Painting mice

Why are we recording a video of my colleague Rita Santos painting the back of a white IKEA mouse with a black marker pen?

The background story is that nearly 50 years ago, in 1974, a US researcher painted real laboratory mice as part of what has become a well known story of scientific misconduct in biomedical research. The story isn’t as well-known among biologists and animal scientists as it is among immunologists (or at least so I assume, based on anecdotal evidence = whom among my contacts and colleagues who were or were not aware of it when I asked), despite being both striking and somewhat sad.

Striking, because once you’ve heard the story, you are likely to remember it. Dr William T Summerlin was doing research into transplantation immunology. He believed that by keeping the tissue in laboratory culture for some time before transplanting it into the recipient animal, it could be transplanted without rejection. His proof-of-concept experiment was to graft skin from black mice to (genetically unrelated) white mice. Did he actually ever transplant skin? I don’t know – this amount of detail is not given in the easily accessible internet sources. But what is very clear is that his demonstration of success was a fraud. An attentive technician discovered that the black patch on the back of the white mouse could be rubbed off with ethanol. As reported in this NY Times account of the case, published only a month later, Summerlin admitted to having painted the mice.

Sad, for a number of reasons, going beyond the actual misconduct itself, which is of course in itself highly lamentable. The painted mice seems not to be a one-off event – when Summerlin was investigated the committee also discovered a seemingly very dubious case of cornea transplant experiments in rabbits. Whereas having their backs painted would hardly have harmed the mice, the failed cornea transplants must have caused the rabbits pain. And none of this was justified. Scientific experimentation is not about simply trying something to see if it works: there has to be a reasonably developed idea of what mechanisms are involved. I don’t find any reference to a theory about mechanisms involved in the purported transformation of a xenograft (from a genetically different individual) into tissue that is not recognised as foreign when transplanted into a recipient. All that is to be found is that Summerlin had claimed for some years that he had a method for laboratory culture of tissues that removed the problem with transplant rejection, and that other researchers were unable to make the procedure work when they tried to repeat it in their own labs (a classic way through which fraudulent or poorly conducted research is discovered). The requirement that an experiment is based on a reasonably developed theoretical framework and previous, related studies is even stronger when the health and well-being of living beings are involved.

Why, then, are we painting mice? As part of the INTEGRITY project, we are developing teaching material into ethics and research integrity issues in animal experimentation, for high school students, ready for road testing in about a month. And please note, we’re not painting mice, we’re painting toy mice. The first R, Replacement, of the 3Rs principle of course. Knowing what mice are like, I actually believe that using real mice for this purpose would not only have been stressful for them but a pain for us!

Reading to update my teaching

Opening a Twitter account has consequences. I’m carefully keeping to scholarly activities, and so the main side effect for me has been an ever expanding list of interesting papers I really need to read. This is mainly a good thing: as a senior researcher and group leader I certainly don’t read enough new papers simply because they are interesting and relevant (as opposed to: because I need to do something with them as I’m either a co-author of, editor for or reviewer of them).

It’s nearly August and I teach in September. My reading list for now is about what I would like to read and think about to update my teaching:

The challenge for me is to find the right balance in helping students to do what they can where they are as researchers using animals, and challenging them to find ways of doing better. Unless I challenge myself, I easily settle too much on the status-quo side!

There’s also Heather Browning’s very recent PhD thesis If I could talk to animals: Measuring subjective welfare, of which I’m very interested in the second part which addresses measuring welfare from a philosophy-of-science perspective. And I want to take Daniel Lakens’ Coursera courses Improving your statistical questions / Improving your statistical interferences.

And then there are a couple of books on my desk…

 

Is a positive attitude to animal research desirable?

The question is motivated by a recently published research paper in JALAAS: Attitudes Toward Animal Research Among Medical Students in the United States by David Q Beversdorf and Nellie R Adams.

The authors recruited student members of the American Academy of Neurology to fill in a questionnaire. 168 students completed the questionnaire, expressing their agreement or disagreement with a set of 14 positively- or negatively-biased statements regarding animal research. After that, they were given the opportunity to watch a video about animal research, and asked to fill in the same questionnaire again, which 108 students did. In the text box to the right, you can see examples of the statements the students were presented with.

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After having watched the video, the students were slightly more positive towards research with animals: the score went from an average of 4.0 to an average of 4.1. (A score near 5 shows a positive attitude to animal research, a score near 1 a negative attitude).

The authors do not make much out of that change (not surprisingly, it’s so small that it doesn’t really matter) but instead discuss the observation that “a substantial number of
medical students express disagreement with statements that
describe essential components of the drug and procedure
development pipeline. As described above, 13.2% disagreed
with the statement ‘New surgical procedures should be tested
on animals before they are used on people,’ and 7.2% disagreed
with ‘New drugs should be tested on animals before
they are used on people.’” They continue to conclude that “The changes in attitudes after observing the video suggests that negative attitudes can be changed, and that medical education may have a role in this setting.” If you want to read the full paper, you can request a copy from the authors at ResearchGate.

Numerous studies with bigger samples and more comprehensive approaches have measured the attitude of different publics to animal research, but the present study is novel in the choice of a very specific public: medical students. My main issue with how the study was conducted is the quite one-sided study design. The video in question is produced by Americans for Medical Progress, an advocacy organisation for biomedical research and in particular the use of animals in such research. While I think the video is quite reasonable, it definitely represents a selective use of information. That’s not surprising, as the organisation behind it is working in favour of public support for biomedical researh with animals.

Should a goal for medical education be to make students more positive to the use of animals in research? I’m really not convinced that this should be a priority. I think it’s crucial that medical students get a reasonable understanding of the role of animals in biomedical research and drug development. But that also includes an understanding of critical challenges to how useful such research is. And an understanding of why the issue is contentious.

There is no equivalent video advocating in a comparable way for the replacement of animals in biomedical research. People such as the authors behind the book Animal Experimentation: Working Towards a Paradigm Change would probably be able to produce one, and it would be as credible (reasonably so) and balanced (not very) as the AMP video.

And sadly, both of them would miss out on the most critical issue in animal-based biomedical research today: how to design studies so that the results are reliable and translate to humans.

Are you going to a conference in 2020?

The beginning of a new calendar year is usually the time to think about conferences. Calls for abstracts are about to open – or in some cases even to close. I haven’t made up my mind yet about which conferences to attend, except for two for which I have an invitation (Scand-LAS and World Congress on Alternatives and Animals in the Life Sciences) which I will honour and one I know I won’t attend (ISAE – I usually attend when it’s in Europe and this year it’s in India).

Here are the ones I’m considering:

Canine Science Forum – Lisbon, Portugal 7-10 July

It would be a first time for me and it would be well justified by my increasing engagement in research with dogs. I would be an attendee only, not a presenter, but I expect that somebody else from our research team will be presenting work. Besides, it’s in Lisbon, a 3-h train ride away, and a place where I always enjoy spending some time. Other than that, I can’t really say much since I’ve never attended before.

World Congress on Alternatives and Animals in the Life Sciences – Maastricht, Netherlands 23-27 August

This is an interesting (in more than one way) conference that I’ve attended twice before (it’s held every 3 years). In my experience, it is more of a conference on Replacement alternatives to animals than Refinement alternatives, and the focus on Replacement has been heavily biased towards animal testing (toxicology etc). Whereas I obviously think it’s great that animal tests are being replaced, I don’t really have any scholarly interest in toxicology tests. But this year I have a particular reason for wanting to be present in a Replacement context: we have a project just starting on organoids and other advanced 3D models for research. This would also be a great chance for our newly hired junior researcher and project manager to get a peek into the alternatives research community. I really should discuss with my team what to present. Maastricht is a lovely city too.

EAAP – Porto, Portugal 31 August – 4 September

Again, this is a conference I’ve never attended but one which I know farm animal researchers in Europe always have on their conference agenda. Farm animal science is my background, and this would be an excellent opportunity for a much needed update, within walking distance from home! What speaks against participation is that I’m not sure what I would be able to present, and since this is a costly conference (even when not having to pay for travel or accomodation) I need to be able to fit it into a research project budget.

Will we be meeting at any of these conferences?

Does cold stress make mothers kill their pups?

This is what a team of researchers claim in a short report published this spring in Journal of Animal Science and Technology. They base this claim on a study in which they housed female periparturient mice in different environmental temperatures (20-23C versus 10-15C) and counted pups.

Based on what I know about on mouse maternal behaviour and pup survival, a topic I have been researching for more than a decade, I don’t believe that cold stress makes females kill their pups. In fact, our research shows that females often eat their already dead pups, but rarely ever do they actively kill pups.

But if presented with convincing data, I would of course change my mind. The important thing about the short report cited in the first paragraph is that it did not present any data on maternal behaviour. The only data presented is about pups – numbers born and numbers surviving.

A few years ago, these reflections would not have left our informal research group discussions, but now there are public fora for this kind of critical discussion. So I just wrote my first PubPeer comment.

Congress blogging Day 1 PM: ISAE, PEI, Canada

The afternoon session the first day started with Temple Grandin’s plenary talk. The title captures what Temple Grandin has really spent her entire career doing: Crossing the divide between academic research and practical application of ethology on farms. As always, her talk was an energizing tour-de-force full of practical advice. She started by underlining that people in The Field don’t know that applied ethology exists, and that to reach them you need to simplify, simplify, simplify (not because they are not intelligent but because there is very little time to convey whatever concept). As an example of how to be clear, simple and specific in formulating guidelines: don’t write “sufficient space”, write “enough for all pigs to lie down at the same time”. Equally worth sharing is her final reflections which paraphrased went something like “I’ve spent my entire life designing slaughter houses, now they’re busy developing artificial meat, in 20 years people may be thinking that my work was unethical”. (I hope many of us are making that kind of reflection at least occasionally, because it is an important one. It is also one I hope to be able to explore in a future, ISAE-related project).

In the afternoon parallel session period, I of course followed the laboratory animal science strand. It started in an unusually coherent way, with two talks reporting work on the same cohort of mice, by Aimée Marie Adcock And Emma Nip. Their work is based on housing female mice in trios in either standard or enriched cages

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As the photos show, this study brings back the environmental enrichment methodology that is hardly seen in laboratory animal welfare studies today, the one where the animals are given a radically richer environment, rather than the minimal-improvement-within-what-is-still-standard. Another interesting approach in this study is that the animals were housed in trios of three different strains (DBA, C57Bl6, Ball/c), which since they are distinctly coloured means that individual animals can be followed on videos without the need for individual marking.

The two subsequent talks reported studies done in collaboration with other animal users. Sophie Brajon reported results from our work on perinatal mortality in laboratory mouse breeding, where we followed mice in two different breeding facilities. Brianna Gaskill’s work took up recommendations made a decade ago by Pascalle van Loo for reducing aggression in mice and applied them on mice in a toxicology study. Once this work which is still under analysis, both studies have potential to influence recommendations for housing and managing mice. This may also be true for the last study reported in this session, by Becca Franks on curiosity in zebrafish, because indeed the way this species is kept has very little to do with their natural behaviour.

Curiosity also drove me to take part in the final event of the day, a workshop on Animal Welfare Assessment Contest. I have seen this activity discussed in ISAE since more than a decade, but in all honesty, I never engaged with  this North American concept of competing in how to evaluate animal welfare. It has always seemed rather alien to me: in Europe sports is the only activity we compete in at universities – outside the ongoing silent competition for the best marks of course. In a well organized session we got the taste of a real competition, got to try our hand at evaluating the welfare of working donkeys in two different contexts and watch the impressive performance of students with actual competition experience (Megan La Follette on the photo below).

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What’s the problem with this photo?

I was asked to comment on what the problem is with the stock photo above. RSPCA did well in pointing out that there is a problem, even though they somehow missed the opportunity to explain what the problem is!

The photo shows a mouse female and her litter. Based on that the pups have some fur but don’t yet have their eyes open, I would guesstimate that they are perhaps a week old. Mice are an altricious species, which means that the young are born in an undeveloped stage and are fully dependent on parental care for basically everything during their first week of life. They do develop very fast, and are ready to leave the nest at about a month of age, and sexually mature just a few weeks later. But at the stage of development in this photo, they aren’t capable of doing very much. The place for such young pups to be is the nest that their mother (or parents, males share the parental care if they can, although under laboratory conditions they may or may not be present) have prepared. That provides them with the thermal microenvironment they need – which needs to be warmer than that of an adult mouse given their poorly developed fur.

Judging from own experience of how long it takes to get a good photo of animals, by the time this photo was taken, the pups were no doubt cold. But that is only part of the problem. Anyone who has seen a professional photo session, or having had their photo taken in a studio, knows that this involves lots of bright light and reflectors. Well, this is precisely the kind of environment neuroscientists use to test anxiety in mice – an open and brightly lit space. Mice are nocturnal animals who spend their lives in tunnels and rarely venture out in the open light, and being forced to be in an environment which in nature would be dangerous is stressful for any mouse. Most likely, it is even more stressful for a female when her rather helpless pups are also exposed in this way.

If you want to learn more about our research in this field. check out the Alive Pup Project. But before you go there, let me just reflect on the different ways of making photos and for which purpose. You will see that in the Alive Pup Project we also show photos of pups taken out of their nest. But these photos were taken when the pups were weighed and inspected as part of a research project aiming to elucidate factors behind pup mortality. This is a big problem in laboratory mouse breeding, and research into the problem requires collecting data. In contrast, it is possible to make nice photos of mice in environments which are less stressful for them the one illustrated above

Still, I need to admit that we have also used stock photos of mice in unnatural environments. We simply did not think. Now we will!