Derbyshire is critical of the origins of the paper in Neuroimage, and is quick to point out that it was the product of a meeting of Focus on Alternatives, a group that has tended to take an antivivisectionist approach to the replacement of animals in research.  The consequence being that the paper had an inappropriately negative tone for a scientific paper and concentrated too much with the ethical issues rather than the scientific facts.

The Journal Alternatives to Laboratory Animals (ATLA), available by registration on the FRAME website, then published further correspondence, including a letter from Stuart Derbyshire and a response from Gill Langley.

ATLA also published a letter from two psychologists from the University of Birmingham. In this they agree in some ways with Derbyshire, that the two arguments made in the Langley paper, of scientific usefulness and moral issues, were not adequately separated. In defence of the paper, they agree with most of the points brought up in relation to the efficacy of animal studies into pain. They supported the observation that the resulting medicines from animal studies have proved disappointing over the years. However, they are quick to point out that alternative human methods often tell us little more than the animal models used before them, and that animal models are still invaluable, especially when different approaches are combined.  In terms of morals and ethics they are in disagreement with Langley and the NeuroImage paper, believing ‘that animals are sufficiently different from us that pursuing experimentation with animals to advance human interests is morally justified’. However, they do point out that they have no results from scientifically valid experiments to back this up.

A response to antivivisection claims about the use of non-human primates in research
There is a need for Europe-wide legislation concerning the use of animals in research, and it is timely to update the 1986 Directive. It is vital that a proper revision of the Directive results in appropriate evidence-based regulation that promotes animal welfare, patient benefits, and European wide scientific and economic competitiveness.

In recent months, antivivisection groups have published a number of reports on the use of primates in research. They repeatedly make two claims. Firstly, the antivivisectionists claim that studies using primates have not provided information which is relevant for advancing human health, secondly they claim that there are already alternative methods which give better information and could replace the use of NHPs.

In general, the primary aim of revising the Directive should be to ensure consistency to the approach of Member States, rather than review every matter relating to animals in research. It is a matter for the competent authorities to determine the scientific validity of any animal study on a case-by-case approach. And it is for the researchers to demonstrate that there is no viable alternative to the use of animals for a proposed study.

Nonetheless, researchers and institutions are very willing to discuss the science of animal research, and respond to the sort of suggestions made by antivivisection groups.

The scientific benefits of primate research
The use of primates has been vital for basic biomedical research and the development of new medicines. Several independent reviews, such as the Weatherall Committee report of 2006, have concluded that there is a strong scientific case for maintaining work on primates for carefully selected research questions, at least for the foreseeable future.

Primates are used in relatively small numbers (they make up around 0.1% of research animals). But they have been crucial in many important medical advances; for example the polio vaccine, life support systems for premature babies, and deep brain stimulation for Parkinson’s Disease.

Currently the main areas of primate study are infectious diseases eg to develop vaccines and treatments for HIV/AIDS and malaria; in neuroscience to better understand the brain and treat conditions ranging from Alzheimer’s disease to schizophrenia; and in reproduction, fertility and fetal research. They are also used in safety testing new medicines and vaccines.

Antivivisection critiques of primate use
Much of the antivivisection critique of primate research is based on a misunderstanding of the science. Antivivisectionists appear to believe that just because a method of research has limitations, and does not always give an answer directly transferable to humans, then it is of no value. In fact, all types of research have limitations. We would not even be able to develop cell cultures or computer models without the use of animal models, which are still required to get such alternatives working.

It is correct that a significant number of animals are used in toxicity testing and that alternatives are being developed in this field. But the antivivisectionists are wrong to claim that primates can be replaced in all areas of research and testing.

In these reports, the antivivisectionists take selective information to make their case against primate research, with the result that their conclusions are inaccurate or misleading. We illustrate this through a number of examples from the most recent antivivisection report: ‘Replacing Primates in Medical Research’: An expert report‘ by Dr Hadwen Trust / FRAME / St Andrew Animal Fund.

The authors have selected five areas of medical research into important human conditions, where they claim there has been ‘very limited success in translating to human benefit’. Here are just a few examples of inaccurate or selective use of scientific information.

Stroke
Regarding stroke, the document contains numerous scientifically inaccurate statements. For instance, it states that primate models of stroke have proved to be “scientifically unsound”. It goes on to claim that ’despite decades of using primates in stroke research there has been no demonstrable advance in our understanding of best practice in therapeutic intervention‘. However, the only currently approved treatment for ischaemic stroke, namely thrombolysis, derives directly from experiments done initially in the mid-70s in primates.

The report claims that ’ischaemic stroke is… a uniquely human affliction‘. This is also wrong. It is well documented that stroke affects other mammals, including primates, and in the wild. The antivivisectionists appear to have misunderstood a quote from the scientific literature on this point.

Referenced information is available on this webpage at AnimalResearch.info

HIV/Aids
In the chapter on the search for an AIDS vaccine, the report blames the lack of progress to date on reliance on primate models. However they do not offer, nor is there known to be available yet, an equivalent model. Since the recent ‘failure’ of two vaccines in clinical trials, there has been much discussion within the HIV/AIDS research community, and while they accept that primates are far from perfect, most recognise that they have played a vital role in HIV research. If anything, more research is needed to improve models of the disease in primates, according to a recent commentary in the science journal Nature.

The report then claims ’the most crucial breakthroughs in understanding and treating HIV infection and AIDS have come from studies not involving laboratory animals‘. But it does not elaborate on what these are.

Contrary to these claims, the first clinical trial of an anti-retroviral medicine in 1986, AZT, came directly out of studies in mice and monkeys. By 1996 combination therapy, consisting of cocktails of anti-virals taken together, had increased life expectancy enormously – a great success story, which required the use of animals in the research process.

Referenced information is available on this webpage at AnimalResearch.info

Cognition
In the chapter on cognition, the report suggests that ’imaging studies in human volunteers are highly relevant and can reveal much richer information than invasive primate studies‘. Techniques such as functional magnetic resonance imaging (fMRI) are simply described as ’cutting edge‘, without their significant limitations being properly acknowledged.

A recent report in Nature carried an extensive review of what we can do and what we cannot do with fMRI. This neuro-imaging technique was described as the most important advance since the introduction of x-rays. However, as the review points out, ’fundamental questions concerning the interpretation of fMRI data abound, as the conclusions drawn often ignore the actual limitations of the methodology‘. The author concluded that ’claims that computational methods and non-invasive neuroimaging (that is, excluding animal experimentation) should be sufficient to understand brain function and disorders are, in my opinion, naive and utterly incorrect‘.

This antivivisection report suggests that there is ’a wealth of opportunities to replace primates in research and testing today‘. These opportunities however still require a significant amount of further research and testing to replace primates adequately in important areas of research.

Conclusion
We all aspire to the aim of undertaking life-saving research without the need to use animals. The various reports from the antivivisection groups do illustrate the enormous advances that have been made in developing non-animal methods of research. They also show the progress that has been made in replacing animal use.

However we are not at the point yet when we can phase out primate research. If our efforts to improve animal and human health are to flourish, then we must enable scientists to use animals in research when it is shown to be necessary and appropriate. 

Centralised databases – a long way to go
Databases are often put forward as a way to collect and disseminate information on animal experiments, including information on procedures and welfare issues. The intention was to avoid duplication or repetition of research, and generate information to assist in progressing the 3Rs. However, the extraordinary complexity of such a venture means there is a long way to go before any meaningful attempt can be made on a European central data repository.

There are some examples in biomedical science of centralised informatics activities that have been successful, for example, ENSEMBL, which provides a central portal for a variety of annotations of genome sequences.

But these existing databases draw together datasets that are relatively well-defined. In comparison, data on the outcomes of animal experimentation in animal houses across Europe will be highly diverse. We know, for example, that the outcome of a procedure on a genetically modified mouse will depend upon a number of factors including:
• the precise procedure that is used (the standard operating procedure, SOP) which may vary across institutes
• the environmental conditions in the animal house, including the type of environmental enrichment in the cage
• the precise mutant allele generated in the genetically modified mice

All these factors will impinge upon the phenotype measured and the welfare outcomes. In essence, describing phenotype and welfare in an animal is a complex 3-dimensional matrix including genotype, phenotyping, SOP and animal house environment.

Standardised descriptions or vocabularies for phenotyping procedures and environmental conditions would have to be implemented across Europe if we are to have meaningful database entries in any central data repository. We would need to agree on the parameters and language to apply to the complex data sets, and on the standards that would need to be adopted.

In summary, at the present time, developing the procedures for documentation and data acquisition in animal houses would be an enormous task. The world is full of unfinished databases. We are not ready to build a whole new one just to satisfy a European bureaucratic requirement.

Replacing Primates in Medical Research—a flawed report with flawed conclusions

We would expect the authors of this report to have written the conclusions before they did their literature research. It represents the same old antivivisection arguments, re-drafted in a slightly different way. As such, it contains much the same flaws and contradictions as other antivivisection material.

The report is supposedly ‘a detailed analysis of the extent to which primate experiments have already been replaced by advanced non-animal alternatives such as cell and molecular methods, computer simulations and ethical studies with human volunteers’.

The authors have selected five areas of medical research into important human conditions, where they claim there has been ’very limited success in translating to human benefit‘. Paradoxically, these five areas supposedly illustrate where ‘notable progress has been made in replacing primate experiments with non-animal techniques’. If replacement has already been happening, yet with limited success in translating to human benefit, it rather suggests the replacements have not helped that much!

The authors point out that very few systematic reviews of primate research have been conducted. They claim that without such reviews ‘there is little independent evidence of the value of primate experiments for human medicine’. Here, the authors fail to recognise that while systematic reviews are at the top of the hierarchy of evidence, they are not the only type of independent evidence recognised in the world of science.

Furthermore, the authors fail to acknowledge the two-step required process to assess animal or other pre-clinical studies by systematic reviews. Reviews can be used in two ways: (i) to study the results of a particular method or intervention in an animal (or other) model to give a clear conclusion or (ii) to determine whether the animal model is a good predictor of results in humans. No systematic review can be used for the latter purpose unless the response in humans is known.

Finally, on this point, the authors fail to give evidence that the replacement methods they favour, such as cell and molecular methods, have themselves been subject to independent systematic reviews.

The biggest flaw of this report, however, is the usual fare of antivivisection groups. They simply present selective evidence and quotations to support a predetermined position. For example, much of the report is devoted to highlighting the flaws and limitations in animal studies. Of course, these exist. But to omit any mention of the equivalent flaws and limitations of non-animal methods is simply another form of bias. No report can be taken seriously when it is so one-sided.

In the introduction to the report, the authors quote the Chief Executive of the government’s National Centre for the Replacement, Reduction and Refinement of Animals in Research, as calling on the government and the funders to ‘develop a national strategy that is not just about the continued use of primates, but which has the clear aim of replacing, refining and reducing that used wherever possible’. We couldn’t agree more. We are pleased to say that that is exactly what is happening.

Systematic reviews and project licences in animal research - a contribution from RDS

Recent reviews have exposed the methodological failings of many studies using animals. For example an article examining US animal studies of amyotrophic lateral sclerosis suggested that most of the studies that were published, including some in top-rank journals, had been done with small sample sizes, with no randomisation of treatment and control groups, and without blinded evaluations of outcomes (Nature 2008). If the analysis is correct, then this is clearly unsatisfactory.

There is increasing recognition of the need to improve both the design and analysis of studies using animals. RDS has consistently supported the use of systematic reviews as one such way to improve research. This is highlighted on our website, on our blog, and has been featured in our newsletter. Such reviews could also help further the 3Rs. SABRE does a good job of pointing out where systematic review are appropriate.

The SABRE petition proposes that each licence application includes references to systematic reviews of existing relevant studies. Depending on how exactly this is interpreted, such a proposal could cause difficulties should the referencing of systematic reviews become a ‘requirement’ for the award of a project licence.

The UK project licence is given under the Animals (Scientific Procedures) Act 1986 (ASPA) is a legal document based on a cost-benefit assessment of whether it is permissible to cause harm to an animal. It is a contract between the UK government and an individual researcher. Its award is dependent on a legally-defined process of review designed to ensure that animals are used in research only when there are no other means of obtaining the results and only when the anticipated outcome warrants any harm inflicted on the animals.

Systematic reviews and meta-analyses of animal studies are rare. Only 103 such reviews, comprising 57 systematic reviews, 29 systematic reviews accompanied by a meta-analysis and 17 meta-analyses were identified up to 2005 (Peters et al, 2006). For the vast majority of proposed animal studies, there will be no systematic reviews to refer to when applying for a project licence.

If systematic reviews do not exist in some areas, and cannot be forced into existence, they cannot be made a legal requirement for the award of a project licence. This is particularly the case when novel, groundbreaking research into basic biological mechanisms or structures is proposed, rather than evaluation of an existing drug, compound or procedure. Even where reviews can be carried out, there remains the difficulty of deciding whether the requirement would be for review of the procedures utilised or the substances undergoing test.

Basic animal studies may be designed to study biological mechanisms (rather than outcomes), or species, strain and methodological differences. Animal studies may open completely new fields of research where there is little prior knowledge (for example the creation for the first time of a genetically modified animal model of the human disease). Yet for a systematic review to draw a meaningful conclusion there must be sufficient available primary studies. But repetition in animal studies is strongly discouraged both by regulators and by the publication process. For much basic research, the literature may be neither sufficiently extensive nor sufficiently consistent to permit systematic review.

Additional questions remain about the applicability of systematic reviews to all types of animal research (Lemon and Dunnett, 2005). Some examples may help to illustrate the problems.

Much basic research is shallow and broad in its cover – that is, only one or two research groups world-wide may be studying an area. This is commonly the case in environmental/conservation research. Even flicking a dart into some deer to monitor their populations in the wild, or taking a biopsy or blood sample from a live seal washed up on the shore, may need a project licence, as would the use of guinea pigs as hosts for a study of tick biology.

Furthermore, a project licence is not always awarded for a single experiment or research project, but more often for a series of procedures. Some of the procedures to be carried out may depend on results obtained during an earlier project licence, which have not yet been published. Researchers do not always know in advance which procedures will be carried out within the remit of their project licence. So, for instance, to require application for renewal of a licence to be delayed until a systematic review had been conducted of the results of the current licence, would be totally unworkable.

In the case of toxicology tests, laboratories use well-characterised and relatively standardised techniques. However, it is not known in advance which substances will be tested over the course of the project licence. So it is unclear what would be the nature of the systematic review required. About 10% of all procedures are for the production of biological materials such as tissues, blood products, infectious agents, vectors, neoplasms and antibodies. And in the assay and quality control of biologicals, the models are generally well validated and the studies are already carried out in methodologically consistent ways.

When animals are used as models of humans, systematic reviews can be used in two ways: (i) to study the results of a particular method or intervention in an animal model to give a clear conclusion or (ii) to determine whether the animal model is a good predictor of results in humans. No systematic review can be used for the latter purpose unless the response in humans is known. However, if animal studies indicate that a potential treatment does not work, then that treatment would not normally proceed into human trials. In such cases, it is not possible to determine whether the animal models are predictive of humans, as no human data is available. In the case of nimodipine (Evans et al 2006), a systematic review of animal studies at the pre-clincial stage would not have led to human trials, and would not allow us to know whether an animal model is or is not predictive of the human situation. On the other hand, a comparison of systematic reviews of animal studies against the results of clinical trials can help validate the animal models, but obviously cannot further inform the decision taken to proceed to human trials. These two distinct purposes of carrying out systematic reviews are different, and must be recognised as such.

We are not arguing against systematic reviews here, nor suggesting that the current mechanisms for undertaking animal research are perfect (or even adequate). Animal studies need to be improved, just as improvements are needed for all types of research. Even in clinical trials, where systematic reviews are accepted as the gold standard, there remain improvements to be made.

RDS acknowledges that systematic reviews are at the top of the hierarchy of evidence, and are superseding narrative reviews because of their improved quality. It is said that the point of conducting systematic reviews is to reduce bias as much as possible in the evaluation of animal research (SABRE 2006). If the use of systematic reviews amounts to no more than the use of explicit and reproducible methods to reduce bias, then all reviews should be systematic.

However, most descriptions of systematic reviews (which are usually about assessing the effect of a therapeutic intervention) emphasise the importance of having a clearly defined topic and a specific question to answer, as well as having rigorous inclusion and exclusion criteria. Both present practical difficulties when basic research using animals is founded on a very diverse range of prior research. Sometimes results from in vitro and in silico studies are relevant, as well as information from other fields of animal research, or even knowledge from human epidemiological and clinical studies. This is a concern because it seems the most common reason for discrepancies between systematic reviews is differences in inclusion criteria (Linde 2003). Even proponents of systematic reviews seem to recognise this issue by calling for ‘initial research to discover the best methodological approaches for systematically reviewing animal studies’ (SABRE Charter, May 2008). We support this proposal.

Narrative (critical) reviews are currently common in basic research in which animal-based studies play a part. By definition even narrative reviews require a substantial body of research to have been undertaken, but at least they can overcome diversity and a lack of consistency in approach, or patchy coverage of the initial studies in a field.

A valid criticism of traditional narrative reviews is that they are ‘rarely explicit about how studies are selected, assessed and integrated. Thus the reader is generally unable to assess the likelihood of prior beliefs or other biases clouding the review process’ (Davies). There is no reason for this to continue. RDS considers that good methodology which leads to an objective account of the available data can be applied to both the systematic and narrative review process, and that the most useful analysis of the literature is obtained though use of the most appropriate review method.

Many of the protocols associated with systematic reviews can also be applied to narrative reviews, so that the data is assessed in a reproducible, transparent manner and bias is reduced. This requires that search criteria are broad, inclusion criteria are specified, and methodologies of included studies are evaluated critically. This approach would allow readers to reach more consistent conclusions from narrative reviews.

There are nonetheless many instances when systematic reviews as currently practised could be of great value. For example, they are desirable when a number of papers studying the same intervention have been published without any clear conclusion, or when a candidate medicine is considered suitable to proceed to human clinical trials. RDS continues to support systematic reviews in animal research.

References:
Davies HTO, Crombie, K (2001) What are systematic reviews? http://www.jr2.ox.ac.uk/bandolier/painres/download/whatis/Syst-review.pdf
Evans I, Thornton H, Chalmers I (2006) Testing Treatments: Better Research for Better Healthcare.
Linde K and Willich SN, J R Soc Med (2003) How objective are systematic reviews? Differences between reviews on complementary medicine.96(1) January.
SABRE Charter (2008) http://www.sabre.org.uk/ May.
SABRE personal communication (2006).
Lemon R and Dunnett SB (2005) Surveying the literature from animal experiments. BMJ 330: 977-978.
Nature News feature. Standard model. (2008) 454/7.
Nature Medicine editorial: when less is not more. (2008) 14, Number 8.
Perel P, Roberts I, Sena E, Wheble P, Briscoe C, Sandercock P, Macleod M, Mignini LE, Jayaram P, and Khan KS (2006) Comparison of treatment effects between animal experiments and clinical trials: systematic review. BMJ 15 December.
Peters, J L, Sutton, AJ, Jones, DR, Rushton, L, and Abrams, KR (2006) A systematic review of systematic reviews and meta-analyses of animal experiments with guidelines for reporting. J Environ Sci Health B 41: 1245-1258.

One such example is that of the work of Dr Siouxsie Wiles, who won the NC3Rs prize in 2005 for her work in improving design of bacterial infection experiments, leading to a reduction in the number of mice needed for this research. Dr Wiles found infection was more efficient when mice were infected naturally by putting an infected mouse in a cage with others, compared with artificial infection routes.

Dr Wiles has been awarded funding for a new project, aiming to develop ways of making human disease agents ‘light up’. Traditional disease models can involve animal suffering, and bacteria numbers are determined by killing infected animals at defined points. If the bacteria are genetically engineered to produce light, they can be measured using imaging techniques in real time, in living animals. So this project should reduce the numbers of animals used and the suffering of individual mice.

The government’s commitment to funding this research was announced last year. The innovative projects announced today are a welcome addition to the extensive scientific work already being done - by public bodies and by commercial concerns - to reduce, replace and refine animal use in research.

Not surprisingly, we don’t support the decision. It is baffling because there is no clear distinction between basic and applied research, except at the ends of the spectrum. It is widely appreciated that much basic research can lead to medical advances, and that much applied research can shed light on basic biological functions.

Whilst the research institute is appealing the decision, the final outcome will also be baffling, since its impact more widely is not known. Switzerland is an exceptional case. It is renowned for having held a number of referenda on animal experiments, all of which have gone in favour of continued research using animals, but which have led to pressure to tighten regulations.

Ultimately, there is no reason that the European Union, which is reviewing its own laws on animal experiments, should be unduly swayed by this decision. It is up to the Swiss to decide how their own laws work. 

One scientist quoted by Arron cautioned that the risks of nanoparticles could outweigh the benefits. But any debate about the use of mice in medical research was clearly not considered relevant to the piece.

Of course the antivivisectionists are quick to latch onto statistics such as only 1 in 5000 drugs getting approved for human use. Last week our young friends at Speaking of Research (the US counterpart of Pro-Test) were busy debunking the oft-quoted ‘92% of drugs that pass animal tests fail human trials’ statistic. We have addressed such nonsense several times on this blog.

Tom at Speaking of Research points out that you can use statistics allied with assumptions to ‘prove’ anything. He comes up with his own (hypothetical) case that 90.5% of dangerous drugs have been kept out of human trials thanks to animal safety tests.

A little strange that the antivivisectionists should turn ‘animal research’ into the much narrower ‘animal testing’ which accounts for less than one eighth of all animal procedures in the UK. Unfortunately we do not believe that this is because antivivisectionists support the 85% of basic research and applied research and development using animals that makes up the rest; rather that they find ‘animal testing’ so much easier to attack.

That aside, the Government’s response is again worth reading. I’m not going to reproduce it all here, just the first two paragraphs:

The Government’s policy on the use of animals in scientific procedures is clear and straightforward. There is still a need for the responsible use of animals for experimental and other scientific purposes to continue if improvements in healthcare and veterinary treatment are to be developed with the minimum of delay and to make proper provision to protect man and the environment from health risks and other hazards.

Some campaigners describe animal experiments as ‘outmoded science’ and argue that there are better ways of making medical and scientific progress using modern, non-animal methods, such as computer modelling and microdosing. However, under the Animals (Scientific Procedures) Act 1986, animal experiments cannot be carried out if the same objectives can be achieved using non-animal methods. So, as relevant, new and improved technologies and testing methods are developed, they already have to be used instead of animal methods.

While we welcome this, perhaps it’s time for the government to raise the bar for responding to e-petitions? At the moment it stands at just 200 signatures. I’m sure most people could drum up more than 200 signatures to any weird and whacky idea.