All posts by annej

Renewed concern about the welfare of laboratory primates

PiLAS staff writer

One of the key points about the Three Rs is that, as Russell and Burch emphasised in The Principles of Humane Experimental Technique,1 “the humanest treatment of animals, far from being an obstacle, is actually a prerequisite for successful animal experiments”. Indeed, they said, the “wages of inhumanity” are “paid in ambiguous or otherwise unsatisfactory experimental results”. The PiLAS article by Chandna et al. on the commonplace single housing of primates in US laboratories,2 is therefore a matter of great concern, for both scientific and humanitarian reasons. It even appears that, far from trying to solve the problem, the US Government may be trying to cover it up.

In the UK, the NC3Rs is putting a great deal of effort into improving the welfare of the non-human primates used for research,3 but one has to wonder what fundamental and significant changes have taken place since the Home Secretary of the time, Douglas Hurd, accepted all but one of 17 proposals put to him by FRAME and CRAE (Committee for the Reform of Animal Experimentation) on the day that the Animals Scientific Procedures Act 1986 came into force.4,5 One of the points highlighted by FRAME and CRAE6 was that: “The very nature of a primate is such that you cannot institutionalise it in the laboratory and have a healthy animal. A primate is such that isolating in itself is deleterious.”

The current initiatives of the NC3Rs deserve to be applauded, but why do non-human primates continue to be used in research and testing at all, and how relevant and reliable are the data obtained to the understanding and treatment of diseases? Commenting on a recent report that the cynomolgus macaque is resistant to doses of paracetamol that would be fatal in humans,7 FRAME’s Scientific Director, Gerry Kenna, said in FRAME News that “This new research raises significant concern about the scientific validity to humans of drug safety studies undertaken in primates. The use of non-human primates in non-clinical safety testing is ethically undesirable and, in view of the substantial cost of such studies, can be expected to increase, markedly, the cost of drug development. Such studies should be considered only if they can be shown to be scientifically justifiable and there are no valid alternatives.”

The FRAME News item concluded by saying that “More time and money should be invested in cell-based and computer models that would be more reliable.”

1 Russell, W.M.S. & Burch, R.L. (1959). The Principles of Humane Experimental Technique, 238pp. London, UK: Methuen.
2 Chandna, A., Niebo, M., Lopresti-Goodman, S. & Goodman, J. (2015). Single housing of primates in US laboratories: A growing problem with shrinking transparency. ATLA 43, P30–P33.
3 Anon. (2015). The Welfare of Non-human Primates, 16pp. London, UK: NC3Rs. Available at: https://www.nc3rs.org.uk/welfare-non-human-primates (Accessed 16.07.15).
4 Anon. (1987). The Use of Non-Human Primates as Laboratory Animals in Great Britain, 16pp. Nottingham, UK, and Edinburgh, UK: FRAME and CRAE.
5 Anon. (1987/88). Response of the Home Secretary to the FRAME/CRAE primates report. FRAME News 17, 6−9.
6 Chivers, D. (1984). Comment in discussion session on Laboratory Primates. In Standards in Laboratory Animal Management, pp. 272. Potters Bar, UK: UFAW.
7 FRAME. (2015). Non-human primates and drug testing. FRAME News 74, 3.

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Turning Apples into Oranges? The Harm–Benefit Analysis and How to Take Ethical Considerations into Account

Herwig Grimm

How can expected study benefits and animal harms be weighed
against each other? What is the unit and common currency
that allows this weighing to be performed?

Suppose you are a scientist, working in the field of oncology and using live animals in your studies. Furthermore, suppose you have an excellent track record, you are well respected in the research community, and you regularly publish in high-ranking journals. One day, a person that you have not met before, wants to see you and talk about your work. Despite the fact that your time is extremely scarce, you invite this very person to your office and postpone your work on a follow-up research proposal. It turns out that the person who wants to talk to you is a member of a major animal protection group. She asks you the following question: “I came across a project summary, published according to Article 43 of Directive 2010/63/EU. Knowing your research, I think it is your project. Can you ethically justify the use of animals in your work? What I mean is: Do the benefits really outweigh the harms? And which ethical considerations do you take into account?” The animal protectionist is actually asking something for which you should, in fact, be well prepared. Directive 2010/63/EU1 was transposed into the national laws of the EU Member States. In Article 38(2) of the Directive, it is emphasised that a harm–benefit analysis of any project involving the use of animals must be carried out, in order to assess whether the harms related to the project are outweighed by the expected benefits. Furthermore, the relevant passage stipulates that ethical considerations have to be taken into account in this assessment (emphasis added by the current author):

“The project evaluation shall consist in particular of the following: d) a harm–benefit analysis of the project, to assess whether the harm to the animals in terms of suffering, pain and distress, is justified by the expected outcome taking into account ethical considerations, and may ultimately benefit human beings, animals or the environment.”

Consequently, the question arises as to how the harm–benefit analysis can be carried out, and how the term “taking into account ethical considerations” might be understood in this context. The meaning of this term is of major importance, since it provides a legally binding basis for the approval or rejection of projects. In other words, everyone who aims to secure the authorisation of a project in the EU has to make sure that the expected benefits outweigh the harms, and the justification must take into account ethical considerations, whatever that in fact means.All this has to be done on legal grounds — it is not just some fancy idea of animal protectionists.
The harm–benefit analysis:
A challenge or mission impossible?

At present, and to my knowledge, it is not at all clear how to prove, in a transparent and objective manner, that the expected benefits of an experimental study outweigh the expected harm to the animals to be used. Moreover, the actual meaning of the term “ethical considerations” remains vague, to say the least. How can expected study benefits and animal harms be weighed against each other? What is the unit and common currency that allows this weighing to be performed? And can ‘ethics’ help to turn apples into oranges, so that only comparable weights are on the scales? Since the Directive does not provide any specifications on standards for the harm–benefit analysis and how to take ethical considerations into account, the passage invites the reader to speculate.

The working document on Project Evaluation and Retrospective Assessment (WD 2013),2 from September 2013, is only of limited help. It provides important criteria and ideas, but it leaves the reader without help when it comes to a methodology for transparent decision-making. It refers to the Bateson Cube, which indicates what should be taken into account, but whether its dimensions (i.e. benefit, likelihood of benefit, harm to animals) are ethical in nature, and whether these dimensions are sufficient for the harm–benefit analysis, remains open to question. Furthermore, no measure is provided to allow the various dimensions to be made comparable. Take, for example, ‘benefit’. Here, a set of analytic questions is given (WD 2013, p. 21):
What will be the benefits of the work?
Who will benefit from the work?
How will they benefit/impact?
When (where possible) will the benefits be achieved?

But, even if we had the answers to all these questions, how can they be integrated in the harm–benefit analysis? Does this mean that there should be no research on orphan diseases, because only few people can benefit? Does it mean that it matters who benefits in terms of age? Does this mean that a new cold remedy is more important than a new cancer treatment, since many more people will use it and it therefore has a greater impact? Is research more important, if it will bring about practical benefits sooner, and should this influence the harm–benefit analysis?

Similar questions arise on the harm side. Is the severity classification enough? Shall we add all harms done to individual animals, and if so, how do we deal with harms that are related to the project indirectly, such as harm to animals that were necessary to establish the particular mouse strain used? Is the absolute number of animals used in an experiment something that should be taken into account — or is it acceptable to adhere to the Three Rs criterion of reduction, and to use the minimum number of animals? And if the absolute number should count, what is a ‘high’ number (100 dogs, or 12,000 mice?) and does ‘high’ vary, depending on the research field in question? And if we knew all that and more, how could we bring all these criteria into one methodology, in order to carry out a transparent harm–benefit analysis? At the moment, this seems to be a mission impossible, rather than a challenge to be dealt with.

Steps to tackle the problem: Criteria, methodologies and committees

One could of course go on and on with this list of open questions. In order to answer at least some of them, researchers from various fields — and in particular, ethicists — try to take on the challenge. For example, the Messerli Research Institute (Vienna, Austria) hosted an international symposium in March 2013, in order to discuss possible steps toward overcoming the aforementioned problems. A conference on the harm–benefit analysis was also held in Bergen in 2014, and we debated the issues at the World Congress on Alternatives and Animal Use in the Life Sciences, held in Prague in late 2014. Many well-known experts in the field of ethical evaluation of animal experiments took part. At all these meetings, the aim was to bring together state-of-the-art knowledge with regard to the issues. For example, in Vienna, 22 speakers from eight European countries and the USA discussed their experiences and the current situation surrounding these issues in their respective countries. The lively discussions went to show that many challenges remain, but some issues can be solved.

In the course of the Vienna symposium, not only the criteria and aspects that should go into the harm–benefit analyses were debated. Importantly, different methodologies such as checklists, scoring systems or comparative methodologies, were also introduced. Most of the experts emphasised the importance of independent and well-balanced committees, and the integration of lay people (i.e. non-specialists) and representatives of animal welfare organisations into these committees. Taking into account the lay people’s perspectives and current public opinion would contribute to an up-to-date ethical evaluation of animal experiments. But, by taking all of these factors into account, are we any way nearer solving the problem of how to transparently weigh apples against oranges successfully?

Although no ‘super-theory’ to resolve all of the issues was identified, the challenges became much clearer. Furthermore, things to be avoided came to the table: A particular and major threat that has to be avoided when developing methodologies for the harm–benefit analysis became very clear, and that is over-bureaucratisation. Any methodology for the harm–benefit analysis has to be a user-friendly tool that leads to deeper reflection on individual animal experiments. The different forms of methodologies were summarised in the following three groups:

— comparative methodologies that use positive lists (white-lists) and negative lists (black-lists) of animal experiments, in order to evaluate the project in question;
— scoring strategies that quantify the extent to which relevant criteria are met, and that provide an algorithm for calculating the harms and benefits of projects; and
— check lists that provide binary (yes/no) evaluation methods, e.g. in the form of decision trees.

Whether these methods are used within or without the committee structure makes a big difference, and both scenarios are indeed possible. Ideally, applicants should follow a structured procedure and provide the relevant information according to a set of clear standards and criteria that have to be met. Interdisciplinary committees would then be able to evaluate the projects according to the same standards and criteria. These evaluations could inform the competent authority’s decisions.

Many things could be said, and indeed have been said in the past, about methodologies, and a great deal has also been written on the subject. Needless to say, we did not come to any final conclusions at the symposium in Vienna, nor in Prague, nor in Bergen, on this complex but vital matter.

How to proceed from here?

In order to reach a clearer vision of how the harm–benefit analysis can be brought into a feasible methodology, any ideas are welcome. Exchanging ideas and arguments might inspire and boost the debate. This short article serves as an open invitation to all interested experts in the field to start such a debate. Since this should happen in a focused way, the following topics might be useful to guide the discussion:

Committees and their limitations and advantages: A great number of EU Member States have established local and national committees to support the authorities in decision-making on submitted proposals. Certainly, such committees have the advantage of bringing skilled experts from the sciences, statisticians, representatives of animal protection groups and lay people, to work together in order to formulate a statement on harms and expected benefits. However, these committees often work without explicit methodology or criteria. So the question arises as to how they can safeguard transparent and non-arbitrary decision-making when they carry out harm–benefit analyses. I am sure that many of the readers are experienced members of such committees, and it would be very useful, if they would contribute with their experience, knowledge and ideas.
Methodologies: It would be of great interest to share knowledge on the advantages and disadvantages of methods used. If committees and the national authorities apply consistent methods and explicit criteria, it would be of utmost importance to get into an exchange of views and experience on whether and how such methods can support and improve the decision-making process.
Ethics and Law: A third question relates to the terminology used in the Directive. If ethical considerations should be taken into account, should these considerations exceed existing law or is “ethics” to be understood within legal limits (and is not allowed to exceed existing law)? Here, ethics runs the risk of contradicting the principle of legality in constitutional states. In other words: How is the term “taking ethical considerations into account” interpreted in different countries. It would be great to get some insight into this.
Experience from the past: Generally, since many countries have carried out harm–benefit analyses in the past, knowledge of their experiences could contribute to future developments.
Ideas for the future: Finally, a possible thought experiment is to think about where we are going to be in 20 years’ time. How will the debate look in 2035? Will we still be trying to weigh apples against oranges?

These questions and statements aim to initiate a debate that is relevant to all EU Member States and everybody involved in animal research. It would be very useful, if experts in this forum were willing to find some time to contribute to a lively and future oriented discussion, in order to solve at least some of the open questions mentioned above. The idea is to continue to build up knowledge on the process of harm–benefit analysis in animal research, and maybe improve the situation for both animals and researchers. Perhaps this forum could bring us closer to the point where researchers were able to respond to the question as to whether, and indeed which, projects involving live animals are justifiable, and which are not. Being able to respond to the question as to whether a project is worth carrying out or not, could demonstrate that scientists are able to take on this responsibility in a knowledge-based society and thus can contribute to ethical welfare.

Prof. Dr Herwig Grimm
Messerli Research Institute
Veterinary University of Vienna,
Medical University Vienna,
and University of Vienna
Veterinärplatz 1
1210 Vienna
Austria
E-mail: herwig.grimm@vetmeduni.ac.at

References
1 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, 22.10.2010, 33–79.
2 Anon. (2013). National Competent Authorities for the Implementation of Directive 2010/63/EU on the Protection of Animals Used for Scientific Purposes. Working Document on Project Evaluation and Retrospective Assessment, 42pp. Brussels, Belgium: European Commission.

Download a pdf of the article here: Discussion Grimm.

The Three Rs: The Way Forward

Michael Balls

It is now 20 years since Russell and Burch
last met to discuss, with others,
the way forward for the Three Rs concept

 

Sitting at my computer, faced with the challenge of deciding what I could say in Wisdom 16, I suddenly realised that it was almost 20 years, to the day, since Bill Russell and Rex Burch met in Sheringham, a small seaside town in Norfolk, UK, for the first scientific meeting they had attended together since the publication of The Principles in 1959 (Figure 1). Sadly, it was also to be the last such meeting, as Rex died a few months later.

Rex Burch and Bill Russell 31 May 1995 and Ecvam Workshop report

 

The meeting took place on 31 May to 3 June 1995, in the form of an e, which I chaired in partnership with Alan Goldberg of CAAT. Our reason for being in Sheringham was that Rex was too ill to go more than a few miles from home, so Alan and I decided that we would invite some of those committed to the Three Rs, to travel to meet him. The other participants included Claire Russell, and some of our colleagues from Germany, Italy, The Netherlands, the UK and the USA (Figure 2).

Workshop participants May 1995

 

An opening ceremony was held in the council chamber of Sheringham Town Hall, where Rex had rented space for his microbiology laboratory since the early 1970s. All the participants made a brief statement, and Bill sang a song, as he always did on special occasions. These opening proceedings were recorded on videotape. The rest of the workshop took place at the Links Country House Hotel in West Runton, about a mile from where I now live.

The principal aims of the workshop were to discuss the current status of the Three Rs and to make recommendations aimed at achieving greater acceptance of the concept of humane experimental technique, and, in the interests of both scientific excellence and the highest standards of animal welfare, the more active implementation of reduction alternatives, refinement alternatives and replacement alternatives.

The report of the workshop was published in the November/December 1995 issue of ATLA.1 It reviewed the origins and evolution of the Three Rs concept as originally outlined in The Principles,2 the selection of appropriate animal species, reduction alternatives, refinement alternatives and replacement alternatives, education and training, and certain special considerations (vaccines and immunobiologicals, transgenic animals, special protection for selected animals, benefit and suffering, and the setting of targets). It concluded with 58 conclusions and recommendations, which were preceded by the following remarks: The workshop participants unanimously reaffirmed the principles put forward by Russell & Burch, that humane science is good science and that this is best achieved by vigorous application of the Three Rs: reduction alternatives, refinement alternatives and replacement alternatives.

Thus, the only acceptable animal experiment is one which uses the smallest possible number of animals and causes the least possible pain or distress which is consistent with the achievement of a justifiable scientific purpose, and which is necessary because there is no other way of achieving that purpose. Any proposed experiments on animals should be subjected to prior and effective expert review by an ethics committee or an equivalent body. The Three Rs should be seen as a challenge and as an opportunity for reaping benefits of every kind — scientific, economic and humanitarian — not as a threat.

Many of the conclusions and recommendations are as relevant today as they were in 1995. The workshop was a memorable occasion in many other ways. It was run according to the ECVAM tradition — five days of hard work, interspersed with good food and good wine, with a determination to have the words of detailed conclusions and recommendations down on paper by the end. One of my lasting memories will be witnessing the pleasure shown by Bill and Rex, as they took the opportunity to sit and talk quietly together after a gap of more than 30 years. All I have to show of that is one out-of-focus photograph, taken during the final reception and dinner at Blickling Hall, on 2 June 1995 (Figure 3).

Bill Russell and Rex Burch 2 June 1995

Many developments of many kinds have taken place since 1995, but, as Roman Kolar spells out in the latest issue of ATLA,3 there is still much to be achieved, if the aims of the workshop and of The Three Rs Declaration of Bologna,4 to which it led, are to be achieved, resulting in the revolution in biomedical research and its application which was proposed in The Principles. For my own part, I am concerned that stating allegiance to the Three Rs concept has become a convenient smokescreen, to which lip service can be paid, whilst little is actually permitted to change. I have therefore proposed that the focus should now be more squarely on humane science, which avoids the problem of seeming conflicts between human benefit and animal welfare,5 and I am rather confident that Bill and Rex would approve.

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 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, D.W., Yager, J.D., Zurlo, J. & van Zutphen, B.F.M. (1995). The Three Rs: The way forward. The report and recommendations of ECVAM Workshop 11. ATLA 23, 838-866.
2 Russell, W.M.S. & Burch, R.L. (1959). The Principles of Humane Experimental Technique, xiv + 238pp. London, UK: Methuen.
3 Kolar, R. (2015). How long must they suffer? Success and failure of our efforts to end the animal tragedy in laboratories. ATLA 43, 129-143.
4 Anon. (2000). The Three Rs Declaration of Bologna. ATLA 28, 1-5.
5 Balls, M. (2014). Animal experimentation and alternatives: Time to say goodbye to the Three Rs and hello to humanity? ATLA 42, 27-333.

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.

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Wisdom 16

In Vitro Methodologies in Ecotoxicological Hazard Assessment: The Case of Bioaccumulation Testing for Fish

Helmut Segner

The concerted research efforts undertaken in recent years have
highlighted the potential of in vitro approaches, as part of an
integrated testing strategy, to replace or reduce in vivo bioaccumulation testing in fish

Worldwide programmes for the regulation of chemicals require an assessment of the risks of chemicals to human and environmental health based on three categories of concern: Persistence, Bioaccumulation and Toxicity (PBT). Among these three categories, bioaccumulation refers to the enrichment of environmental chemicals in organisms. It encompasses the absorption, distribution, metabolism and excretion (ADME) of a chemical inside an organism, and ultimately determines the internal toxic dose. For the aquatic environment, the most widely used parameter to estimate the bioaccumulation potential of a chemical is the so-called Bioconcentration Factor (BCF). The BCF represents the ratio of the steadystate chemical concentration in the organism and the chemical concentration in the respiratory medium, i.e. water. For the experimental determination of the BCF, the test procedure as described in OECD Test Guideline 3051 represents the current ‘gold standard’. In this test, fish are exposed to a chemical for 28 days, to reach an equilibrium of chemical concentration between fish and water, followed by a 28-day depuration period to measure the elimination rate. This test, in addition to being lengthy and costly, requires a high number of animals (> 100 fish per test).

Regulatory programmes require bioaccumulation information for chemicals which are lipophilic (for example, those with a log Kow > 3), and which are produced at a certain tonnage (for example, the European Community REACH Regulation requires BCF information for lipophilic chemicals that are produced at > 100 tonnes per year). Experi mentally determined BCF data are not available for the vast majority of existing compounds. For instance,  in a Canadian investigation of 23,000 existing chemicals, it was found that bioaccumulation data existed for less than 4% of them (cf. Nichols et al.2). If the missing BCF data had to be generated by means of the OECD 305 test, this would entail a drastic increase in animal use.3,4 Therefore, there is an urgent need to develop alternative methods to reduce the number of fish used for in vivo bioaccumulation testing. The bioconcentration of chemicals in fish results from the competing rates of chemical uptake via the gills and skin (k1) and chemical elimination via respiratory exchange (k2), faecal egestion (ke) and metabolic biotransformation (km).5 In addition, dilution as a result of growth (kd) can influence bioconcentration.

With the involvement of these different processes, it is clear that non-animal approaches to bioconcentration assessment cannot be based on one single method, but have to rely on an array of methodologies.2,4,6 An initial non-animal based approximation of the bioconcentration potential of an organic chemical in aquatic organisms can be obtained from an in silico hydrophobicity model, which considers bioconcentration as a passive partitioning process resulting from the competing uptake and elimination processes. In this model, bioconcentration can be predicted from the lipophilicity of a chemical, as estimated from its octanol–water partition coefficient, Kow.5 Also, it can actually be measured by using artificial membranes which simulate the passive diffusion processes across the respiratory epithelia.7

The development of in vitro methods

Diffusion-based methodologies have proven instrumental in the prediction of the BCF values of lipophilic chemicals that undergo no endogenous metabolism in the organism. However, as they are not able to take into account chemical loss due to biotransformation (km), they overestimate the BCF values of metabolisable xenobiotics. To correct for the influence of biotransformation on fish BCF values, a possible approach is the use of metabolically competent in vitro assays that show which chemicals are biotransformed, and at what rates. In mammalian toxicology, in vitro assays for the analysis of xenobiotic metabolism largely rely on liver preparations such as subcellular liver fractions (S9, microsomes) and isolated hepatocytes, as the liver is the organ with the highest metabolic activity. Corresponding technologies are also available for fish, and it has been demonstrated that they are suitable for determining biotransformation parameters (see Segner & Cravedi8 and Fitzimmons et al.9). However, their reproducibility and their capability of predicting in vivo BCF values remain to be demonstrated.

In recent years, intensive efforts have been undertaken — largely coordinated by the Health and Environmental Sciences Institute (HESI) — to advance the development of piscine in vitro assays for regulatory purposes. After an initial phase of reviewing the available knowledge and technologies,2,6 in the next step, standardised protocols for liver S9 preparations and isolated hepatocytes from rainbow trout were established.10,11 A major drawback experienced in these studies, particularly with freshly-isolated hepatocyte suspensions, was the between-isolate variability of metabolic capabilities, which is related to factors such as seasonal oscillation, and the nutritional status, gender or genetic background of the donor fishes. Here, a major step forward was the introduction of a cryopreservation method for fish hepatocytes,12 enabling the year-round provision of uniform batches of metabolically characterised hepatocytes to laboratories worldwide. By using a standardised assay protocol, Fay et al.13 recently performed an international ring study with cryopreserved rainbow trout hepatocytes, and were able to demonstrate the good interlaboratory and intra-laboratory reproducibility of the metabolic rate values obtained with the in vitro hepatocyte assay.

To be able to extrapolate from the metabolic rate values measured in the isolated fish hepatocytes to the metabolic rate value (km) in the intact fish, physiologically-based prediction models were developed. 14,15 These models initially scale from the clearance rate of the isolated liver cells to that of the whole liver, and from there to the metabolic transformation rate of the whole fish. The predicted km values are then used to calculate the in vivo BCF value of the test chemical. Currently, the availability of data on BCF values  predicted from in vitro assays is still limited, and it is still too early to come up with a conclusive statement on the predictability of the in vitro approach, partly also because of the variable quality of the in vivo BCF data; however, the existing results look promising.

Looking to the future

There are lessons to be learned from the recent development of in vitro assays as components of alternative integrated testing strategies for the assessment of bioaccumulation in fish. Although a broad spectrum of in vitro assays and methods have been available in ecotoxicology for a while,16 they have never made their way to regulatory implementation. Partly, this is due to the fact that they were considered to be technically not ready nor sufficiently standardised. In the case of the piscine in vitro metabolism assays, this obstacle has been overcome through targeted and internationally concerted research efforts on the standardisation and harmonisation of the assay protocols. Another constraint to the regulatory acceptance of in vitro assays is that they were considered not to be appropriate for the protection goals of ecotoxicology, which are ecological entities such as populations and communities. However, ecotoxicological hazard assessment largely relies on classical toxicity tests for measuring organism-level endpoints such as lethality (cf. Segner17), and these endpoints may well be predictable by in vitro assays, provided that: a) the in vitro assays are rationally selected to represent the critical toxicological processes; b) the assays are standardised; and c) valid extrapolation models are available. These requirements are fulfilled in the case of bioaccumulation assessment — i.e. the in vitro assays measure biotransformation as the critical toxicokinetic process, they are standardised, and there exist physiologically-based models for the scaling of the in vitro metabolic rate values to the in vivo metabolic rates. As ecotoxicology deals with a huge diversity of species, the interspecies scaling of metabolic rates is another critical issue, but this question is also currently under investigation. In conclusion, the concerted research efforts undertaken in recent years have substantially moved the field ahead, and the results obtained highlight the potential of in vitro approaches, as part of an integrated testing strategy,4 to replace or reduce in vivo bioaccumulation testing in fish.

Acknowledgements

The financial support of Stiftung Forschung 3R, ünsingen (Switzerland) and the Health and Environmental Sciences Institute (HESI) is gratefully acknowledged.

Prof. Dr Helmut Segner
Centre for Fish and Wildlife Health
Department of Infectious Diseases and Pathobiology
Vetsuisse Faculty
University of Bern
PO Box 8466
CH 3012 Bern
Switzerland
E-mail: Helmut.segner@vetsuisse.unibe.ch

References

1 OECD (2011). OECD Guideline for Testing of Chemicals No. 305. Bioaccumulation in Fish: Aqueous and Dietary Exposure, 72pp. Paris, France: Organisation for Economic Co-operation & Development.
2 Nichols, J.S., Erhardt, S., Dyer, M.J., Moore, M., Plotzke, K., Segner, H., Schultz, I., Thomas, K., Vasiluk, J. & Weisbrod, A. (2007). Use of in vitro Absorption, Distribution, Metabolism, and Excretion (ADME) data in bioaccumulation assessments for fish. Human & Ecological Risk Assessment 13, 1164–1191.
3 de Wolf, W., Comber, M., Douben, P., Gimeno, S., Holt, M., Léonard, M., Lillicrap, A., Sijm, D., van Egmond, R., Weisbrod, A., & Whale, G. (2007). Animal use replacement, reduction,  and refinement: Development of an integrated testing strategy for bioconcentration of chemicals in fish. Integrated Environmental Assessment & Management 3, 3–17.
4 Lombardo, A., Roncaglioni, A., Benfenati, E., Nendza, M., Segner, H., Fernández, A., Kühne, R., Franco, A., Pauné, E. & Schüürmann, G. (2014). Integrated testing strategy (ITS) for bioaccumulation assessment under REACH. Environment International 69, 40–50.
5 Arnot, J.A. & Gobas, F. (2006). A review of bioconcentration factor (BCF) and bioaccumulation factor (BAF) assessments for organic chemicals in aquatic organisms. Environmental Reviews 14, 257–330.
6 Weisbrod, A.V., Sahi, J., Segner, H., James, M.O., Nichols, J., Schultz, I., Erhardt, S.,  Cowan-Ellsberry, C., Bonnell, M. & Hoeger, B. (2009). The state of   science for use in bioaccumulation assessments for fish. Environmental Toxicology & Chemistry 28, 86–96.
7 Kwon, J.H. & Escher, B.I. (2008). A modified parallel artificial membrane permeability assay for evaluating bioconcentration of highly hydrophobic chemicals in fish. Environmental Science & Technology 42, 1787–1793.
8 Segner, H. & Cravedi, J.P. (2001). Metabolic activity in primary cultures of fish hepatocytes. ATLA 29, 251–257.
9 Fitzsimmons, P.N., Lien, G.J. & Nichols, J.W. (2007). A compilation of in vitro rate and affinity values for xenobiotic biotransformation in fish, measured under physiological conditions. Comparative Biochemistry & Physiology 145C, 485–506.
10 Han, X., Nabb, D., Mingoia, R. & Yang, C. (2007). Determination of xenobiotic intrinsic clearance in freshly isolated hepatocytes from rainbow trout (Oncorhynchus mykiss) and rat and its application in bioaccumulation assessment. Environmental Science & Technology 41, 3269–3276.
11 Johanning, K., Hancock, G., Escher, B., Adekola, A., Bernhard, M.J., Cowan-Ellsberry, C., Domodoradzki, J., Dyer, S., Eickhoff, C., Embry, M., Erhardt, S., Fitzsimmons, P., Halder, M., Hill, J., Holden, D., Johnson, R., Rutishauser, S., Segner, H., Schultz, I. & Nichols, J. (2012). Assessment of metabolic stability using the rainbow trout (Oncorhynchus mykiss) liver S9 fraction. Current Protocols in Toxicology 53, 14.10.1–14.10.28.
12 Mingoia, R.T., Glover, K.P., Nabb, D.L., Yang, C.H., Snajdr, S.I. & Han, X. (2010). Cryopreserved hepatocytes from rainbow trout (Oncorhynchus mykiss): A validation study to support their application in bioaccumulation assessment. Environmental Science & Technology 44, 3052–3058.
13 Fay, K.A., Mingoia, R.T., Goeritz, I., Nabb, D.L., Hoffman, A.D., Ferell, B.D., Peterson, H.M., Nichols, J.W., Segner, H. & Han, X. (2014). Intra- and inter-laboratory reliability of a cryopreserved trout hepatocyte assay for the prediction of chemical bioaccumulation potential. Environmental Science & Technology 48, 8170–8178.
14 Nichols, J.W., Schultz, R.I. & Fitzsimmons, P.N. (2006). In vitro–in vivo extrapolation of quantitative hepatic biotransformation data for fish. I. A review of methods, and strategies for incorporating intrinsic clearance estimates into chemical kinetic methods. Aquatic Toxicology 78, 74–90.
15 Cowan-Ellsberry, C.S., Dyer, S., Erhardt, S., Bernhard, M.J., Roe, A., Dowty, M. & Weisbrod, A. (2008). Approach for extrapolating in vitro metabolism data to refine bioconcentration factor estimates. Chemosphere 70, 1804–1817.
16 Castano, A., Bols, N.C., Braunbeck, T., Dierickx, P., Halder, M., Isomaa, B., Kawahara, K., Lee, L.E.J., Mothersill, C., Pärt, P., Repetto, G., Sintes, J.R., Rufli, H., Smith, R., Wood, C. & Segner, H. (2003). The use of fish cells in ecotoxicology. The report and recommendations of ECVAM workshop 47. ATLA 31, 317–351.
17 Segner, H. (2011). Moving beyond a descriptive aquatic toxicology: The value of biological process and trait information. Aquatic Toxicology 105, 50–55.

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Current Dilemmas Segner

Coffee in Class: An Alternative to Animal Experiments in Pharmacology?

Anoop Kumar Agarwal, Syed Ilyas Shehnaz, Razia Khanam and Mohamed Arifulla

The stimulant effect of coffee on psychomotor performance was
introduced as a potential alternative clinical pharmacology
experiment for medical and pharmacy students

Animal experiments have been designed and standardised to demonstrate the effects of certain drugs on body organs, as part of undergraduate health professional education. However, the logistics of animal availability, the expenses incurred, increasing awareness of concerns about animal welfare1–3 and the ‘Three Rs’ concept (i.e. Replacement, Refinement and Reduction),4 have often either reduced these experiments to tutor demonstrations or have resulted in their complete withdrawal from the undergraduate curriculum.5

As an alternative to satisfy the ethical concerns of animal rights activists, Computer Assisted Learning (CAL) was introduced.6–8 Although CAL is an effective means of fulfilling the educational objectives of laboratory sessions, the lack of hands-on experience with living tissues, the lack of practical experience to facilitate the future application of the procedures in research, as well as the absence of biological variation, are the major limitations of CAL.9

In view of the current scenario, we considered it necessary to investigate alternative exercises which would expose the students to experimental methodology with scientific explanation. Clinical pharmacology experiments, such as dosage calculations, rational drug selection, evaluation of drug information, and the analgesic effect of NSAIDs, have been used to supplement CAL.10 In an effort to identify alternatives to animal experiments and CAL at the undergraduate level, the Department of Pharmacology, Gulf Medical University, Ajman, United Arab Emirates, introduced a new experiment in the teaching curriculum of Bachelor of Medicine/Bachelor of Surgery (MBBS) and Pharm D (Doctor of Pharmacy) programmes. The aim of the experiment was to demonstrate the stimulant effect of coffee on psychomotor performance in students, by using simple paper and pencil tests, namely, the Six-Letter Cancellation Test (SLCT) and the Digit/Letter Substitution Test (DLST). These tests objectively assess the psychomotor functions of an individual, and integrate different mental functions, such as perception, recognition, integration and reaction, in the assigned task. These tests are not meant for assessing memory or intelligence. Since they are speed tests, performance is influenced by mental alertness, concentration and coordination abilities. Both tests consist of three sections: instructions, the key (target) letters, and the working-out part. In the SLCT, the subject identifies the key letters in the working-out part, whereas in the DLST, the numbers in the working-out part have to be substituted by the corresponding letters given in the key. The duration of each test is 90 seconds. In both the tests, the extent of the working-out part exceeds the potential for completion in the stipulated time. The maximum and minimum scores for these tests vary in different subgroups. Parallel worksheets (with a different key) are used on each occasion (Figure 1 and Figure 2), to nullify the effect of memory.11

Minimal materials are required

The equipment required for the experiment is readily available at low cost. It comprises three sets of parallel worksheets for the SLCT, three sets of parallel worksheets for the DLST, a stop watch, an office bell, and standard hot coffee (2g instant coffee/200ml).

Figure 1

The protocol

The experimental protocol is divided into five individual stages:

1. The practice session: The tutors familiarised themselves with the tests and planned the experiment to ensure smooth implementation. Ethical approval was granted from the Institutional Ethics Committee prior to the conduct of the experiment. The tests were administered during the pharmacology laboratory sessions to seven batches of 25–30 medical and pharmacy students. The students were told about the importance and relevance of the tests, and were issued with the instructions necessary for performing the tests. Written consent was obtained from those who volunteered to participate in the experiment. One practice session was organised, in order to familiarise the participants with both the tests.

2. The pre-coffee session: The worksheet for the SLCT were distributed, and the students were asked to write their names on the back of the sheet. This was fllowed by the first bell, indicating the beginning of the ‘working-out time’, which ended with a second bell after 90 seconds. The students were asked to start and stop immediately when the bell rang, and strict monitoring of time was ensured. The sheets were randomly exchanged among the students for score calculations. The second test, the DLST, was administered in a similar manner, after an interval of 5 minutes.

3. The post-coffee session: A 20ml cup of standard coffee was served to each student. After 20 minutes, the two tests were re-administered as before, with parallel worksheets, i.e. a new key was used in each session, to nullify the effect of memory.

4. Interpretation of scores: The students were asked to record the scores of the two tests in the three sessions (practice, pre-coffee and post-coffee) in their record books, and to draw conclusions based on a pharmacological explanation. The mean scores of both the tests for the practice, precoffee and post-coffee sessions were expressed as the mean ± standard deviation (SD). Comparison of scores was done by using the Wilcoxon signed-rank test. The significance level was set at 0.05.

5. Student feedback: Feedback on the experiment was obtained by using a structured, content-validated and pre-tested questionnaire on a five-point Likert-like scale (strongly agree to strongly disagree). The statements enquired about the ease of understanding and performing of the tests, the appropriateness of the time allowed and the methodology, the understanding of the concepts and the generation of links between theory and actual effects of CNS stimulants, the interest levels generated, and the willingness to perform similar experiments in the future. In addition, open-ended responses about the students’ experiences were also encouraged. Voluntary participation was emphasised, and full confidentiality of the data was ensured to all the
participants.

Agarwal table 1Figure 2

Results

Of a total of 180 medical and pharmacy 162 participated in the experiment (response rate 90%). The mean scores (± SD) of the SLCT for the practice, pre-coffee and post-coffee sessions are given in Table 1. Although there were no significant differences between the practice and pre-coffee session scores for both the tests, a statistically significant difference (p < 0.05) was observed between the pre-coffee and the post-coffee scores. Furthermore, a small number of students obtained a low score (5%), or had no change in their scores after coffee intake (6%).

The students gave a very good feedback on the experiment, as reflected in the questionnaire (Table 2). However, in the free responses, a few students (12%) reported that they lost interest by the third session, due to the simplicity and repetitive nature of the test, and that, as a result, they felt that they did not perform at their best.

Table 2

Discussion

We have endeavoured to introduce a clinical pharmacology experiment as an alternative to CAL and animal experiments, for use in undergraduate health professional education. This experiment has very few requirements, and can be easily performed within laboratory session time-frames. The experiment also reinforces the concept of the mild stimulant effect of coffee. However, test performance is affected by various factors, including motivation, understanding, interest, mood, environment, quality of the worksheet, and personality type, so scores obtained may vary for different groups and sub-groups. Moreover, a clear understanding of the principle of the paper and pencil test among the faculty is important, in order to plan the experiment in an organised manner. Although these tests had been used earlier as a teaching tool among medical students, the perceptions of the students had not been obtained.11 The experiment was reintroduced in our setting with certain modifications, and student feedback was obtained with regard to its acceptability and relevance. This experiment had been introduced for the first time in the Pharm D curriculum to demonstrate the effects of the drug.

The test scores of both the tests (SLCT and DLST) showed a significant increase in psychomotor performance after coffee intake, suggesting a stimulant effect. However, a number of sources of variation were identified: differences in quantity of coffee consumed, loss of interest due to repetitive nature of the test, and anxiety after coffee intake. The latter is a known side-effect of coffee in certain individuals. Students who did not show an increase in score stated that they frequently consumed coffee during the day.

The student feedback revealed that the majority found the experiment interesting and informative. This could probably motivate them to learn more about the drugs and their effects. However, in the open-ended responses, a few students felt that the tests were too simple and that a higher degree of complexity was necessary to keep up their interest.

Author for correspondence:
Dr Syed Ilyas Shehnaz
Department of Pharmacology
Gulf Medical University
PO Box 4184
Ajman
United Arab Emirates
E-mail : shehnazilyas@yahoo.com

References

1 Gitanjali, B. (2001). Animal experimentation in teaching: Time to sing a swan song. Indian Journal of Pharmacology 33, 71.
2 Solanki, D. (2010). Unnecessary and cruel use of animals for medical undergraduate training in India. Journal of Pharmacology & Pharmacotherapeutics 1,59.
3 Desai, M. (2009). Changing face of pharmacology practicals for medical undergraduates. Indian Journal of Pharmacology 41, 151–152.
4 Guhad, F. (2005). Introduction to the 3Rs (refinement, reduction and replacement). Contemporary Topics in Laboratory Animal Science 4, 58-59.
5 Setalvad, A.R.N. (2009). Medical Council of India, New Delhi, Amendment Notification of 8 July 2009 to the Minimal Standard Requirements for Medical Colleges with 150 Admissions Annually, Regulations 1999. Available at: http://www.mciindia.org/helpdesk/how_to_start/STANDARD%20FOR%20150.pdf (Accessed
01.08.14).
6 Dewhurst, D. (2004). Computer-based alternatives to using animals in teaching physiology and pharmacology to undergraduate students. ATLA 32, 517–520.
7 Badyal, D.K., Modgill, V. & Kaur, J. (2009). Computer simulation models are implementable as replacements for animal experiments. ATLA 37, 191–195.
8 Wang, L. (2001). Computer-simulated pharmacology experiments for undergraduate pharmacy students: Experience from an Australian University. Indian Journal of Pharmacology 33, 280–282.
9 Kuruvilla, A., Ramalingam, S., Bose, A.C., Shastri, G.V., Bhuvaneswari, K. & Amudha, G. (2001). Use of computer assisted learning as an adjuvant to practical pharmacology teaching: Advantages and limitations. Indian Journal of Pharmacology 33, 272–275.
10 Gitanjali, B. & Shashindran, C.H. (2006). Curriculum in clinical pharmacology for medical undergraduates of India. Indian Journal of Pharmacology 38, Suppl., 108–114.
11 Natu, M.V. & Agarwal, A.K. (1997). Testing of stimulant effects of coffee on the psychomotor performance: An exercise in clinical pharmacology. Indian Journal of Pharmacology 29, 11–14.

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Discussion Agarwal

Likelihood Ratios in Assessing the Safety of New Medicines

Robert A. Coleman

Until we know the true predictive value of animal-based methods
for predicting clinical safety issues, it is impossible to assess
the advantage or otherwise of non-animal based approaches

The use of animals in the discovery and development of new medicines has generated debate for decades. For much of this time, contrasting views have been primarily polarised on the basis of practicality versus ethics, with proponents arguing that for the development of new medicines to treat human disease, the end justifies the means, while the opponents’
chief objection has been the associated animal suffering. Commercial and public health pressures over the years have ensured that those in the ‘practicality camp’ have held sway.

More recently, however, the issue has become increasingly complex, with growing concerns that, irrespective of ethical considerations, data generated in animal (i.e. non-human) models are not necessarily or sufficiently relevant to human patients.1–3 There is now general consensus that inter-species variability is a real issue, and that animal models are far from perfect for the purpose of ensuring either the efficacy or the safety of potential new medicines intended for human subjects. Supporters of the continued use of animals argue that, while they do not provide an absolute indication of either efficacy or safety, in the absence of any other approach, one that is somewhat unreliable is better than none at all.

Such an argument has some merit, if indeed it is valid. However, in this field, all may not be entirely as it seems. Firstly, human-based in vitro and in silico alternatives are becoming ever more sophisticated,4 thus overcoming many of the criticisms originally directed toward them. For example, it has long been held that it would be impossible to model the complexity of the intact patient through a study of isolated cells and tissues, and while this problem may never be wholly overcome, the gap gets ever smaller. Secondly, it is important to understand that we really don’t know how good existing animal-based methods are. In the field of efficacy, there is a wealth of evidence that results obtained by using experimental animals can be hugely misleading.5–11 Here, we have the advantage that drugs that promise efficacy in patients on the basis of animal data can advance into clinical testing, and their utility can be directly assessed. For the majority of these drugs, the clinical outcome has been disappointing. With safety, the issue is different, as drugs with identified safety issues in animals will seldom, if ever, advance to clinical testing, thus the relevance of the animal data to safety in humans may never be determined. However, what we do know is that many drugs identified as safe in pre-clinical profiling eventually prove to cause serious and use-limiting side effects in human subjects. 12 The key question is, “Could such failures have been avoided, had we relied on human-based test methods?” Until we know how frequently non-animal methods could have identified safety issues that were missed by animal tests, it is impossible to assess the advantage or otherwise of those methods. It is a fact that, despite the continued use of animals as human surrogates in pharmaceutical research, there has never been a solid, published, peer-reviewed study demonstrating fitness for purpose, whereas reviews identifying the shortcomings are abundant.

Assessing the value of animal studies

It is for this reason that any information that sheds light on the actual value of animal-based testing for its intended purpose is of inestimable worth. Until recently, much evidence, while valuable, has been indirect. For example, a recent telling study demonstrated that pre-clinical fast-tracking (i.e. abbreviated safety testing) of potential new medicines resulted in no increase in the proportion of candidates that subsequently proved toxic in human subjects.
13 In another study, the ability of animal studies to detect serious post-marketing adverse events was demonstrably poor.14 While such reports add to the volume of data providing witness to the shortcomings of animal-based approaches to ensuring clinical safety, they do not provide a robustly measurable metric of predictive efficiency. In view of the colossal amount of data generated over the years in pre-clinical safety studies on thousands of new potential medicines, many of which have progressed to clinical testing and even to market, it is amazing that, until recently, no comprehensive analysis of such data has been applied in order to explore the value of the current approach to safety testing.

Likelihood ratios

In the light of such a background, it is of considerable significance that serious attempts are now being made to extract intelligence from the wealth of information available in publicly accessible sources, in order to shed more light on the actual predictive power of animal-based safety testing. A particular example is the utilisation of the Safety Intelligence Programme (SIP),15 which overcomes semantic issues to extract valid information from all available data sources. SIP has been used, for example, to explore the predictive power of animal models for the detection of liver toxicity associated with a wide range of human medicines,16 highlighting the highly variable efficiency of different models in combination with different drug toxicities. More recently, SIP has been used to particular effect in two studies that have explored directly the value of dogs, mice, rats and rabbits in predicting safety issues in human subjects.17,18 While most previous studies have relied on determining ‘concordance’ between animal and human data, that tells only a part of the story, and is too simplistic a measure to be of much real value. Its
problem is that it only deals with positive correlation, i.e. the frequency that toxicity in experimental animals and in human subjects coincide, ignoring the issue of true prediction. What is needed is a determination of likelihood ratios (LRs),19 both positive (PLRs) and negative (NLRs), to gain a more complete picture. What emerged when LRs were determined was that, although there was indeed some measure of concordance between positive toxicity data between animals and humans, in terms of LRs, none of the species proved to offer any useful level of real predictive power. Although the studies and their conclusions did not escape criticism from some quarters,20 the suggested limitations, real or perceived, are arguably irrelevant to its overall validity.21

What did emerge from the application of this approach were absolute values for both PLRs and NLRs for a wide range of specific drugs. The importance of this is that, for the first time, such measures can provide a robust yardstick against which to evaluate the relative merits of alternative approaches to toxicity testing.

Conclusions

To summarise, the use of more-rigorous approaches to the evaluation of animal models as predictors of the likelihood that any chemical will be similarly toxic or non-toxic in human subjects provides not only a realistic measure of their actual fitness for purpose, but crucially, also a basis by which the efficiency of other, ideally human-based, approaches can be evaluated through their exposure to the same range of drugs. The use of the same drugs ensures that any criticisms related to potential bias, or other potentially confounding factors, are negated. Such a prospective study would be of inestimable value. Dare we hope that the government and pharmaceutical companies will take up the challenge and fund
such a study?

Dr Robert A. Coleman
Independent Consultant
UK
E-mail: robt.coleman@btinternet.co
m

References

1 Wall, R.J. & Shani, M. (2008). Are animal models as good as we think? Theriogenology 69, 2–9.
2 Hartung, T. (2013). Food for Thought… Look back in anger — What clinical studies tell us about preclinical work. ALTEX 30, 275–291.
3 Pippin, J.J. & Sullivan, K. (undated). Dangerous medicine:
Examples of animal-based “safety” tests gone wrong. Washington, DC, USA: The Physicians Committee for Responsible Medicine. Available at: http://www.pcrm.org/research/animaltestalt/animal testing/dangerous-medicine-examples-of-animalbased-tests (Accessed 03.11.14).
4 Ashton, R., De Wever, B., Fuchs, H.W., Gaça, M., Hill, E., Krul, C., Poth, A. & Roggen, E.L. (2014). State of the art on alternative methods to animal testing from an industrial point of view: Ready for regulation? ALTEX 31, 357–363.
5 Pound, P., Ebrahim, S., Sandercock, P., Bracken, M.B. & Roberts, I. (2004). Where is the evidence that animal research benefits humans? British Medical Journal 328, 514-517.
6 Kaste, M. (2005). Use of animal models has not contributed
to development of acute stroke therapies: Pro. Stroke 36, 2323–2324.
7 Pippin, J.J. (2005). The Need for Revision of Pre-Market Testing: The Failure of Animal Tests of COX-2 Inhibitors, 23pp. Washington, DC, USA: The Physicians Committee for Responsible Medicine. Available at: pcrm.org/pdfs/research/testing/exp/COX2Report.pdf
(Accessed 04.11.14).
8 Hackam, D.G. & Redelmeier, D.A. (2006). Translation of research evidence from animals to humans. Journal of the American Medical Association 296, 1731–1732.
9 Knight, A. (2008). Systematic reviews of animal experiments
demonstrate poor contributions towards human healthcare. Reviews on Recent Clinical Trials 3, 89–96.
10 Matthews, R.A.J. (2008). Medical progress depends on animal models — doesn’t it? Journal of the Royal Society of Medicine 101, 95–98.
11 Seok, J., Warren, H.S., Cuenca, A.G., Mindrinos, M.N., Baker, H.V., Xu, W., Richards, D.R., McDonald-Smith, G.P., Gao, H., Hennessy, L., Finnerty, C.C., López, C.M., Honari, S., Moore, E.E., Minei, J.P., Cuschieri, J., Bankey, P.E., Johnson, J.L., Sperry, J., Nathens, A.B., Billiar, T.R., West, M.A., Jeschke, M.G., Klein, M.B., Gamelli, R.L., Gibran, N.S., Brownstein, B.H., Miller-Graziano, C., Calvano, S.E., Mason, P.H., Cobb, J.P., Rahme, L.G., Lowry, S.F., Maier, R.V., Moldawer, L.L., Herndon, D.N., Davis, R.W., Xiao, W., Tompkins, R.G. & Inflammation and Host Response to Injury, Large Scale Collaborative Research Program (2013). Genomic responses in mouse models poorly mimic human inflammatory diseases. Proceedings of the National Academy of Sciences of the USA 110, 3507–3512.
12 Li, A.P. (2004). Accurate prediction of human drug toxicity: A major challenge in drug development. Chemico-biological Interactions 150, 3–7. 13 Arnardottir, A.H., Haaijer-Ruskamp, F.M., Straus, S.M., Eichler, H.G., de Graeff, P.A. & Mol, P.G. (2011). Additional safety risk to exceptionally approved drugs in Europe? British Journal of Clinical Pharmacology 72, 490–499.
14 van Meer, P.J., Kooijman, M., Gispen-de Wied, C.C., Moors, E.H. & Schellekens, H. (2012). The ability of animal studies to detect serious post marketing adverse events is limited. Regulatory Toxicology & Pharmacology 64, 345–349.
15 Berry, S. (2012). Safety Intelligence Program Provides Insight into Drug-Induced Cardiac Effects. Stone, UK: Instem plc. Available at: http://www.biowisdom.com/content/safety-intelligence-program (Accessed 04.11.14).
16 Fourches, D., Barnes, J.C., Day, N.C., Bradley, P., Reed, J.Z. & Tropsha, A. (2010). Cheminformatics analysis of assertions mined from literature that describe drug induced liver injury in different species. Chemical Research in Toxicology 23, 171–183.
17 Bailey, J., Thew, M. & Balls, M. (2013). An analysis of the use of dogs in predicting human toxicology and drug safety. ATLA 41, 335–350.
18 Bailey, J., Thew, M. & Balls, M. (2014). An analysis of
the use of animal models in predicting human toxicology and drug safety. ATLA 42, 181–199.
19 Altman, D.G. & Bland, J.M. (1994). Diagnostic tests 2: Predictive values. British Medical Journal 309, 102.
20 Brooker, P. (2014). The use of second species in toxicology
testing. ATLA 42, 147–149.
21 Bailey, J. (2014). A response to the ABPI’s Letter to
the Editor on the use of dogs in predicting drug toxicity
in humans. ATLA 42, 149–153.

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The Cost of Standing Strong for Replacement

Katy Brown

To what extent does maintaining a stand against
the use of animals in experiments harm the
career progression of a young researcher today?

The experience of young researchers choosing to avoid animal experiments varies greatly, depending on the situation in which they find themselves. A series of interviews with young researchers, that were carried out as part of a Lush Prize Background Paper, pointed to the importance of centres or research groups dedicated to animal alternatives. It is clear that those students lucky enough to find themselves at one of these institutions have a much easier time in pursuing this career route. That said, there are still obstacles and challenges, including funding issues and resistance from those working outside these institutions. Those who choose this often difficult path outside specialist institutions may face a very tough time — in fact, two of the young people interviewed had no option but to decide on a career change, in large part due to issues around animal use. Individuals from a number of organisations involved in promoting alternatives to animal experiments and/or animal rights were asked to give their opinions on the feasibility of this career path choice by young researchers. What clearly came out of these interviews is that young researchers who wish to go into a career without ever being expected to use animals need to be determined, resourceful and tenacious. This is illustrated by comments such as:
— “You’ve got to get used to the fact that it may not always be an easy route.”
— “Forging an entire career in toxicology where you’re never involved in animal testing, is much more challenging.”
— “Some budding scientists may well be put off entering science, particularly toxicology, because of the issue of animal welfare.”

Interviews with young researchers

The issues surrounding this area were investigated in more depth by talking to six young researchers, not just from the UK, but also from Portugal, Denmark, Italy and the USA. Four were the recipients of the 2012 Lush Prize, and the two other individuals entered the life sciences, but chose to change career path, at least in part due to issues concerning animal use. Their full stories can be found in the Young Researchers Lush Prize Background Paper on the dedicated website (see here).

Sofia — the early-exiter — Portugal

Sofia started to study for a Biology BSc, but dropped out in the first year. At the point she decided to leave, she had not been asked to dissect or vivisect, but knew that before the end of the year she would be asked to do so. Sofia was concerned that the lecturers weren’t discussing these matters with the students and that the students were, in fact, apparently indifferent to the subject and perceived themselves as powerless. She felt that the general opinion of the student body was that dissection and vivisection were scientifically mandatory, and, even if they didn’t want to do it, there was nothing they could do to prevent it — and indeed that conscientious objection was something the great majority of students would not consider. In the end, she felt excluded from the course.

Joe — the committed conscientious objector — UK

Joe studied for a life science degree, then a related PhD, and finally took up a post-doctoral position. He avoided the undergraduate degree modules that would have involved dissection. During his PhD studies, there was increasing pressure on him to become involved in rodent studies, but he resisted this pressure, having stated at the start of his PhD that he would not be willing to carry out animal experiments. Initially, he was able to guide his own research in a way that avoided animal testing, by using non-animal methods. This did, however, get harder and harder as the research progressed, and he was encouraged “from the inside more and more to use non-human models to look at some of the key areas that he was investigating. He said that the position he ended up in, “essentially had me boxed into a corner from which changing career or carrying out animal experimentation (either directly or indirectly) were the only options.” He added that “the alternatives were pushed to one side, or not seen as being able to give the whole picture, by the people leading my research group.” Joe has changed his career path, and now works in the field of conservation.

Chiara — the well-supported enthusiast — Italy

Chiara works for Anna Maria Bassi, at the Analysis and Research Laboratory in Pathophysiology (LARF), in the Department of Experimental Medicine of the University of Genova, Italy. This laboratory is involved in the development and validation of in vitro models for use as alternatives to animal testing. Despite being based in this institution, with its non-animal research stance, Chiara’s major difficulty has been fundraising and finding partners with whom to develop new and competitive projects. She explained that, in Italy: there are very few research groups that promote in vitro models as alternatives to animal testing; the Italian Parliament has only very recently banned certain animal tests and endorsed funds for research on in vitro models with particular attention to the policy of the Three Rs and the European legislation; there are very few academic courses on alternative methods; and not very many research groups are devoted exclusively to working on in vitro models.

Felix — the motivated human research-focused scientist — USA

Felix has, since being an undergraduate, found it more useful to focus his research on a human-based approach, rather than an animal-based approach. He strongly believes that a human-based approach can provide more-relevant information, adding that he feels that “there was, and still is, some type of resistance by those that argue that much more information can be gathered from animal-based research.” He thought that “the most difficult steps are in the very early stages, such as undergraduate and post-graduate studies, where our scientific freedom is limited. However, for me, the post-doctoral stage has been most challenging, because our non-animal system has unfortunately been viewed with some level of resistance by some of our colleagues.”

Line — the animal-free testing convert — Denmark

Line started with the intent of working with animals, because she thought that she could help the animals — i.e. so that the animals used would be as few as possible, and would be as well taken care of as possible.” She then changed her career path, because she felt that it was stressful to be around animal suffering every day. She now works entirely on human tissue, which she feels is “very interesting from a scientist’s point of view”, because “when researching human health, the use of animal models will always be far less accurate than using humans.” She thought that the undergraduate level can be the most challenging when it comes to avoiding animal use, because “often you are not able to choose, as the institutions are streamlined and every student has to do the same courses.”

Liz — the slow-burner — UK

Liz studied A-level Biology, which involved a rat dissection. She was concerned about animal welfare and didn’t want to do the dissection, but she wanted to pass the course and go to university. She added, “It was said that we could apply to dissect a plant, but that we wouldn’t get as good a mark!” When she went to university, she didn’t study Biology. This was, in part, due to advice at her college that it would involve a lot of dissection. Having completed a work placement in the chemical industry and enjoyed it, she decided to go down that career path, but during the course of her degree she was more and more drawn to the options that involved a biological element.
For her MSc course, she chose Toxicology, with the long-term aim of working in a hospital and thus with humans, not animals. Her friend, who had completed the course, ascertained that there was no practical animal work involved.

She obtained a graduate job in a hospital toxicology laboratory, and then in a cancer prevention unit. Here, she was involved in examining human samples (breast tissue, etc.) from biopsies and surgery, for biomarkers of cancer, and she was also required to test organs from experimental animals. Liz said, “I was still training and learning, but felt that the experiments were not always well planned and the animal data derived were not always very informative.” She then decided to take more proactive action, and since then has completed a PhD in neurotoxicology, based on developing an in vitro brain model from human cells. This was followed by a postdoctoral position funded by the Humane Research Trust. After this first post-doctoral position, despite the promising nature of the brain cell model, Liz was unable to secure further funding, despite that fact that she “really felt that I had to get back to my original work, not only for myself, but also for the sake of those who’d supported me thus far.” Thankfully, due to the Lush Prize and funding from the British Brain Research Fund, she has been able to get her research aims back on track, at least for the time being. Liz added, “Because I have now chosen a career that specifically deals with developing replacement models, with a supervisor specifically involved in this field, avoiding animal use will not be an issue until I find myself unemployed. But due to it still being an emerging field, employment opportunities and funding are an issue, as is convincing our traditionalist colleagues of the worthiness of our research.”

Summary

The testimonies of these individuals largely speak for themselves. The responses point to the importance of specific institutions or research groups that focus on the development and use of alternatives, and these should, of course, be better supported. Those who find themselves outside such institutions or teams, are more likely to feel stranded and isolated. Then again, Liz did have the support of a research group dedicated to replacement, but she has still had a significant struggle to find funding. The interviews with some of these particular young researchers indeed pointed toward a tangible ‘cost’ in terms of having to steer their career on the often difficult path toward the use of non-animal based methods.

Katy Brown
Ethical Consumer Research Association
Unit 21, 41 Old Birley Street
Manchester M15 5RF
UK
E-mail: craig@lushprize.org

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The Use of Animals in Experiments — Not Because of Lack of Empathy?

Jolanta Zwolinska

The choice of an individual to use animals in experiments
is influenced by a wide range of social, religious and sometimes
career-driven factors, rather than a lack of empathy

Representatives of various religions and philosophical ideologies frequently make reference to the well known belief that one’s attitude toward the disabled, the sick, old people and children, is a measure of the humanity or moral value of the person. Yet, disputes arise when animals are included into the group of living beings entitled to the same type of consideration. The fact that they are used in scientific experiments is a highly controversial matter, and conflicting views are held by people both within and without the scientific community. This article presents a number of factors which might influence the decision of an individual to stand for or against animal use in experiments, including arguments voiced by representatives of the various sciences, both in support of or against the  continuation of animal experimentation.

Some historical and religious background

The approach adopted by ancient ethicists, in assuming  the dichotomy of human body and soul, resulted in man’s alienation from nature. Aristotle proclaimed a hierarchical structure of the world and the existence of essential differences between humans and animals, the latter being considered as inferior to the former. The intellect, according to that philosopher, was the main determinant of moral values.1 Adopted by the Judeo–Christian and Islamic traditions, the dogma of the immortal soul inherent in humans but not in animals, served to create a vast chasm between mankind and the animal world. Accordingly, Man rules over the world in which animals play an ancillary function.2–4 The cultures of Hinduism and Buddhism are based on the principles of respect for life and the protection of every living creature from suffering. Considered as being similar in their essence to humans, it is dictated in these religions that animals deserve to be protected and treated with reverence.3,5,6

Shaped throughout the ages, our stereotypical opinion of animals has been encoded into our collective consciousness, and cannot be easily overcome by newly emerging social concepts and ideas. For centuries, our attitude toward animals has been based on domination and power.4  In contemporary Christian culture, the majority of ethologists, psychologists and lawyers sympathise with an anthropocentric model of the biosphere, and take a negative stand with regard to animal rights. According to Bialocerkiewicz,5 animal rights reflect our attitude toward life and suffering and our appreciation of the universal principle of humanitarianism. Humans are not entitled to treat nature barbarically — i.e. to kill or mutilate, inflict pain or suffering. Bialocerkiewicz does not find any reason to recognise a unique role of mankind in the grand scheme of things, and emphasises a lack of religious, ethical or economic justification for awarding humans the right to take arbitrary decisions concerning the lives of other species.

The Catholic Church also acknowledges problems related to animal suffering. It speaks for an absolute ban on the mass breeding of animals and for the abandonment of procedures of animal testing used for cosmetics and various types of stimulants.7 According to Kozuchowski,8 a priest, it is our respect for ourselves and our claim to be perceived as more evolved beings that forbid us to treat animals as ordinary objects. A negative attitude toward animals is inevitably linked with a negative attitude toward other human beings.

The concept of ‘animal rights’ and morality

Cohen believes that rights result from contracts which are binding between members of a given community, and that rights, unavoidably, have inherently associated duties. Animals cannot undertake such obligations, and therefore they are not entitled to any rights in this sense (as cited in Mukerjee6). Guzek9 also points to the relativity of the concept of ‘animal rights’. He emphasises that rights can only be awarded to members of communities which are able to comply with commonly recognised ethical norms, so animals are not eligible to have such rights. Guzek believes that extremist activists of ‘animal right movements’ expect that animal rights should be similar
to, or identical to, human rights. Yet, evidence derived from observations shows that, whenever there is a conflict between animal rights and human interests, the latter always win. In Guzek’s opinion, human and animal rights are not, and cannot be, equal.9 Mukerjee, however, points out that children and mentally ill individuals cannot assume any obligations, nor do they comply with any norms, and yet they are not deprived of rights.6
According to Kotowska,4 the protection awarded to animals by the legal system of a given community depends on the attitude generally adopted by its members toward animals. If animals are treated as objects by the majority, then they will also be treated as objects by the adopted customary law, because there would, of course, be no one in such community to protest in their defence.
Many contemporary philosophers are reluctant to admit that it is pointless to extend our system of morality to include animals, opposing the claim that animal research does not constitute a moral problem. They emphasise the fact that speciesism is the cause of cruelty committed by man toward laboratory animals. Other philosophers take a less radical approach, accepting only some methods of animal use, and expressing favourable opinions about the banning of the most abusive research methods.6 Frey, a philosopher, emphasised that he was not an antivivisectionist, but that he accepted only those experiments with animals which yielded significant benefits and could also be conducted with human subjects.10 Singer, author of Animal Liberation,11 recognised by publicists as “the bible of the animal liberation movement”, believes that animal experimentation is acceptable only in the case of trial tests for life-saving drugs.

A contradiction in definition

The contradiction in the fact that people use animals as experimental models to acquire information pertaining to humans, and yet they refuse to acknowledge that animals have qualities recognised as human, is noted by Pisula.12 According to Griffin,13 the belief that no animal is capable of suffering or worthy of sympathy cannot be supported by any contemporary scientific evidence, and Spaemann14 emphasises that animals are not able to give meaning to, or control, their suffering. They are, indeed, doomed to suffer, in that it is particularly hard to endure, if they cannot respond to it with aggression or by escape. As a result of scientific progress, it is more and more difficult to justify the claim about the uniqueness of our species. Birmelin and Arzt, in their book, entitled Haben Tiere ein Bewusstsein [Do Animals Have Consciousness?],15 wrote: “…in terms of their mentality and emotions animals are more similar to us than we used to believe…”. What differs between us and animals, however, is not these qualities per se, but their intensity. Animals use senses which have become blunted in human beings. After long-term observations of social behaviour in elephants, zoologists assume that certain forms of morality and selfawareness may occur in more-highly evolved animals.16 Today, we also know that primates are able to experience emotions such as anger, fear, boredom, longing and loneliness.6

Opinions at the laboratory bench

It was emphasised by Mukerjee that scientists often decide to use animals, only if they are convinced that this is the only way to help people, and that sympathy for animals frequently affects this deliberation. Researchers try to reconcile the dictates of science with a humane approach — in fact, many of them love animals and volunteer to work for their benefit.6 Szyszko believes, however, that the choice of research method does not depend on sympathy for animals, or the need to acquire knowledge necessary for saving human health and life. Instead, it is proposed that senior academic staff members might sometimes encourage younger researchers to conduct animal experimentation, in order to contribute to the scientific accomplishments of the given institution.
As a result, animal experimentation is conducted all too often, and its purpose is not always justified by the needs of science. According to Szyszko, many higher-order animals suffer and die needlessly, frequently only to fulfil the excessive ambitions of young academics.17 In addition, Bialocerkiewicz highlights the fact that, in order to advance their careers and scientific outputs, some researchers are ready to carry out even the cruellest experiments, and gives an example of Baltimore, a physiologist awarded the Nobel Prize, who does not believe that “animal testing poses any moral problems”.5 As a result of such explicit approval by high-profile individuals, animals used in research can become perceived to be merely instruments — i.e. objects which can be exposed to any manner of tests.18 We see this in the fact that animals are often referred to as “experimental models”, “bioreactors”, or “source of replacement parts”, and this inevitably reinforces that idea that they have no rights and that they can be readily exposed to suffering and extermination.19 Feinberg insists that animals should not be treated as objects, although undoubtedly, they cannot be perceived in the same category as humans.20

Conclusion

Mukerjee points out that we are all morally responsible for the appropriately humane treatment of animals.6 The choice of an individual to use animals in experiments is influenced by a wide range of social, religious and sometimes career-driven factors, rather than a lack of empathy on the part of the researcher. Indeed, it is commendable that sensitivity to human pain and suffering defines the course of action for people professionally involved in medicine. What must be emphasised is that this sensitivity should be manifested as empathy for beings which are weaker and subordinate to humans, and the right choices should be made accordingly.
We should not make people suffer for the sake of animal welfare, but we also should not sentence animals to terrible suffering which leads to questionable benefits for people, not least in terms of the scientific validity of the results obtained. Due to progress in science, it is more and more difficult to justify the claim about uniqueness of our species, and being human is not only a privilege, but also an obligation to the creatures with which we share the Earth.

Jolanta Zwolińska
Faculty of Medicine
University of Rzeszów
Rzeszów
Poland
E-mail: jolantazwolinska@op.pl

References

1 Serpell, J. (1996). In the Company of Animals: A Study of Human–Animal Relationships, 2nd revised edition, 316pp. Cambridge, UK: Cambridge University Press.
2 Tatarkiewicz, W. (2004). History of Philosophy, 21st edn, 376pp. Warsaw, Poland: PWN.
3 Lejman, J. (2006). Animal ethics in the light of the idea of sustainable development. Problemy Ekorozwoju 1, 99–105.
4 Kotowska, M. (2011). Selected aspects of animal protection
according to criminal law. National and international perspectives. In Criminology of Contemporary Ecological Threats (ed. M. Kotowska & W.Pływaczewski), pp. 94–105. Olsztyn, Poland: Katedra Kryminologii i Polityki Kryminalnej, Uniwersytet Warmińsko-Mazurski.
5 Białocerkiewicz, J. (2005). Legal status of animals. Animal rights or legal protection of animals, 319pp. Toruń, Poland: Dom Organizatora.
6 Mukerjee, M. (1997). Trends in animal research. Świat Nauki 4, 68–76.
7 Krenzer, F. (2004). Morgen wird man wieder glauber. [You will believe again tomorrow. A Catholic faith information book.], 41st edn, 380pp. Limburg, Germany: Lahn-Verlag.
8 Kożuchowski, J. (2011). Ethical responsibilities of man toward the world of animals. Robert Spaemann’s Vision. Studia Ecologiae et Bioethicae UKSW 9, 29–48.
9 Guzek, J.W. (2005). Outline of Human Pathophysiology, 699pp. Warsaw, Poland: PZWL.
10 Frey, R.G. (1983). Vivisection, morals and medicine. Journal of Medical Ethics 9, 95–104.
11 Singer, P. (1995). Animal Liberation, 368pp. London, UK: Pimlico.
12 Pisula, W. (2001). Introduction to the monograph. In
Animal Minds
(ed. D.R. Griffin), pp. 16–17. Chicago, IL, USA: University of Chicago Press.
13 Griffin, D.R. (2004). Animal Minds, 320pp. Chicago, IL, USA: University of Chicago Press.
14 Spaemann, R. (2001). Grenzen: Zur Ethischen Dimension des Handelns [Borders: On the Ethical Dimension of Actions], 427pp. Stuttgart, Germany: Klett-Cotta.
15 Arzt, V. & Birmelin, I. (2001). Haben Tiere ein Bewusstsein [Do Animals Have Consciousness?], 279pp. Warsaw, Poland: Bertelsmann Media.
16 Vetulani, J. (2014). Bright prospects for thinking. Interview conducted by Rafał Romanowski, Żyjmydłużej 2, 10–13.
17 Szyszko, S. (2005). Epitaph for a dog (a few comments on ‘vivisection’). Przegląd Medyczny Uniwersytetu
Rzeszowskiego
1, 95–98.
18 Kornas, S. (2005). Animal experimentation. In Encyclopedia of Bioethics. Christian Personalism. The voice of the Church (ed. A. Muszal), pp. 128–132. Radom, Poland: Polskie Wydawnictwo Encyklopedyczne.
19 Żukow-Karczewski, M. (2013). Medical experiments and research involving animals. Polska: Wolnemedia.net. Available at: http://wolnemedia.net/historia/eksperymenty-i-doswiadczenia-medyczne-nazwierzetach/ (Accessed 21.05.13).
20 Feinberg, J. (1978). Human duties and animal rights. In On the Fifth Day: Animal Rights and Human Ethics (ed. R. Knowles Morris, R. & M.W. Fox), pp. 11–38. Lancaster, UK: Gazelle Book Services Ltd.

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Comparative Substitution

Michael Balls

The scientifically-justifiable choice of species is a crucial issue in animal
experimentation, which should not be based on ignorance and habit,
or 
on slavish compliance with political expectations
and regulatory requirements

 

 

The Principles of Humane Experimental Technique1 was published a year after it was written, and Russell and Burch added an Addendum, because they felt that year had “seen much activity in several parts of the field”.

One point they made in the Addendum was that “the comparative substitution of lower for higher animals raises difficult issues”, but, “where great severity is concerned … we must be glad to see lower forms substituted for mammals”. Unfortunately, they said nothing more about the “difficult issues” to which they referred, and I wish I had asked them about it, while I had the chance to do so.

The discussion on comparative substitution in the chapter on Replacement in the main part of the book focuses on the use of non-sentient material (plants, micro-organisms), degenerate metazoan endoparasites and free-living metazoan invertebrates. They regarded such use as a “limited gain”, and considered that “to shed obsessional tears over the fate of these organisms would bring the whole concept of humanity into contempt”. They preferred to concentrate on “the wholly desirable progress and prospects of replacement proper”, i.e. the use of any scientific methods which “replace methods which use conscious living vertebrates”.

Russell and Burch considered that not enough was being done with lower vertebrates, and saw the predominant use of mammals as “yet another expression of the high-fidelity fallacy”.2 Here, Russell’s experiences as a zoology student at Oxford came to the fore, since “our ignorance of the behaviour of common laboratory mammals is offset by a wealth of knowledge about that of numerous lower vertebrate species”. He had worked on mating behaviour in Xenopus laevis, the South African clawed toad, alongside the Nobel-prizewinning work of Tinbergen and his colleagues on the behaviour of birds and fish. In a section on The Choice of Species, Russell and Burch argued that, in terms of humanity, the “subtle matching of procedure to species, and species to objectives, is more significant that it appears at first sight”. They said that “a formal or informal training in zoology has again and again proved its value in the progress of medical research”, and lamented attempts to “correct the mistaken choice of a wrong species by forcing it to conform to the requirements of the investigation”.

They marvelled at “the present large-scale choice of laboratory species”, but regretted that, “out of the almost astronomical number of vertebrate species, only a minute selection are employed … this includes about 20 mammal species, three bird species, about four reptile species, half a dozen or so amphibians, and half a dozen or so fish”. Of the mammalian species, they said, “only about half the species are used in numbers over 1,000 per annum, the overwhelming bulk being made up of the four chief species (mouse, rat, guinea-pig and rabbit), and, of these, more than two-thirds are mice”.

Lack of sufficient understanding of the animals used for experimental purposes is still rife today. For example, the customary feeding ad libitum of caged rats and mice fundamentally alters their endocrinological, neurological and behavioural status. In real life, they spend most of their time searching for food, not eating it, while keeping alert because of the threat of predators. Also, the feminisation of male fish and amphibians by so-called endocrine disruptors, was taken as a warning of threats to human masculinity, whereas it is a normal part of the adaptability of these lower vertebrates. Even worse are attempts to genetically humanise laboratory animals, in order to make them better models for humans, without sufficient understanding of the cascade of complications likely to result from the consequent distortions of the very nature of the animals concerned.

Russell and Burch said rather little about choosing between the higher mammals, except to say that they were pleased to note that the Indian Government had “imposed salutary regulations” on the shipment of monkeys to provide kidney cells for vaccine production, and that the Medical Research Council had “issued recommendations on humane shipment”, which had been “adopted by all the British airlines concerned with livestock transport”. They welcomed “such action being taken on behalf of animals, which, although our near relatives, receive none of the privileges accorded by the Home Office to cats, dogs and the equidae” (the commonest animals to be encountered in Victorian England, when the Cruelty to Animals Act 1876 became law).
Over the years, I have had many discussions with veterinarians about whether there should be a hierarchy of laboratory mammals in the terms of the need to justify their use, with rats and mice near the bottom, and dogs and non-human primates at the top, or whether all the species should be afforded the same standards of consideration and care. I take the former view, and that is the position laid down in the Animals (Scientific Procedures) Act 1986, as amended to comply with Directive 2010/63/EU.

But what about the substitution of one species high in the hierarchy with another species high in the hierarchy. This point arose when somebody occupying a high position in the Three Rs movement, commenting on our study on the use of tests in dogs for predicting human toxicology and drug safety,3 warned us that “dropping the dog on the basis of your existing evidence could result in an increase in the use of nonhuman primates”. That would present advocates of the routine use of a second, ‘high-fidelity’, nonrodent species in toxicity testing with a dilemma and a challenge. How could they justify replacing pointless tests in one highly-protected species with pointless tests in another highly-protected species?

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. (2013). The wisdom of Russell and Burch. 3. Fidelity and discrimination. ATLA 41, P42–P43.
3 Bailey, J., Thew, M. & Balls, M. (2013). An analysis on the use of dogs in predicting human toxicology and drug safety. ATLA 41, 335–330.
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, can be obtained from FRAME.

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Laboratory Animal Science and the Use of Scientific Knowledge

PiLAS Staff Writer

The World Conference on Science for the Twenty-first Century: A New Commitment, took place on 26 June to 1 July 1999 in Budapest, Hungary, under the auspices of the United Nations Educational, Scientific and Cultural Organisation (UNESCO) and the International Council for Science (ICSU). The participants produced a very interesting 7-page, 46-paragraph Declaration on Science and the Use of Scientific Knowledge, which is well worth reading.1 There were no specific references to laboratory animal experimentation, but a number of the general points made have implications in terms of this particular form of scientific activity, and deserve to be given careful consideration. For example:

– Preamble para 4: Today, whilst unprecedented advances in the sciences are foreseen, there is a need for a vigorous and informed democratic debate on the production and use of scientific knowledge. The scientific community and decision-makers should seek the strengthening of public trust and support for science through such a debate. Greater interdisciplinary efforts, involving both natural and social sciences, are a prerequisite for dealing with ethical, social, cultural, environmental, gender, economic and health issues. Enhancing the role of science for a more equitable, prosperous and sustainable world requires the longterm commitment of all stakeholders, public and private, through greater investment, the appropriate review of investment priorities, and the sharing of scientific knowledge.

– Consideration para 21: That scientists with other major actors have a special responsibility for seeking to avert applications of science which are ethically wrong or have an adverse impact.

– Consideration para 22: The need to practise and apply the sciences in line with appropriate ethical requirements developed on the basis of an enhanced public debate.

– Consideration 23: That the pursuit of science and the use of scientific knowledge should respect and maintain life in all its diversity, as well as the life-support systems of our planet.

– Proclamation 31: The essence of scientific thinking is the ability to examine problems from different perspectives and seek explanations of natural and social phenomena, constantly submitted to criticalanalysis. Science thus relies on critical and free thinking, which is essential in a democratic world.

– Proclamation 40: A free flow of information on all possible uses and consequences of new discoveries and newly developed technologies should be secured, so that ethical issues can be debated in an appropriate way.

– Proclamation 41: All scientists should commit themselves to high ethical standards, and a code of ethics based on relevant norms enshrined in international human rights instruments should be established for scientific professions. The social responsibility of scientists requires that they maintain high standards of scientific integrity and quality control, share their knowledge, communicate with the public and educate the younger generation.

The challenge to all stakeholders in laboratory animal science — be they scientists, research organisations, industries, industry associations, funding bodies, medical research charities, patient associations, politicians, governments, animal welfare activists or antivivisectionists — is to strive to work, not as isolated parties, but together, to live up to the expectations of those who met in Budapest in 1999. The question that all these stakeholders must not be allowed to avoid is this: In all honesty, is this challenge being met?

1 Anon. (1999). World Conference on Science Declaration on Science and the Use of Scientific Knowledge, 7pp. Paris, France: UNESCO. Available at: http://www.unesco.org/science/wcs/eng/declaration_e.htm

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