Tag Archives: Three Rs

The choice of procedures

Michael Balls

The scientifically-justifiable choice of procedure
is a crucial issue in animal experimentation,
in the interests of both humanity and efficiency

In the opening of their chapter on Reduction and Strategy in Research in The Principles of  Humane Experimental Technique,1 Russell and Burch pointed out that “one general way in which great reduction can occur is by the right choice of strategies in the planning and performance of whole lines of research”.

They referred to the way Charles Hume had put it “in a searching essay”,2 that is, that “the central problem is that of choosing between trial and error on a grand scale and deductively inspired research”. The second type of choice can “take the form of testing deductions from well and consciously formulated hypotheses, or it may involve working from hunches — really the same thing, for where hunches are of any value, they are found to be based on equally precise hypotheses”. They said that the essence of the strategy “is that particular experiments are selected on some basis … from a larger set of experiments that  could have been performed”, and, as Hume had pointed out, “insighted” research “must be vastly less wasteful of animals”, where animals are to be used in the experiments.

I discussed this in an earlier comment on the Wisdom of Russell and Burch in relation to Reduction,3 with particular reference to experimental design and statistical analysis, but I now want to consider their discussion on The Choice of Procedures, in their chapter on Refinement.

Russell and Burch said that almost any research question “can always be answered in principle by a number of different procedures”, and the mark of distinction of “the great experimenter is the knack of choosing the most rapid, elegant and simple one”. But they then ask whether there are any simple rules in this context. One “general principle, important for both humanity and efficiency, is that of avoiding elaborate and roundabout methods”. Another rule is “the very careful formulation of questions”. One approach is to “first ask the question, then draw up, at least mentally, a list of procedures by which it could be answered”. If the list is long enough, consideration can be given to choosing the best procedure. If the list is too short, the question may need to be reformulated, to permit a wider range of procedural choice.

The greater the experimenter, in terms of ability and quality, the easier and better will be the choice of procedure. In addition, the humane experimenter will be careful to take account of Hume’s point about the wastage of animals, and Russell and Burch’s emphasis on the need for humanity and efficiency. This should be obvious, in terms of the use of resources, even if it were not a requirement of the laws under which animal experimentation is now permitted. It is a vital aspect of the education and training which must be regarded as essential for all those who are to undertake research, particularly if there is any risk of causing animal suffering, but also in the interests of the humans for whose benefit the research is being conducted.

It is encouraging that, in its response to the European Citizens’ Stop Vivisection Initiative,4,5 the European Commission has proposed four actions, one of which is to “analyse technologies, information sources and networks from all relevant sectors with potential impact on the advancement of the Three Rs”, in order to “present by end 2016, an assessment of options to enhance knowledge sharing among all relevant parties. The assessment will consider how  systematically to accelerate knowledge exchange through communication, dissemination, education and training.”

In order to aid researchers in their thought process when designing any project that could involve experimental animals, the FRAME Reduction Steering Committee designed a Strategic Planning Poster.6 The poster, which is available in several languages, guides the scientist through the decision points and steps needed when designing a whole programme of work, including the individual experiments within it. This resource, and the FRAME Training Schools in  Experimental Design and Statistical Analysis,7 encourage researchers to think about how to design their sequence of experiments, in order to minimise the number of animals that are exposed to the most severe procedures and to contemplate whether animals are needed at all.

Professor Michael Balls
c/o FRAME
Russell & Burch House
96–98 North Sherwood Street
Nottingham NG1 4EE
UK
E-mail: michael.balls@btopenworld.com

References
1 Russell, W.M.S. & Burch, R.L. (1959). The Principles of Humane Experimental Technique, xiv + 238pp. London, UK: Methuen.
2 Hume, C.W. (1957). The strategy and tactics of experimentation. The Lancet, 23 November, 1049–1052.
3 Balls, M. (2013). The Wisdom of Russell and Burch. 4. Reduction. ATLA 41, P24–P25.
4 Anon. (2015). Commission Response to the European Citizens’ Initiative “Stop Vivisection”. Brussels, Belgium: Intergroup on the Welfare and Conservation
of Animals. Available at: http://www.animalwelfare intergroup.eu/2015/06/05/european-commissiondoesnt- go-far-enough-to-meet-citizens-demands-toimprove-
animal-welfare/ (Accessed 08.07.15).
5 Balls, M. (2015). The European Citizens’ Stop Vivisection Initiative. ATLA 43, 147–150.
6 Gaines Das, R., Fry, D., Preziosi, R. & Hudson, M. (2009). Planning for reduction. ATLA 37, 27–32.
7 Fry, D., Gaines Das, R., Preziosi, R. & Hudson, M. (2010). Planning for refinement and reduction. ALTEX 27, Special Issue, 293–298.

 

Rex Leonard Burch: Humane Scientist and Gentle Man

Michael Balls

Rex Burch played a vital role in collecting background information
about practical animal experimentation, and was the first person
to use the term ‘alternatives’ in that context. The senior author of
The Principles, William Russell, greatly valued their collaboration

 

Whereas The Principles of Humane Experimental Technique1 reveals a great deal about its first author, it gives us very little insight into the character and contributions of its second author, Rex Burch. As a result, his role in the origins of the Three Rs concept could be underestimated. Happily, that can be avoided, not least because his co-author, William Russell, was solicitous in ensuring that his partner received the credit properly due to him.2 In addition, a few of us had the privilege of knowing Rex,3-5 and he recorded his own thoughts in a report produced under contract to ECVAM and published in ATLA.6

Soon after the UFAW project began, Russell informed Charles Hume that he would need some assistance, and the UFAW Council somewhat reluctantly agreed to the appointment of Burch. The plan was that Burch would “discuss with existing research workers, their techniques and attitudes to modifications, in terms of humaneness, and the idea of replacement techniques which did not require animals”. 6 To facilitate this, a letter of introduction was prepared (not without a struggle2) and sent to 20 individuals in December 1954. The response was positive, so Burch was able to begin his visits. He carried a tape recorder with him, then sent the interview tapes to Russell, who had their contents transcribed and placed in the records on which their eventual report and The Principles would be based.

Rex Burch in the lab

Russell later commented that, “As fieldwork for our project, Rex travelled widely over Britain, interviewing well over a hundred experimentalists, and every single one of them was cooperative. This shows, I think, not only the goodwill in the British scientific community, but the confidence Rex must have inspired by his friendly approach and his real understanding of the issues. These interviews were, of course, vitally important for our project, but I feel that there was more to them than that. Rex sowed many seeds, and helped to channel the goodwill of scientists into the more systematic approach to humane technique which has produced so many advances in recent years.”4 That was an immense achievement, since it must not be forgotten that, in those days, before the publication of The Principles with its introduction of the Three Rs concept, there was virtually no activity in the middle ground between the pro-vivisectionists and the anti-vivisectionists. There were only two positions on animal experimentation — for or against.

Burch’s success in gaining the confidence of research scientists undoubtedly stemmed from his own considerable experience in laboratory work, even though he was only 28 years old when he joined the UFAW project. He put it like this,6 in his own words: “In the early part of the Second World War, I worked in the North Riding Laboratory of Pathology and Public Health at Scarborough in North Yorkshire. One of my duties was to test milk for tubercle bacilli, and I had a constant stock of 200 guinea-pigs for this purpose. After their intraperitoneal injection of a centrifuged deposit of the milk sample (two guinea-pigs were used for each sample, one being killed after three weeks, the other after six weeks), their inguinal glands were examined for tubercle bacilli. My concern, before taking charge of this work, was about the humane killing. I was shown the method — a short, sharp blow to the nape of the neck with a specially made wooden instrument, which certainly caused instant death. I was made to practice on dead animals, and my first live guinea-pig did die instantly. From here I joined the army, serving in the Middle East and still using guinea-pigs for this diagnostic work. After the war, I read medicine at Guy’s Hospital Medical School in London (UK), but had to abort my studies and find paid work. I was offered work in the research department of Boots Pure Drug Company, where a large variety of laboratory animals were used. My appointment was that of an assistant in tropical medicine. At an early stage, disease broke out in a large mouse unit which supplied the various laboratories, and all of the mice had to be destroyed. I suggested that there should be a diagnostic laboratory for all of the laboratory animals, in order to provide early warning of disease. Within a matter of days, the Head of Research instructed me to set up such a unit. This work gave me wide experience in animal husbandry and experimental procedures, because I became deeply involved with work in the breeding units and with all those working experimentally in different disciplines. After a few years, I resigned from my appointment to set up my own research unit and breeding unit, in Huntingdonshire. My bank manager soon made me aware that there was a Professor Alastair Worden, only a few miles away, carrying out what he thought was similar work to my own. I soon made my acquaintance with Alastair Worden and, at our first meeting, he invited me to carry out histological and microbiological work for him on a casual basis. In the course of what seemed a comparatively short time, he told me that UFAW was appointing a very talented zoologist from Oxford University, Dr William Russell, to carry out a laboratory animal survey.” Worden knew that Russell needed an assistant, and suggested that Burch should contact Hume about the possibility of an appointment.

It is clear that, in 1954, Burch had greater, and wider, experience in practical laboratory work than his senior partner. Nevertheless, Burch’s crucial role in the project was not recognised at first, even by UFAW’s President, Edward Hindle FRS, who refused to support his application for membership of the Institute of Biology. Happily, largely as a result of Russell’s vigorous support, Burch became an MIBiol in mid-1959, at the time of the publication of The Principles. He was elected a Fellow of the Institute in 1987.2

For many years, there was confusion about when the term ‘alternatives’ was first used in relation to experiments on living animals. For a long while, the credit was given to Terry Hegarty, a FRAME Trustee and the son of FRAME’s Founder Chairman, Dorothy Hegarty. However, on a visit to the Russell Archive at the University of Nottingham, I discovered that the fact is that it was Burch himself who first used the term ‘alternatives’ in the context of the Three Rs — in October 1954, even before he had joined William Russell to work on the UFAW project! The story is as follows.2 Russell produced a draft of the letter of introduction to be sent to heads of university departments and research institutions, which was amended by Burch. In a sentence in which Russell wrote, “Mr. Burch is on a fact-finding mission, and with the information we hope to be able to suggest improvements in routine methods of research…”, Burch crossed out “improvements”, and suggested, as indicated in a footnote, that “some possible alternatives” should be used instead. Russell rejected that suggestion, crossed out “to suggest possible alternatives to techniques used at present” and suggested that “to produce a review of progress in the development of humane techniques” should be used. His position was supported by Charles Hume, Head of UFAW, who said, “The only thing that worries me a little is the reference to suggesting possible alternatives. Of course we do hope to do this, but if we say so bluntly I fear there may be a counter action — ‘Who are you to tell me how to do my job?’, I would suggest deleting the line ‘to suggest …at present’ and substituting something of this sort: ‘to present a review of progress in the development of humane techniques’.”

Happily, this temporary difficulty did not have a lasting effect on the excellent relationship between Russell and Burch, which was crucial to the development of the Three Rs concept. However, the publication of The Principles was not without controversy, largely because Russell rejected any external attempts to edit his text, but correspondence with Hume again bears witness to his respect for Burch. The original agreement with Methuen, publishers of The Principles, was that 80% of any royalties would go to UFAW and 20% would go to Russell, as its author, who would also own the copyright. Russell asked that his 20% should be shared equally with Burch, whereupon Hume impatiently commented that 10% of not very much would not be very much.2

Friday 5 August 1994, was a very special day for me, as I went to Sheringham, Norfolk, UK, to meet Rex Burch for the first time. As I said later,4 “I was totally captivated by his enthusiasm, his genuine excitement about the activities of FRAME and ECVAM, and his kindness. Within a few minutes, I felt as if I had known him for many years, and I knew that I would never forget him for the rest of my life”. That meeting had two important practical consequences. Firstly, the ECVAM contract, which led to a fascinating article in ATLA, The Progress of Humane Experimental Technique since 1959: A Personal View.6 That is the closest we will ever get to the autobiography he had hoped to write. It tells of his life before the UFAW project, gives his perspective on what went on between 1954 and 1959, and comments briefly on what had happened since the publication of The Principles.

Secondly, since Rex was too ill to travel far from home, I went back to Ispra, and arranged with Alan Goldberg that we should organise a workshop on the Three Rs, to be held in Sheringham, and with Russell and Burch as special participants. This was the first and, sadly, the last scientific meeting which the two authors of The Principles had attended since 1959. It resulted in a report with 58 far-reaching recommendations,7and it was a memorable event of very special significance to Rex Burch, humane scientist and gentle man.
Professor Michael Balls
c/o FRAME
Russell & Burch House
96–98 North Sherwood Street
Nottingham NG1 4EE
UK
E-mail: michael.balls@btopenworld.com

References
1 Russell, W.M.S. & Burch, R.L. (1959). The Principles of Humane Experimental Technique, xiv + 238pp. London,UK: Methuen.
2 Balls, M. (2009). The origins and early days of the Three Rs concept. ATLA 37, 255–265.
3 Balls, M. (1994). A day with Rex Burch. FRAME News 36, 1.
4 Russell, W.M.S., Sainsbury, D., Self, K. & Balls, M. (1996). Editorial. Rex Leonard Burch (1926–1996). ATLA 24, 313–316.
5 Goldberg, A. (2010). The Principles of Humane Experimental Technique: Is it relevant today? ALTEX
27, Special Issue, 25–27.
6 Burch, R.L. (1995). The progress of humane experimental technique since 1959: A personal view. ATLA
23, 776–783. Reprinted in ATLA 37, 269–275.
7 Balls, M., Goldberg, A.M., Fentem, J.H., Broadhead, C.L., Burch, R.L., Festing, M.F.W., Frazier, J.M., Hendriksen, C.F.M., Jennings, M., van der Kamp, M.D.O., Morton, D.B., Rowan, A.N., Russell, C., Russell, W.M.S., Spielmann, H., Stephens, M.L., Stokes, W.S., Straughan, .W., Yager, J.D., Zurlo, J. & van Zutphen, B.F.M. (995). The Three Rs: The way forward. The report and recommendations of ECVAM workshop 11. ATLA 23, 838–866.
The Principles of Humane Experimental Technique is now out of print, but the full text can be found at
http://altweb.jhsph.edu/pubs/books/humane_exp/het-toc. An abridged version, The Three Rs and the
Humanity Criterion, by Michael Balls (2009), can be obtained from FRAME.

‘One R’ is the new ‘Three Rs’

Craig Redmond

The Lush Prize, which rewards initiatives to end animal testing,

believes that more attention needs to be given to the ‘One R’

of absolute replacement, and that research methods that

exploit animals in any way (including tissues and cells)

should not be considered as ‘alternatives’

Introduction
Replacement of animal experiments is one of the Three R concepts (alongside Reduction and Refinement) first put forward by Russell and Burch in 1959.1 However, this can be either absolute replacement (i.e. methods that do not involve animals or animal tissues) or relative replacement (e.g. methods that use only cells or tissues of animals in vitro or ex vivo).

It has become accepted by many in the research community that some uses of animals can be classed as ‘alternatives’. In particular, the use of whole animals of species thought to either not experience pain or to have a lower level of sentience (e.g. fruit flies, nematodes and zebrafish), or of animal parts (including tissues, embryos, sera and cells). The use of these methods is reinforced by regulatory bodies, making it more difficult to reach a time when no animal use will occur in scientific research. André Ménache, of Antidote Europe, believes that in the region of 80% of ‘alternatives’ validated by ECVAM (the European Union Reference Laboratory for alternatives to animal testing) still use animals or animal tissues (personal communication, 04.09.13).

Founded in 2012, the Lush Prize rewards global initiatives to end animal testing, particularly in the area of toxicology. A total of £250,000 is shared annually between five prize categories covering science, training, young researchers, lobbying and public awareness. Lush Prize believes that more attention needs to be given to the ‘One R’ of absolute replacement, and that research methods that exploit animals in any way (including tissues and cells) should not be considered as ‘alternatives’.

Alternatives’ that still exploit animals
The United States Department of Agriculture refers to ‘alternatives’ as “a term that has different meanings to different people, and this difference largely depends on which side of the issue one is found”.2 So, for example, animal researchers might use relative replacement methods in addition to their use of animals (or look to refine existing animal tests), whereas abolitionists see ‘alternatives’ in terms of absolute replacement. Other examples of where the use of the term ‘replacement’ serves to reinforce the idea that relative replacement is routinely acceptable are:

— where Russell and Burch defined a replacement technique as “any scientific method employing non-sentient material which may in the history of animal experimentation replace methods which use conscious living vertebrates”.3 The words ‘non-sentient’, ‘conscious’ and ‘vertebrates’ ensure that the use of invertebrates and species considered as ‘lower organisms’ continues to be accepted.
— when, in its current “step-by-step approach to an alternatives search”, the Johns Hopkins University Center for Alternatives to Animal Testing (CAAT) suggests that, in addition to cell culture, tissue culture, models, simulations, etc., researchers “might look for a non-mammalian animal model
— fish or invertebrates, for example — that would still give you the data you need”. 4

Perhaps as a direct result of this widespread inherent acceptability of relative replacement alternatives, researchers at the University of British Columbia, looking into people’s acceptance of the use of particular species in laboratories, found that species such as fish and invertebrates “are typically rated below mammals, and, as such, are often considered an appropriate replacement for mammals in research”. 5

The philosopher Joel Marks notes that “developing alternatives to the use of animals can mean simply using a different animal” and considers that “the characterisation of the other animal […] as ‘lower’ on a phylogenetic ‘scale’ is arbitrary and disputed. The alternatives movement is therefore at risk of becoming a bait-and-switch con”.6 By this, Marks means that ‘alternatives’ are advertised as one thing (i.e. absolute replacement), but often turn out to be something completely different (i.e. simply the use of another species).

Some examples of relative replacement alternatives are:
Invertebrates: The horseshoe crab, Limulus polyphemus, is used in the Limulus amoebocyte lysate (LAL) assay. This method replaces the rabbit pyrogen test for the detection of endotoxin in, for example, hepatitis B vaccines. 7 The rabbit test involves injecting the test substance into a marginal vein of the ear of each of three rabbits.8 However, the LAL assay uses blood cells from the horseshoe crab, with up to 30% mortality resulting from the bleeding procedure.9

Fish: Zebrafish (Danio rerio) are widely used in research, including research on genetics, cancer and, increasingly, toxicology. The maintenance costs of zebrafish are less than one thousandth of the costs of maintaining mice,10 and they can produce 100–300 eggs per week, making their embryos useful for high-throughput screening.11 It is widely acknowledged that fish can feel pain, 12 with as much evidence for this as there is for birds and mammals.13 Other studies have shown that they have conscious awareness.14

Tissues: Hundreds of thousands of animals are bred and killed each year in Britain alone, solely to provide tissues for research.15 Human tissues to be preferred, due to species differences, yet animal tissue is often used on the grounds of cost and availability.16 Human tissue can be obtained from patients during diagnosis, removed as ‘waste’ during surgical operations, placentas or ‘afterbirth’, or tissues obtained after death.15

People can voluntarily donate blood or other tissue for transplantation or research, or their organs or bodies after death. Human tissue removed from the body in the course of disease diagnosis or treatment is the main source.17 However, although upwards of 600,000 residual surgical tissues are generated each year in England and Wales, only a tiny fraction of them are made available to researchers.18

The use of fetal calf serum
The move toward the use of in vitro cell culture to provide both human and animal cells for alternative methods is a step in the right direction. However, the use of fetal calf serum (FCS) as a cell culture media supplement is unacceptable, in the light of the availability of serum replacements and serum-free culture methods.19 collected for FCS production is obtained by cardiac puncture, performed by inserting a needle directly into the heart of the non-anaesthetised fetus.20 according to Jochems et al.20 it is very likely that the fetus is alive at the time of blood collection, and “will experience pain and/or suffering at the moment of heart puncture for blood collection and possibly for a period after that, until it actually dies”. The scientific validity of using FCS has been questioned. Risk of contamination is an issue,21 with the potential presence of viruses, bacteria, mycoplasma, yeast, fungi, immunoglobulins and endotoxins.20

Conclusions
These are just a few examples of animal use that some see as ‘alternatives’. The use of sentient animals, such as fish and horseshoe crabs, should not be accepted by those working in the field of alternatives to animal testing, despite the entrenched position within the research community and regulatory bodies. Neither should cruel processes such as the collection of FCS be condoned. In addition to greater humanity and greater acceptability, there are a multitude of clear scientific benefits to avoiding the use of animals or animal products.

The Lush Prize promotes the ‘One R’ of Replacement over all of the ‘Three Rs’, believing that the  true absolute replacement of animals is essential for ethical and scientific progress.

Craig Redmond
Lush Prize
Unit 21
41 Old Birley Street
Manchester M15 5RF
UK
E-mail: craig@lushprize.org
References
1 Russell, W.M.S. & Burch, R.L. (1959). The Principles of Humane Experimental Technique, 238pp. London, UK: Methuen.
2 Taylor Bennett, B. (1996). Alternative methodologies. In Essentials for Animal Research. A Primer for Research Personnel, pp. 9–17. Darby, PA, USA: Diane Publishing.
3 Balls, M. (1994). Replacement of animal procedures: Alternatives in research, education and testing. Laboratory Animals 28, 193–211.
4 Altweb Project Team (undated). Search for Alternatives. Baltimore, MD, USA: CAAT, Johns Hopkins
University. Available at: http://altweb.jhsph.edu/resources/searchalt/index.html (Accessed 22.10.14).
5 Ormandy, E.H., Schuppli, C.A. & Weary, D.M. (2012). Factors affecting people’s acceptance of the use of zebrafish and mice in research. ATLA 40, 321–333.
6 Marks, J. (2012). Accept no substitutes: The ethics of alternatives. The Hastings Center Report 42, Nov–Dec (Special Report), S16–S18.
7 Park, C.Y., Jung, S.H., Bak, J.P., Lee, S.S. & Rhee, D.K. (2005). Comparison of the rabbit pyrogen test and Limulus amoebocyte lysate (LAL) assay for endotoxin in hepatitis B vaccines and the effect of aluminum hydroxide. Biologicals 33, 145–151.
8 WHO (2013). The International Pharmacopoeia. Preface: 3rd Supplement. Geneva, Switzerland: World Health Organisation. Available at: http://apps.who.int/phint/en/p/docf/ (Accessed 07.09.13).
9 Leschen, A.S. & Correia, S.J. (2010). Mortality in female horseshoe crabs (Limulus polyphemus) from biomedical bleeding and handling: Implications for fisheries management. Boston, MA, USA: Massachusetts Division of Marine Fisheries. Available at: http://www.mass.gov/eea/docs/dfg/dmf/publications/mortality-in-female-horseshoe-crabsabstract.
pdf (Accessed 25.10.14).
10 Reed, B. & Jennings, M. (2011). Guidance on the housing and care of zebrafish Danio rerio, 64pp. Horsham, West Sussex, UK: Research Animals Department, Science Group, RSPCA.
11 van Vliet, E. (2011). Current standing and future prospects for the technologies proposed to transform toxicity testing in the 21st century. ALTEX 28, 17–44.
12 FAWC (1996). FAWC report on the welfare of farmed fish, 43pp. London, UK: Farm Animal Welfare Council.
13 Braithwaite, V. (2010). Do Fish Feel Pain?, 208pp. Oxford, UK: Oxford University Press.
14 Cottee, S.Y. (2012). Are fish the victims of ‘speciesism’? A discussion about fear, pain and animal consciousness. Fish Physiology & Biochemistry 38, 5–15.
15 Focus on Alternatives (undated). Focus on Human Tissue in Research, 4pp. Available at: http://www.frame.org.uk/dynamic_files/foa_humantissue.pdf (Accessed 03.09.13).
16 EMA (2012). Committee for Advanced Therapies (CAT) Scientific Workshop: Reducing the number of laboratory animals used in tissue engineering research — 11th October 2012 — European Medicines Agency, London [EMA/CAT/708346/2012], 4pp. London, UK: European Medicines Agency. Available at: http://www.ema.europa.eu/docs/en_GB/document_library/Report/2012/12/WC500136419.pdf (Accessed 25.10.14).
17 Nuffield Council on Bioethics (1995). Human Tissue: Ethical and Legal Issues, 182pp. London, UK: Nuffield Council on Bioethics. Available at: http://nuffieldbioethics.org/project/human-tissue/ (Accessed 25.10.14).
18 Bunton, D. (2011). The use of functional human tissues in drug development. Cell & Tissue Banking 12, Issue 1, 31–32.
19 Newman, C. (2003). Serum-free cell culture — the ethical, scientific and economic choice. The Biomedical Scientist, September 2003, 941–942.
20 Jochems, C.E., van der Valk, J.B., Stafleu, F.R. & Baumans, V. (2002). The use of fetal bovine serum: Ethical or scientific problem? ATLA 30, 219–227.
21 Focus on Alternatives (2009). Serum-free Media for Cell Culture, 52pp. http://www.frame.org.uk/dynamic_files/foa_fcs_free_table_may09.pdf
(Accessed 03.09.13).
 

Beyond the Three Rs

For a long while after the Three Rs were first proposed by Russell and Burch, anti-vivisectionists rejected the concept, on the grounds that experiments on living vertebrates which cause them pain, suffering, distress or lasting harm, were ethically unacceptable and scientifically unnecessary, so there was no point in reducing, refining or replacing them. In recent decades,
however, some organisations, such as the BUAV and PETA, have moved tentatively into the middle ground, and have made positive contributions toward the Three Rs, without comprising their fundamental beliefs.

The ultimate goal of Russell and Burch themselves was replacement, which they said, “is always a satisfactory answer”, with reduction and refinement merely being steps along the way. That was also the position of the founders of FRAME, the Fund for the Replacement of Animals in Medical Experiments, although the charity has made many contributions in support
of the other two Rs, since its foundation in 1969.

The latest issue of ATLA contains two important articles about the future of the Three Rs, as well as the latest in an important series of outstanding exposures of the insurmountable limits of laboratory animals as models of humans.

In this issue of PiLAS, Craig Redmond argues the case for replacing the Three Rs with One R (Replacement),1 but goes further in saying that only what Russell and Burch defined as absolute replacement (where “animals are not required at all at any stage”) should be considered acceptable, since relative replacement can still involve suffering, as in the use of invertebrates,
less-sentient vertebrates, or cells and tissues taken from protected animals and used in vitro or ex vivo.

Michael Balls goes further in his ATLA Comment,2 proposing that “the time has come to plan for a future where the Three Rs will have served their purpose, animal experimentation will have been consigned to history, and humane biomedical science in research, testing and education will have become the norm, for the benefit of humans and animals alike”.

Finally, the article by Jarrod Bailey in the latest ATLA issue, on monkey-based research,3 demonstrates that major molecular differences, revealed by comparative
genomics and molecular biology, underlie inter-species phenotypic disparities. The collective effects of these differences are striking, extensive and widespread, and show that the superficial similarity between human and monkey genetic sequences is of little benefit for biomedical research.
Therefore, the extrapolation of biomedical data from monkeys to humans is highly unreliable, and the use of monkeys must be considered of questionable value, particularly given the breadth and potential of alternative methods of enquiry that are currently available to scientists.

References
1 Redmond, C. (2014). ‘One R’ is the new ‘Three Rs’. ATLA 42, P50–P52.
2 Balls, M. (2014). Animal experimentation and alternatives: Time to say goodbye to the Three Rs and hello to humanity? ATLA 42, 327–333.
3 Bailey, J. (2014). Monkey-based research on human disease: The implications of genetic differences. ATLA 42, 287–317.

Personal Reflections on Veterinary Science Training and the Three Rs

Rosemary Elliott

Students against the thoughtless use of animals
in veterinary education need  support
to be true to their values, to use
science to uphold them, and to never
give up on advocating for the
highest standards of animal welfare

This opinion piece is something I needed to write personally, and it shocks me to notice the ten years that have passed since I enrolled as a mature-age student in veterinary science. It was a privilege to have this opportunity, and I am grateful that I can now advocate for animals in a way I never could have done before, particularly through my work with Sentient. By the time I enrolled in veterinary science, the faculty where I studied had made huge advances in the ethical use of animals in teaching, most notably through the banning of ‘terminal surgeries’. Since my time as a student, things have improved even more, and the faculty is leading the way in developing a national curriculum for animal welfare and ethics in veterinary teaching. Yet, despite this, some attitudes and practices ‘die hard’, and many of my experiences could only be described as vicarious trauma — hence the passing of so much time before I felt ready to give life to them with words.

Although the animals I speak of were used primarily for educational purposes, I do hope that those of you working in the laboratory setting with research animals, will find that my reflections will stimulate your own thinking and resonate with some of your experiences. We have much in common in our struggle to do our work and to do the best we can by the animals in our care.

Anatomy classes

Admittedly, I grew up with an idealised view of veterinary practice, starting with Dr Doolittle as a very small child, then moving on to All Creatures Great and Small and the other James Herriot books. Although 60 years had passed since the setting of those stories, with the onset of intensive farming, I went on assuming it was primarily about supportive teamwork and above all, empathy for animals.

The first wake-up call came in the form of Anatomy 1A. The laboratory, a huge room with grimacing dead greyhounds lying on metal trays, became a hothouse of anxieties, sometimes played out by the larking about that involved the throwing of body parts. The smell of formalin was so powerful that I can still conjure it up. In groups of three or four, we learned anatomy the traditional way, which some of us described as “being thrown a dead dog and a textbook”. There was minimal instruction, so we were left to hack away in the pursuit of identifying a lengthy page of anatomical features, ‘all examinable’, with the pressure to clean up before our class ended.

I don’t remember being told why they were always greyhounds, or how they were sourced. There was certainly no time for debriefing, and we learned early on that it was unacceptable to appear emotional. I soon became desensitised to this horrible scene until the 4th year, when I was initiated into surgical training on freshly-killed pound dogs of various breeds — soft and floppy and somehow, not being greyhounds, they seemed more individual, more like pets. I remember struggling with sad moments of wondering about their lives and whether they had been loved, and feeling ashamed of my tacit acceptance of greyhounds as production animals.

Now, this raises for me the question of respect for life, even after a life is over. If we are serious about honouring the dignity of animals and safeguarding their welfare, veterinary training must provide as many opportunities as possible for effective learning that reduces the number of animals used, such as initial skills training through videos and silicone simulator anatomical models. At Sentient, we also call for the veterinary profession to supply cadavers from ethical sources, rather than colluding with the widespread disposal of racing greyhounds by ‘convenience euthanasia’.

Practical animal husbandry training

Chickens and eggs
The Animal Husbandry practical classes on production animals were another shock. I expected state-of-the-art facilities for the animals, but the picnic atmosphere was killed for me by the sight of housing systems more typical of factory farming. The laying hens were all kept in battery cages, rows of little prisons with no natural lighting. Questioning this was frowned upon — we were there to show our enthusiasm and tick the boxes for the mastery of required skills. I was told these were working farms, the message to first year students from some staff members being that animal welfare is, by default, secondary to industry profit.

One of my saddest memories was being tested on my ability to remove a hen from a battery cage, restrain and examine her, then to return her to the cage, head first. She flapped with gusto and resisted going back inside. I felt I had performed an act of cruelty, giving her a tiny taste of what her body could do, but perhaps never would again. Then there was the class where chicks were deliberately infected with coccidiosis, so that we could observe the characteristic droppings. And the class on egg production, where the tutor smilingly broke fertilised eggs — to prove what, I don’t understand anymore — but leaving me with the image of an embryonic bird with a throbbing heartbeat, destined for the sink.

Compassion for birds was a disadvantage in veterinary training, where speciesism was very much alive and well. Most student complaints about animal treatment focused on mammals. In the 4th year, we were taught to euthanase chickens humanely, but were given needles of the wrong gauge, which caused their wing veins to blow. I felt murderous at the sight of students who laughingly persisted, while the ‘spent’ hens blinked helplessly as they were repeatedly traumatised by needles that failed to bring an end to their joyless lives. The demonstrator did not intervene. I learned nothing useful from this class — because after observing the other students, I decided not to even try. When I complained, the response was that they had run out of the correct-sized needles and would replace them at the next class, as if the problem had been purely a practical one.

This class could have formed a foundation for trainee veterinarians in the careful preparation and respect for birds during euthanasia. Instead, it reinforced the view of chickens as somehow less sentient, due to their status as production animals. We must consider how invoking the Three Rs would have helped here. Perhaps by the initial replacement of live birds by video footage of how to correctly perform euthanasia? But when it came to the use of live birds, which was essential, what was needed was a commitment by staff to the humane treatment of animals, and an expectation that students would demonstrate this attitude through their own behaviour. Instead, the atmosphere was cavalier and callous, and not at all conducive to any form of refinement.

Tail-biting management classes
The university’s pig farm was another indictment, with sows in sow stalls, and non-breeding pigs in dark little pens where we practised catching them with snares. This was the scene of my own most shameful memory, where, despite my opposition to unnecessary invasive procedures, I performed teeth-cutting, tail-docking and ear-notching on a piglet. I remember feeling terrified at the thought of this practical class, because of the expectation to ‘do as farmers do’ — which was ostensibly to improve the welfare of the piglets by preventing tail biting. I was unaware of any conscientious objection policy at that stage of my training. So I stalled for time, trying to look useful without actually doing anything, until the tutor presented me with my own piglet and a pair of what looked like pliers. He was clearly annoyed by my questions and my need for reassurance about avoiding the ear vein or how to create the least degree of trauma — a ridiculous question, in view of the fact that there was no analgesia.

I will always remember holding this warm, pink little being with the racing heart, who I just wanted to protect, but instead, I brutalised him. I try to cope with this memory by knowing that I worked as quickly as I could, and then rubbed his body all over to distract him from the pain, as I carried him back to his mother and made sure he found a teat to suckle on. I also try to cope by using this as the basis for my commitment from that day onwards to never again perform such an atrocity. But I will never forget standing there, praying the pain would soon end. I will never forget being inconsolable for days. I still cry at the memory of what I did. I betrayed my own values out of fear of failure and not being confident enough to stand up to pressure, aided by false reassurances that I was doing the right thing.

That night, I researched teeth cutting, and found a recent article in a veterinary journal that documented how the procedure predisposes to injury and infection in the mouth and gums. I have since visited free-range pig farms, where the ‘cannibalism’ we were indoctrinated with was not an issue. All these pigs kept their tails, and their baby teeth. I had been fed a lie, pressured into performing unnecessary and inhumane procedures on a piglet that would never be allowed on a puppy. And I was an educated adult, who had already considered these welfare issues and was not relying on this to earn a living.

My pathetic attempt at reduction, by limiting my actions to one piglet, did nothing to safeguard the welfare of that individual. Like so much of what we were expected to do in veterinary training, it simply should not have been allowed, because the procedure itself was unethical. In such cases, I believe we can be guided by the Three Rs to give priority to replacement, by showing students a video of these procedures, not least so that they know what to expect on farm visits. This can go hand-in-hand with advice about how these practices can be avoided through less-intensive husbandry practices. When invasive procedures are required, the focus should then be on initial video or simulator learning, to reduce the number of animals used, followed by refinement — i.e. teaching students to always use anaesthesia and/or analgesia to minimise stress and suffering.

Other lessons
There were multiple instances, where students at the university farm were expected to perform unnecessary invasive procedures on production animals as part of their learning experience, with routine farming practices cited as the ‘gold standard’. My question was always this: Why are we following the ways of the farmers, rather than offering something more as potential veterinarians? But most students, particularly those from rural backgrounds, accepted the status quo, citing the need for ‘real-world’ practice. It was also regarded as essential preparation for extramural farm placements, which brought further horrors that I will not elaborate on here. Common sights during my training were cattle being dehorned without analgesia, in some instances leading to maggot-infested wounds; the repeated attempts to lasso a terrified cow on a hot summer’s day, which led to her jumping a fence and almost breaking her hip; exsanguination of a sheep to prove the procedure is humane; and tail-docking and castration of lambs, who were pinned down on their backs in a ‘cradle’, all without analgesia, while students delayed the procedure by joking about who would remove which testicle.

Promotion of the Three Rs and alternatives
My reason for writing this, apart from my own need to debrief and reflect, is to think about the context that must be created, if the Three Rs are to be of use in veterinary education. Adherence to the Three Rs will only come about within a culture of empathy and respect for animals, which should also be extended to students. Offering a transparent conscientious objection policy, and reinforcing in every unit of study that there are alternatives to ethically-contentious procedures, is a crucial part of this. The way I coped was to find like-minded students, involve myself in the student-run animal welfare association, use the conscientious objection policy and formal avenues within the faculty to lodge complaints, and seek the support of staff members. And I will always be grateful to several academics, who were wonderful role models on how to uphold ethics in teaching, who had witnessed and objected to far worse in their own veterinary education, and who encouraged me and my fellow students to be true to our values, to use science to uphold them, and to never give up on advocating for the highest standards of animal welfare.

E-mail: rosemary.elliott@sentient.org.au

The 9th World Congress on Alternatives and Animal Use in the Life Sciences

The 9th in this series of congresses which began in 1993, was held at the Hilton Prague Hotel, Czech Republic, on 24–28 August 2014. Its organisation was co-chaired by Dagmar Jirová (Prague) and Horst Spielmann (Berlin), on behalf of the Alternatives Congress Trust, with the administrative support of Guarant International.

The Congress was attended by about 1,100 participants, and the programme consisted of seven plenary lectures, more than 450 oral presentations, and about 500 posters. In addition, there was an exhibition with 60 booths, plus a number of satellite meetings and workshops, and many private discussion sessions.

The overall focus of the Congress was Humane Science in the 21st Century, as represented by nine main themes: new technologies; predictive toxicology; the Three Rs in academia and education; communication, dissemination and data sharing; efficacy and safety testing of drugs and biologicals; human relevance; ethics; refinement and welfare; and global co-operation, regulatory acceptance and standardisation.

The congress facilities provided by the hotel were superb, which helped to make this a particularly friendly congress. Many of the participants in the 1993 Congress were present, but it was also good to see a great number of younger scientists, 41 of whom had been specifically invited due to generous sponsorship.

It would be impossible to say much in detail about the Congress, given the enormous variety of topics covered. However, it is worth noting that two of the plenary lecturers gave contrasting insights into the state of humane science and the Three Rs as it is today.

Uwe Marx (Berlin) described the breathtaking progress being made toward developing a “human-on-a-chip”, as means of providing information of direct relevance to humans, replacing the need to resort to laboratory animal models. Early organ-on-a-chip versions — comprising artificial lungs, liver, kidneys, heart and gut — are already in use.

By contrast, Roman Kolar (Neubiberg) warned that many apparent commitments to the Three Rs have proved to be no more than lip-service, and political initiatives to avoid or replace animal experimentation have either failed dramatically, or have been watered down in the political decision-making process.

Of the Three Rs, it appeared that Reduction was rarely mentioned in Prague, and Refinement, however welcome, pales into insignificance in the face of the huge ethical and logistical dilemmas involved in maintaining animals under laboratory conditions. Replacement took the centre stage in most of the sessions, but, given the year-on-year increase in the production and use of, in particular, genetically-modified animals, there is a lot more to be done before humane science becomes more than just a dream. The 10th Congress will be held in Seattle in 2017 — it is to be hoped that much more progress will have been made by then.

Responsibility for Animal Experiments: Where the Buck Stops

bunny and arm

Sadly, just after the Animals (Scientific Procedures) Act 1986 had been amended1 to bring into force in the UK the new European Directive 2010/63/EU on the protection of animals used for scientific purposes,2 allegations were made about the abuse of laboratory animals at one of the country’s leading academic research establishments.3 Nothing more must be said about that, pending the outcome of various investigations. However, it raises the question of who is responsible for ensuring that animals are used properly, and who is culpable, when things go wrong.

In the UK, we have long experience of the clearly-defined responsibilities of the Home Office ministers and officials, including the Animals in Science Regulation Unit (ASRU) Inspectorate, and various named persons in the breeding or user establishment, including the certificate holder, the day-today care person, the veterinary surgeon, the project licensees and the personal licensees. The former EU Directive, Directive 86/609/EEC, said relatively little about specific responsibilities, but the new Directive says far more in several of its articles, as follows:

Article 23 requires Member States to ensure the competence of all staff, by seeing that all staff are adequately educated and trained before they perform any of their functions.

Article 24 spells out the specific requirements for personnel to be responsible for overseeing the welfare and care of the animals in the establishment, ensuring that the staff dealing with animals have access to information specific to the species housed in the establishment, and ensuring that the staff are adequately educated, competent and continuously trained, and supervised until they have demonstrated the requisite competence, whilst also ensuring that projects are carried out in accordance with the project authorisation.

Article 25 requires Member States to ensure that each establishment has a designated veterinarian with expertise in laboratory animal medicine, charged with advisory duties in relation to the well-being and treatment of the animals.

Article 26 lays down the requirement for an animal welfare body, while Article 27 outlines its tasks in relation to the welfare of the animals, the application of the Three Rs, the internal establishment and review of operating procedures with regard to monitoring, reporting and follow-up, including the development and outcomes of projects and their further contributions to the Three Rs.
Article 30 requires the Member States to ensure that all establishments keep records of the number and species of animals involved, their origins, their use and their death or re-homing.

Article 34 specifies that the competent authorities of the Member States must carry out regular inspections, to verify compliance with the requirements of the Directive, taking into account the number and species of animals involved, the record of the establishment in complying with the requirements of this Directive, the number and types of projects carried out, and any information that might indicate noncompliance.

Article 35 states that, when there is due reason for concern, the Commission shall undertake controls of the infrastructure and operation of national inspections in Member States, and the competent authority of the Member State concerned shall undertake measures to take account of the results of the control.

There are, of course, other individuals and organisations with responsibilities. These include the research funding bodies and the peer reviewers who assess research grant proposals, as discussed in a useful booklet, Responsibility in the use of animals in bioscience research,4 published by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs). Also of importance are the editors and editorial boards of the journals which publish articles which involve the use of laboratory animals, as was considered in a thoughtful article by Helen Galley.5

Given the clarity of the legislation and the specified responsibilities of so many different kinds of people, how can things go so wrong, as happens, as far as we know, on relatively rare occasions? First of all, human beings are involved, and whether those involved are doctors, lawyers, airline pilots, other professionals or even parents, we are a fallible
species and sometimes do wrong things, accidentally or deliberately. We need rules within which to work, and people in place to try to ensure that we do so, but they can’t watch all of us all of the time. In the end, it all depends on personal integrity, and, in the case of performing potentially-harmful experimental procedures on living vertebrates, the ultimate and inescapable responsibility must rest with those who actually perform the procedures. That is where the buck stops.

References
1 Anon. (2012). The Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012. Statutory Instrument No. 3039, 57pp. London, UK: The National Archive. Available at: http://www.legislation.gov.uk/uksi/2012/3039/contents/made (Accessed 23.05.13).
2 Anon. (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, 20.10.2010, 33–79.
3 Woolf, M. & Jacobson, S. (2013). Inquiry into ‘abuse’ of lab rats at college. The Sunday Times, 14 April 2013, p. 8.
4 Anon. (2013). Responsibility in the use of animals in bioscience research, 24pp. London, UK: NC3Rs. Available at: http://www.nc3rs.org.uk/downloaddoc.asp? id =719 (Accessed 23.05.13).
5Galley, H.F. (2010). Mice, men and medicine. British Journal of Anaesthesia 105, 396–400.