Shaping the Future of Amphibians in Research

Issue 2, February 2019

Providing animal technicians with the latest news from the NC3Rs Tech3Rs European Xenopus Research Centre Welcome to the second edition of Tech3Rs. In each issue, we share updates on recent advances in the 3Rs and highlight new resources, research and events.

This newsletter will help animal technicians working in research establishments to identify opportunities to embed the 3Rs in practice and ensure high standards of animal welfare. If you have Shaping the future of any ideas for future issues or are working on a 3Rs approach you would like us to feature, please get amphibians in research in touch – we would love to hear from you! You can email us Do you work with amphibians? Share your experiences at [email protected] to help improve knowledge on optimal welfare standards. In this issue we take a look at the future use of amphibians A recent report, published in Lab Animal, A few of the unresolved questions in research, focus on recent identifies key issues and important but are summarised below. The full set of publications in zebrafish welfare unresolved questions on amphibian welfare. questions can be found in the report. and discuss our plans for the Knowledge on amphibian husbandry Can Xenopus superovulation / general 2019 IAT Congress. and welfare remains limited, with little breeding protocols be refined? established guidance or evidence-based What is the ideal nutritional composition SUBSCRIBE – Don’t want refinements for their captive care. of a regulated amphibian diet? to miss the next issue? The report provides a summary of a Is live food a necessity in an Subscribe online at workshop jointly hosted by the NC3Rs amphibian diet? www.nc3rs.org.uk/tech3rs and ZSL (Zoological Society of London) How does water quality and to receive free hard copies of to discuss the welfare of amphibians in composition affect amphibian welfare? the newsletter (UK institutes only). research. The event brought together If you work with amphibians, we invite experts from across academic and zoo you to review and consider these research communities to identify the unanswered welfare questions and welfare challenges facing amphibians in share any insights you may have to research and to discuss the steps that need finding the answers. Please send any to be taken to improve conditions for this correspondence to Dr Samuel Brod diverse group of animals. ([email protected]).

Read the paper: Brod S et al. (2018). Discussing the future of amphibians in research. Lab Animal 48(1):16-18 doi:10.1038/s41684-018-0193-6 3Rs champions We would like to hear from you and help you share your ideas with 3Rs papers of interest others. In every issue of Tech3Rs we will be highlighting animal technicians who are championing the 3Rs at their establishments. Each issue we feature recent 3Rs publications of interest, James Ward is a Departmental Colony solutions on that initial study. We have Manager at the University of Oxford. around 160 technicians currently working providing summaries and links Here he describes the approach he with mice, so the next step was to pick a to the full articles for further takes to reduce the number of mice long-term study. Our training officer helped information. This issue we used for breeding. me to develop a training plan consisting of a presentation and a theory test, followed focus on the welfare of fish in What 3Rs ideas are you developing? by a practical assessment. We wanted to the laboratory, in particular the Our idea has been to be more active in make sure that everyone understood why commonly used zebrafish. keeping breeding colonies under tight we were introducing the cupping technique control through several related services and its intended benefits. The majority Is heightened-shoaling a good Effects of environmental enrichment Welfare challenges influence and it is my job to help make that happen. of our technicians are now trained and candidate for positive emotional on survivorship, growth, sex ratio and the complexity of movement: When we audited our mouse numbers for our standard operating procedures have behavior in zebrafish? behaviour in laboratory maintained fractal analysis of behaviour in 2015, we found that 57.5% of mice culled been updated to reflect the new handling (Franks et al., 2018) zebrafishDanio rerio (Lee et al., 2018) zebrafish (Deakinet al., 2019) – had been surplus to breeding. While some techniques. We are still gaining experience NC3Rs funded research surplus is unfortunately inevitable, we Kiya Robinson is an Associate with different strains and ages of mice. Research on positive emotions in Environmental enrichment can help wanted to reduce this number. Licensee at Covance, Harrogate. However, whenever possible, new mouse laboratory animals is becoming increase the complexity of a fish’s Despite being the second most In this interview, Kiya shares her studies will now be cup handled. increasingly recognised as fundamental environment and improve their welfare. commonly used species in research, experiences of implementing What are your future plans? to research animal behavior and welfare. Despite this, the majority of zebrafish welfare assessment in fish falls refined handling techniques. Good welfare is not simply the absence used in research are usually kept in far behind rodents. Many routine A few more technicians still need to of negative experience (e.g. pain and barren tanks due to a lack of clear procedures, such as fin clipping, are This helps to reduce What 3Rs idea are you developing? complete their initial training. Then suffering) but also the presence of evidence in this area. carried out in zebrafish with no clear our focus will shift to improving their positive emotions (e.g. pleasure). indication of the pain the fish are In recent years there has been a wealth breeding numbers” handling techniques with experience. Despite this, relatively little is known The aim of this study was subjected to. of scientific information to support the We are also evaluating how to help about positive emotions and affiliated to investigate the effects of use of refined handling techniques in I help researchers set up enough breeders technicians stay engaged with this behaviors in fish. environmental enrichment (in the form The aim of this study was to determine laboratory mice. The use of tunnel or cup for their needs through our breeding method whilst it is still being rolled out. of aquarium gravel and aquatic plants) if fractal analysis, a method to study handling methods versus tail handling is a calculation service and conduct a review Monitoring the impact on welfare and The aim of this study was to characterise on the health and welfare of zebrafish. complex patterns, could be used to large focus, and so I was intrigued to see of each colony every three weeks to the unit will be very important. and evaluate heightened-shoaling in analyse zebrafish movement behaviour how these methods would work in our This study showed that tank ensure that this remains at the right zebrafish and its potential as an indicator following painful procedures. institution. I undertook a small trial and environmental enrichment had level. I also run the training on colony of positive emotional behavior. the difference in behaviour with the mice positive effects on larvae survivorship, Fractal analysis showed less complex management as well as genetics, covering surprised me. Heightened-shoaling is defined growth and behaviour in zebrafish. movement patterns in fish following topics like backcrossing and when the as discrete periods of tight group The study also found monopolisation painful procedures (e.g. fin clipping). breeding stock needs refreshing. This How did you develop your idea? I undertook a small trial cohesion and increased behavioral of resources by dominant fish and The administration of the local helps to reduce breeding numbers and synchrony in groups of fish. The associated aggression in enriched anaesthetic lidocaine reversed the Covance is deeply committed to animal surplus animals without affecting the and the difference in authors characterise and describe this tanks, which may have negative changes in movement patterns welfare and continuous improvement. science being conducted. behavior and provide supplementary effects on welfare. associated with fin clipping, providing As part of this commitment, I looked behaviour with the videos to help demonstrate. further evidence that fin clipping is a In our annual audit for 2017, we found that through the information on the NC3Rs mice surprised me.” The authors showed that fish from painful procedure requiring pain-relief. we had got the surplus down to 48.1%. website and then attended the mouse The study extends research on the enriched tanks displayed lower levels While this is still high, and we want to get handling workshop in 2017. I was sceptical So far, our measurement has mainly social behavior of zebrafish and of anxiety behaviour compared with The authors also demonstrated a it even lower, this already represents a at first but hearing the experience of taken the form of technician feedback. heightened-shoaling appears to be a fish from barren tanks. The challenge decrease in movement complexity with reduction of thousands of animals, so it other technicians at the workshop was Once technicians are used to the good candidate for future research into is to design enrichment that provides increasing strengths of acetic acid (a has been a real success. convincing. There is strong scientific data method they confirm that the mice are positive emotional behavior in zebrafish. measurable welfare benefits and can standard pain test), which allowed the to support these techniques. I initially What are your future plans? easier to handle, appear less anxious, also be practically adopted without development of a scale of pain intensity handled some health screen mice for This article featured in a special issue and that it does not require a large compromising husbandry procedures which could be adopted as a new tool two weeks. We then identified a four- on Animal Emotion. Further articles We are hoping to start offering training in increase on their time. Once we have a or the research question of interest. for assessment and monitoring of week repeat dose study, obtained our about current progress in understanding colony management to researchers as standard and measured approach, an zebrafish welfare. client’s approval and selected a team of animal emotions to help improve animal Lee CJ et al. (2018) Effects of environmental well as the animal care staff so that those important future goal of the project is to 10 technicians that would work in the are available at www.mdpi.com/journal/ enrichment on survivorship, growth, sex ratio and Deakin AG et al. (2019) Welfare challenges academics who want to have a more active share the refined handling techniques role can do so in a productive way. I have room and offer feedback. This study animals/special_issues/Anima_Emotion behaviour in laboratory maintained zebrafish influence the complexity of movement: fractal with our colleagues around the world. Danio rerio. Journal of Fish Biology 94:86-95. analysis of behaviour in zebrafish.Fishes 4(1):8. also started advertising these services so demonstrated its value when future Franks B et al. (2018) Is heightened-shoaling a doi: 10.1111/jfb.13865 doi: 10.3390/fishes4010008 that any groups that are not aware of them technicians would talk to me about good candidate for positive emotional behavior have the chance to use them, so there their concerns, because we had already Please email [email protected] in zebrafish?Animals 8(9). doi: 10.3390/ani8090152 might be more work in the future! considered these concerns and found to be featured in our next issue. Animal Technician Hub The NC3Rs has a broad range of projects and resources directly relevant to the work of animal technicians in laboratories around the world. Whether it’s the latest refinements in mouse handling, or practical advice on blood sampling, these resources can help support animal technicians to apply best practice in their work. This poster highlights the relevant web resources available on the hub. Further information can be found here: www.nc3rs.org.uk/animaltech

Tech3Rs: Laboratory mouse How to pick up Housing and Blood sampling A newsletter for aggression study a mouse husbandry animal technicians

Our regular newsletter Crowdsourcing data Non-aversive methods Refining the housing Refining blood sampling providing animal project for technicians for handling mice. and husbandry of from laboratory animals technicians with the to identify patterns and common laboratory – advice on technique, latest news. triggers of aggression animal species. route and volumes. in laboratory mice.

Procedures Rodent welfare hub Welfare of Grimace scales Events calendar with care non-human primates

A dedicated website Resources and outputs Opportunities to refine Posters and other The latest workshops, of online tutorials and from our programmes the use of non-human resources on the use conferences and guidance to support related to laboratory primates. of grimace scales events from the NC3Rs the adoption of best rodent welfare. to assess pain in and other related practice for commonly laboratory animals. organisations, many of used procedures in which will be relevant animal research. for animal technicians.

IAT Congress 2016 NC3Rs/IAT E-learning Contemporary 3 Minute 3Rs 2017 workshop Animal Technicians’ resources training on the 3Rs podcast summary: Playtime Symposium for Rats presentations

Summary report on our Video presentations Free online tutorials A video presentation A monthly podcast workshops to explore on refinement designed to support demonstrating the produced with approaches to providing opportunities and the training of 3Rs in principle and LabAnimal and the laboratory rats with a approaches to technicians and practice. NA3RsC summarising complex environment improving the welfare others working with the latest research and and increased of a range of laboratory laboratory animals. news in 3Rs science opportunity to exercise. animal species. and technology.

We recommend that you pull out this poster and display it in your animal unit to highlight the resources available to all. Highlights from our Solutions news and blog New Solution – Assessing The NC3Rs blog is a is a wireless tablet training system aquatic organism health attached to the home enclosure that platform to talk about status non-invasively overcomes many of the disadvantages the research we champion linked to the traditional approaches. Researchers at the University of Naples and the issues we care Federico II have developed a non-invasive For further information about method to detect and identify potential about. Recently we automated training systems and pathogens (viruses, bacteria, protozoa, highlighted a new approach a video demonstration, visit: metazoa, parasites) in aquatic facilities. to training macaques. www.nc3rs.org.uk/mymou The technology uses environmental DNA (eDNA) extracted from water samples, Welfare-friendly, coupled with DNA sequencing technology (metabarcoding). The improved identification tablet-based method of pathogens in local and imported animals, for training macaques prevents potential pathogens spreading to The monkeys we‘ve healthy animals, and reduces the number Training non-human primates (NHPs) trained using the of sentinel fish used. to perform cognitive tasks in the laboratory is a time-consuming system have learnt The Solution provider is keen to work process with significant welfare and with animal care staff interested in experimental limitations. To refine the to use it quickly and providing water samples to help assess practice, Drs James Butler and Steve we are excited at the and validate this eDNA metabarcoding Kennerley from University College technology against current practice. London developed Mymou, a novel prospect of other labs system for automated training of NHPs using the system” Further information can be found here: within their home cage rather than in www.crackit.org.uk/aquaticassessment the laboratory. The Mymou system Drs James Butler and Steve Kennerley

Listen to the 3 Minute Webinar – NC3Rs mouse Spotlight on 3Rs Podcast aggression study guinea pig

We’ve teamed up with LabAnimal and the North American Did you participate in the NC3Rs Mouse husbandry 3Rs Collaborative to bring you 3 Minute 3Rs, a monthly Aggression Study? podcast that gives you the lowdown on the latest 3Rs We have updated our housing and research in just a few short minutes. If you did and would like to find out more about the results, husbandry page for guinea pigs. a recording of the webinar hosted by the NC3Rs on 11 You can find3 Minutes 3Rs anywhere you get podcasts December 2018 is available upon request. Please email Check out the new page for by searching for LabAnimal or visit www.nc3rs.org.uk/ [email protected] information on social behaviour, podcast to listen to all of our past episodes. for further information. considerations for enclosures and environmental enrichment. If you have any ideas or photos of guinea pig enrichment you would like to share with us. Please email enquiries@ nc3rs.org.uk

Further information: www.nc3rs.org.uk/guinea-pigs Upcoming events

NC3Rs workshop, IAT Congress Tuesday 9 - Wednesday 10 April, North West England

NC3Rs staff will be leading a workshop on ‘Fundamental concepts in experimental design: randomisation and blinding’ at IAT Congress.

Recent studies have highlighted that the results from animal experiments are often not as reliable as they should be. The role of an animal technician in improving experimental design is often underestimated. During this two-hour workshop, we will cover some approaches that are proven to improve experimental quality and which animal technicians are likely to be involved in – randomisation and blinding. We will explain what they are, why they are important and when they are needed. The workshop will also provide practical advice and guidance on how to blind and randomise.

This workshop is for technicians involved in supporting and running experiments (e.g. setting up experiments, performing procedures, preparing animals for studies). Whilst we will focus on common species such as rodents and fish, the principles will apply across all species. This interactive workshop will include talks, voting, group work and hands-on activities.

Other events at IAT Congress:

We will have a stand in the congress exhibition hall; Dr Katie Lidster will be presenting an update on the please come and say hello. NC3Rs mouse aggression study on Thursday 11 April.

Animal Tech Month NC3Rs/IAT Animal Technicians’ March 2019 Symposium 2019

As part of the IAT’s Animal Technologist Month in March to Wednesday, 2 October 2019, central London celebrate the hard work of technicians, the NC3Rs will be Save the date for this year’s NC3Rs/IAT Animal Technicians; tweeting about how technicians contribute to the success of Symposium. A free, informative meeting to provide animal 3Rs science. Follow us @NC3Rs and look out for the hashtag technicians with an opportunity to find out more about the #animaltechmonth – why not join in the conversation and latest advances in the 3Rs. More details to follow. share your own experiences?

NC3Rs T +44 (0)20 7611 2233 www.nc3rs.org.uk Gibbs Building F +44 (0)20 7611 2260 www.crackit.org.uk 215 Euston Road [email protected] @NC3Rs London NW1 2BE