motivation of female laboratory rabbits to reach a platform social contact and food Animal Behaviour 75 1 31 42 Mellen J and Sevenich MacPhee M 2001 Philosophy of environmental enrichment Past present and future Zoo Biology 20 3 211 226 Verwer C M van der Ark A van Amerogen G van den Bos R and Hendriksen C F M 2009 Reducing variation in a rabbit vaccine safety study with particular emphasis on housing conditions and handling Laboratory Animals 43 1 155 164 European Regulations 2010 Sixth Report on the Statistics on the Number of Animals used for Experimental and other Scientific Purposes in the Member States of the European Union Sec 1107 European Commission Sixth 8 12 2010 6 6 2013 Home Office Code of Practice for Animals housed in Scientific Procedure Establishments Hawkins P Hubrecht R Buckwell A Cubitt S Howard B Jackson A and Poirier G M 1993 Refining rabbit care a resource for those working with rabbits in research RSPCA UFAW Mykytowycz R and Hesterman E R 1972 An Experimental Study of Aggression in Captive European Rabbits Orctolagus cunniculus L Behaviour 52 1 2 104 123 Selzer D and Hoy S 2003 Comparative investigations on behaviour of wild and domestic rabbits in the nestbox World Rabbit Science 11 1 13 21 Krohn T C Ritskes Hoitinga J and Vendsen P 1999 The effects of feeding and housing on the behaviour of the laboratory rabbit Laboratory Animals 33 2 101107 Manser C E Broom D M Overend P and Morris T H 1998 Operant studies to determine the strength of preference in laboratory rats for nest boxes and nesting materials Laboratory Animals 32 36 41 Blom H J M Van Tintelen G Van Vorstenbosch C J A H V Baumans V and Beynen A C 1996 Preferences of mice and rats for types of bedding material Laboratory Animals 30 234 244 Van Loo P L P and Baumans V 2004 The importance of learning young the use of nesting material in laboratory rats Laboratory Animals 38 17 24 Podberscek A L Blackshaw J K and Beattie A W 1991 The behaviour of group penned and individually cages laborator y rabbits Applied Animal Behaviour Science 28 353 363 Ferrante V Verga M Canali E and Mattiello S 1992 Rabbits kept in cages and in floor pens reactions in the open field test J Appl Rabbit Res 15 700 707 Gunn D and Morton D B 1995 Inventory of the behaviour of New Zealand White rabbits in laboratory cages Applied Animal Behaviour Science 45 277 292 Held S D E Turner R J and Wooton R J 1995 Choices of laboratory rabbits for individual or grouphousing Applied Animal Behaviour Science 46 81 91 Dixon L M Hardiman J R and Cooper J J 2010 Assessing the spatial needs of pet rabbits Oryctolagus cuniculus Journal of Veterinary Behaviour 5 303 308 Stauffacher M 1994 Improved husbandry systems an ethological concept In Welfare and Science Proceedings of the Fifth FELASA Symposium Bunyan J ed 68 73 Royal Society of Medicine Press London UK

Enhanced housing for male laboratory rabbits a pilot study investigating potential benefits of floor pens 19 20 21 22 23 24 Whary M Peper R Borkowski G Lawrence W and Ferguson F 1993 The effects of group housing on the research use of the laboratory rabbit Laboratory Animals 27 4 330 341 Lidfors L 1997 Behavioural effects of environmental enrichment for individually caged rabbits Applied Animal Behaviour Science 52 157 169 Hansen L T and Berthelsen H 2000 The effect of environmental enrichment on the behaviour of caged rabbits Oryctolagus cuniculus Applied Animal Behaviour Science 68 2 163 178 Lockley R M 1965 The private life of the rabbit Book Club Associate 86 87 Hansen L T and Berthelsen H 2000 The effect of environmental enrichment on the behaviour of caged rabbits Oryctolagus cuniculus Applied Animal Behaviour Science 68 2 163 178 Welsh Assembly 2006 Code of practice for the welfare of rabbits following the code ISBN 978 0 7504 5356 1 Crown copyright September 2009 17

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April 2015 Animal Technology and Welfare Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting PENNY HAWKINS SECRETARY 1 MANUEL BERDOY2 CHARLOTTE BUR3 DEBBIE BURSNALL4 JOANNA CRUDEN5 WANDA MCCORMICK6 AMY MILLER7 HELEN PROCTOR8 DAVID WHITTAKER 2 MAGGY JENNINGS1 and ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Department of Veterinary Services c o University Laboratory of Physiology University of Oxford Oxford Oxfordshire OX1 3PT Centre for Animal Welfare The Royal Veterinary College Hawkshead Lane North Mymms Hertfordshire AL9 7TA Division of Biomedical Services University of Leicester Lancaster Road Leicester Leicestershire LE1 7HB GSK Research and Development Park Road Ware Hertfordshire SG12 0DP Moulton College West Street Moulton Northampton Northamptonshire NN3 7RR School of Agriculture Food and Rural Development University of Newcastle Newcastle upon Tyne NE1 7RU World Animal Protection 222 Grays Inn Road London WC1X 8HB UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Corresponding author penny hawkins rspca org uk Introduction The RSPCA UFAW Rodent Welfare Group holds a oneday meeting every autumn so that its members can discuss current welfare research exchange views on rodent welfare issues and share experiences of the implementation of the 3Rs of replacement reduction and refinement with respect to rodent use A key aim of the Group is to encourage people to think about the whole lifetime experience of laborator y rodents ensuring that every potential negative impact on their wellbeing is reviewed and minimised Our 21st annual meeting was held on 23rd October 2014 attracting 90 delegates from a wide range of universities and pharmaceutical companies throughout the UK Presentation topics included animal sentience reducing suffering during procedures assessing rodent health and welfare and how to ensure the right decisions are made when providing environmental enrichments such as running wheels The day ended with a discussion on the Culture of Care and how this can be recognised promoted and maintained within institutions This report summarises the meeting and ends with a list of action points for readers to raise at their own establishments Animal sentience what do we know and why does it matter Helen Proctor World Animal Protection Animal sentience can be defined as the ability to feel both positive and negative emotions and to be aware of a variety of states and sensations 1 Research into animal sentience is constantly expanding so that we can now infer more than ever about the subjective minds of animals 2 In recent years research has shown that some animals grieve 3 that decapod crustaceans can feel pain4 and that mice and rats can be empathetic 5 6 This fascinating area of science provides us with insights into the emotional lives of animals with important implications for how we utilise and interact with them 1 However because animal sentience is concerned with the inner mind of our fellow animals studying sentience may be viewed as controversial due to its apparently subjective nature 1 7 Critics argue that it is impossible to measure animal emotions objectively or even attribute any meaningful experience to them 8 But in a recent systematic review of the scientific literature we found that much research using animals does assess and use the subjective states of animals 19

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting objectively and scientifically 2 Furthermore it uses these states to evaluate the effectiveness and safety of drugs for human therapy Our systematic review included over 2 500 papers published between 1990 and 2012 selected on the basis of their inclusion of keywords specific to animals and animal sentience 2 We found that knowledge of animal sentience comes largely from laborator y research given that over 79 of relevant studies were conducted in the laboratory The majority of studies 69 were conducted for human benefit e g pharmaceutical research and development rather than for the purpose of gaining insights into animal welfare or behaviour Almost all studies assumed the existence of sentient traits such as pain fear and pleasure Rodents were the subject of most of the papers in our review and as a result we can infer a lot about their subjective minds To give just three examples studies have shown that rodents are capable of Regret defined as recognising that you made a mistake and that if you had done something differently there would have been a better outcome Researchers studying decision making in rats found that animals who skipped the chance to have a highvalue treat so they ended up with a lower value reward looked back at the location of the high value treat On the basis of the animals behaviour the implication was that they regretted their decision Neurological studies showed that the orbitofrontal cortex of the rat brain was active when the animals looked back which is the same area that is active in the human brain when we are feeling regretful 9 Empathy or the ability to understand and share the feelings of another has been examined in laboratory rats by placing a free rat into an arena containing a cagemate who is trapped in a restrainer 5 After several sessions the free rat will learn to open the restrainer and free the trapped animal but they do not open restrainers that are empty or contain objects Given a choice between opening two restrainers containing a cagemate or chocolate respectively rats preferred to open the restrainer with the cagemate inside first then open the second restrainer and share the chocolate This provides strong evidence of empatheticallymotivated helping behaviour in the rat Laughter in the form of ultrasonic vocalisation patterns of around 50 kHz which have been recorded in rats in response to play with other rats or tickling by humans These chirps are widely accepted to indicate positive affect or mood and are increasingly believed to be analogous to laughter in humans 10 See video at http www sciencemag org content 334 6061 1427 suppl DC2 Article and video at http www wired com 2013 09 tickling ratsfor science 20 Studies such as these have clear implications for those using or caring for laboratory animals They may simply confirm what empathetic staff have already observed or indicate potential issues with respect to data quality e g if social animals capable of empathy are housed individually or help to identify ways of refining housing husbandry and care Of course some of this research presents an ethical dilemma if regulated procedures are used to generate data that can successfully improve the lives of other animals Ultimately encouraging wider recognition that animals are sentient beings and that their feelings matter both to them and to us can provide a driver to replace animal use If you are interested in learning more about the science of animal sentience then join the discussion Visit the Sentience Mosaic www sentiencemosaic org where you can have your say in virtual debates read inspiring interviews and learn about all the great scientific research taking place around the world Skin to skin contact looking at refinements in skin closure techniques Debbie Bursnall University of Leicester Surgical embr yo transfer is a ver y commonly conducted procedure So ensuring that the most refined techniques are used will have a significant impact on laboratory mouse welfare Skin closure at the end of the procedure is an important area to consider Many options are available for closing the skin all of which aim to produce healing by primary intention which is directly opposing the skin layers to facilitate quick natural healing Commonly used skin closure methods have developed from medical and veterinar y practice but there is little published information about the quality of the wound closure in mice A new study involving the use of CD1 mice for embr yo transfer prompted a study to compare different skin closure methods to see which was best tolerated and provided the most effective healing as we wanted to ensure that we were observing good practice and minimising suffering To avoid generating additional animal use the mice used in the evaluation study were undergoing embryo transfer anyway as part of another project The study compared four skin closure methods in surgical embr yo transfer mice tissue adhesive GLUture Abbott Animal Health absorbable suture Vicr yl 6 0 Ethicon 7mm Autoclips Har vard Apparatus and staples Proximate 35 Ethicon Each of the four methods was used to close a single lateral dorsal skin incision in 124 CD1 mice at 0 5 dpc in a randomised study conducted over 15 days

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting All mice were anaesthetised with isoflurane 2 to 2 5 in oxygen and subcutaneous carprofen was administered at 10 mg kg on induction for pain relief A local anaesthetic bupivacaine was also administered at the incision site after shaving and then a scalpel was used to make a lateral dorsal skin incision of 5 to 6 mm followed by a dorsal ventral muscle incision An infundibulum embryo transfer was performed The wound was then closed using either i a single blanket suture with 6 0 Vicryl 35 mice ii a thin line of tissue adhesive 37 mice iii two staples 19 mice or iv a single 7 mm Autoclip 33 mice Wound closure took the least time with Autoclips 25 seconds and tissue glue took the longest time 140 seconds The mice were closely monitored at least daily for 14 days post operatively focusing on skin condition and whether the closure device was retained and the wound still sealed If a mouse had removed the device they were carefully examined to see whether remedial action was necessary to alleviate pain or the risk of infection Two skin samples from each closure method were taken post mortem and sent for histology at 4 and 9 weeks post op to evaluate skin healing The results of the study are summarised in Table 1 Method Advantages Sutures Good skin to skin contact Does not need to be removed Excellent wound healing Tissue glue Staples Autoclips Disadvantages Good skin to skin contact but some mice removed the glue Does not need to be removed Excellent wound healing Skin needs to be dry Care is needed to control the amount applied Time consuming Can be removed by mouse Non skin penetrating Good wound healing even if mice remove staples Difficult to control on application Removed by all mice Quick to apply Needs to be removed with a separate tool Skin penetrating Skin puckers and is red after removal Moderate wound healing larger scar than other methods Table 1 Comparison of four different wound closure techniques for embryo transfer in CD1 mice Although the advantages and disadvantages for the animals were given top priority when deciding which technique was to be preferred we also reviewed the financial cost of the different methods Sutures and glue worked out as 2 70 and 2 33 per animal respectively while staples and Autoclips were significantly cheaper but required outlay on equipment 8 70 for the staples and over 400 in the case of Autoclips We concluded that sutures were the best tolerated and most effective method of skin closure for embryo transfer with our CD1 mice If the closure using sutures failed it would heal well with no additional intervention which is another advantage because it means that further wound repair procedures are not necessary Sutures are also the most cost effective method of wound closure Sutures will therefore be used for skin closure in CD1 mice undergoing embryo transfer at our facility and other studies are planned to compare different suture types and patterns We will also evaluate the optimum technique for skin closure for different strains to help ensure that each surgical procedure is fully refined from beginning to end A comparison of abdominal and scrotal approach methods of vasectomy and the influence of analgesic treatment in laboratory mice Amy Miller University of Newcastle Like embr yo transfer vasectomy is a commonly conducted procedure in the production of genetically altered GA animals which also means that it is very important to ensure that it is fully refined so as to minimise suffering There has been some debate as to which surgical approach is preferable from an animal welfare aspect The BVAAWF FRAME RSPCA UFAW Joint Working Group on Refinement11 recommends that vasectomy is performed via an incision in the scrotal sac rather than via laparotomy arguing that the former could be less painful due to minimal tissue trauma This recommendation was made on the basis that the abdominal musculature bears the weight of the abdominal contents and the scrotal approach would avoid trauma to this supporting musculature and requires a smaller opening reducing the risk of infection Incising the scrotal sac also allows the vas to be exposed without exteriorising the testis and only one suture is required for wound closure People working in 21

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting the field had also commented that mice appeared less hunched following surgery via the scrotal route The recommendation was thus made on the basis of current thinking on good practice but empirical studies were needed to assess whether the scrotal sac route actually is less painful than laparotomy We conducted a study to evaluate this for CD1 mice funded by BBSRC and Pfizer which has been published in Laboratory Animals12 so a brief summary will be presented here All of the mice used in the project were required for the University s transgenics production programme In an initial pilot study groups of mice underwent vasectomy via either abdominal or scrotal approach surgery All animals received carprofen for pain relief because it is known that abdominal vasectomy affects behaviour in CD1 mice so a control group without analgesia was not necessary Mice were filmed for 15 minutes before surgery and at 1 24 and 48 hours post surgery and data were obtained using automated behaviour recognition software HomeCageScan Cleversys Inc Behaviour changes after surgery were compared between groups at each time point Exploratory behaviours such as rearing walking and sniffing were most greatly reduced one hour after surgery whereas the duration of grooming increased By 48 hours these changes had largely subsided Behaviours associated with pain occurred significantly more frequently one hour after abdominal surgery than with the scrotal approach although there was an increase in belly pressing in animals who had undergone surgery via the scrotal sac However these differences were very small so it was not possible to draw any definitive conclusions with respect to which method was better from an animal welfare aspect The subsequent main study evaluated the responses of the mice to different levels of drug treatment on the basis that more painful procedures would require more aggressive therapy so this should help to better identify any differences in the effects of the two surgical approaches Mice received three analgesic treatments i meloxicam ii meloxicam plus paracetamol or iii saline subcutaneously We always consider the justification for withholding pain relief very carefully and in this case it was judged by our local ethical review process to be necessary in order to achieve meaningful data on the effectiveness of the analgesics Unfortunately the results of the main study showed that neither meloxicam nor meloxicam plus paracetamol had any demonstrable beneficial effects for the animals This means that either the analgesia was ineffective or the behavioural scoring was not sufficiently sensitive to pick up subtle indicators of the beneficial effects These results did show us that the 22 control group was necessary however as without the control we may have concluded that both analgesic treatments were equally effective when in fact both may have been equally ineffective Our study and other recent data on the effects of nonsteroidal anti inflammatory drugs NSAIDs in mice suggest that either considerably larger doses of these or more potent analgesics more precise monitoring of surgical outcomes or a combination of both these factors is needed to determine the true extent of pain experienced by mice undergoing vasectomy Meanwhile the skill of the surgeon may be more important than the surgical approach so immediate welfare gains can be made by benchmarking the performance of surgeons and ensuring that they are skilled and competent Development and validation of a body condition score for Guinea pigs Wanda McCormick Jenna Catlin and Kate Leslie Moulton College John Lowe Dodson Horrell Ltd Body condition scores BCS have successfully been applied to a wide range of animal species in different contexts as tools for health and welfare assessment However to date no BCS system exists for use in Guinea pigs This project aimed to create and validate a suitable BCS scale which can be used in a range of captive environments for Guinea pigs This species was chosen because it is estimated that around half a million Guinea pigs are kept as pets in the UK accounting for 1 1 of households but their owners are often inexperienced in assessing and maintaining animal health so these Guinea pigs often develop obesity and dental problems due to improper feeding Although the BCS was devised with companion animals in mind it will also be useful for assessing Guinea pigs in a laboratory setting whether at a breeder or user establishment We used an initial sample of 24 Guinea pigs group housed in pens at Moulton College to obtain a range of body measurements alongside the body mass of each animal Measurements of girth chest and length were found to be highly significantly correlated to body mass p

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting ribs spine and pelvis were used to create a BCS To validate the new BCS body measurements were recorded from over a hundred Guinea pigs housed in a range of settings including owners homes pet stores nurseries and rescue centres Each Guinea pig was assessed for BCS independently by both an assistant and a researcher then these results were compared There was no significant difference p 0 452 between the two sets of BCS data suggesting the scale is reliable for use by both trained and untrained persons The measurements of body mass in this larger sample were again highly significantly correlated p

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting this is not meant to be an exhaustive list but an illustration of some typical problems No clear question or hypothesis What is the question exactly How was benefit decided How was benefit measured Is it this particular shelter or shelters in general There was no control condition additional data should have been obtained from cages without added shelters Each mouse was chosen by the observer which would have introduced bias e g they may have selected dominant or submissive animals The animals should have been chosen randomly there are plenty of easy ways to do this Animals were only observed during their inactive phase during the human working day behaviour at night was not recorded and is likely to have been different No observations were made over the weekend when animals may have behaved differently because fewer staff were in and noise levels were lower Insufficient number of animals the sample size was actually just 6 not 180 because they are not independent No accounting for the effect of the observer video cameras could have been used instead and could also have been used to obtain more data No baseline data were taken before the shelters were added Only females were used although this may not have been an issue if the aim was to evaluate the benefits of a shelter for a project involving only female mice No time apparently allowed for the mice to habituate to the shelter All cages were at the same level and in groups of 3 cages should have been randomly selected to allow for differences in height above the floor and light levels Although Norman s study showed that the chosen mice were interacting with the shelter when he made the observations it is not robust enough to demonstrate a significant benefit or to help make a decision about the best type of shelter to provide for the animals Evaluation studies like this need to have a hypothesis asking a clear question that can be tested by the experiment Incidentally was Norman s question really about benefits to the animals or about what the animals want Both questions are good ones but they do not mean the same thing animals like humans don t always want what is good for them Evaluation studies also need adequate and independent sample sizes and valid controls with potential sources of variation included in the design none of this was evident here Sources of bias should also be recognised and addressed Biases may occur in 24 selection or allocation e g no randomising of animals cages self selection or in failing to take account of the behaviour of the species e g response to novelty circadian rhythms or the influence of the observer e g the subject may be stressed by or attracted to the observer A further source of bias can be failure to implement blinding when obtaining or analysing data Ideally the observer should not know which animals have received the treatment and nor should the person analysing the data This may not always be possible and would not have been feasible in our hypothetical example but studies should always be blinded if they can be There is currently much emphasis on the importance of good experimental design in the life sciences in general and studies to evaluate enrichment are no exception It is always a good idea to obtain advice from someone with exper tise in statistics and experimental design when planning your study and the first port of call for this would usually be the local AWERB which should have access to a source of statistical advice Some useful publications are also listed in the reference section of this paper 13 14 15 16 Animal technologists have a great deal to offer with respect to evaluating implementing and reviewing enrichment and you should be able to access the same level of support as the researchers at your establishment The running wheel debate Charlotte Burn Royal Veterinary College Running wheels are often provided as an enrichment but there has been debate regarding their benefits Here I discuss two questions Are running wheels good for rodent welfare do they keep rodents healthy and do rodents actually want them and Do they make for better science In rodent cages oppor tunities for exercise are extremely limited so unsurprisingly the provision of a wheel brings about many of the general health and cognitive benefits expected from regular exercise These benefits include enhanced heart function18 and cognitive function 19 Rodents also do seem to want running wheels indicated by the fact that they are prepared to expend considerable energy e g by pressing a lever many times to gain access to a wheel 20 The rewarding effects of wheel running have been linked to the release of opioids21 and rats show conditioned place preference for places that they associate with just having been for a run 22 However another behavioural study has shown that rats avoid The NC3Rs Experimental Design Assistant will also soon be available see https www nc3rs org uk experimental design last viewed 2 January 2015

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting places that predict wheel access23 suggesting that rodents experiences of wheel running may not be straightforward pleasure Despite the potential health benefits of wheel running to most rodents individuals who wheel run excessively can develop physical deformities such as arching of the spine lordosis or hyper flexion of the tail resembling Straub tail a response to morphine treatment in rodents These morphological changes may or may not be associated with pain and can also affect data quality e g in behavioural tests requiring good motor skills or balance 24 More research is needed into the effects of wheel design and length of time spent running on both welfare and science 24 There has also been some debate as to whether wheelrunning is a stereotypy Stereotypic behaviours have been defined as repetitive invariant and apparently functionless or goalless 25 and wheel running does fit this description in some but not all individual rodents On the other hand providing stereotypic rodents with a wheel often reduces stereotypies but this could mean that either i the wheel is an enrichment improving welfare and satisfying a previously frustrated motivation or ii wheel running is a redirected stereotypy leaving welfare largely unchanged The fact that both stereotypies and wheelrunning are reduced by fluoxetine and naloxone26 27 seems to support ii but it is not that conclusive because these substances tend to reduce per formance of all rewarding behaviours Wheelrunning occurs in diverse environments and animals use wheels in a variety of ways e g jumping on and off or building nests inside which is less supportive of the idea that it is a stereotypy It is perhaps more likely that wheel running exists in both stereotypic and nonstereotypic forms 26 Similarly extreme use of running wheels has parallels with addiction 24 For example some individuals are seemingly unable to regulate their wheel use despite the onset of adverse effects e g becoming physically deformed or emaciated they spend such a large proportion of their day wheel running that normal social and or maintenance behaviours become disproportionately reduced and they become aggressive if wheel running is prevented Again not all wheel use follows this pattern Turning to the implications of wheel running for science if performed to moderate levels the many benefits of exercise in an other wise restrictive environment could probably lead to more physiologically normal animals Also wheel running itself can be a useful tool for assessing treatment effects in specific cases e g where treatments are Want as defined by Dawkins 2004 17 hypothesised to affect activity levels However if performed to excess the behavioural and physical effects of wheel running could lead to welfare problems as well as abnormal treatment responses that may affect standardisation Huge strain sex and individual differences in propensity towards wheel running exist 24 so evaluation of the welfare and scientific harms and benefits may require a case by case approach In conclusion excessive wheel use can be harmful to health welfare and science but moderate use can confer benefits keeping rodents healthier allowing them to do something they want to do and making them more physiologically normal models In strains known to run to excess wheels should be avoided and other enrichment provided to encourage exercise Use of wheels should be monitored but care is needed if removing wheels from excessive users due to the potential for withdrawal similar to drug withdrawal that could cause suffering 24 Safe wheel design is important to avoid entrapment and deformity plus other enrichments should always be provided to allow choice and encourage a range of activities Building a nationwide NACWO exchange initiative Jo Cruden and Sam Izzard GSK Stevenage Requirements within the revised Animals Scientific Procedures Act 1986 ASPA such as actively ensuring adequate Continuing Professional Development CPD and keeping formal records of this provide strong encouragement for people in named roles to develop and learn However over the past few years we have noticed a gap in the education of Named Animal Care and Welfare Officers NACWOs not in terms of training per se but with respect to having opportunities to build a strong network and learn about other facilities The revisions to the ASPA prompted us to explore the potential for a scheme that would enable and support NACWOs to meet spend time with each other and share their knowledge and ideas We created a proposal for an exchange in which a NACWO will spend the day with another NACWO at a different facility and vice versa We envisage three main benefits insight into day to day work in other facilities opportunities to share ideas and good practices and the ability to build up a network of contacts Both the host and visiting establishment will gain Continuing Professional Development CPD credits based on the number of hours spent actively visiting and discussing roles responsibilities and how things are done at each establishment A tick list has been developed to help structure and prepare for visits and a post exchange report is completed and submitted to the Named Training and Competency Officers NTCOs at both 25

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting facilities The exchange protocol can also be used for internal exchanges for NACWOs working at different sites within the same establishment to help promote cross site communication A successful pilot study has been run in collaboration with Imperial College London which was used to further refine the scheme in the light of feedback from the participants A second exchange is well underway with MRC Harwell and there has been a lot of interest from other groups interested in taking part The scheme is now being rolled out nationally in partnership with the IAT including a secure section on the Institute s website for NACWOs to keep records of visits and ideas to share inviting participants to present at IAT Council and highlighting the scheme on the IAT website Section of ASPA Guidance mentioning a culture of care Establishment Licence Holder ELH Section 3 13 2 NACWO Sections 8 8 1 8 8 2 New participants will be led through the process by a guidance document detailing how the scheme works what is expected of them and what they will gain from the exchange Ideally they will link up with someone who has already completed an exchange and contact exchange monitors will be listed within the secure section of the IAT website Our vision is a network of NACWOs communicating and sharing ideas as well and gaining CPD although there are differences in the way our respective roles are structured we all have the same goals when it comes to animal care and welfare If you are interested in taking par t please contact Andy Cunningham ac572 le ac uk and see the announcement in the back of this issue David Whittaker University of Oxford A Culture of Care is much easier to say than it is to define deliver and sustain This now often over quoted sound bite first appeared formally in print in the 2000 edition of the Guidance to the ASPA and is directly referred to in several places within the 2014 Guidance28 as listed in Table 2 Fulfilling these requirements and recommendations requires careful thought about who and what the Culture of Care is for who is responsible for its delivery what it means in practice inputs and whether it can be measured in terms of outputs or deliverables The culture should also be sustainable and sustained in the long term Simply put the Culture of Care should demonstrate caring respectful attitudes and behaviour towards animals and encourage acceptance of responsibility and 26 G G G Training Local Module Section 9 11 AWERBS Section 10 5 A culture of care a personal experience Requirement recommendation G G You will need to be proactive and provide effective leadership You will need good management and communication skills and the commitment to nurture a culture of care in your establishment NACWOs should have appropriate personal authority to promote high standards and will need good communication and diplomacy skills to champion a culture of care amongst both scientific and husbandry staff The NACWO should champion a culture of care at your establishment acting as a role model for all those who care for and use animals We recommend that each establishment should prepare a local module information on the functions and processes of the local AWERB and how the local culture of care is promoted should be included More generally AWERBS should help to promote a culture of care within the establishment and as appropriate in the wider community Table 2 References to a culture of care in the 2014 ASPA Guidance document28 accountability 29 This goes beyond just meeting the minimum requirements of the legislation While each organisation s culture will depend upon the values and attitudes of its staff and the local processes in place that determine how people work and behave all establishments should have a vision of what their own Culture of Care means how we do things around here Caring for and about animals is of course central to the concept but it should also encompass caring for the equipment facilities and each other including team members users customers clients and other internal and external stakeholders The specified roles and tasks of the ELH NACWOs trainers and AWERBs are set out in Table 2 but everyone has a part to play

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting including and especially the animal technologist at the cageside Information has to flow effectively in all directions within a culture that supports openness between and learning from one another but has zero tolerance for poor practice or noncompliance Think about what you can see hear and smell and consider whether it extends to or goes beyond compliance and how it makes you feel If something does not feel right you should be able to express your concerns safely and effectively It is vitally important for all establishments to have a policy and procedure in place for any member of staff to raise concerns about any aspect of animal care or use This should include clear communication channels that staff are confident to use without fear of negative consequences either professionally or socially The University of Oxford has a dedicated system for raising concerns in place as do other establishments see the report of the workshop on Raising Concerns about Laboratory Animal Welfare held at the 2014 IAT Congress 30 The new LASA RSPCA Guiding Principles for AWERBs29 and the revised RSPCA resource book for lay members31 also discuss practical ways of developing and maintaining a culture of care and these will both be good sources of ideas and inspiration for you and your establishment List of action points based on all of the presentations and discussions If embr yo transfers are conducted at your establishment suggest a review of wound closure techniques to see whether the most effective welltolerated technique is being used If males are vasectomised suggest that the protocol is reviewed using the relevant section of this report and reference 12 as a basis to see whether further refinements could be implemented and evaluated Think about other commonly conducted procedures which are done according to standard protocols at your establishment Would any of these benefit from a review You could suggest this to your AWERB Named Persons or other relevant local committee If you care for Guinea pigs try using the size ometer and provide some feedback to the contact above When designing studies that aim to evaluate husbandry refinements including environmental enrichment obtain advice from someone with expertise in statistics and experimental design to ensure that your data will be significant and robust Ensure that you have identified and minimised sources of bias including those due to the behaviour of the species sex and or strain If running wheels are routinely provided at your establishment ask for the AWERB or relevant animal care committee to look at the section of the report outlining the debate There may be a case for altering the wheel design setting up a protocol for monitoring levels of use or using alternative enrichments if running is excessive If you are a NACWO participate in the exchange scheme or if you are not make sure your NACWOs are aware of it Think about your role in your establishment s Culture of Care whether or not you are a NACWO trainer or sit on the AWERB What does it mean to you and could you become more active in maintaining a positive culture Acknowledgements Thank you to all the speakers and delegates for the talks and discussions We are also grateful to members of staff at the University of Leicester for giving up their time to take a group of delegates on a tour of their facility References All the URLs below were last viewed on 2 January 2015 21 22 23 24 25 26 27 28 29 10 Proctor H 2012 Animal sentience Where are we and where are we heading Animals 2 628 639 Download at http www mdpi com 2076 2615 2 4 628 Proctor H Carder G and Cornish A R 2013 Searching for animal sentience A systematic review of the scientific literature Animals 3 882 906 Download at http www mdpi com 2076 2615 3 3 882 King B 2013 When animals mourn Scientific American 309 62 67 Elwood R W Barr S and Patterson L 2009 Pain and stress in crustaceans Applied Animal Behaviour Science 118 128 136 Bartal I B A Decety J and Mason P 2011 Empathy and pro social behavior in rats Science 334 14271430 Download at http tinyurl com knfsflw Langford D J Crager S E Shehzad Z Smith S B Stocinal S G Levenstadt J S Chanda M L Levitin D J and Mogil J S 2006 Social modulation of pain as evidence for empathy in mice Science 312 1967 1970 Bekoff M 2005 Animal emotions and animal sentience and why they matter Blending science sense with common sense compassion and heart In Animals Ethics and Trade The Challenge of Animal Sentience Turner J and J D Silva J Earthscan London 27 40 Dawkins M S 2012 Why Animals Matter Animal consciousness animal welfare and human well being Oxford University Press Oxford UK Steiner A P and Redish A 2014 Behavioral and neurophysiological correlates of regret in rat decisionmaking on a neuroeconomic task Nature Neuroscience 17 995 1002 doi 10 1038 nn 3740 Panksepp J and Burgdorf J 2003 Laughing rats and the evolutionar y antecedents of human joy Physiology Behavior 79 533 547 http dx doi org 10 1016 S0031 9384 03 00159 8 27

Report of the 2014 RSPCA UFAW Rodent Welfare Group meeting 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 28 Robinson V Morton D B Anderson D Carver J F A Francis R J Hubrecht R Jenkins E Mathers K A Rosewell I Wallace J and Wells D J 2003 Reduction and refinement in production of genetically modified mice Laboratory Animals 37 Suppl 1 1 51 Miller A L Wright Williams S L Flecknell P A and Roughan J V 2012 A comparison of abdominal and scrotal approach methods of vasectomy and the influence of analgesic treatment in laboratory mice Laboratory Animals 46 304 310 BAP BNA ESSWAP and LASA 2013 Guiding Principles for Behavioural Laboratory Animal Science Free download at http tinyurl com on86a3g Bate S T and Clark R A 2014 The Design and Statistical Analysis of Animal Experiments Cambridge University Press Cambridge Festing M Overend P Gaine Das R Cortina Borja M and Berdoy M 2002 The Design of Animal Experiments Reducing the Use of Animals in Research Through Better Experimental Design Royal Society of Medicine Press London Field A and Hole G 2003 How to Design and Report Experiments SAGE Publications Ltd London Dawkins M S 2004 Using behaviour to assess animal welfare Animal Welfare 13 S3 7 Werner C F rster T Widmann T P ss J Roggia C Hanhoun M Scharhag J B chner N Meyer T Kindermann W Haendeler J B hm M and Laufs U 2009 Physical exercise prevents cellular senescence in circulating leukocytes and in the vessel wall Circulation 120 2438 2447 Ehninger D and Kempermann G 2003 Regional effects of wheel running and environmental enrichment on cell genesis and microglia proliferation in the adult murine neocortex Cereb Cortex 13 845 851 Sherwin C M 1998 Voluntary wheel running a review and novel interpretation Anim Behav 56 11 27 Lett B T Grant V L and Koh M T 2001 Naloxone attenuates the conditioned place preference induced by wheel running in rats Physiol Behav 72 355 8 Belke T W and Wagner J P 2005 The reinforcing property and rewarding aftereffect of wheel running in rats a combination of two paradigms Behav Processes 68 165 172 Masaki T and Nakajima S 2008 Forward conditioning with wheel running causes place aversion in rats Behav Processes 79 43 7 Richter S H Gass P and Fuss J 2014 Resting is rusting a critical view on rodent wheel running behavior The Neuroscientist 20 313 325 Mason G J 1991 Stereotypies a critical review Anim Behav 41 1015 37 Latham N and W rbel H 2006 Wheel running a common rodent stereotypy In Stereotypic Animal Behaviour Fundamentals and Applications to Welfare Second Edition Mason G and Rushen J CABI Wallingford England 91 92 Vargas Perez H Sellings L H Paredes R G PradoAlcala R A and Diaz J L 2008 Reinforcement of wheel running in BALB c mice role of motor activity and endogenous opioids J Mot Behav 40 587 93 Home Office 2014 Guidance on the Operation of the Animals Scientific Procedures Act 1986 Download at https www gov uk research and testing using animals 29 30 31 RSPCA and LASA 2015 Guiding Principles on Good Practice for Animal Welfare and Ethical Review Bodies A report by the RSPCA Research Animals Department and the LASA Education Training and Ethics section Jennings M Download at www rspca org uk laymembers Hawkins P Ryder K Mortell N and Patten D 2014 Raising concerns about laborator y animal welfare report of a workshop at IAT Congress 2014 Animal Technology and Welfare 13 81 85 Jennings M and Smith J 2015 A Resource Book for Lay Members of Ethical Review and Similar Bodies Worldwide Third Edition RSPCA Southwater Download at www rspca org uk laymembers

April 2015 Animal Technology and Welfare PAPER SUMMARY TRANSLATIONS INHALTVERZEICHNIS Verbesserungen im Einsatz Haltegeschirr ersetzt Kopfhalterung beim Legen von Kan len bei Laborratten CLARE SIMS Dorothy Hodgkin Building University of Bristol Whitson Street Bristol BS1 3NY Korrespondierende autorin svces bristol ac uk Preistr ger des 2015 Andrew Blake Tribute Award Abstract Ratten wird eine Kan le in die Jugularvene gelegt damit Blutproben entnommen und mithilfe eines automatischen Messsystems ABS System analysiert werden k nnen Anhand dieser Blutproben wird der Hormonspiegel im Blut ber einen bestimmten Zeitraum hinweg gemessen Beim urspr nglichen Protokoll wird die Jugularvenenkan le durch einen Zugang am Kopf der Ratte gelegt Anschlie end wird sie durch eine Schutzfeder aus Metall gef hrt und an das ABS System angeschlossen Die Metallfeder wird mit zwei im Sch del der Ratte implantierten Metallschrauben befestigt Die Verbesserung des Protokolls besteht darin dass der Zugang der Kan le au en am R cken der Ratte gelegt wird und dass das Haltegeschirr des vaskul ren Zugangs die Schutzfeder in Position h lt Die Ergebnisse belegen dass durch diese Verbesserungen im Protokoll das Wohlergehen der Tiere sowie die Tierhaltung verbessert wurden und dass die Anzahl der f r die Studie ben tigten Tiere insgesamt verringert wurde Dar ber hinaus wurde ersichtlich dass der Spiegel des Stresshormons Corticosteron im Blut beim Einsatz des Haltegeschirrs nicht steigt Stichw rter Ratten Jugularvenenzugang Corticosteron Haltegeschirr Verbesserung 29

Paper Summary Translations Verbesserte Haltung von m nnlichen Laborkaninchen eine Pilotstudie zu m glichen Vorteilen der Bodenhaltung JOANNA CRUDEN1 JONATHAN COOPER2 und OLIVER BURMAN 1 2 GlaxoSmithKline Park Road Ware Hertfordshire SG12 0DP Animal Behaviour Cognition and Welfare Research Group School of Life Sciences University of Lincoln Joseph Banks Laboratories Lincoln LN7 6TS Korrespondenzautor joanna l cruden gsk com Einleitung In der gegenw rtigen Einrichtung werden m nnliche Kaninchen blicherweise in Einzelk figen gehalten mit Zugang zu einer 2 5 m2 gro en Bodenfl che zweimal pro Woche Nach berpr fung der Kaninchenhaltung wurde die Entscheidung getroffen eine Gruppe von m nnlichen Kaninchen dauerhaft in Bodengehegen unterzubringen Hierdurch ergab sich die M glichkeit nderungen im Verhalten der Kaninchen zu beobachten wenn sie von K figen in Bodengehege verlegt werden Vor der Verlegung in die Bodengehege wurde eine Kohorte von 4 m nnlichen Kaninchen ber einen Zeitraum von 24 Stunden in ihren Einzelk figen gefilmt dieser Vorgang wurde wiederholt nachdem sie zwei Wochen lang im Bodengehege untergebracht waren Die Videoaufnahmen wurden anhand der Fokusmethode mit feststehenden Intervallen analysiert Dazu wurde alle 15 Minuten die Aktivit t K rperhaltung und Position im K fig bzw im Bodengehege protokolliert Die Kaninchen verbrachten erheblich weniger Zeit in der Mitte des K figs verglichen mit dem mittleren Bereich des Bodengeheges p 0 012 sie fra en weniger p 0 002 und im Bodengehege war eine signifikante Verringerung der Fellpflege p 0 002 und des Sitzens p 0 019 zu verzeichnen Die Kaninchen schienen dar ber hinaus weniger Zeit im hinteren Bodenbereich sowie in Interaktion mit Anreicherungen des Bodengeheges zu verbringen Allerdings schienen Erkundungsverhalten Aufrichten auf den Hinterpfoten sowie die Zeit die sie vollst ndig ausgestreckt liegend verbrachten zuzunehmen Keiner dieser Unterschiede war jedoch bei der in dieser Studie verwendeten Stichprobengr e 0 1 p 0 05 statistisch signifikant Diese Pilotstudie legt nahe dass eine Bodenhaltung von m nnlichen Kaninchen aufgrund des erweiterten Raumes und der vermehrten Verhaltensm glichkeiten verhaltensbezogene Vorteile mit sich bringt Weitere Studien sind jedoch erforderlich um zu untersuchen ob diese Ergebnisse f r die gesamte Laborkaninchenpopulation repr sentativ sind Stichw rter Laborkaninchen Bodengehege Verhalten und Wohlbefinden 30

Paper Summary Translations Bericht zur Tagung der RSPCA UFAW TierschutzGruppe f r Nager 2014 PENNY HAWKINS SECRETARY 1 MANUEL BERDOY 2 CHARLOTTE BUR 3 DEBBIE BURSNALL 4 JOANNA CRUDEN 5 WANDA MCCORMICK 6 AMY MILLER 7 HELEN PROCTOR 8 DAVID WHITTAKER 2 MAGGY JENNINGS 1 und ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Department of Veterinary Services c o University Laboratory of Physiology University of Oxford Oxford Oxfordshire OX1 3PT Centre for Animal Welfare The Royal Veterinary College Hawkshead Lane North Mymms Hertfordshire AL9 7TA Division of Biomedical Services University of Leicester Lancaster Road Leicester Leicestershire LE1 7HB GSK Research and Development Park Road Ware Hertfordshire SG12 0DP Moulton College West Street Moulton Northampton Northamptonshire NN3 7RR School of Agriculture Food and Rural Development University of Newcastle Newcastle upon Tyne NE1 7RU World Animal Protection 222 Grays Inn Road London WC1X 8HB UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Einleitung Die RSPCA UFAW Tierschutz Gruppe f r Nager veranstaltet jeden Herbst ein eint giges Treffen bei dem die Mitglieder ber Themen aktueller Tierschutz Forschung diskutieren und Erfahrungen und Meinungen ber nagerrelevante Tierschutz Probleme sowie ber die drei Prinzipien Vermeidung Reduktion und Verfeinerung des Einsatzes von Nagern austauschen Ein Hauptziel der Gruppe ist es zum Nachdenken ber die Gesamtheit der Lebenserfahrungen von Nagern als Laborversuchstiere anzuregen und sicherzustellen dass jede potenzielle Beeintr chtigung ihres Wohlbefindens gepr ft und auf ein Minimum gesenkt wird An unserer 21 j hrlichen Tagung am 23 Oktober 2014 nahmen 90 Teilnehmer von einer Vielzahl von Universit ten und Pharmazieunternehmen aus dem ganzen Vereinigten K nigreich teil Die Pr sentationen drehten sich um Themen wie die Empfindungen von Tieren das Lindern von Schmerzen bei Eingriffen die Einsch tzung des Gesundheitszustandes und von Tierschutzma nahmen sowie das Sicherstellen von richtigen Entscheidungen im Bezug auf die Ausgestaltung des Lebensumfeldes z B durch das Bereitstellen von Laufr dern Die Tagung endete mit einer Diskussion ber Culture of Care und wie diese in Instutionen erkannt gef rdert und aufrecht erhalten werden kann Dieser Bericht fasst die Tagung zusammen und enth lt am Ende eine Liste von Aktionspunkten die die Leser in Ihren eigenen Einrichtungen ansprechen k nnten 31

Animal Technology and Welfare April 2015 CONTENU DE LA REVUE Am lioration en action Du cylindre au harnais pour la canulation de rats de laboratoire CLARE SIMS Dorothy Hodgkin Building University of Bristol Whitson Street Bristol BS1 3NY Auteur ressource svces bristol ac uk Gagnant du prix Andrew Blake Tribute Award 2015 R sum Une canule est introduite l int rieur de la veine jugulaire des rats afin que les chantillons de sang soient pr lev s au moyen d un syst me automatis de pr l vement sanguin Ces chantillons de sang sont utilis s afin de mesurer les niveaux d hormones contenues dans le sang au fil du temps Dans le protocole de d part la canule introduite dans la veine jugulaire ressort du corps du rat par une incision op r e au sommet de la t te Le rat est ensuite aliment au travers d un ressort m tallique protecteur et reli au syst me automatis de pr l vement sanguin Le ressort m tallique est maintenu en place par deux vis en m tal fix es au cr ne du rat L am lioration de ce protocole consiste faire en sorte que la canule ressorte dans le dos du rat et qu un harnais d acc s vasculaire maintienne le ressort protecteur en place Les r sultats d montrent qu am liorer ainsi le protocole permet une am lioration du bien tre des animaux et des pratiques d levage ainsi qu une r duction du nombre total d animaux utilis s tout au long de l tude Ils prouvent galement que le taux de corticost rone s cr t e en r ponse au stress prouv n augmente pas lors de l utilisation du harnais d acc s vasculaire Mots cl s Rats canulation jugulaire corticost rone harnais d acc s am lioration 32

Paper Summary Translations H bergement am lior pour lapins de laboratoire m les tude pilote enqu tant sur les avantages potentiels des enclos au sol JOANNA CRUDEN1 JONATHAN COOPER2 et OLIVER BURMAN2 1 2 GlaxoSmithKline Park Road Ware Hertfordshire SG12 0DP Animal Behaviour Cognition and Welfare Research Group School of Life Sciences University of Lincoln Joseph Banks Laboratories Lincoln LN7 6TS Auteur ressource joanna l cruden gsk com R sum Dans l installation actuelle les lapins m les sont g n ralement h berg s dans des cages individuelles avec acc s deux fois par semaine un enclos au sol de 2 5 m2 Apr s un examen des conditions d h bergement des lapins la d cision a t prise de transf rer d finitivement une population de lapins m les dans des enclos au sol Cela permettait d observer les changements de comportement des lapins lorsqu ils sont transf r s de leurs cages des enclos au sol Avant leur transfert dans des enclos au sol une cohorte de 4 lapins m les h berg s individuellement a t film e pendant 24 heures dans les cages d h bergement cela a ensuite t r p t apr s qu ils aient t transf r s dans des enclos au sol pendant deux semaines Les enregistrements vid o ont t analys s en utilisant un chantillonnage instantan focalis intervalle fixe avec un enregistrement d activit de posture et de position effectu toutes les 15 minutes dans l enclos ou la cage Les lapins passaient beaucoup moins de temps dans la zone centrale de la cage par rapport la zone centrale de l enclos au sol p 0 012 mangeaient moins p 0 002 et on constatait une r duction significative du toilettage p 0 002 et de la position assise p 0 019 dans les enclos au sol Dans l enclos au sol les lapins semblaient galement passer moins de temps l arri re et moins de temps interagir de mani re enrichissante mais il semblait y avoir une augmentation du comportement exploratoire et du temps pass se dresser sur les pattes arri re ou tre enti rement tendus Aucune de ces diff rences n tait toutefois statistiquement significative l chelle des chantillons utilis s dans cette tude 0 1 p 0 05 Cette tude pilote sugg re qu h berger les lapins m les dans des enclos au sol serait avantageux pour leur comportement en raison de l augmentation des possibilit s spatiales et comportementales que cela conf re mais d autres tudes seraient b n fiques pour valuer si ces r sultats sont repr sentatifs de la population des lapins de laboratoire en g n ral Mots cl s Lapins de laboratoire enclos au sol comportement et bien tre 33

Paper Summary Translations Rapport de la r union du Groupe sur la bientraitance des rongeurs de la RSPCA UFAW 2014 PENNY HAWKINS SECR TAIRE 1 MANUEL BERDOY 2 CHARLOTTE BUR 3 DEBBIE BURSNALL 4 JOANNA CRUDEN 5 WANDA MCCORMICK 6 AMY MILLER 7 HELEN PROCTOR 8 DAVID WHITTAKER 2 MAGGY JENNINGS 1 et ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Department of Veterinary Services c o University Laboratory of Physiology University of Oxford Oxford Oxfordshire OX1 3PT Centre for Animal Welfare The Royal Veterinary College Hawkshead Lane North Mymms Hertfordshire AL9 7TA Division of Biomedical Services University of Leicester Lancaster Road Leicester Leicestershire LE1 7HB GSK Research and Development Park Road Ware Hertfordshire SG12 0DP Moulton College West Street Moulton Northampton Northamptonshire NN3 7RR School of Agriculture Food and Rural Development University of Newcastle Newcastle upon Tyne NE1 7RU World Animal Protection 222 Grays Inn Road London WC1X 8HB UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Introduction Le Groupe sur la bientraitance des rongeurs de la RSPCA UFAW organise une r union d une journ e chaque automne afin que ses membres puissent discuter des recherches en cours sur la bientraitance changer des id es sur les probl mes de bientraitance des rongeurs et partager leurs exp riences relatives la mise en place des 3 R remplacement r duction et perfectionnement en ce qui concerne l utilisation des rongeurs L un des objectifs cl s du groupe consiste encourager une r flexion sur la vie des rongeurs de laboratoire et faire en sorte que chaque impact n gatif potentiel sur leur bien tre soit corrig et minimis Notre 21e r union annuelle du 23 octobre 2014 a regroup 90 d l gu s venus d un grand nombre d universit s et de soci t s pharmaceutiques du Royaume Uni Parmi les sujets pr sent s citons la sentience des animaux comment r duire la souffrance pendant les proc dures comment garantir la sant et le bien tre des rongeurs et comment faire en sorte que les bonnes d cisions soient prises lorsque des am liorations environnementales telles que des roues d exercice sont mises en place La journ e s est termin e par une discussion sur la culture des soins et sur la fa on dont celle ci peut tre reconnue encourag e et maintenue dans les institutions Ce rapport r sume la r union et se termine par une liste de mesures envisager par les lecteurs dans leur propre tablissement 34

April 2015 Animal Technology and Welfare INDICE DE LA REVISTA Nos unimos a la sofisticaci n Pasamos de usar dispositivos para la cabeza a arneses para la canulaci n de ratas de laboratorio CLARE SIMS Edificio Dorothy Hodgkin Universidad de Bristol Whitson Street Bristol BS1 3NY Autor para correspondencia svces bristol ac uk Ganador del premio Andrew Blake Tribute Award de 2015 Resumen Para tomar muestras de sangre a ratas mediante un sistema de tomas de muestras de sangre automatizado ABS por sus siglas en ingl s se les introduce una c nula en la vena yugular Estas muestras se utilizan para calcular los niveles de hormonas presentes en la sangre con el paso del tiempo En el protocolo original la c nula de la vena yugular se coloca fuera del cuerpo de la rata mediante una incisi n en la parte posterior de la cabeza Se le alimenta a trav s de un surtidor de metal protector y se conecta a un sistema ABS El surtidor de metal se ancla con dos tornillos met licos implantados en el cr neo de la rata La sofisticaci n que presenta este protocolo consiste en que la c nula se encuentra ahora en el exterior del lomo de la rata y un arn s de acceso vascular ancla el suministro protector en su sitio Los resultados demuestran que mediante la sofisticaci n del protocolo se ha mejorado el bienestar y la cr a de los animales y ha disminuido el n mero total de animales utilizados en el estudio Del mismo modo tambi n se ha demostrado que el nivel de la hormona del estr s la corticosterona en la sangre no aumenta con el uso de arneses de acceso vascular Palabras clave ratas canulaci n yugular corticosterona arn s de acceso sofisticaci n 35

Paper Summary Translations Conejeras optimizadas para conejos macho de laboratorio estudio piloto de investigaci n de los posibles beneficios de las jaulas de cercado hasta el suelo JOANNA CRUDEN1 JONATHAN COOPER2 y OLIVER BURMAN2 1 2 GlaxosmithKline Park Road Ware Hertfordshire SG12 0DP AnimalBehaviour Cognition and Welfare Research Group Grupo de investigaci n del comportamiento conocimiento y bienestar animal Facultad de Ciencias de la Vida Universidad de Lincoln Laboratorios Joseph Banks Lincoln LN7 6TS Autora para correspondencia joanna l cruden gsk com Resumen En las instalaciones actuales los conejos macho normalmente se alojan en jaulas individuales y tienen acceso dos veces por semana a jaulas de cercado hasta el suelo de 2 5 m2 Tras replantearse el alojamiento para conejos se tom la decisi n de transferir los grupos de conejos macho a jaulas de cercado hasta el suelo de manera permanente De esta forma se tuvo la oportunidad de observar los cambios de comportamiento de los conejos al trasladarlos de jaulas individuales a jaulas de cercado hasta el suelo Antes del traslado a cajas de cercado hasta el suelo se grab en v deo durante 24 horas a un grupo de cuatro conejos macho alojados individualmente Se repiti el proceso despu s de haberlos alojado en jaulas de cercado hasta el suelo durante dos semanas Se visualiz el metraje grabado con muestreo focal instant neo de intervalo fijo gracias a lo cual se obtuvo informaci n sobre la actividad la postura y la posici n de los conejos en la jaula individual o en la jaula de cercado hasta el suelo cada 15 minutos Los conejos pasaron mucho menos tiempo en la zona central de la jaula individual en comparaci n con la zona central de la jaula de cercado hasta el suelo p 0 012 comieron menos p 0 002 y se produjo una reducci n significativa del aseo p 0 002 y del tiempo que pasaban sentados p 0 019 en las jaulas de suelo Se pudo percibir que los conejos pasaban menos tiempo en la parte trasera de la jaula y dedicaban menos tiempo a la interacci n con los accesorios del entorno en la jaula de cercado hasta el suelo aunque se apreci un incremento en la exploraci n la posici n erguida y el tiempo que pasaban completamente estirados No obstante desde el punto de vista estad stico ninguna de estas diferencias se ha considerado significativa a tenor del tama o de las muestras recogidas en el presente estudio 0 1 p 0 05 Este estudio piloto apunta a que existen beneficios de comportamiento al alojar conejos macho en jaulas de cercado hasta el suelo debido a un espacio m s amplio y a las oportunidades de comportamiento no obstante unos estudios complementarios ser an beneficiosos para valorar si estos resultados son representativos de la poblaci n de conejos de laboratorio en general Palabras clave conejos de laboratorio jaulas de cercado hasta el suelo comportamiento y bienestar 36

Paper Summary Translations Informe sobre la reuni n del Grupo para el bienestar de roedores RSPCA UFAW 2014 PENNY HAWKINS SECRETARIA 1 MANUEL BERDOY 2 CHARLOTTE BUR 3 DEBBIE BURSNALL 4 JOANNA CRUDEN 5 WANDA MCCORMICK 6 AMY MILLER 7 HELEN PROCTOR 8 DAVID WHITTAKER 2 MAGGY JENNINGS 1 y ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Department of Veterinary Services c o University Laboratory of Physiology University of Oxford Oxford Oxfordshire OX1 3PT Centre for Animal Welfare The Royal Veterinary College Hawkshead Lane North Mymms Hertfordshire AL9 7TA Division of Biomedical Services University of Leicester Lancaster Road Leicester Leicestershire LE1 7HB GSK Research and Development Park Road Ware Hertfordshire SG12 0DP Moulton College West Street Moulton Northampton Northamptonshire NN3 7RR School of Agriculture Food and Rural Development University of Newcastle Newcastle upon Tyne NE1 7RU World Animal Protection 222 Grays Inn Road London WC1X 8HB UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Introducci n El Grupo para el bienestar de roedores RSPCA UFAW celebra una reuni n de un d a cada oto o para que sus miembros puedan tener un debate 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 Uno de los objetivos primordiales del Grupo es fomentar que las personas piensen sobre la experiencia vital de los roedores de laboratorio garantizando a su vez que cualquier impacto negativo posible en su bienestar sea revisado y reducido Nuestra vig simo primera reuni n anual tuvo lugar el 23 de octubre de 2014 en la que participaron 90 delegados de un amplio n mero de universidades y farmac uticas de todo el Reino Unido Los temas de presentaci n incluyeron conciencia animal reducci n del sufrimiento durante procedimientos evaluaci n del bienestar y salud de roedores y c mo garantizar que se toman las decisiones adecuadas al suministrar mejoras ambientales como ruedas para correr La jornada finaliz con un debate sobre la cultura de atenci n y sobre esta puede ser reconocida promocionada y mantenida con la instituciones Este informe resume la reuni n y finaliza con una lista de puntos de acci n para los lectores para presentar en sus propias instalaciones 37

Animal Technology and Welfare April 2015 INDICE DELLA REVISTA Perfezionamenti in azione dal casco alla bardatura per l incannulamento dei ratti da laboratorio CLARE SIMS Dorothy Hodgkin Building University of Bristol Whitson Street Bristol BS1 3NY Autore corrispondente svces bristol ac uk Vincitore del premio Andrew Blake Tribute Award 2015 Riassunto Sui ratti viene eseguito l incannulamento della vena giugulare per consentire il prelievo di campioni di sangue mediante sistema automatico ABS Questi campioni servono a misurare i livelli di ormoni nel sangue nel corso del tempo Nel protocollo originale la cannula per la vena giugulare viene esteriorizzata dal corpo del ratto mediante un incisione sulla parte superiore del cranio Quindi viene inserita lungo una molla protettiva in metallo e collegata al sistema ABS La molla viene mantenuta in posizione con due viti metalliche fissate nel cranio del ratto Il perfezionamento di questo protocollo prevede che la cannula venga esteriorizzata sulla parte posteriore del ratto e che la molla protettiva sia mantenuta in posizione con una bardatura per accesso vascolare I risultati mostrano che questo perfezionamento del protocollo migliora sia il benessere dell animale che la prassi zootecnica e porta a una riduzione nel numero complessivo di animali utilizzati nello studio Indicano inoltre che l uso della bardatura per l accesso vascolare non porta a un aumento della risposta ormonale allo stress corticosterone nel sangue Parole chiave ratti incannulamento giugulare corticosterone bardatura di accesso perfezionamento 38

Paper Summary Translations Alloggi potenziati per conigli maschi da laboratorio studio pilota di valutazione dei potenziali vantaggi di box a terra JOANNA CRUDEN1 JONATHAN COOPER2 e OLIVER BURMAN2 1 2 GlaxoSmithKline Park Road Ware Hertfordshire SG12 0DP Animal Behaviour Cognition and Welfare Research Group School of Life Sciences University of Lincoln Joseph Banks Laboratories Lincoln LN7 6TS Autore corrispondente joanna l cruden gsk com Riassunto Nella struttura attuale solitamente i conigli maschi vengono alloggiati singolarmente in gabbie con accesso bisettimanale a box a terra di 2 5 m2 A seguito di una rivalutazione degli alloggiamenti per conigli stato deciso di trasferire una popolazione di conigli maschi nei box a terra in maniera permanente Tale decisione ha permesso di osservare i cambiamenti comportamentali dei conigli trasferiti dalle gabbie ai box Prima del trasferimento a quest ultimi sono state effettuate registrazioni video all interno delle gabbie di una coorte di 4 conigli maschi alloggiati singolarmente per un periodo di 24 ore tale operazione stata ripetuta a due settimane dalla sistemazione nei box a terra I comportamenti sono stati registrati con campionamento focale istantaneo a intervalli fissi raccogliendo dati sulle attivit la postura e la posizione all interno della gabbia o del box ogni 15 minuti I conigli hanno trascorso molto meno tempo nell area centrale della gabbia rispetto a quella del box a terra p 0 012 e hanno consumato meno cibo p 0 002 oltre a dedicare meno tempo alla pulizia p 0 002 e a stare seduti p 0 019 nei box a terra Si notato inoltre che i conigli hanno trascorso meno tempo sulla parte posteriore del pavimento o a interagire con l arricchimento del box a terra bench si sia osservato un comportamento esploratorio pi accentuato e un periodo di tempo prolungato in posizione eretta o sdraiata Tuttavia nessuna delle differenze rilevate pu considerarsi statisticamente significativa per la dimensione dei campioni utilizzati nello studio 0 1 p 0 05 Questo studio pilota conferma la presenza di vantaggi comportamentali nell alloggiamento a terra di conigli maschi derivanti da uno spazio pi ampio e da opportunit di cambiamento del comportamento anche se sarebbe utile condurre ulteriori studi per valutare se i risultati ottenuti sono rappresentativi della popolazione complessiva di conigli da laboratorio Parole chiave conigli da laboratorio box a terra comportamento e benessere 39

Paper Summary Translations Relazione sull incontro del RSPCA UFAW Rodent Welfare Group Gruppo sul benessere dei roditori del 2014 PENNY HAWKINS SEGRETARIA 1 MANUEL BERDOY 2 CHARLOTTE BUR 3 DEBBIE BURSNALL 4 JOANNA CRUDEN 5 WANDA MCCORMICK 6 AMY MILLER 7 HELEN PROCTOR 8 DAVID WHITTAKER 2 MAGGY JENNINGS 1 e ROBERT HUBRECHT9 1 2 3 4 5 6 7 8 9 Research Animals Department Science Group RSPCA Wilberforce Way Southwater West Sussex RH13 9RS Department of Veterinary Services c o University Laboratory of Physiology University of Oxford Oxford Oxfordshire OX1 3PT Centre for Animal Welfare The Royal Veterinary College Hawkshead Lane North Mymms Hertfordshire AL9 7TA Division of Biomedical Services University of Leicester Lancaster Road Leicester Leicestershire LE1 7HB GSK Research and Development Park Road Ware Hertfordshire SG12 0DP Moulton College West Street Moulton Northampton Northamptonshire NN3 7RR School of Agriculture Food and Rural Development University of Newcastle Newcastle upon Tyne NE1 7RU World Animal Protection 222 Grays Inn Road London WC1X 8HB UFAW The Old School Brewhouse Hill Wheathampstead Hertfordshire AL4 8AN Introduzione Durante il periodo autunnale RSPCA UFAW Rodent Welfare Group Gruppo sul benessere dei roditori indice annualmente un incontro della durata di un giorno permettendo ai suoi membri di discutere degli attuali studi di ricerca sul benessere di scambiare opinioni sulle problematiche inerenti al benessere dei roditori e di condividere esperienze sull applicazione del principio delle 3R sostituzione replacement riduzione reduction e perfezionamento refinement nell utilizzo di roditori a scopi scientifici Uno degli obiettivi principali del Gruppo incoraggiare le persone a considerare l esperienza di vita dei roditori da laboratorio nel suo insieme assicurando il controllo e la riduzione di ogni possibile impatto negativo sul loro benessere Il 21 incontro annuale tenutosi il 23 ottobre 2014 ha attirato 90 delegati provenienti da diverse universit e case farmaceutiche di tutto il Regno Unito Gli argomenti trattati hanno incluso la sensibilit animale la riduzione del grado di sofferenza durante le procedure la valutazione della salute e del benessere dei roditori e come assicurare che vengano prese le giuste decisioni sugli arricchimenti ambientali come ad esempio le ruote L incontro si concluso con un dibattito sulla cultura della cura e su come questo elemento pu essere riconosciuto promosso e mantenuto all interno degli istituti La presente relazione riassume l incontro e si conclude con un elenco di provvedimenti che i lettori possono mettere a punto presso le proprie strutture 40

April 2015 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 Improving animal welfare reducing energy cost and increasing flexibility PREBEN LEONHARDT and ANDY DOMONE Scanbur Technology A S Silovej 16 18 DK 2690 Karlslunde Denmark Corresponding author preben leonhardt scanbur eu Introduction The Full Economic Costing System FEC for Animal Units came into use at Universities and Government establishments in 1998 It provides a focus of the costs involved in animal research and what researchers are then charged for using the animal unit facilities Energy costs are increasingly a large part of the FEC The required number of air changes and a need to maintain a relative humidity of 55 make animal unit buildings some of the highest energy consumers changes hour compared with 15 20 for a room Typically the total air volume that needs to be humidified is reduced by 80 or more In the United States of America USA biotechnology industry it is commonplace to connect the IVC racks directly to the buildings central ventilation system In Europe an air handling unit placed in the animal room supplying pressure and ventilation of the IVC racks is more usual This USA system of direct supply does mean that the contained micro environment can be treated independently Over the last decade there has been a move away from open cages towards containment systems such as ventilated cabinets or Individually Ventilated Cages IVCs These containment systems provide a superior micro environment for rodents but pose the question Why not treat this micro environment independently from the room The disadvantages are that it requires additional ducting more space in the technical area and allows little flexibility in the use of the building The rooms are dedicated to IVC racks in a specific layout Full investment in this central system is required from day one Capacity cannot be adjusted at a later stage Immediately we can see that an advantage would be that the total air volume required for the contained animals would be much lower than that for the room This despite contained systems needing 60 80 Consequently there are many advantages to using a mobile air handling and humidification supply unit that can control the micro climate in contained systems as good as a centralised system if not better 41

Tech 2 Tech New technology available Energy savings The Danish company Scanbur A S has developed a new product named ScanClime Mini It is specifically designed for use with IVC racks and has the same features as a standard air handling unit e g High Efficiency Par ticulate Air HEPA 14 filters positive negative pressure variable and can run 4 single or 2 double sided racks In addition it supplies a controlled humidity for the air within the IVC cages As described earlier it is possible to save energy by humidifying only the air of the IVC cages compared with centrally humidifying the air of a complete room e g a standard animal room often is 3 x 5 metres and 3 metres high and the room volume of air is app 50 m3 In such a room with up to 20 air changes per hour 1 000 m3 of air per hour is needed to be humidified centrally Alternatively a standard IVC rack contains 0 6 m3 of air and is ventilated up to 80 times per hour When connecting 4 racks to an IVC it is only 200 m3 of air per hour need to be humidified and consequently it is possible to obtain an 80 reduction in volume Animal welfare The ambient weather conditions of Northern Europe dictate that the relative humidity around the research models is controlled up to 80 of the year to keep it within the 45 65 range required for animal welfare 1 Most central ventilation systems in an animal unit have a variation of 10 of the relative humidity and a poor old or badly maintained system will vary up to 15 With these variations it is always difficult to operate safely within the 45 65 range as this would require a set point of 55 in relative humidity Many systems cannot achieve 55 as a set point but will due to lack of steam capacity have a set point at 45 With these variables many animal units will have shorter or longer periods where the relative humidity is considerably below the stipulated threshold in the current legislation 1 Too low humidity will have a negative effect on animal welfare and ringtail is one of the most significant examples of compromised welfare due to low relative humidity Another example is nu nu athymic mice being prone to dry skin conditions The cost of centrally humidifying the above room is GBP 2 574 per year The cost of humidifying 4 IVC racks would be GBP 439 On a larger scale an animal unit with 8 rooms and annual increase in energy cost of 5 would mean a saving over 10 years of GBP 205 000 by changing from central to de central humidification The above calculations are based on an animal room at maximum capacity of racks In many units rooms also contain a cage changing station working tables etc Some units also humidify laboratories and corridors In these situations the savings using decentral humidification will be even higher For the Full Economic Costing FEC the advantage is that the running cost goes hand in hand with the number of cages in use when choosing a decentralised solution The Scanclime Mini provides humidity control with a variation of 3 from a chosen set RH point of 55 The humidity will remain close to the set level all year round This new technology has been unavailable earlier and is standardising studies focused in current eye skin and respiratory system research providing a stable relative humidity that is controlled and repeatable within a tight narrow range See pink graph line Flexibility The Scanclime Mini is simply adjustable to 65 RH for specific work with nu nu mice avoiding skin conditions When the building then is needed for another purpose e g as a laboratory the cost of rebuilding is much Figure 1 Humidity in IVC cage connected to ScanClime Mini 42 Research today is moving faster than ever Research groups are often reorganised split off and moved around a university campus on a frequent basis This creates an even greater need for buildings that can be flexible and changeable to the user With these decentralised units it is now possible to build more standardised laboratory buildings and use them quickly as animal units Capital building costs are reduced

Tech 2 Tech lower than today With a decentralised humidification solution the animal unit can bring their equipment to a new facility when moving Flexibility will also be practical in daily use as the basic room condition for laboratories and animal rooms will be identical Only the mobile equipment in the rooms defines the rooms use The room s layout can change and adapt to the changes in research Flexibility also applies for the capital investment required With the current central ventilation systems from day one it should be an installed system providing for a maximum capacity even when only working at less than half full The decentralised system needs only be utilised where and when needed and expanded as expansion happens at the pace of the research For the refurbishment of existing animal units a decentralised system can be installed in stages one room or area at a time whilst the animal unit is occupied Investment can be spread over several years whereas the current central systems require 100 investment from the start of refurbishment and in all probability the facility will have to be closed for the duration Conclusion New technologies have always influenced the work in animal units Contained systems have improved the protection of vulnerable animal strains Walk In cage changing systems have improved the protection of staff against allergens and robotic systems have improved the ergonomics in the wash area With this new development of decentralised humidification for IVC caging a new standard for stable humidification has been set This brings about an improved cage environment reduced energy costs and greater flexibility within the animal unit This will assist animal units to meet the increasing demands for good animal welfare standards It will improve cost effective research and bring about true flexibility of use of buildings and rooms adapting to the ever changing needs of the research groups References 1 European Commission 2010 Directive 2010 63 EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes Official Journal of the European Union L276 33 79 43

Animal Technology and Welfare April 2015 AS ET SPECIAL TRAVEL BURSARY 2014 ESSAYS Advantages and disadvantages of animal technologists performing regulated procedures in research ANGELIKA KOSCIOLEK University College London Biological Services Unit Cruciform Building Gower Street London WC1E 6AU Corresponding author waymak ucl ac uk AS ET Special Travel Bursary 2014 entry Instructions to applicants Many animal technical staff have a dual role they are responsible for the care and welfare of animals in their charge but also licensed to carry out regulated procedures on those animals Discuss the advantages and disadvantages of animal care staff performing procedures on animals in their care explaining ways in which any potential conflict between the two roles can be minimised Some men see things as they are and say why I dream things that never were and say why not George Bernard Shaw 1 It is with this same determination and passion that animal care staff make a difference in the world of animal science They play one of the most important roles in animal research Their work which is often underestimated makes a great impact within the industry and this is of notable importance for the welfare of laboratory animals They are not only responsible for the care and well being of the research animals but they also make sure the suffering of animals undertaking experiments is reduced to a minimum They closely follow the 3Rs requirements and Home Office guidance and constantly strive to improve animal comfort and welfare Moreover animal technical staff gain sufficient knowledge about the animals under their care through attending regular courses equivalent lectures and seminars and by continuously improving their experience within the field They always try to stay up to date with current affairs 44 in the industry which in turn benefits experimental animals and scientists working alongside the technologists As with any career there are both good and bad sides The goal is to find a happy medium where the good overpowers the bad and all the disadvantages are diminished To start with animal technologists have a great advantage over the scientists since they work closely with animals on a regular daily basis They tend to know these animals much better than a scientist who may perform a procedure This is one of the advantages that need highlighting since getting to know the animal will enable the animal technologist to per form procedures with a greater care and compassion and will allow them to take extra care to ensure that animal welfare is to the highest standard Consequently this will lead to a less stressful and more comfortable environment for the animal where it can relax and feel at ease Furthermore the knowledge about particular animals may help animal technologists in identifying any abnormal behaviour before during or after a procedure For instance an animal that feels threatened or alarmed may behave unpredictably and will try to defend itself In this situation it is important for a technologist to get to know the animal well before performing any procedure Getting to know an animal under care can also have its disadvantages There are cases of technical staff becoming attached to the animal resulting in an ethical dilemma that could

AS ET Special Travel Bursary 2014 essays affect the technologists ability to perform procedures This emotional distress may lead to inaccurate results The technologists can try to minimise attachment with research animals by taking into account the overall result of their work For example using one laboratory animal may help or even save many other lives of both humans and other animals If a technologist takes this into consideration and accepts the fact that they are doing a job for a greater good the distress may lessen These circumstances may especially apply to higher animals belonging to such orders as primates carnivora or artiodactyla where getting to know the animal well before starting any experiment is usually essential On the other hand learning about different species and the way they behave may have a great impact in understanding how animals react to different situations and how they manage to survive in the wild This can also be very helpful in planning and maintaining future welfare management For example by encouraging animal technologists to participate in a variety of inhouse welfare studies one can learn how to handle different species and strains This may be very useful during procedures since diverse strains of the same species may exhibit a completely different type of behaviours and reaction to stress Controlling animal diet and keeping record of its consumption can serve as an example in that the way food is eaten varies amongst different strains of mice This may indicate that some strains are more likely to get stressed more easily than others It also may suggest that levels of boredom across diverse strains differs Taming of newly arrived animals can be yet another example of inhouse studies rabbits for instance arriving from a designated breeding establishment need some time to adjust to the new environment and surroundings During this time technicians can make a difference by spending at least 15 30 min a day to try to make these animals less stressed nervous or frightened Another example was given by Hall2 at the 2014 IAT Congress when she discussed the importance of good welfare and knowledge of the laboratory housed dogs while refining a technique of oral gavage During this study a Welfare Assessment Framework WAF was developed where patterns of behaviour cardiovascular nociceptive and affective measures showed improvements in many parameters in dogs that were previously trained and whom technicians spent more time with All the above examples benefit from creating a calmer environment for the animals technicians and even scientists Hence once again studying and observing animal behaviour may have a great impact towards experiments research and procedures being performed As discussed the disadvantage to this may be an emotional attachment and distress to the laboratory animals Another aspect is that at times performing a procedure or a surgery can become stressful or even disturbing to both the animal care staff and their animals This is why a constant practice improvement and refinement of one s ability to perform procedures using the 3Rs are essential One such example may be illustrated by performing intravenous or intraperitoneal injections If an animal technologist is not experienced enough to perform such injections it may result in distress to the animal under procedure as well as to the technologist This problem can be overcome by gaining confidence and practice in doing different kinds of injections on cadavers and by improving understanding of the impor tance of safeguarding animal welfare the integrity of studies and compliance with ASPA A disadvantage from the animal technologists point of view may be that they will have to euthanise the animals to practise procedures although usually animals euthanised for other reasons will be used rather than a novel animal However an advantage to this will be a highly experienced and confident technologist who is able to perform procedures with ease and confidence Consequently this conflict can be simply justified on the grounds that the animal technologists will save more lives and improve welfare of future laboratory animals under their care Another great advantage of being an animal technologist qualified to perform procedures is the involvement in experiments and research Knowing that one can be part of discovering cures new medications and assisting in a breakthrough in combating diseases is certainly very satisfying and rewarding This aspect can be mutually beneficial to humans and other animals as many drugs and treatments developed for human medicine go on to be used in veterinary medicine and much is learnt about animal anatomy during post mortems etc This is also a great way to obtain experience and knowledge in carrying out procedures or small surgeries on different species A disadvantage to this may be that even after applying the 3Rs on occasion more animals may be euthanised than a technologist would like Balance is key here in the sense that one must be a realist yet also compassionate with regard to the research and the animals Promoting balanced approaches towards present research will help the general public to understand the significance of the work being carried out whilst also addressing their concerns Following the previous dilemma another disadvantage to the animal technologist may arise The fact that even nowadays the general public knows very little about the work of the animal technologist which can be very discouraging at times On this basis animal care staff are usually persecuted for what they do and where they work After years of intimidation from animal rights extremists technical staff and researches are becoming more open about the use of animals in 45

AS ET Special Travel Bursary 2014 essays research The best approach to this problem is by animal technologists and scientists to constantly encourage the public sector to learn more about animals in research in the form of a variety of seminars and free events and advertise the industry through the media Further according to Deeny3 more than 70 scientific organisations such as universities charities research councils and learned societies have now joined the Concordat on Openness on Animal Research Leech4 also informs that The European Animal Research Association EARA has been established in response to the need expressed by the research community to inform the European public on the continued need for and benefit of the human use of animals in biomedical research This development may increase public awareness and understanding which in turn will help research and technical sectors to promote and develop their work to the highest standards To sum up the animal care staff will always encounter predicaments and emotional discomfort However they should not to be discouraged but be proud of what they do and the difference they make in the world Their work is outstanding and exemplary and has been and will continue to be a great contribution to humanity and the animal kingdom References 1 2 3 4 46 George Bernard Shaw 2012 Back to Methuselah Act 1 Selected Plays and Prefaces CreateSpace Independent Publishing Platform 25 August 2012 ISBN 10 1479194476 Hall L 2014 Refining oral gavage assessing and improving animal welfare in the laboratory housed dog IAT Congress 2014 abstracts PRC Associates Ltd Deeny A 2014 Times Higher Education At the heart of the higher education debate Closed minds the media and animal research Available at http www timeshighereducation co uk comment opinio n closed minds the media and animalresearch 2013836 article Accessed 27 06 2014 Leech K 2014 Animal research Time to talk IAT Congress 2014 abstracts PRC Associates Ltd

April 2015 Animal Technology and Welfare Advantages and disadvantages of animal technologists performing regulated procedures in research ADAM JOHNSON The University of Manchester BSF 4th Floor Stopford Building Oxford Road Manchester M13 9PT Corresponding author adam johnson manchester ac uk AS ET Special Travel Bursary 2014 entry It is the intention of this essay to discuss the advantages and disadvantages of animal technologists being responsible for the care and welfare of animals in their charge and performing procedures on these same animals The essay will also attempt to explain how if potential conflict exists between these two roles it may be minimised This is an issue that some laborator y animal technologists may have to deal with on a daily basis whilst others may not par ticipate in per forming regulated procedures on the animals they care for For those that do the issue may sometimes be multifaceted with both obvious and less obvious advantages and disadvantages These technologists will be used to balance these points against each other when considering how to approach the given task These pros and cons will often be specific to the species their individual behaviour traits and the personal opinions of the technologist This short essay will aim to illustrate some of the most common aspects of the issue also been used to great effect with other species including rats 4 In practice the trainer is often the person undertaking the daily husbandry responsibilities and the actual regulated oral dosing may be undertaken by a different person In the author s personal experience marmosets can become quite unsettled when a person other than the regular husbandry staff enters the housing room A further refinement to the above would be to ensure where possible that the technical staff member that has trained the marmosets to take the apple juice voluntarily and is present on a daily basis for substantial amounts of time is the same person that offers the actual oral dose The marmosets would ordinarily be more trusting of this person and be more relaxed about taking the oral dose than if it were offered by a less familiar face or indeed one that was only present for regulated procedures and whom the marmosets may therefore associate with negative situations An example of a situation in which it may be advantageous for technical staff to both perform procedures and attend to the daily husbandry duties may be the oral dose training of marmosets In marmoset models of Parkinson s disease it is typical that treatments are given via the oral route 1 2 However if it is accepted that some animals may form negative associations with people that are performing regulated procedures it must also be accepted that the technologist could also be associated with stressful situations In the case of the oral dosing of marmosets this is unlikely but it could certainly be true for other scenarios It is a common refinement to train the marmosets to receive the oral treatments by periodically offering them a desirable treat such as apple juice via syringe for a period prior to offering the treatments 3 This method ser ves the purpose of acclimatising the marmosets to the procedure and therefore reducing the stress that it may cause It also eliminates the need for handling which generally induces a stress response in marmosets Similar styles of training have If this becomes a concern the solution may vary between species With the marmosets spending extra time with the affected animals is almost always beneficial Simply sitting in the enclosure and offering occasional treats often has a positive effect on the way that the animals view their carer Introducing new environmental enrichment and simple games will also keep the marmosets occupied and less concerned about the presence of the technologist 47

AS ET Special Travel Bursary 2014 essays Whilst our primary focus is providing the highest standards of welfare and care for the animals that we work with we may sometimes put our own emotion to one side A potential disadvantage of technical staff per forming regulated procedures with the same animals that they care for on a daily basis is the emotional effect this may have on that person Although this may be true it often appears to be the case that animal technologists prefer to carry out the procedures themselves The reason for this is usually that they have a knowledge of each individual animal that is difficult for another person to understand A technologist may have a greater understanding of how different animals are likely to respond to a given procedure and be able to take subtle measures to further reduce the potential impact that the procedure may have upon the welfare of the animal A typical example of this might simply be the approach towards the initial handling of the animal prior to the procedure The animal technologist will often be the person most capable of calming the animal prior to the procedure as they handle the same animal on a regular basis and will be familiar with the personal behaviours of each individual As scent is an extremely important sense to many animals the familiar scent of the technologist is also likely to reassure an otherwise apprehensive animal In attempting to minimise the emotional impact upon the technical staff one of the most important factors to consider is the quality of training they receive If a technologist is confident that they can perform the procedure well and minimise to the highest degree possible the level of suffering that the animal is likely to experience they are likely to be more comfortable in that situation Of course under the Animals Scientific Procedures Act 19865 it is a legal requirement that any person carrying out a regulated procedure must be certified as fully competent to perform that procedure but the important point with regards to this discussion is that the technologist feels confident enough in their technique that they are able to assure themselves there is a benefit to the animals welfare in them performing such a procedure A technologist should not be forced to perform a procedure that they were unhappy with so it is important that they consider each new procedure carefully before committing to it Anecdotally in Japan it is customary for some of those that work with animals in biomedical research to hold a grieving ceremony for the animals that they have worked with This is partly due to cultural traditions regarding animals and whilst commendable may not be a solution that directly translates quite as appropriately within our own society However it does address an issue that is sometimes overlooked within our own culture It is important to nurture a culture of support between colleagues within facilities to ensure 48 that the challenges of animal care staff performing regulated procedures are met with maximum confidence and minimum stress for both the animal and the technologist In conclusion it is clear that some advantages and disadvantages do exist where technical staff are performing regulated procedures on the same animals that they care for on a daily basis The most significant advantage being that technologists are often able to reduce stress and suffering to a level that might not be possible for someone less familiar with the animals to achieve The disadvantages of animals creating negative associations with their carer or the emotional effect the situation may have upon the technologist are sometimes unavoidable but we are able to minimise the impact of these factors with high quality training the support of colleagues and recognising the needs of individual animals From the wider ethics of laboratory animal research to maintaining quality of research whilst simultaneously prioritising animal welfare technical staff are not strangers to problems that have no clear right or wrong answer When confronted with an issue such as the one addressed in this essay laborator y animal technologists are typically well equipped to find a solution that benefits the animals in their care and also importantly a solution in which they can have confidence References 1 2 3 4 5 Pearce R K et al 2002 The monoamine re uptake blocker brasofensine reverses akinesia without dyskinesia in MPTP treated and levodopa primed common marmosets Movement Disorders 17 5 877886 van Vliet S A et al 2006 Neuroprotective effects of modafinil in a marmoset Parkinsonmodel behavioural and neurochemical aspects Behavioural Pharmacology 17 5 6 453 462 Erkert H G E 1999 in Poole T and English P eds The UFAW Handbook on the Care and Management of Laboratory Animals 7th ed Terrestrial Vertebrates Vol 1 pp 574 590 Blackwell Science Oxford Atcha Z et al 2010 Alternative method of oral dosing for rats Journal of the American Association for Laboratory Animal Science 49 3 335 343 Home Office 1986 Animals Scientific Procedures Act 1986 HMSO London

April 2015 Animal Technology and Welfare Advantages and disadvantages of animal technologists performing regulated procedures in research MELANIE DUNSCOMBE Biomedical Research Facility University of Southampton Faculty of Medicine Mail Point 828 Southampton General Hospital Tremona Road Southampton SO16 6YD Corresponding author m s dunscombe soton ac uk AS ET Special Travel Bursary 2014 entry Many animal technical staff have a dual role they are responsible for the care and welfare of animals in their charge but are also licensed to carry out regulated procedures on those animals Discuss the advantages and disadvantages of animal care staff performing procedures on animals in their care explaining ways in which any potential conflict between the two roles can be minimised Animal technology has always been a controversial and highly emotive subject Over the past few decades animal research has gone from being demonised in the media to an almost underground issue because of this only recently has it started being portrayed in a positive light Over the years the nature of the work has changed dramatically from Louis Pasteur s work on the germ theor y of disease in 1881 to the Roslin Institute s cloning of sheep in 1996 1 The wide variety of procedures that animal technical staff carry out are potentially stressful both for the animals and for the technicians involved themes in the field of animal experimentation All data is valuable when the experiment has been condensed and compacted down to the absolute minimum number of animals in order for it to be viable It will have reached the point where each animal is vital and the procedures need to be performed precisely so that the experiment is worthwhile Technologists regularly carry out a large number of experimental procedures developing specific skills and ultimately achieving an extremely high level of competence Another advantage is that there will be a good continuity of care if the same technologist is able to carry out a procedure on an animal and follow it up with all the appropriate aftercare Technologists routinely spend a lot of time observing the behaviour of the animals in their care as well as their physical condition and as a result will generally be able to pick up on a slight deviation from normal much faster than another technologist who is not the primary carer for the animals in question Researchers may not necessarily be able to check on their experimental animals daily they may have to delegate to another member of their team who may not know the animals so well One of the main advantages of animal care staff being licensed to carry out regulated procedures on the animals in their charge is that the animals interests will be their priority Researchers on the other hand could be preferentially considering the financial costs of their reagents or equipment ahead of animal welfare The cost of reagents cannot be ignored completely for example an antibody which has taken months to acquire and is consequently very valuable will obviously be of consideration but as a general rule the cost of the procedure will always be weighed in favour of the cost in terms of suffering that the animal will pay Also some researchers have little or no experience with animals and as a result have no idea about how to approach them whereas with animal technicians it is very common for them to have had a great deal of experience with a wide variety of animals Animal technologists by the nature of their choice of career have a great deal of love and respect for animals and have generally built this up throughout their lives by having all kinds of different pets and hobbies involving animals Researchers are taught a lot about careful experimental design the three Rs reduction refinement and replacement are constant recurring It is beneficial to involve animal care staff in the more technical side of the experiments that are happening with the animals in their care It is helpful if 49

AS ET Special Travel Bursary 2014 essays technologists are able to take an interest in what the researchers are trying to find out about then the role of licensee has more meaning A better understanding of the nature of the research being done also helps with aftercare knowing about specific symptoms to look for and normal levels of deterioration to expect prior to recovery are essential The application of local anaesthetic prior to blood samples being taken has been a topic of discussion in my experience Some researchers could neglect to factor in the time needed for local anaesthetic to be applied and left to take effect There are time constraints involved with sample processing and subsequent data analysis when working in a laboratory with a lot of researchers keeping to the bookings of analytical equipment could mean that they neglect the welfare of the animal The technologist might have to attempt to enforce the application of anaesthetic and allow time for it to be effective The fact that animal technologists by the nature of their choice of career have a great deal of love and respect for animals can also be a disadvantage when it comes to carrying out experimental procedures on the animals in their care There will undoubtedly be a strong personal conflict regarding the humane treatment of animals in captivity such questions as What gives us the right to decide that animals are experimental equipment for humans Is there any justification that can be deemed good enough to warrant the restraint and environmental manipulation of surgical or genetic interference with infection and treatment and ultimately the killing of a living sentient animal Is the benefit for the health and economic growth of humans a fair justification It is possible that a technologist could be asked to perform a procedure that while detailed in project and personal licences they do not feel comfor table performing They could feel their level of competence is not at a high enough level or it could be that they feel the cost to the animal outweighs the benefit of the research Legislation exists in order to ensure that experimentation is only permitted when there is no alternative means of achieving a specific set of experimental results The expected benefits must outweigh any possible pain suffering distress or lasting harm that the animals may experience The existence of this legislation is encouraging but the possibility exists for details to be confused due to differing interpretations The legislation helps to minimise the conflict between the roles of carer and licensee There are clearly defined severity levels for all procedures and regular monitoring of these severity levels on project licences Likewise there are clearly defined endpoints on project licences Good communication between researchers 50 and technologists regarding what to expect for each experiment is essential if there is ever any question about procedures whether an animal needs to be culled from an experiment or whether a person is competent at a specific procedure there are always the Home Office named persons available to consult the NACWO Named Animal Care and Welfare Officer the NVS Named Veterinary Surgeon the NTCO Named Training and Competency Officer and the NIO Named Information Officer The AWERB Animal Welfare and Ethical Review Body meets several times a year in order to discuss all aspects of experimental work being carried out at an establishment considering project applications and monitoring their progress on a local scale There needs to be conflict between the dual roles of carer and licensee without it the whole concept of humane animal experimentation would be compromised Minimising this conflict is essential for the wellbeing of the technicians as well as the animals A good technologist cares deeply and strongly for their animals it is this that makes them so genuinely thorough when considering how to approach all aspects of procedures to potentially be per formed Technologists are fully aware of the ethics involved in what they do There is a fine balance between caring for the animals and being able to willingly do things to them that cause them discomfort in the pursuit of medical advancement It could be so easy to simply switch off mentally and blindly carry on performing what could be interpreted as violent procedures against animals so many times when asked how they could do what they do all day every day have technologists responded that they just try not to think about it too much The point is that they do think about it A good technologist will always have in the back of their mind an everlasting respect for the animals in their care and will always be considering on some level whether or not what they are doing is justified The balance comes when the benefits to so many are shown the treatments and procedures we have discovered and perfected thanks to the animals that technologists have carefully performed procedures on References 1 Animal Research info Timeline online Animal Research info Available from http www animal research info en medical advances timeline accessed 25th June 2014

April 2015 Animal Technology and Welfare POSTER PRESENTATIONS Originally presented at IAT Congress 2014 Introducing an objective animal health and welfare assessment system into the biological resources unit of Cancer Research UK London Research Institute DANIELLE COX VALERIE BOREL VANNIER CLAIRE HARPER GARY MARTIN MERCEDES SANCHEZ and GARY CHILDS Cancer Research UK Clare Hall Blanche Lane South Mimms Potters Bar Hertfordshire EN6 3LD Corresponding author gary martin cancer org uk Introduction An objective system was required by staff at the Cancer Research UK CRUK London Research Institute LRI to allow the lifetime experience of an animal under procedure to be assessed A team consisting of animal technologists researchers and the Named Veterinary Surgeon NVS devised and developed a scoring system so that when assessing a sick mouse there are two specific areas to be considered the mouse welfare and the science The development and introduction of a scoring system has many advantages G G G G defines patterns of individual strain phenotypes allows Health and Welfare monitoring of the mice against specific criteria provides a detailed plan of action when mice exhibit signs of ill health provides standardised data records for individual animals How the scoring was developed The Biological Resources Unit BRU staff at the LRI identified a need to develop a system to evaluate observable patterns in mice with unknown phenotypes A system was suggested that would score the following points of concern when evaluating a sick mouse G G G is the health and welfare of the animal being compromised will there be any additional information gained that can benefit the research has the humane endpoint been reached A group consisting of a researcher the NVS and an animal technologist looked at previous and existing scoring systems developed and practised in other establishments A scoring system of particular interest had been developed1 using body condition scoring and behaviour to assess endpoints for sick mice This scoring system had two levels of assessment firstly sick mice were identified by their body condition this however appeared to be very subjective and open to personal opinion The second part assessed the animals behaviour and used numbers to determine the level of 51

Poster Presentations sickness but again was deemed to be subjective The group felt that this system had a complex scoring system and required a veterinary surgeon and two technologists to score each animal and as such was deemed to be impractical It was therefore decided to develop our own scoring system which could be adapted for individual project licence requirements and used clinical signs that were grouped within a four tier section each with its own action plan Developing the scoring system Once it was decided that we would develop our own scoring system we documented the clinical signs and observations of mice that would be relevant to the study However some mice were of unknown phenotype and as these phenotypes became apparent they were noted and added to the list as the work progressed We divided the signs and observations into four categories dependent on the severity of the phenotype or condition white green yellow and red and produced colour coded cage tags as a visual aide for anyone working in the room The specifics of the four categories were tailored to the individual project licence selected for the trial Each of the categories has its own agreed action plan which is easy to follow and allows for standardised outcomes and enhanced communication between technologists and the researcher Using Microsoft Excel we designed a form that allowed the capture of all relevant data relating to a specific mouse It provided information regarding the date of initial and subsequent assessments observed signs using a tick system allowed re scoring of previously noted conditions and the identification and scoring of additional conditions G G G G G G G pale extremities hyperactivity tremors agitation aggression abnormal breathing seizures when handled loss of co ordination mild It was agreed that these were mild signs and could be the early indicators of a mouse becoming sick so requiring closer monitoring A record of how long it took a mouse to progress to a yellow level sign would be taken The yellow level consists of the signs G G G G G G sustained hunched posture nasal discharge lethargic significant abnormal breathing prolonged seizures weak and stiff limbs It was agreed that if any of these signs were exhibited action should be taken and a decision made regarding closer monitoring or removal of the mouse from the experiment The most serious signs were classed as red G G G G G G G G hind limb paralysis abdominal palpable mass visible solid mass ulcerated tumor ulcerated wound 20 weight loss ataxia complete loss of co ordination unresponsive to stimuli This is the defined endpoint for a mouse and immediate action must be taken White level The common health problems that occur in mouse strains used by this laboratory were noted and included G G Red overgrown teeth dermatitis Yellow We felt there was a need for a separate scoring sheet for this level which linked to the main scoring sheet and specifically showed the different levels of common issues such as dermatitis thus ensuring that animal welfare is maintained Green level signs are G G G G G 52 unusual gait piloerection ungroomed appearance eyes become dull un sustained hunched posture Green White

Poster Presentations weight loss Please fill weight record sheet once the mouse has loss 10 it will be weighed twice a week for the 20 loss 0 1 2 3 4 5 Sick mouse assessment sheet 53

Poster Presentations How the scoring system works All mice within the quarantine unit are checked in their home cage on a daily basis If upon checking there is any cause for concern the cage is removed from the Individually Ventilated Cage IVC rack and placed in the cleaning station for further observation If a health or welfare issue is noted with any animal healthy cage mates are removed into a clean cage leaving the sick mouse mice in their own environment At this point with the cage in the cleaning station the assessment process begins the mouse being observed for at least 3 5 minutes Notes are taken during the assessment but it is very important to observe the animal before referring to the Sick Mice Assessment Sheet as you may find yourself being led by the points noted on the form and adapting the actions of the mouse to this rather than the other way round This provides a more accurate interpretation of the observation you may overlook a phenotype if using the form as a checklist All information is entered onto the assessment form and given a unique case number The form is evaluated and appropriate action as detailed below is taken 1 If a section in white is ticked for overgrown teeth we trim them as required up to a maximum of two occasions and give wet mash diet If no sign of improvement is seen the mouse is removed from the experiment If dermatitis is ticked the specific scoring system is used and the Named Veterinary Surgeon is contacted to discuss appropriate treatment 2 If there are two or more boxes ticked in the green section the researcher is contacted by email further monitoring is discussed and agreed with the animal being placed on special care on the CRUK mouse database 3 If one or more boxes are ticked in the yellow section either on its own or in combination 4 With any ticks in the green section the researchers are contacted by phone and email to review the mouse immediately At this point the appropriate action must be taken and discussed with the researcher and then recorded on the database 5 If any box is ticked in the red section immediate action must be taken all researchers within the team are informed by phone and email and the situation discussed but if a response is not immediately received the NACWO or NVS is informed to make the final decision A video clip is taken and full post mortem performed Conclusions We have successfully introduced a robust and detailed scoring system that provides a standardised method to assess mice with unknown phenotypes Its use directly minimises the amount of pain and distress a mouse may experience whilst clearly identifying humane endpoints 54 The system is user friendly and by placing specific phenotypes in coloured bands each with its individual plan of agreed actions it can be adapted to all appropriate project licences The scoring system has been in place for a year and has been responsible for improved communication between the BRU and the research team along with improved response times for urgent issues Following the successful trial period it is intended to adapt and use the Sick Mouse Assessment Sheets with other similar projects throughout the BRU References 1 Paster E V Villines K A and Hickman D L 2009 Endpoints for mouse abdominal tumor models refinement of current criteria Comp Med 2009 Jun 59 3 234 41

April 2015 Animal Technology and Welfare The development of hormone loaded diets to promote xenograft growth ALISON RITCHIE PAM COLLIER NIOVI NICOLAOU and ANNA GRABOWSKA Cancer Biology Division of Cancer and Stem Cell Sciences School of Medicine The University of Nottingham City Hospital Campus Nottingham NG5 1PB Corresponding author alison ritchie nottingham ac uk Introduction Materials and methods In our facility we use several tumour models which are dependent for growth on hormone supplementation delivered via slow release subcutaneously implanted pellets These include prostate and breast tumours which rely on 5 a DHT and 17 b Estradiol E2 respectively Administration of E2 results in welldocumented side effects such as bladder calculi and urine scald Figures 1 and 2 Therefore to improve animal welfare and to avoid steroid supply problems we developed a new way to provide hormone supplementation via the diet Following appropriate formulation of 5 a DHT we initially tested this approach using the LnCap prostate cell line 1 The success of this led us to develop a further diet containing E2 using the MCF 7 breast line 2 Male and female MF 1 NuNu mice were implanted with bioluminescent and wild type cells of LnCap and MCF 7 respectively at a concentration of 2 x 106 cells per site males being implanted subcutaneously females into the mammary fat pad Pellets were implanted using a 10 gauge trochar IRA Figure 3 Figure 3 Implant trochar and pellet The mice were divided into the following treatment groups Figure 1 Bladder calculi Figure 2 Urine scald 1 No hormone supplementation 2 Supplementation via slow release pellets IRA i Males were implanted sub cutaneously with 5 aDHT 0 5mg 21 day release pellets Only a wild type group was set up due to shortage of pellets ii Females were implanted sub cutaneously with 17 b Estradiol 0 1 mg 21 day release pellets 3 Supplementation via diet ssniff i Males were fed 5 a DHT 2 4mg kg ii Females were fed 17 b Estradiol 2mg kg Figure 4 5 a DHT diet Figure 5 17 b Estradiol diet 55

Poster Presentations The doses were calculated as being equivalent to that received daily from the pellets and dyes were incorporated into the diets to avoid confusion Tumours were measured weekly using vernier calipers to calculate volumes and bioluminescent lines were imaged using the IVIS Spectrum Caliper Life Sciences to measure both tumour size and viability Figure 6 C Figure 6 IVIS Spectrum Mice were also weighed daily to ensure they were eating the test diets and that their weight gain was comparable with the control diet group Results As can be seen from both the imaging results and growth curves Figures 7 and 8 the supplemented diet facilitated tumour growth in both cases With the LnCap the increase was obvious from the start but with the MCF 7 the effects were more marked from day 14 onwards The MCF 7 growth data showed an increase with diet but not as much as with the pellets while both the control groups showed a lower growth rate LnCap or no growth at all MCF 7 However the slower growth rate of MCF 7 could allow a longer window of opportunity for treatment effects when carrying out therapy studies A i ii B i ii D Figure 8 Growth curves C LnCap D MCF 7 Conclusions Delivery of hormones required for establishing and maintaining tumour growth via the diet rather than via slow release pellets is a major refinement in welfare terms by reducing the need for an invasive implant and possible removal procedure and deleterious side effects while still promoting tumour growth While growth was slightly slower with the E2 dietar y supplement we are currently testing diet with a slightly higher Estrogen content to see if we can improve growth while keeping the unwanted side effects at bay iii Acknowledgements Figure 7 Bioluminescent images Ai LnCap no DHT day 35 Aii LnCap DHT diet day 35 Bi MCF 7 no E2 day 14 Bii MCF 7 E2 pellet day 14 Biii MCF 7 E2 diet day 14 Equally importantly we also obser ved a marked reduction in side effects in mice treated with the E2 dietary supplement compared with pellets with only some slight urinary retention which was eliminated by taking the mice off the supplemented diet for a few days which is much easier and less invasive than removing a subcutaneous pellet 56 Thanks to ssniff Spezialdi ten GmbH for their help and expertise in developing the diet To Phil Clarke and Marian Meakin for their technical assistance and to Allan Thornhill for Figures 1 and 2 References 1 2 Horoszewicz J S et al 1983 LNCaP Model of Human Prostatic Carcinoma Cancer Research 43 1809 1818 Nils Brunner et al 1992 IGF I and IGF II expression in human breast cancer xenografts Relationship to hormone independence Breast Cancer Research and Treatment 22 39 45

April 2015 Animal Technology and Welfare Managing emotion in the workplace for those working with research animals ALISON HOPKINS Monkey Puzzle Training and Consultancy Ltd The Waggon House Dean Street Farm Dean Somerset BA4 4SA www monkeypuzzletraining co uk Corresponding author alison monkeypuzzletraining co uk Introduction It is becoming increasingly recognised that working closely with animals par ticularly in a laborator y environment can be both physically and emotionally challenging Because of the unique nature of the work in this area and the context many work related issues can become exacerbated and this leads to increased levels of sickness stress and burnout for staff Why can we no longer ignore this Emotions are considered to be the foundation of a person s ability to understand themselves and relate to others When you are in control of your emotions you can think clearly and creatively manage stress and challenges communicate well with others display trust empathy and confidence Without emotional control you will spin into confusion isolation and negativity By recognising and harnessing your emotions you can gain control over the way you react to challenges improve your communication skills and enjoy more fulfilling relationships This is the power of developing emotional awareness and managing emotion particularly within the workplace In the workplace it is harder to express emotion as we may normally do so through fear of judgment and the necessity to conform The expectation to be emotionally strong combined with an emotionally challenging role can lead to high levels of stress increased sickness and burnout for staff What s coming to light for the first time is that euthanasia of their charges triggers feelings of guilt remorse and grief in many carers but most suffer in silence because the subject is taboo and they feel they have no way to unburden themselves Few receive any formal training on how to cope with their feelings or practical or emotional support whether from the institutions and scientists they work for or from colleagues and family members To add to their distress laborator y animal technicians are often portrayed by animal rights extremists as torturers and murderers Andy Coghlan New Scientist March 2008 2 There are several labels associated with positive and negative emotions The six basic ones as identified by Paul Ekman3 are anger disgust fear happiness sadness and surprise These can all be felt in varying degrees by individuals at any one time In a research laboratory environment particularly involving the use of animals all of the above mentioned emotions could be felt in the same day creating a rollercoaster effect How can managers and individuals better understand emotional conflict and deal with it effectively to reduce stress and burnout Can this be as simple as creating an environment of openness and understanding If it is how do we do this and how do we know when we have this What the research says There are some basic psychological techniques that can be introduced and deployed within the workplace Such techniques can be taught and encouraged within the workplace to allow individuals to gain control over the emotion they are experiencing and deal with it positively reducing intensity and stress It has been identified Davis 2008 1 that those working with animals employ coping mechanisms to deal with the emotional demands of balancing caring with euthanasia particularly focussing on the guilt that this may trigger Thought re framing is one good example This technique helps to neutralise negative emotion by allowing the individual to experience it in a different context one that is more empowering 57

Poster Presentations Let us take guilt as an example Dealing with guilt can be fairly complex and often the trigger for guilt is a result of a conflict in our beliefs We are all brought up within society rightly so to believe that killing is wrong However we are not taught that within this belief there are levels of euthanasia that may be acceptable at an individual level By putting a new frame around a context it can be seen in a different way Do the two sentences below give you a different sense of emotion I had to euthanise a batch of mice today because they were not needed anymore I euthanised a batch of mice today that had been bred specifically for scientific purposes and had made a significant contribution to scientific advancement This is just one example where a re frame can create greater meaning and empowerment Staff can be trained to employ these techniques for themselves and leaders can develop the ability to coach their teams to use them What if you were proud to be doing your job recognising the higher purpose and feeling happy to go into work each day What if you were confident that you could cope with whatever challenges lay ahead by knowing the value of your work and its greater impact on society Introduction With emotional understanding and the employment of some simple psychological techniques this is possible both for yourself and others within the workplace References 1 2 3 Davis K 2008 Lab Animal Europe March 2008 Coghlan A 2008 New Scientist March 2008 p8 9 http www helpguide org toolkit developing_emotional_ awareness htm INLPTA NLP Techniques 58

April 2015 Animal Technology and Welfare A simple solution to prevent the abdominal migration of temperature loggers and to facilitate their smooth retrieval post study in macaques LEO VAN GEEST MEREI KEEHNEN RIANNE KLOMP JACCO BAKKER ROBIN VAN DER SCHILT and JAN LANGERMANS Animal Science Department Biomedical Primate Research Centre Rijswijk The Netherlands Corresponding author geest bprc nl Introduction Before being infected with an Influenza virus 12 cynomolgus monkeys Macaca fascicularis were implanted with temperature data loggers These devices are small implantable recorders that measure temperature with a high degree of accuracy and store the data in their internal memory All measurements are in real time and can be accessed after the logger has been retrieved at the end of the study After retrieval of the data the logger can be reprogrammed and re used for as long as the battery lasts The transmitters upper surface is very smooth and has no ridge or other affixing possibility to attach the logger to the abdominal wall In previous experiments we experienced that such loggers migrated through the entire abdomen Figure 1 which made surger y to retrieve the loggers difficult necessitating almost an explorative laparotomy In order to refine this retrieval surgery a simple homemade solution was devised a bled knot of nonabsorbable suture material with needle was created around the logger and the needle was used to attach the logger to the abdominal wall during wound closure Five months after insertion the temperature loggers were removed from all 12 animals All loggers were still fixed in the position where they had initially been stitched The retrieval surgery was scored as a minor discomfor t All measured data was uploaded successfully into a PC The implanted loggers did not have an adverse effect on the animal s health which was checked daily After retrieval all animals were alive The bled knot of non absorbable suture material with needle around the logger showed to be a simple solution to prevent the abdominal migration of temperature loggers and to facilitate their smooth retrieval post study in macaques Figure 1 Abdominal X ray lateral view a non attached logger that had migrated to the vertebral column is clearly visible It is conceivable that surgery to retrieve the logger would be difficult almost necessitating an explorative laparotomy Materials and methods Animals housing and care This study included 12 adult healthy male cynomolgus monkeys Macaca fascicularis that were housed at the Biomedical Primate Research Centre BPRC Rijswijk the Netherlands aged 6 13 years and weighing 5 611 3kg All macaques were pair housed same sex couples in the experimental facility The monkeys had no history of abdominal surgery The procedures performed in this study were in agreement with the regulations for animal handling as described in the EU Directive 63 2010 and in accordance with the Weatherall report 2006 2 During the study the monkeys were kept under close veterinary supervision The animals were fed with 59

Poster Presentations commercial monkey pellets ssniff Spezialdi ten GmbH Soest Germany supplemented with fruit and vegetables and drinking water was available ad libitum Food was removed 16 hours prior to sedation but water intake was never restricted 2 0 and the skin incision was closed with single knot sutures 2 0 The position of the logger is shown in Figure 4 During the course of the study animals were checked at least twice daily for appetite general behaviour stool consistency and local side effects of the surgery Surgery Anaesthesia was achieved by means of 10 mg kg ketamine Ketamine 10 Alfasan Nederland BV Woerden NL 100 mg ml and medetomidine IM Sedastart AST Farma B V Oudewater NL 1 mg ml As analgesia 1h prior to surger y 0 20 mg kg meloxicam Metacam Boehringer Inhelheim Alkmaar The Netherlands was administered together with 0 02 mg kg buprenorphine Buprecare AST farma B V Oudewater The Netherlands After surgery animals received meloxicam 0 10 mg kg PO once daily for two days Figure 3 The pre op prepared logger was inserted into the abdominal cavity and the needle was then used to attach the logger to the abdominal wall during linea alba wound closure Subsequently the abdomen was shaved and prepared with chlorhexidine and povidone iodine The macaques were placed on a heated blanket in order to stabilise body temperature and were breathing room air spontaneously throughout the surgery After pre operative preparation a 3cm skin incision below the umbilicus was made along the midline Subsequently the skin was separated from the abdominal muscles The abdomen was opened the sterile transmitter with a bled knot of non absorbable suture material FS 26 18 45cm size 2 0 around it Figure 2 was inserted into the abdominal cavity and the needle was used to attach the logger to the abdominal wall during linea alba wound closure Figure 3 The abdomen was closed with a continuing suture Figure 4 Abdominal X ray lateral view The logger was inserted into the abdominal cavity and the needle was then used to attach the logger to the abdominal wall during linea alba wound closure Results The temperature loggers were removed smoothly from all 12 animals 5 months after insertion All loggers were still fixed in the position where they had initially been stitched Figure 5 The retrieval surger y was scored as a minor discomfor t All measured data was uploaded successfully into a PC Figure 6 Figure 2 The pre operative prepared logger with suture material and bled knot around it 60 The implanted loggers did not affect the animal s health which was checked daily

Poster Presentations Figure 5 Minor surgery to retrieve the transmitter Figure 6 An example of part of the temperature data collected from an animal during this study A clear night day pattern is visible Acknowledgements Euprim net 2 EC grant agreement no 262242 References 1 2 Directive 2010 63 EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes Official Journal of the European Union L 276 79 The use of non human primates in research A working group report chaired by Sir David Weatheall FRS FMedSci https royalsociety org media Royal_Society_Content policy publications 2006 Weatherall Report pdf 61

Animal Technology and Welfare April 2015 Welfare challenges in high disease containment agricultural animals HUGH SIMMONS Animal Sciences Unit APHA Weybridge Woodham Lane Addlestone Surrey KT15 3NB Corresponding author hugh simmons apha gsi gov uk Infectious organisms are characterised legally in 3 ways by the 1 Specified Animal Pathogens Order on how contagious they are to animals 2 COSSH ACDP on how transmissible they are to humans 3 Genetic Modification Order on what effect they would have on the environment All of these pieces of legislation categorise organisms in to categories 1 to 4 with 3 to 4 being high containment A typical design for a building which can undertake category 3 agricultural animal work is shown in Figure 1 Figure 1a Design of cat 3 4 building Roof Space Air Handling Units with HEPA filters also controls temperature Figure 1c Animal room Several barriers to entry staff entering room wear PPE These facilities are extremely expensive to build and once constructed can be impossible to modify To try and get the best animal welfare possible when designing and managing them it is helpful to consider the Five Freedoms as an analysis tool Infectious organisms are 1 2 3 4 5 Figure 1b Basement Effluent Treatment Plant 62 Freedom Freedom Freedom Freedom Freedom from hunger and thirst from discomfort from pain injury and disease to behave normally from fear and distress Even when trying to assure freedom from hunger thirst and discomfort there is a significant interplay between

Poster Presentations biosafety and animal welfare For example long stem fibre is key to good ruminant digestion and bedding is a traditional way of providing comfort for all animals If the building effluent treatment plant cannot deal with any solids these cannot be used and alternative approaches must be found In the field of high containment disease research freedom from pain injury and disease is the main conflicting freedom due to the nature of the work To minimise the conflict there needs to be a whole team approach involving animal care staff the NVS the NACWO the PIL and associated scientists The AHVLA experience with Classical Swine Fever has shown that with this team the management of adverse effects can move on from welfare score charts and humane endpoints originally based on field experience from the 2000 outbreak in the UK to the use of real time white blood cell WBC count and PCR for viraemia for predictive endpoints group D Figure 2 Experiment AJ White blood cell counts 65536 Group Group Group Group Cells l 32768 16384 A B C D 8192 4096 10 5 0 Days post challenge 5 Figure 2 White Blood Cell counts after CSF challenge The freedom to behave normally should not differ between low and high security containment as the ASPA minimum space allocations do not distinguishing between the two It should be possible to provide environmental enrichment The freedom from fear and distress is more of a challenge in this environment due to reduced staff contact therefore it is important to develop the emotional resilience of the animals through temperament selection acclimatization and training to maintain this freedom The aim of this work is to develop the 4Ps Positive cultural attitude toward identifying issues use of pre start and wash up meetings facility user group Proactive ongoing development of welfare score sheets pre emptive endpoints comfort Biosafety involved from the beginning Preventative design of buildings design of experiments developing emotional resilience in animals Productive Good animal welfare good science 63

Animal Technology and Welfare April 2015 Choosing a system for managing training records ANGELA KERTON1 ALISON HOPKINS2 and ANDREAS STAUBI2 1 2 CBS Department South Kensington Campus Faculty of Medicine Imperial College London SW7 2AZ a tune software AG Julius Reiber Strasse 15 64293 Darmstadt Germany Corresponding author ahopkins a tune com Introduction There is no question that well trained competent personnel are key to a successful laboratory animal research establishment and exceptional animal welfare Within the European Union EU the directive 2010 63 EU1 aims to harmonise regulations and puts emphasis on the management of training and competence to the Establishments In the UK the concept of the Procedure Individual Licence PIL has been modified but generally maintained and combined with the requirement for appropriate training and record management programmes internal to the establishment Functional and non functional requirements Decision makers have a great range of tools and systems to choose from when looking at data management systems for managing training records Therefore it is ver y impor tant to identify core requirements for any training management system 1 Logging The data to be recorded is reliant on legal requirements and organisational policies This can be simple information name and degrees achieved or information about authorisations PIL details training courses and lists of procedures techniques etc The latter type of data requires the system to support complex data types which combine techniques or procedures with species competence levels and anaesthesia codes It is important to know all data requirements legal organisational operational before assessing any system 2 Tracking Training records are not static Information should be recorded and tracked This is especially true for reminders and expiration dates 64 relating to training as well as competence checks for internal and external courses A system should show the impor tant deadlines and send notifications to the people involved Only with a complete set of data is it possible to check whether an individual is properly trained and respectively competent to per form a cer tain procedure for a species required under a given protocol 3 Linking and checking It is insufficient to simply name the procedure technique The training record is only complete with a reference to the species and the personal and project licenses under which the individual is approved to work Additional information about competency level is required by law as well as the identity of the trainer date and expiration date 4 Repor ting The retrieval transformation and presentation of information The system has to deliver any legally required annual statistics operational reporting needs including accounting and controlling aspects such as charges for trainings and management of licences 5 Security Controlling access encrypting data and communication channels 6 Consistency Provision of mechanisms to minimise error e g drop down menus 7 Validity Audit trail Demonstrating the change history including the old new value timestamp and record of the person performing the amendment

Poster Presentations 65

Poster Presentations Conclusion Domain specific systems combine all the advantages of central databases with the possibility to model the complex relationships between species techniques procedures competencies and if applicable Personal Licences Training administrators supervisors and principal investigators and any member of the research staff then have the opportunity to check personal training records and ask questions on the availability of cer tain competencies within the team or the organisation If the system also manages Project Licences Protocols it is possible to have a quick automated cross check helping all individuals involved to contribute to compliance Domain specific systems combine all the advantages of central databases with the possibility to model the complex relationships between species techniques procedures competencies and if applicable Personal Licences Training administrators supervisors and principal investigators and any member of the research staff then has the opportunity to check personal training records and ask questions on the availability of cer tain competencies within the team or the organisation If the system also manages Project Licences Protocols it is possible to have a quick automated cross check helping all individuals involved to contribute to compliance Considerations There are more aspects to be considered like technical infrastructure requirements and the financial and timemanagement costs for initial implementation and operation Within the different categories the bandwidth of solutions and their impact is massive Nevertheless we hope that this over view has provided a basic framework for the assessment and selection of the right tool for the job within your establishment 66 Reference 1 Ar t 23 Directive 2010 63 EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes

April 2015 Animal Technology and Welfare Experimental and husbandry procedures impact results and animal well being LAURA ANNE ROBERSON ANNA KARIN GERDIN NATALIA IGOSHEVA OZAMA ISMAIL NATASHA KARP MARK SANDERSON CARL SHANNON EMMA CAMBRIDGE DAVID SUNTER RAMIRO RAMIREZ SOLIS JAMES BUSSELL and JACQUELINE K WHITE1 Wellcome Trust Sanger Institute Hinxton Cambridge CB10 1SA UK Corresponding author lr4 sanger ac uk Purpose The minimisation of variability through standard experimental procedures is an important part of optimising animal use given that the quality of experimental results translates to improved data quality statistical power and reduced animal use In this study we investigated the impact of standard animal husbandr y and common experimental procedures on data variation and the well being of laboratory mice to describe the effect of relatively simple procedures on data variation Methods All procedures were performed under the prevailing principles and authority of the Animals Scientific Procedures Act 1986 Male and female C57BL6 NTac mice underwent Telemetry Implantation surgical implantation of a radiotelemetry transmitter with a weight of 1 4g TA11PA C10 Data Sciences International USA was performed under anaesthetic by inhalation of isofluorane and oxygen and eyes were protected from dr ying with ointment Analgesia was administered Buprenorphine 0 1 mg kg s c immediately after anaesthesia Further doses of buprenorphine were given immediately and 24 h post surgery Post operatively mice were placed in a recovery rack pre heated to 30 C for 2h When mice were fully ambulant they were housed in their individual cages In addition to normal food and water a dietary supplement Complan Complan Food Ltd UK was given for 14 days to support recovery Visual examination of the overall condition of all mice and body weight checks were performed daily for 14 days post surgery Cage changing Individual housing Overnight fasting Blood sampling via tail tip excision Body temperature measurement using a rectal probe Intraperitoneal injection Body weight cardiovascular activity blood glucose and body temperature were measured as parameters that reflect stress and physiological responses to it Results Blood pressure heart rate locomotor activity body temperature and blood glucose were elevated in response to a range of experimental procedures This has impor tant implications for the design and implementation of multi component experiments such as phenotyping pipelines where the lasting effects from previous tests may modify the outcomes of subsequent ones and also on long term animal health and welfare This study describes simple refinements to working practices which minimise method related stress to reduce data variability and the numbers of animals required in an experimental protocol in accordance with the 3Rs Cage Changing Both sexes n 6 n 7 locomotor activity blood pressure heart rate 67

Poster Presentations More prolonged response than was elicited by more invasive procedures Blood Sampling Both sexes n 6 n 7 blood pressure heart rate Repeat measurement response to the same magnitude as the first Female response more prolonged than male response 50 min vs 40 min blood glucose body temperature 1 hour post sampling 72hr Individual housing bodyweight both sexes 3 loss 6 loss During recovery from surgical procedures this weight loss was more profound Upon return to group housing weight loss was reversed blood glucose body temperature 1 hour after separation Recommendations Group house animals whenever possible To aid rehousing and minimise aggression individually house all animals from the group when one is removed G seed individual cages with olfactor y cues nesting material from group cage G daily transfer of olfactor y cues between individual cages aspen bedding substrate G rehouse animals with original cage mates G rehouse animals into original group cage G Do not perform cage clean before experimental procedures Female response more prolonged than male response 105 min vs 75 min Reference 1 68 Gerdin A K Igosheva N Roberson L A Ismail O Karp N et al 2012 Experimental and husbandry procedures as potential modifiers of the results of phenotyping tests Physiol Behav 106 602 611

April 2015 Animal Technology and Welfare An evidence based approach to determine the environmental requirements for animal holding rooms STEVEN CUBITT FINTAN LYONS PETER BARRINGTON and GENE MURRAY CCTECH St John s Innovation Centre Cowley Road Cambridge CB4 0WS Corresponding author info cctech eu Introduction The standardisation of scientific animal model physiological processes such as growth maturation and reproduction are influenced by environmental factors such as temperature relative humidity and ventilation rates This can effect G G G G For these reasons modern animal facilities are designed to meet high standards of control and stability in order to minimise any possible adverse influence on experimental investigations Research has shown that Biomedical buildings in the UK are not energy efficient Fertility lactation and teratogenesis Variation of food and water intake Variation of drug activity Incidence of disease Building performance optimisation Each building has its own unique profile G G G G G G Figure 1 Biomedical Facility Total Energy Usage G building design age of building type of mechanical ventilation building management type of research housing systems species sex strain age etc 69

Poster Presentations Process 1 Detailed survey of all the services 2 Fingerprint commissioning survey 3 Evidence based modelling using environmental data on the environment for both animals and people 4 For example ammonia carbon dioxide temperature humidity sound lab animal allergens 5 Monitor how changes in the Building Performance effect the environment energy and water consumption Understanding how your building works is vital for research outcomes Summary Biomedical research is very important to the UK We need to maintain standards of welfare quality competitiveness and sustainability whilst improving or maintaining environmental quality Utilising the highly skilled resources we have within the UK we can meet these challenges Outcome From the Evidence Based Model optimise the buildings performance to provide the best environment for the research animals people maintenance and energy Identify target savings and share opportunities for success Establishing a representative team helps to secure active support for change It is the essential prerequisite to develop low carbon cost effective animal facilities 70 This will help maintain the UK as one of the global leaders for biomedical research into the future We have shown significant savings in energy carbon and cost are possible whilst maintaining a high standard of environment for research animals and people

April 2015 Animal Technology and Welfare The Siberian hamster breeding and husbandry techniques MELISSA BEAN University of Nottingham Biomedical Services Unit Medical School Queens Medical Centre Nottingham NG7 2UH Corresponding author melissa bean nottingham ac uk Abstract For the animal technologist who wishes to learn more about the animals they work with there is limited information available on the Siberian hamster Phodopus sungorus At Nottingham University we have a breeding colony of Siberian hamsters that have been with us for several years The stock animals are used primarily for studies involving photoperiodism and the colony is maintained on a continuous tick over breeding system which consists of six breeding pairs improved so that fewer breeding pairs were needed This would save the researcher money and reduce the numbers of animals being used thus promoting the 3Rs Most textbooks covering laboratory animals group the different breeds of hamster into one The guidelines and advice that they give is aimed more at the Syrian hamster so it is difficult to find accurate breeding data on the Siberian hamster Husbandry information on the Siberian that can be found on the internet indicates that The young become sexually mature soon after weaning or at a couple of month s age which helps to give this species an impressive capacity for reproduction Macdonald 1984 Nowak 1991 Parker 1990 Yet this is not what we were seeing at Nottingham Origins and husbandry Figure 1 Siberian hamsters Photo Gregory Demas Indiana University A new pair will typically take 2 3 months before the first litter is produced Often they lose this litter usually due to cannibalism There was a desire for the breeding performance of the Siberians at Nottingham to be The Siberian hamster originates from Mongolia Siberia They are dark brown in colour with a thick dorsal stripe Coat changes to near white during winter months In the wild they dig burrows of up to 1m where they build a nest and raise young Hamsters at Nottingham housed in conventional small rat cages RB3 s NKP Given sawdust sizzle nest chew stick and cardboard tube Full clean out every other week breeding cages left longer between clean outs Room temperature 19 23 C as per Home Office guidelines Humidity around 55 Filtered water using long spouted bottles Can be difficult to handle as very active wriggly To handle scoop hamster between both hands Hands can be swivelled to expose underside to check sex Method 9 breeding pairs were observed These animals varied in starting age from 30 days weaning age to 120 days Some pairs were given standard enrichment 71

Poster Presentations which includes sizzle nest chew stick and cardboard tube while the others were also given a cardboard house Stock animals were also given cardboard houses All houses were given after the animals had been placed in clean cages Equal amounts of sizzle nest was placed inside the house and outside The following day the hamsters were observed to see if they had built a nest under the house or under the hopper This method was repeated each time the hamsters were cleaned out The breeding pairs were observed for how long it took them to have their first litter and number of pups born within the first four months Below are examples of the old style cage with sizzle chew stick and tube and the new cage with cardboard house Figure 4 New fully furnished cage Observations and results Graphs 1 2 and 3 show hamster preferences and breeding data Figure 2 Old style cage with sizzle chew stick and tube Graph 1 Nest building prefrence in stock Siberian hamsters Figure 3 New cage with cardboard house 72 Graph 2 Days to First litter House v No House

Poster Presentations Graph 3 Pups born House v No House Discussion It is clear from the data in Graph 1 that the hamsters enjoy the house as a form of enrichment By the second clean out 90 of the stock animals had chosen to build a nest under the house rather than under the food hopper which clearly indicates their preference The breeding hamsters all chose to build a nest under the house Graph 2 shows that those hamsters with a house took an average of 43 days to have their first litter and those without a house took an average of 64 days This would indicate that the presence of the house does encourage the animals to breed more quickly Graph 3 shows that an average number of 11 pups were born within the first 4 months from those pairs with no house and an average number of 5 pups were born to the pairs with a house This would indicate that the presence of the house does not result in more pups reaching weaning age and may indicate that it does not reduce the number of pups being cannibalised It can be concluded that the cardboard house is a beneficial source of enrichment for the Siberian hamster but more research needs to be undertaken to confirm whether breeding efficiency is improved by the presence of the house The house shall continue to be used regularly at Nottingham Observations will continue to be recorded in the future Figure 5 Siberian Hamsters 73

Animal Technology and Welfare April 2015 Instructions to Authors Subjects considered for publication may include original articles technical notes and reviews pertaining to all aspects of animal science and technology management and education The Editorial Board wishes to offer particular encouragement to papers leading to improvements in environmental enrichment the general care and welfare of the animals used in particular those species and strains exhibiting harmful genetic defects and papers describing refinements in techniques a reduction in the number of animals that need to be used or alternatives to animal use Papers describing experimental procedures will only be accepted for publication if authors clearly state that the procedures conform to the prevailing principles and Codes of Practice of the Animals Scientific Procedures Act 1986 Papers submitted from outside the U K should state what legislation and or ethical approval the work has been carried out under In addition authors who describe surgical techniques with recovery should include details of post operative care and any analgesic therapy provided All submissions should follow the ARRIVE Animal Research Reporting of In Vivo Experiments guidelines Kilkenny C Browne WJ Cuthill IC Emerson M Altman DG 2010 Improving Bioscience Research Reporting The ARRIVE Guidelines for Repor ting Animal Research PLOS Biol 8 6 e1000412 doi 10 1371 journal pbio 1000412 The Editorial Board reser ves the right to seek independent advice on any aspect of the content of an article but the final decision on acceptance or rejection remains with the Board Submission Material submitted for publication will be considered provided that it is contributed exclusively to Animal Technology and becomes the property of the Institute of Animal Technology The relevant ar ticle must clearly indicate where photographs and or graphs are to be inserted Address for submission atw iat org uk Hard copy The original manuscript plus two copies should be sent to the address below together with a copy on disk CD or DVD All sheets should be typewritten on one side in double spacing and serially numbered Any photographs or graphs should be supplied as originals and conform to the format in 4 below Address for submission Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL No responsibility will be accepted for loss or damage to such articles Electronic files of submissions are required together with separate files of photographs and any graphics that appear in the manuscript Electronic submissions should be sent via email via atw iat org uk alternatively manuscript plus two copies may be sent as hard copy to the address below All sheets should be typewritten on one side in double spacing with 4 cm margins and serially numbered Additionally a copy on disk should be provided or sent by email via atw iat org uk Articles for submission should be sent to Journal Editorial Board Chairman 5 South Parade Summertown Oxford OX2 7JL No responsibility will be accepted for loss or damage to such articles Format Articles may be submitted either electronically or by hard copy as follows 1 The first sheet of the article should contain the following Electronic i the full title of the paper ii the initials and last name of the author s iii the full address of the depar tment s and institution s where the work was carried out iv the address for correspondence if different to above Articles should be submitted in Word format with double spacing to the lines and all pages serially numbered Any photographs or graphs must be submitted as separate files and conform to the format in point 4 below 74 2 For the remainder of the paper the text should be

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

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

Attention ALL NACWOs We are pleased to announce the launch of two exciting initiatives for Named Animal Care and Welfare Officers NACWOs NACWO section on IAT website www iat org uk A dedicated NACWO section on the members area of the IAT website will be available soon Folders containing information and references relating to Enrichment Training and CPD Experimental procedures Surgical procedures Animal behavioural information and NACWO exchange documentation see below will be available Access to this area of the website can be requested through the IAT Website Co ordinator website iat org uk NACWO exchange programme In order to provide NACWOs with opportunities to develop strong networks learn about different facilities and programmes of work share best practice and gain valuable Continuing Professional Development the IAT is keen to promote a NACWO exchange programme Information on the initial pilot visits for the NACWO exchange programme were presented at the IAT Congress in March and the RSPCA Rodent Welfare Group meeting in October The programme gives interested NACWOs and Institutions the opportunity to host and arrange visits to other facilities Checklists are provided as a guide to the visits and a flow chart on the process has been prepared The exchange programme provides a great opportunity for people to meet and talk about their own experiences and to develop a network of NACWOs to support and learn from each other and further enhance the wellbeing of animals in our care CPD points will be allocated for both visiting and hosting NACWOs The Institute would also like to use information from these visits to feed into the Council Animal Welfare group If you are interested please contact Andy Cunningham ac572 le ac uk for further details

INDEX TO ADVERTISERS April 2015 3Rs Bedding xx AAALAC Fellowship Award 2015 xi Allentown Inc OBC Bell Isolation Systems xv Harlan Laboratories IFC Institute of Animal Technology xiv xvi xviii xix IPS Product Supplies Ltd xvii IBC LBS v PFI Systems iv ssniff Spezialdi ten GmbH iii SCANBUR xii Special Diets Services viii Surrey Diagnostics vi Sychem Ltd x Tecniplast UK xiii Vet Tech Solutions vii xx