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Institute of Animal Technology Bulletin G G Let s Talk Coping with Change Tunnel Handling Paper Review plus Qs and As G G NC3Rs Evaluating Enrichment Situations Vacant Vol 56 No 6 June 2020

Bulletin Vol 56 No 6 June 2020 EDITOR Carole Wilson bulletineditor iat org uk ASSISTANT EDITOR Paul Barwood bulletinassistant iat org uk ADVERTISEMENT MANAGERS PRC Associates Ltd mail prcassoc co uk Published monthly by the Institute of Animal Technology Registered Office 5 South Parade Summertown Oxford OX2 7JL CONTENTS Dear Readers Let s Talk Coping with Change The Practicalities of Tunnel Handling Taming Anxiety in Laboratory Mice a Paper Review A Question and Answer Session with the Paper s Authors 5 6 7 9 14 15 18 19 21 ISSN 0263 2861 For enquiries other than Bulletin related contact IAT ADMINISTRATOR admin iat org uk or 0800 085 4380 Final copy date for July Bulletin 23rd May The opinions expressed in the Bulletin do not necessarily reflect those of the Editor or the Institute AS ET 10th Anniversary 22 Name that Animal Crossword Solution 24 Dispatches 25 26 NC3Rs Evaluating 27 32 Environmental Enrichment Diary Dates 35

Bulletin June 2020 De ar Re aders Finally there seems to be a small glimmer of light at the end of the tunnel Depending on whereabouts you are in the UK you will have seen some lifting on the lockdown restrictions If you are not in the UK you have probably also seen some changes as I write this there are currently no new COVID 19 cases in New Zealand and some aspects of life are returning to normal In later editions of the Bulletin we are going to be looking at the impact of this epidemic and what and how you our readership have been coping in these very strange times Please feel free to get involved and send me your stories of managing during this time so I can share them with everyone else use my email address on page 3 Remember to keep social distancing Lynda and I show you how on the front cover However for this edition I thought we would look at the innovations that make life better for the animals in our care On pages 27 32 we are reprinting an NC3Rs blog on evaluation of environmental enrichment This is a very interesting article and well worth a read especially for those of you trying to determine the type of enrichment that your animals would most benefit from The second piece I would like to draw your attention to is actually in three parts Coming from the IAT Animal Welfare Group it includes a review of Jane Hurst s paper on Taming anxiety in laboratory mice and was the paper which really introduced the concept of tunnel handling read the review on pages 15 18 The review of the paper raised several questions with the reviewers which were put to the author You can read those questions and the author s responses on page 19 21 Finally John Waters who worked with Jane Hurst introduces himself and answers questions specifically about tunnel handling on pages 9 14 With that I would like to remind everyone to take care and Keep Reading Carole 5

Bulletin June 2020 9 The practicalities of tunnel handling from a technical perspective by John Waters My name is John Waters and I am currently the NACWO for the Mammalian Behaviour Evolution group MBE based at Leahurst University of Liverpool I have been an Animal Technician for 30 years now and it is a profession I am proud to be a part of We are some of the most dedicated professionals I have ever worked with Outside of the service not many other professions work 365 days of the year They should be very proud of what they do as their role is the foundation of many advances within the fields of science research and development Head of the MBE group is Professor Jane Hurst who received the 2010 NC3Rs prize for her contribution to animal welfare on publishing her paper Taming anxiety in laboratory animals highlighting alternative methods for handling mice However the use of alternative methods to handle mice in the laboratory will only be of benefit if carried out correctly Incorrect implementation could result in high levels of stress reducing the impact or even having the reverse effect

10 June 2020 Bulletin of increased stress and anxiety Following the publication of Taming anxiety in laboratory animals Hurst West 2010 there was uncertainty about how the alternative handling methods would impact upon the daily running of animal units if there was widespread adoption It became clear that a few issues needed to be addressed in order to facilitate a smooth transition to the new methods of handling and allow laboratory mice and the technical research community to benefit from the new findings The largest impact would be on Animal Technicians responsible for the day today care of animals and who would be responsible for the changeover of methods For this any negative impacts would have to be as small as possible The technicians needed guidance from somebody who had implemented the method recognised common mistakes made and could offer guidance on how to overcome any problems My own contribution has thus focussed on understanding the range of concerns and potential issues across a broad range of Animal Technicians and facilities and the provision of simple clear and practical advice To understand the possible problems I organised a workshop on mouse handling at the IAT Annual Congress 2012 In addition to implementing the new methods in my own local animal facility where I am NACWO I conducted two small studies to gain some objective data on potential issues of concern I then helped to put together a tutorial on mouse handling both by providing the technician s perspective on practical handling issues and by filming appropriate video clips with associated commentary to demonstrate good practice in different scenarios and how to avoid common problems that we had identified Some of the topics covered were G G G G making instructional video clips practicalities of tunnel handling practicalities of cup handing cage transfer Being involved with this work shows that Animal Technicians can play an important role within the research sector We are dedicated individuals who not only hold animal welfare in high regard but also have a valuable part to play being the foundations of scientific advances I am hoping my experience within the area of non tail handling will encourage others to get involved with research projects The most satisfying part for me as an Animal Technician has been to be able to help my fellow professionals implement these techniques without potentially having the same difficulties I had Hopefully this will aid in a smoother transition over to one of the most important advances in laboratory animal welfare

Bulletin June 2020 11 These were some of the questions that were addressed 1 Do the tunnel and cup handling methods work on all strains of mice The positive response to tunnel handling appears to be very general Tunnel handling works very well for all strains of laboratory mice and for wild house mice By varying the size of tunnel it also works very well for harvest mice wood mice voles rats and even edible dormice It seems unlikely that there will be a strain of laboratory mouse that it does not work with Establishment protocols may sometimes impinge on the practicality of tunnel handling Strains do differ in their habituation to sitting on the hand without restraint C57 mice are better being handled by a tunnel as they can be more unsettled on the hand and try to get off as soon as possible unless they have extensive experience of cup handling Other strains such as BALB c and outbred ICR CD 1 habituate much more quickly and show settled behaviour with both methods Mouse in tunnel Rama CC BY SA 2 0 FR https creativecommons org licenses by sa 2 0 fr deed en

12 June 2020 Bulletin Once mice are comfortable with tunnel handling they are more amiable to being picked up on the hand with less emphasis on the habituation period that requires a little bit of time investment Once mice lose the fear factor associated with tail handling they will interact with the handler regardless of method choice 2 If using a tunnel do I have to remove enrichment from the cages Once competent in capturing and transferring mice using a tunnel it makes no difference and can help if the enrichment is left in How much enrichment in the cage will depend if it impedes safe transfer using the tunnel It is advisable to put a tunnel in each cage which can also be used as enrichment as the mice habituate themselves to the tunnel as it is part of their environment The mice will readily enter the tunnel and climb over it in their active periods With clear tunnels the mice can still be observed through the tunnel thus not needing to remove the tunnel from the cage 3 Will the tunnel and cup method increase the amount of time needed for husbandry procedures Initially when learning the tunnel and cup method there will be an increase in time This extra time is no more than the time needed to learn any new procedure within Animal Technology As with any new technique there is a period for the user and the mice to become efficient Time studies have shown that the initial time investment when starting out is overcome once the technicians and mice are accustomed to the method For beginners it is favourable to start with tunnel handling as the availability of a tunnel within the home cage enables the mice to familiarise themselves with the tunnel thus easing the process of getting the mice into the tunnel There is an initial increase in the time needed for cupping but this can be easily carried out during a clean Willingness to interact with the handler starts to increase noticeably after about 5 cage cleans or handling events Once mice are familiarised with either of these non aversive methods they are much less likely to try and avoid being picked up making them easier to handle efficiently When changing methods it is advisable to begin with a small number of cages this has the benefit of allowing the mice and technician to become efficient without too much impact on the daily running of an animal unit

Bulletin June 2020 13 4 What size of tunnel should I use Tunnels can be supplied in different sizes according to what type of cage they are to be used in The sizes range from 10cm to 13cm with a diameter of 5cm mice are very wary of entering tunnels with a much smaller diameter than this Where possible use a tunnel that is present in the home cage as mice enter these most readily but where that isn t possible mice will still respond well to an external tunnel after one or two experiences of being picked up It is important to ensure if a home tunnel is used that the correct length is considered to prevent the home tunnel turning side on thus trapping the mouse inside Tunnels can be sourced from numerous suppliers and it is advisable to consult with the suppliers to see if the tunnels supplied meet your requirements 5 Are red tunnels better than clear There seems to be no preference to tunnel colour by the mice The clear tunnels have the benefit of being able to inspect the mice through the tunnel during a cage change or routine health checks The colour choice comes down to personal preference 6 Can any other type of tunnels be used Of course if preferred cardboard tunnels often used for enrichment can be utilised but it has the disadvantage of the mice being able to grip the inside of the cardboard tunnel thus making it more difficult to get the mice out for inspection or for use in a scientific procedure Shaking of the tunnel is to be avoided always as this will stress the mice and all your good intentions will be undone 7 Where can I find a supplier of clear tunnels that are suitable for autoclaving The tunnels can be sourced from numerous suppliers throughout the industry You must source the tunnels that fit your requirements i e autoclavable size and colour The size of the cages that the mice are housed in will dictate the size so it is advisable to ask for a sample before purchasing

14 June 2020 Bulletin 8 What is the best method of handling when working with young mice Young mice are more active and exploratory therefore tunnel handling of mice at weaning age can be a little more difficult Habituation of young mice can be helped by placing a handling tunnel in the breeding cage so the weaners are used to it Experience has shown that weanling mice naturally follow each other and maybe more than one mouse will be in the tunnel Ensure your hand is covering the exits from the tunnel and gently place the tunnel on the base of the cage and allow the weaners to exit the tunnel Some weaners can be difficult due to being more active In this instance a little more persuasion is needed Without too much force direct the weaners into the tunnel and once inside quickly cover one end with the palm of your hand and lift from base of cage Ever so slightly tip the tunnel towards your hand that is covering the exit so the mice will naturally be at the blocked end of the tunnel With your free hand open the new cage and gently tip the mouse backwards just above the cage base The sex of the mice can be determined using a tunnel Once the mouse is contained by placing one hand over the end gently tilt the tunnel so that the mouse naturally drops to the bottom of the tunnel Then the tail can be restrained and lifted to examine the genital area We find we are also able to move weaner mice from transport boxes to cages using a tunnel even when they are unlikely to have had previous experience of a tunnel at the suppliers The key is to make slight adjustments to the protocol for each scenario Weaner mice are not suited to sitting on the hand without restraint using the cup method due to their hyperactivity and lack of habituation to the hand Although they can be moved between cages between closed hands we would recommend use of a tunnel for about 3 weeks post weaning to habituate them to handling then they can usually be picked up by cupping on the hand without any problems

Bulletin June 2020 15 IAT Animal Welfare Group AWG Paper review Taming anxiety in laboratory mice Jane Hurst and Rebecca West Nature Methods Vol 7 No 10 October 2010 pp826 826 Some of the words information of this paper review has come directly from the paper with knowledge of the original author Published as a Brief Communication in Nature Methods The introduction states that routine laboratory animal handling has a profound effect on anxiety and stress responses It outlines that the researchers set out to assess the impact of handling methods The group found that picking up by the tail induced aversion and high anxiety This was compared to using a home cage tunnel or open hand which the group describe as methods that can minimise a widespread source of anxiety in laboratory mice Species Mouse Strains BALB C C57BL 6 and ICR CD 1 Methods The experiment had five separate groups of mice These were used to address different questions G G G strain differences in response groups 1 4 variation between handlers with different experience groups 1 2 handling in the light or dark phase of the light cycle groups 1 4 dark phase group 5 light phase Supplier of mice Harlan now Envigo groups 1 4 In house mice group 5 Caging 45 x 28 x 13 cm polycarbonate cages MB1 North Kent Plastics lined with corn cob absorb 10 14 bedding shredded paper nest material ad libitum food water and clear acrylic tunnels Identifications method Hair dye Clairol Nice n Easy BALB c ICR CD1 Natural black C57BL 6 Brilliant blond

16 June 2020 Bulletin Handling methods When testing started mice were handled only by their designated method During this the nest material and home cage tunnel were removed Tail pick up Grasped at the base of the tail between the thumb and forefinger and the mouse was gently lifted onto the opposite gloved hand or laboratory coat sleeve and held there by the tail for 30 seconds before release back into cage Tunnel handling The authors describe that the mouse was guided into the home cage tunnel soiled with familiar scent as it was brought toward it and the tunnel lifted above the cage and held for 30 seconds The handler s hands were sometimes loosely cupped over the tunnel s ends to prevent escape on the first day or two before mice became accustomed to tunnel handling Cup handling A cupped mouse was scooped up on one or both open hands and allowed to sit or walk over the hands for 30 seconds without other physical restraint To prevent mice leaping from the hands for the first experience of cupping the hands were closed loosely around the mouse for a maximum of 30 seconds until attempts to escape diminishes after which the hands were opened Combined cup and tunnel handling A fourth handling method combining tunnel and cup handling was tested using C57BL 6 mice to establish an alternative way to prevent mice initially leaping from the hands during cup handling The paper suggests that inexperienced handlers may at first find cup handling of untamed mice quite difficult for some strain and age combinations that are more likely to jump from the hand or handlers may be concerned about being bitten when untamed mice are between the hands No cupped mice attempted to bite in this study Common handling procedure For this study in each handling session the handler picked up each mouse in turn by the designated method and held it for 30 seconds Handlers wore laboratory coats contaminated with mouse scent from previous handling sessions and clean close fitting nitrile surgical gloves that were rubbed in bedding soiled by the same strain and sex of mice before the start of each handling session During the first nine handling sessions the occurrence of any urination or defecation was recorded each time a mouse was picked up and held Handling Test schedule The first batch of BALB c females was handled by a single handler There were eight cages per handling method for 16 separate handling sessions To assess any variability in response across handlers the

Bulletin June 2020 17 second batch of BALB c females was handled by eight different handlers that varied widely in their prior experience of handling rodents Each individual handler handled three cages of mice using one cage per handling method cages were assigned randomly in a balanced design Handlers experience Two handlers no prior experience Three handlers research staff with 3 4 years experience working with rodents Three handlers full time animal care staff and a researcher with substantial rodent handling experience Handlers were given a practice session using stock mice before the start of the experiment after demonstration of each method and test procedure Response toward the handler was assessed over nine daily handling sessions using all batches of mice with responses tested during handling sessions 1 5 and 9 Response to the handler Behaviour in anticipation of handling was assessed by voluntary approach and interaction with the handler before and after handling on specified test days After removal of the cage lid nest material and home cage tunnel the handler stood immobile for 60 seconds directly in front of the cage A gloved hand tail and cup methods or gloved hand holding the home cage tunnel tunnel method was then held resting on the substrate in the front half of the cage without moving for an additional 60 seconds to assess voluntary interaction Both mice were then handled by the designated method Trials were recorded on video or DVD for transcription by a single observer using an event timing program blinded with respect to test on the second batch of mice For each test the time spend in the front half of the cage by each mouse and time spent interacting at close contact with the introduced hand or hand and tunnel time spent sniffing the handling device paws on climbing on inside tunnel or chewing the glove were summed for analysis was measured Anxiety tests Mice were tested using an elevated plus maze after seven or nine handling sessions This was undertaken by a familiar handler and handling method Mice were delivered to the centre of the maze facing an open arm for five minutes Mice handled by the combined tunnel and cup method were delivered to the maze in their home cage tunnel The number of entries and total time when all four feet were in each arm of the maze the frequencies of protected stretched attend postures into the open arms from the central hub or closed arms and unprotected stretched attends on the open arm were scored

18 June 2020 Bulletin Tail manipulation for abdominal inspection To assess the response to lifting the mouse by its tail to examine its ventral surface and anogenital area The investigators captured BALB c females group 2 and outbred ICR CD 1 of both sexes group 3 using their familiar method They were placed on the handler s gloved hand the base of the tail was then grasped between the thumb and forefinger of the other hand and the back legs of the mouse lifted off the hand to expose the ventrum and held for 10 seconds Response to the handler was assessed immediately before and after handling Scruff handling To assess response to the experience of being restrained by the loose skin of the scruff ICR CD 1 BALB c and C57BL 6 mice in batches of 3 and 4 were captured using their familiar method after 11 previous handling sessions and placed on the bars of a clean cage top The mouse was then held by the scruff above the home cage for 10 seconds before being released back into the cage Response to the handler was assessed immediately before and after scruff handling and before and after handling by their normal method the following day to assess how quickly the mice recovered from scruff handling Response to cup handling The final test of BALB c batch 1 females assessed the response to familiar handler before and after all mice were handled using the cupping method This was designed to test how mice previously handled indirectly using a tunnel would respond to direct physical contact with the hand Response to the handler was assessed before and after cup handling by inserting a gloved hand only into the cage for all mice tested on day 45 after 15 prior handling sessions with the familiar handling method Data Analysis Data was analysed using parametric or nonparametric ANOVAs or Mann Whitney U tests Additional information The paper has several supplementary figures and tables that demonstrate clearly the data collected This corresponds with the finding as described in the text of the paper Non aversive handling is a current topic of discussion The work of the authors of this paper has been expanded and a number of further papers have been written on the subject Most of the information can be found on the NC3R s website https www nc3rs org uk how to pick up a mouse In addition to the papers the website also has a video tutorial video clips tips for implementation and much more If you would like to investigate further to see if non aversive handling would work in your establishment this would be the place to visit Review IAT Animal Welfare Group lead Matt Bilton

Bulletin June 2020 19 A question and answer session with the paper authors From an Animal Technologist perspective this paper raised several questions which the reviewers put to the authors Below are the answers the authors provided to the reviewer s questions 1 This paper does not describe very clearly if gloves are worn by all handlers If so did gloves get changed between boxes If they were not changed were the gloves disinfected between boxes and if so was this seen as a possible aversion Was there no prolonged voluntary interaction due to possible astringent scent on gloves As the review has correctly quoted from the original paper Handlers wore laboratory coats contaminated with mouse scent from previous handling sessions and clean close fitting nitrile surgical gloves that were rubbed in bedding soiled by the same strain and sex of mice before the start of each handling session Gloves were not changed or disinfected between cages as that is not necessary for biosecurity in our facility for cages in the same room However gloves were changed between cages housing mice of the opposite sex see handling and test schedule as attraction to opposite sex scent could have introduced a confound for some cages and we recommend that group house males are not given contact with female scent as this can stimulate aggression Because mouse scent would be on gloves after handling the first cage and was present on all home cage tunnels gloves were briefly rubbed in appropriate soiled bedding prior to handling the first cage to make sure that all were equivalent There was no astringent scent from disinfectants and importantly gloves and associated scents were identical for all handling methods We carried out an additional experiment to test the effect of mouse soiled protective clothing on voluntary interaction using the first batch of mice mentioned in Methods but there was insufficient space to include data Nature Methods have strict limits This found that mice spent more time interacting when the handler wore soiled gloves and lab coat sleeve compared to clean clothing across all three methods but the effect of handling method on interaction was considerably greater than the effect of soiled clothing

20 June 2020 Bulletin 2 If disinfection of the gloves took place was the same cage handled first every time for example cage 1 was handled first then cage 2 etc Were the trials all performed at the same time during the working day eg all test started at 1pm Gloves weren t disinfected As indicated in Methods order of testing for cages assigned to the same handling method which includes all handling was randomised and the order was balanced across the three different methods throughout In other words we always started with a cage selected at random using random number tables and one cage per method was handled tested followed by the next cage per method Testing always started at the beginning of a light or dark period but not within the first 30 minutes and generally took several hours to complete given the number of mice and duration of tests 3 Was any consideration given to the impact of reverse light cycle and hormone behaviour changes when study started and during the study Did the animals undergoing reverse light cycle go into this at 3 4 weeks of age so that the variable was minimal during the test phase The answer to both questions is yes As indicated in our Methods mice were maintained under either a reversed or a non reversed light schedule throughout This meant that mice bought in from suppliers were housed under a reversed light cycle from entering our facility at 3 4 weeks of age and remained under reversed lighting throughout the test phase batches 14 Mice bred in house were bred under reversed lighting but housed under non reversed light conditions from weaning at 3 4 weeks of age and throughout the test phase 4 When animals were divided up into groups they are referred to as batch 1 4 from commercial supplier and batch 5 from in house it s not very clear if each batch was then subdivided into all test categories for example that batch 5 was both light and dark phase tested Supplementary Table 5 in the paper provides a very useful summary of the handling and test schedule for each batch of mice including the lighting schedule for each All batches compared the three handling methods with a similar handling and test schedule at the start for each but each batch differed in the additional questions they addressed Batches 1 and 2 allowed us to show that neither individual differences between handlers nor their

Bulletin June 2020 21 prior handling experience had a major effect on the responses of BALB c female mice Instead mice showed a major difference in response to the three handling methods regardless of who was handling them Batches 3 and 4 allowed us examine responses to the handling methods using three different strains and both sexes All four of these batches were handled and tested in the dark phase of the light cycle to ensure that mice were active during testing so that any lack of interaction or low activity was not due simply to inactive mice Batch 5 used both sexes of two strains C57BL 6 and BALB c under non reversed lighting conditions that were handled and tested in the light phase of the cycle For this batch I also came in overnight to test voluntary interaction during the dark phase among mice that were normally handled in the light phase This confirmed that the slightly lower interaction among some tunnel and cupped mice immediately prior to handling but not after handling in the light phase was most likely because they were not fully aroused before handling It should be noted that there is no attempt to compare responses directly between batches in this study which may have differed in a number of ways We compared responses to the different handling methods within batches where everything was kept equivalent and appropriately randomised 5 Were handlers randomised or were the same handlers involved daily with their set group e g no change in handler Were handlers encouraged to shower so no perfumes deodorants could interfere with the study Where more than one handler was involved batches 2 and 3 cages were assigned randomly to each handler in a balanced design so that each handler had one cage of females per handling method batch 2 or two cages of each sex per handling method batch 3 There was no change in handler in any batch with the same handler responsible for handling testing and cage cleaning No restrictions were applied to the use of perfumes or deodorants and handlers were not encouraged to shower more than they usually would which is why it is not mentioned This is because we were interested in whether normal variation between handlers would influence mouse responses However we found no evidence of major differences in response towards different handlers but substantial differences between handling methods 6 Was the study carried out in the holding room or in a procedure room The study was all carried out in the holding room

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24 June 2020 Name that Animal Solution from May Bulletin

Bulletin June 2020 Dispatches Dr Birte Nielsen joins the team at UFAW The Universities Federation for Animal Welfare UFAW is pleased to announce the appointment of Dr Birte L Nielsen as Assistant Scientific Director of both UFAW and its sister charity the Humane Slaughter Association HSA Birte will join the charities in September Birte brings a wealth of experience and in depth knowledge to the role having spent nearly 30 years carrying out research in basic and applied ethology the latter in relation to animal welfare mainly in farm species She has worked with and has published papers on rats growing pigs dairy cows broiler chickens and broiler breeders as well as horses sows laying hens and even ostriches 25

26 June 2020 Bulletin She spent the first 8 years of her career in Edinburgh Scotland completing a PhD on pig feeding behaviour and working as a post doc on metabolic stress in dairy cows She then moved to Aarhus University Denmark where she was in charge of the research unit for Behaviour and Stress Biology This gave her a broad footing in basic and applied ethology and allowed her to partake in various expert panels and working groups such as chairing the Danish Animal Welfare Council 2005 2010 and as a member 2012 2018 of the Animal Welfare working group at Anses which is the French Agency for Food Environmental and Occupational Health and Safety She is a former President of the International Society for Applied Ethology ISAE Since 2010 she has been based in Paris France working on the links between olfaction behaviour and neuroscience More recently she has been concentrating on the study and modelling of feeding behaviour this time with special emphasis on ruminants and in the context of Precision Livestock Farming UFAW s Chief Executive and Scientific Director Dr Huw Golledge said We are delighted that Birte will be joining the charities As we grow our influence and focus on rigorous evidence based approaches to animal welfare Birte will bring significant additional expertise to our science teams

Bulletin June 2020 27 Approaches to evaluating environmental enrichment News page summary How technicians across the UK introduce new environmental enrichment into their facilities and what barriers they face Earlier this year NC3Rs staff met with Animal Technicians from different universities around the UK to discuss the ways they introduce new environmental enrichment into their facilities In this blog post we summarise the discussions what the common practice seems to be barriers staff face when it comes to evaluating the impact of enrichment on animal welfare and potential solutions to facilitate more formal studies in this area Introduction Providing an appropriate environment for laboratory animals is essential for their wellbeing A suitable home environment provides sensory and motor stimulation and allows the animals to perform a range of species typical behaviours such as locomotion nest building foraging and exploring Giving animals the opportunity to display a wide behavioural repertoire can also improve monitoring of their welfare as it gives technicians and researchers the ability to recognise deviations from normal behaviour that could indicate distress or suffering e g poor nest building in mice Bedding and nesting material refuges and gnawing items are the most commonly used enrichment resources for mice and rats

28 June 2020 Bulletin As champions for animal welfare technicians play a key role in generating ideas for new enrichment items and in assessing their impact on animals as illustrated by submissions to the Janet Wood Innovation Award and Andrew Blake Tribute Award Provision of environmental enrichment should not be a random process whereby different objects are included in cages based merely upon their availability because laboratory animal species show preferences for different resources e g nesting and foraging materials and these can differ depending on sex strain age reproductive state and group size It is therefore important to evaluate whether the animals are using and benefiting from the enrichment provided and what their preferences are Ideally good scientific practice should be used when evaluating different types of enrichment for their effect on animal welfare Well designed and correctly analysed experiments allow reliable findings to be made that can be used to make a robust case for providing the enrichment and shared with other animal facilities to encourage wide adoption Implementing appropriate experimental design with the help of other staff such as researchers and statisticians allows reproducible results that can lead to a scientific publication or a poster presentation at a conference A well designed experiment should not be influenced by bias such as the expectations of the investigator or the position of the cage on the rack It should also use sufficient animals and be applicable to different sexes and strains How is new enrichment currently evaluated Based on our discussions the try and fail approach appears to be the most common way of trying out new enrichment items mainly from samples provided by commercial suppliers Initially new items are introduced to a couple of cages most often to sentinel or surplus animals and their success as enrichment is evaluated based on visual checks during routine husbandry procedures In these informal evaluations technicians Apparatus for investigating the preference of assess whether the mice for different coloured cages C Sherwin enrichment has been used

Bulletin June 2020 29 by the animals for example by looking for signs of chewing or faeces on the enrichment They check how many cages show signs of enrichment use and whether the enrichment item raises any practical issues e g being too big to comfortably fit within the cage or too difficult to clean and autoclave Whilst this approach is useful in assessing whether a new enrichment might be beneficial and needs to be investigated further it doesn t enable a good understanding of how the enrichment is being used and by which animals whether their welfare is improved as a consequence and to what degree It can also miss subtle effects positive or negative Some facilities test new enrichment items with a formal and systematic process incorporating good experimental design for more robust and unbiased observations Time is set aside to make behavioural observations e g on a pre defined behaviour checklist or physiological measurements and to record these data for analysis For example technicians at University College London UCL are supported by the Welfare Trials Group to conduct robust enrichment trials through independent review of study design and a mentoring scheme which has led to successful trials for chew stick preference in mice and the safety of nesting materials This approach provides a better evidence base for convincing researchers and senior management to invest in the new enrichment across the animal unit In many cases these studies have had wider impact by providing valuable information and guidance to other Enrichment for zebrafish includes live food e g rotifers plastic shelters and gravel substrate

30 June 2020 Bulletin facilities for example via publication in the IAT Journal Animal Technology and Welfare presentation at the NC3Rs IAT Animal Technicians Symposium or on a public website such as this UCL site on zebrafish feed trials What are the barriers to formal evaluation studies The technicians we talked to were all enthusiastic about getting involved with projects on environmental enrichment but highlighted a number of reasons why some animal facilities and staff are not able to assess the suitability of new enrichment items and their effect on animal welfare in a more formal and scientific manner The most common issue is lack of time as most technicians have to fit any enrichment evaluation studies around other daily tasks that they perform as part of their job Some may have to use their personal time not only to collect but also to analyse the data This is the main reason why many facilities would rather assess enrichment informally as part of the everyday husbandry routine instead of committing time and resources to more formal assessment Solutions to this problem include asking managers for dedicated project time collecting data at quieter times in the week and sharing of workloads between technicians The cost to purchase new enrichment to run a trial study can also a prohibiting factor for properly evaluating new items Often suppliers will provide some free samples for technicians to try in a couple of cages However if a wider evaluation study is to take place more enrichment items will have to be purchased and facilities may be reluctant to spend money for something that might not end up being used if it does not show any welfare benefits Apart from the limitations posed by facility resources another limiting factor is the availability of animals for prolonged periods of time to test new items Most commonly sentinel animals or surplus animals are used for enrichment evaluation studies However the number of sentinel animals is often small and the strain used for sentinels might not always overlap with the strains that are routinely housed in the facilities As such observations from sentinel animal may not always be relevant to those mice the new enrichment is intended for Occasionally non experimental animals may be available However as these belong to the researchers the length of time for which they are available can be unpredictable or subject to change if the experimental needs change which can force technicians to stop their evaluation activities earlier than planned Additionally as they have to perform their studies on

Bulletin June 2020 31 whichever animals are available at the time it is often difficult if not impossible to control in the experimental design for different animal characteristics e g sex age and strain One of the ways this has been successfully addressed is via technicians collaborating with researchers who have a professional interest in environmental enrichment and the potential benefits for their science e g behavioural neuroscientists to design more comprehensive studies Researchers can sometimes be reluctant for technicians to introduce new environmental enrichment to their animals as they want to ensure that all conditions remain the same throughout their experiments This is also a reason why some animal facilities tend to be conservative with the enrichment they provide and only supply some standard items such as nesting material a cardboard tunnel and chew block In such cases it may be possible to perform pilot studies to assess whether the researcher s concerns are well founded and to introduce beneficial enrichment in gaps between studies once the impact on experimental outcomes is known Apart from the practical difficulties mentioned above lack of knowledge and expertise on how to design a study analyse the data and obtain robust results might also prevent some technicians from conducting an organised and systemic evaluation of new enrichment Advice and guidance can be sought from statisticians or researchers as was the case for a technician led study evaluating different types of nesting material for singlyhoused head plated mice Zoe Windsor a Research Support Technician at the UCL Institute of Neurology conceived and designed the experiments and collaborated with Simon Bate a statistician at GSK who analysed and interpreted the data For those universities that have them NC3Rs Regional Programme Managers are another good source of support and advice Lack of recognition that better welfare means better science can still be a barrier to formal evaluation studies Ideally technicians would be given the time resource and advice they need to evaluate enrichment because of the potential benefits for science animal welfare and the Culture of Care It may be helpful to ask named persons the AWERB or internal 3Rs groups for support to enable robust enrichment evaluation Let us know what you think Some technicians indicated that ready to use study protocols including score sheets similar to those we provided for our mouse aggression study with clear advice on data analysis would be useful to those considering evaluating the impact of enrichment on welfare To address this the NC3Rs RSPCA and IAT are considering developing an online resource to provide technicians with

32 June 2020 Bulletin practical useful tools and knowledge so they can design and conduct formal and organised evaluation studies of novel environmental enrichment items This online resource could also serve as a hub for technicians to share their experiences with different types of enrichment that they have tried and tested in their facilities If you have been involved in enrichment evaluation studies or if you have any suggestions about what you would like to see in an online resource on this topic please get in touch with the NC3Rs Animal Welfare team to share your experiences and ideas Reprinted from the NC3Rs Blog with permission BSF Research Technician Medical Research Council Brain Network Dynamics Unit at the University of Oxford Salary 22 417 25 941 p a with discretionary range up to 28 331 p a pro rata for part time We seek a Research Technician to provide specialist technical support for research at the MRC Brain Network Dynamics Unit https www mrcbndu ox ac uk that uses animals The post holder will be required to play a central role in the daily organisation and smooth running of the Biomedical Services Facility BSF of the MRC Unit Candidates should have a minimum IAT Level 2 Diploma or equivalent training experience have a good track record in animal technician work with small rodents and be fully conversant with all aspects of A SP A This is a fixed term 36 month post in the first instance to start from 1 August 2020 or as soon as possible thereafter The post is full time although part time will be considered For more details see https www mrcbndu ox ac uk vacancies Informal enquiries can be addressed to peter magill ndcn ox ac uk Closing date for applications is 12 00 noon on Wednesday 8 July 2020

hello agendalifesciences co uk www agendalifesciences com Project Licence Holder Leicestershire Salary Competitive Our client a world leading institute working towards challenges in oncology LQ DPPDWLRQ FDUGLRYDVFXODU DQG PHWDEROLF GLVHDVH GUXJ GLVFRYHU DUH ORRNLQJ for a Project Licence Holder Senior Research Scientist to manage two home RI FH SURMHFW OLFHQFHV 7KH VXFFHVVIXO FDQGLGDWH ZLOO DOVR EH UHTXLUHG WR PDQDJH in vivo studies and support the Study Director team for oncology studies Junior Research Technician Leicestershire Salary Competitive Our client a leading CRO specialising in pre clinical cancer research are looking to recruit a Junior Research Technician to join their in vivo laboratory team The successful candidate will be required to carry out daily animal husbandry duties DQG UHJXODWHG SURFHGXUHV LQ DFFRUGDQFH ZLWK RPH 2I FH UHJXODWLRQV 7KH company provides appropriate training and offers a career pathway so you can continue to progress and learn new skills in the laboratory animal industry Senior Animal Technician County Mayo Ireland Salary DOE On behalf of our client we are looking to recruit a Senior Animal Technician to assist with setting up a team in a newly established Germ Free egg facility to ensure that it is ready to go to commercialisation in 2021 The successful candidate will be a key member of the business responsible for the day to day RSHUDWLRQ RI WKH RFNV EULQJLQJ WKHP XS WR FRPPHUFLDO OHYHO DQG OHDGLQJ D WHDP of technicians Senior Animal Technician Nottingham 23 754 28 331 On behalf of our client we are looking to recruit a Senior Animal Technician to oversee the day to day organisation of experimental animal holding areas within a biological services facility The successful candidate will be required to supervise mentor and train technical staff and utilise their expertise in animal technology to conduct experimental procedures Licensed Animal Technician Cheshire 20 404 28 000 On behalf of our client we are looking to recruit a Licensed Animal Technician to provide technical support for a range of in vivo oncology models within their drug discovery unit and other research teams as required Register with us today and we ll start the search for your ideal role

Bulletin June 2020 35 DIARY Dates Events Dates subject to change due to the current quarantine situation and possible ongoing restrictions 20 October AS ET Congress Bursary Closing date 20 October See page 19 Details contact as et org uk 1 3 December 2020 AFSTAL Congress Marseille Details https www colloqueafstal com 202 Front cover Carole and Lynda WEBINARS 23 June Biosecurity workshop 24 June It s about you and your animals 25 June Automation for all Details info tecniplastuk com Photo Paul Barwood