Have the non-technical summaries of animal experiments in Europe improved? An update

Following a review of Directive 2010/63/EU on the protection of animals used for scientific pur­poses in the European Union (EU), non-technical project summaries (NTS) of all approved projects must be published in a central database using a standard template. Our initial review of the NTS reported in ALTEX in 2018 had found the NTS to be deficient in their accessibility and quality, notably the "adverse effects" section where the harms to the animals are meant to be described. Here we repeat our review to see if these legislative changes have improved the accessibility and quality of the NTS. As before, we focused on the NTS from the United Kingdom (UK) and Germany; even though the UK has left the EU, it is using the same template. We found significant improvement in the reporting of five of the six elements we identified as essential to the "predicted harms" section. However, there was no significant improvement in the reporting of adverse effects. Only 41% of German NTS and 48% of UK NTS are fully reporting this important element of the "predicted harms" section. In our view, researchers need support in describing the impact of their research on the animals and to assist here we include a checklist for competent authorities and a list of suggested terminology for standard administration and sampling procedures. Unless the NTS improve further, their utility as a tool for sharing of good practices in the 3Rs or to support evidence-based policy­making will remain limited.

All countries of the European Union (EU) are required to publish "non-technical summaries" (NTS) of research projects that use animals. To improve transparency, the public must have access to NTS and understand their content. Our previous review found that the information provided in the NTS was lacking in many cases. This is preventing a full understanding of what animals experience during experiments. In particular, NTS often failed to fully describe what procedures the animals would be subjected to, how often they would take place, how long they would last, and the harm they would cause. Here we repeat our review to see if recent legislative changes, including the requirement for NTS to be published in a central database using a standard template, have made a difference. While there has been some improvement in reporting, many NTS still fail to adequately describe the harm that animals will experience.

Evaluating the impact of the metastatic spinal cord compression coordinator in a regional cancer network: a prospective, non-randomized pilot study.

BACKGROUND: Extradural metastatic spinal cord compression (MSCC) is a debilitating and potentially irreversible complication of cancer. Delay in treatment could lead to irreversible neurological damage, adverse quality of life and a burden on health care resources. Lack of effective communication between teams has been identified as one of the reasons for delay in treatment. The MSCC coordinator (often a nurse, radiotherapy radiographer or a doctor) is responsible for coordinating the diagnosis and management of patients with MSCC. The role has been shown to streamline service, ensure timely decision-making and improved survival outcomes. However, available data are anecdotal or from limited series presented as abstracts in conferences. In this study, we assessed the impact (time to treatment) of the newly introduced role on the treatment pathway compared to similar period in the preceding year. METHODS: This was a multi-centre, prospective, pilot study carried out in Kent, UK between 1st April to 30th June 2021. Patients were considered eligible if they had magnetic resonance imaging (MRI)-confirmed cauda equina or cord compression. The data prospectively collected include: (I) time from diagnostic imaging to radiotherapy treatment; (II) number of referrals to hospital palliative care (HPC), occupational/physiotherapy (OPH) and community hospice referrals (CHP). A comparative retrospective data for (I) was collected for the same time period in the preceding year. The study outcome assessed was reduction in time from radiological diagnosis of MSCC to receiving radiotherapy. RESULTS: Fifty-eight patients in 2020 and 24 patients in 2021 were included in the dataset. The MSCC coordinator role (introduced in 2021) led to reduction in the time from imaging to treatment (P=0.045). Compared to 2020, there was a shorter mean/median time to treatment, seeing more patients being treated within 24 hours. All hospitals except East Kent Hospitals saw more patients being treated within 24 hours. 7 referrals each made to HPC, OPH and CHP respectively. CONCLUSIONS: Introduction of MSCC coordinator role led to improved time from imaging to radiotherapy treatment. The new service led to engagement with rehabilitative and palliative services. Future work should be done to assess the long-term impact of this role on utilization of support services and patient recovery.

Incorporating new approach methodologies into regulatory nonclinical pharmaceutical safety assessment.

New approach methodologies (NAMs) based on human biology enable the assessment of adverse biological effects of pharmaceuticals and other chemicals. Currently, however, it is unclear how NAMs should be used during drug development to improve human safety evaluation. A series of 5 workshops with 13 international experts (regulators, preclinical scientists, and NAMs developers) was conducted to identify feasible NAMs and to discuss how to exploit them in specific safety assessment contexts. Participants generated four "maps" of how NAMs can be exploited in the safety assessment of the liver, respiratory, cardiovascular, and central nervous systems. Each map shows relevant endpoints measured and tools used (e.g., cells, assays, platforms), and highlights gaps where further development and validation of NAMs remains necessary. Each map addresses the fundamental scientific requirements for the safety assessment of that organ system, providing users with guidance on the selection of appropriate NAMs. In addition to generating the maps, participants offered suggestions for encouraging greater NAM adoption within drug development and their inclusion in regulatory guidelines. A specific recommendation was that pharmaceutical companies should be more transparent about how they use NAMs in-house. As well as giving guidance for the four organ systems, the maps provide a template that could be used for additional organ safety testing contexts. Moreover, their conversion to an interactive format would enable users to drill down to the detail necessary to answer specific scientific and regulatory questions.

An Estimate of the Number of Animals Used for Scientific Purposes Worldwide in 2015.

Few attempts have been made to estimate the global use of animals in experiments, since our own estimated figure of 115.2 million animals for the year 2005. Here, we provide an update for the year 2015. Data from 37 countries that publish national statistics were standardised against the definitions of 'animals' and 'procedures' used in the European Union (EU) Directive 2010/63/EU. We also applied a prediction model, based on publication rates, to estimate animal use in a further 142 countries. This yielded an overall estimate of global animal use in scientific procedures of 79.9 million animals, a 36.9% increase on the equivalent estimated figure for 2005, of 58.3 million animals. We further extrapolated this estimate to obtain a more comprehensive final global figure for the number of animals used for scientific purposes in 2015, of 192.1 million. This figure included animals killed for their tissues, normal and genetically modified (GM) animals without a harmful genetic mutation that are used to maintain GM strains and animals bred for laboratory use but not used. Since the 2005 study, there has been no evident increase in the number of countries publishing data on the numbers of animals used in experiments. Without regular, accurate statistics, the impact of efforts to replace, reduce and refine animal experiments cannot be effectively monitored.

Botulinum toxin testing on animals is still a Europe-wide issue.

There have been significant developments in the use of animals to test Botulinum toxin products in Europe in recent years. This paper summarises and discusses these from the perspective of the animal protection organisation. A cell-based assay has been validated by Allergan and is now being used for the replacement of the mouse bioassay for the batch testing of their Botulinum toxin A products. Two further companies (Merz and Ipsen) have recently validated similar cell-based assays to replace animals in their batch testing. However, the number of animals being used in batch tests across Europe remains at record levels; an estimated 400,000 animals per year, based on official statistics and non-technical summaries. There are concerns from animal protection organisations about the authorisation of animal testing for Botulinum toxin products that are to be used for aesthetic purposes. Furthermore, should testing for companies that have not yet implemented the alternative method continue to be permitted under the EU Directive 2010/63 on the use of animals for scientific purposes? Whilst we are on the cusp of an era where the mouse bioassay has been replaced for the potency testing of Botulinum toxin A for injection, it is important that Europe sees a reduction of animal testing in real terms.

Recommendations to improve the EU non-technical summaries of animal experiments.

Under the new Directive 2010/63/EC, member states have to publish non-technical summaries (NTS) of the projects involving animals that they authorise. These summaries must include information on the objectives of the project including the predicted harm and benefits and the number and types of animals to be used. Summaries should also demonstrate compliance with the 3Rs. The intention was that NTS would help increase the transparency of animal research in the EU. In this article, we review the status of the publication of NTS across member states and give some general observations on publication speed, identification, accessibility and quality. We also review in more detail the quality of reporting in a selection of NTS from Germany and the UK. We consistently found that NTS from Germany and the UK were deficient in their description of what is being done to the animals and what they might experience as a result. Using examples taken from specific NTS we highlight what we view to be good and bad examples to assist member states and researchers in producing better NTS in the future. The NTS can also be an important tool in sharing of best practice in the 3Rs and the avoidance of duplicative animal testing. For this to happen however, member states need to publish timely, ensure that NTS are accurate and, ideally, there needs to be some centralisation of the NTS.

Ten years of REACH - An animal protection perspective.

It has now been 11 years since the EU's new chemicals legislation (Regulation No. 1907/2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals [REACH]) came into force. Two important statements in the REACH Regulation in relation to animal testing and alternatives are: Article 1(1), which states that one of its purposes is to promote alternative methods; and Article 25(1), which states that animal testing should be used as a last resort. This review looks at the mechanisms that were put in place within REACH to achieve these aims and asks, not only if they are being implemented properly, but also if they have been sufficient. Whilst the chemical industry has heavily used data-sharing and read-across, this review concludes that nevertheless over 2.2 million animals have already been used in new tests for REACH registrations. This equates to an annual average of 275,000 animals; 58,000 more per year than the best-case estimate made by the European Commission in 2004. The use of in vitro and (Q)SAR approaches as standalone replacements for animal tests has been relatively low. The levels of funding for research into alternative methods remain low, and there are concerns over the speed of formal adoption of those that have been validated. In addition, there have been issues with the recognition that testing as a last resort and the promotion of alternative methods applies to all parties, including the Commission, Member States and the agency responsible, the European Chemicals Agency. This review provides ten recommendations for better implementation of these two key aspirations, as well as lessons to be learned for future similar legislation.

The added value of the 90-day repeated dose oral toxicity test for industrial chemicals with a low (sub)acute toxicity profile in a high quality dataset: An update.

A previous retrospective analysis of substances in the ECHA CHEM database concluded that, for industrial chemicals with a 'low (sub)acute toxicity profile', further testing in the 90-day study is unlikely to change this profile (Taylor et al., 2014). We have further tested this hypothesis by assessing the outcome of substances with testing proposals for which a prediction was made in that paper that the NOAEL based on the 90-day study would be 1000 mg/kg bw/d. Indeed, for seven out of ten substances for which data was available, the profile was shown to be held. For three substances, the reduced NOAEL was explained by renal effects in the rats, two of which had been seen in the 28-day study but had been dismissed by the study submitter. We conclude that the low toxicity profile will be even more protective if the NOEL is used from the 28-day study and an independent expert view is taken of the human relevance of any effects reported in the 28-day study.

Wider recommendations for institutions made in the Brown Report following the BUAV investigation into the use of animals at Imperial College London.

In December 2013, a group of experts produced a report on the management of an animal unit at Imperial College London, following a BUAV investigation that found evidence of systematic failures in the care and monitoring of animals used in procedures there. The Brown Report looked at four areas: the animal welfare and ethical review body (AWERB); the operation of the unit; training; and overall culture. The report made 33 recommendations to improve practices at Imperial College, many of which were relevant to other institutions. In this report, we identify the recommendations that are applicable to all animal facilities, and redraft them as a checklist with supporting information to assist those reviewing their animal care policies. We support the Brown Report's recommendation that institutions should have a vision statement and an action plan, as well as a 'champion' for the Three Rs. We encourage all institutions that use animals to, as a first step, review the performance of their animal units against this checklist.

The added value of the 90-day repeated dose oral toxicity test for industrial chemicals with a low (sub)acute toxicity profile in a high quality dataset.

A survey conducted on the EU Notification of New Substances (NONS) database suggested that for industrial chemicals with a profile of low toxicity in (sub)acute toxicity tests there is little added value to the conduct of the 90-day repeated dose study. Avoiding unnecessary animal testing is a central aim of the EU REACH chemicals legislation; therefore we sought to verify the profile using additional data. The OECD's eChemPortal was searched for substances that had both a 28-day and a 90-day study and their robust study summaries were then examined from the ECHA CHEM database. Out of 182 substances with high quality 28-day and 90-day study results, only 18 reported no toxicity of any kind in the (sub)acute tests. However, for 16 of these there were also no reported signs of toxicity at or close to the limit dose (1000mg/kgbw/d) in the 90-day study. Restricting the 'low (sub)acute toxicity in a high quality dataset' profile to general industrial chemicals of no known biological activity, whilst allowing irritant substances, increases the data set and improves the prediction to 95% (20 substances out of 21 substances). The low toxicity profile appears to be of low prevalence within industrial chemicals (10-15%), nevertheless, avoidance of the conduct of a redundant 90-day study for this proportion of the remaining REACH phase-in substances would avoid the use of nearly 50,000 animals and save industry 50million Euros, with no impact on the assessment of human health.

Experiences of the REACH testing proposals system to reduce animal testing.

In order to reduce animal testing, companies registering chemical substances under the EU REACH legislation must propose rather than conduct certain tests on animals. Third parties can submit 'scientifically valid information' relevant to these proposals to the Agency responsible, the European Chemicals Agency (ECHA), who are obliged to take the information into account. The European Coalition to End Animal Experiments (ECEAE) provided comments on nearly half of the 817 proposals for vertebrate tests on 480 substances published for comment for the first REACH deadline (between 1 August 2009 and 31 July 2012). The paper summarises the response by registrants and the Agency to third party comments and highlights issues with the use of read across, in vitro tests, QSAR and weight of evidence approaches. Use of existing data and evidence that testing is legally or scientifically unjustified remain the most successful comments for third parties to submit. There is a worrying conservatism within the Agency regarding the acceptance of alternative approaches and examples of where registrants have also failed to maximise opportunities to avoid testing.

EU member state government contribution to alternative methods.

Article 47 of the new EU Directive 2010/63/EU on the protection of animals used for scientific purposes requires national governments to contribute to the development and promotion of alternative methods. A recent survey of EU member states found that reported funding of alternative (3Rs) methods totalled € 18.7 million in 2013, provided by only seven countries (Austria, Belgium, Denmark, Finland, Germany, Sweden, and the UK). There were indications that the contributions of some of these countries have increased since the implementation of the new Directive. However, funding of alternatives is between 0 and 0.036% of national science R&D expenditure and nearly half of the countries that responded reported that they do not specifically contribute. Data (and, by assumption, financial contribution) remains unavailable from half of the member states across the EU, regardless of the method of collection.

A critique of the EC's expert (draft) reports on the status of alternatives for cosmetics testing to meet the 2013 deadline.

The 7th Amendment to the EU's Cosmetic Directive (now recast as Regulation 1223/2009) bans the testing of cosmetic ingredients and products on animals, effective 2009. An extension until 2013 was granted, for marketing purposes only, for three endpoints: repeated dose, toxicokinetics, and reproductive toxicity. If the European Commission determines that alternatives for these endpoints are not likely to be available, it can propose a further extension. To this end, the Commission has instructed experts to produce reports on the status of alternatives for the 2013 deadline. We criticized the draft reports on a number of issues. First, the experts fell into the "high fidelity fallacy trap," i.e. asserting that full replication of the in vivo response, as opposed to high predictivity, is required before an animal test can be considered useful for regulatory purposes. Second, the experts' reports were incomplete, omitting various methods and failing to provide data on the validity, reliability, and applicability of all the methods discussed, regardless of whether the methods were in vivo, in vitro, or in silico. In this paper we provide a summary of our criticisms and provide some of the missing data in an alternative proposal for replacement of animal tests by 2013. It is our belief that use of the Threshold of Toxicological Concern (TTC) will be a useful method to mitigate much animal testing. Alternative approaches for carcinogenicity and skin sensitization could be considered sufficient in the very near future, even though these tests are not listed under the 2013 extension. For repeated dose, toxicokinetics, and reproductive toxicity a combination of in vitro methods may be able to provide appropriate protection for consumers, especially when viewed in the context of the poor predictivity of the animal models they replace. We hope the revised report will incorporate these comments, since a more thorough and positive review is required if the elimination of animal testing for cosmetics in Europe and beyond is to be achieved.

Reporting the implementation of the Three Rs in European primate and mouse research papers: are we making progress?

It is now more than 20 years since both Council of Europe Convention ETS123 and EU Directive 86/609/EEC were introduced, to promote the implementation of the Three Rs in animal experimentation and to provide guidance on animal housing and care. It might therefore be expected that reports of the implementation of the Three Rs in animal research papers would have increased during this period. In order to test this hypothesis, a literature survey of animal-based research was conducted. A randomly-selected sample from 16 high-profile medical journals, of original research papers arising from European institutions that featured experiments which involved either mice or primates, were identified for the years 1986 and 2006 (Total sample = 250 papers). Each paper was scored out of 10 for the incidence of reporting on the implementation of Three Rs-related factors corresponding to Replacement (justification of non-use of non-animal methods), Reduction (statistical analysis of the number of animals needed) and Refinement (housing aspects, i.e. increased cage size, social housing, enrichment of cage environment and food; and procedural aspects, i.e. the use of anaesthesia, analgesia, humane endpoints, and training for procedures with positive reinforcement). There was no significant increase in overall reporting score over time, for either mouse or primate research. By 2006, mouse research papers scored an average of 0 out of a possible 10, and primate research papers scored an average of 1.5. This review provides systematic evidence that animal research is still not properly reported, and supports the call within the scientific community for action to be taken by journals to update their policies.

The SCHER report on non-human primate research - biased and deeply flawed.

The European Commission's Scientific Committee on Health and Environmental Risks (SCHER) recently issued an Opinion on the need for non-human primate (NHP) use in biomedical research, and the possibilities of replacing NHP use with alternatives, as part of the Directive 86/609/EEC revision process. Here, we summarise our recent complaint to the European Ombudsman about SCHER's Opinion and the entire consultation process. It is our opinion that the Working Group almost entirely failed to address its remit, and that the Group was unbalanced and contained insufficient expertise. The Opinion presumed the validity of NHP research with inadequate supporting evidence, and ignored substantial evidence against the need for NHP research and examples of valid alternatives that could replace the use of NHPs. Because the European Commission and others might base their approach to NHP research directly on the inquiry's findings during the revision of Directive 86/609/EEC, the implications of a flawed analysis of the efficacy of NHP research are extremely serious, both for animal welfare and for human health and safety. The conduct of the SCHER inquiry, and its published Opinion, should therefore be of major and widespread concern, and should not be given any political, scientific or legislative credibility.

Estimates for worldwide laboratory animal use in 2005.

Animal experimentation continues to generate public and political concern worldwide. Relatively few countries collate and publish animal use statistics, yet this is a first and essential step toward public accountability and an informed debate, as well as being important for effective policy-making and regulation. The implementation of the Three Rs (replacement, reduction and refinement of animal experiments) should be expected to result in a decline in animal use, but without regular, accurate statistics, this cannot be monitored. Recent estimates of worldwide annual laboratory animal use are imprecise and unsubstantiated, ranging from 28-100 million. We collated data for 37 countries that publish national statistics, and standardised these against the definitions of 'animals', 'purposes' and 'experiments' used in European Union Directive 86/609/EEC. We developed and applied a statistical model, based on publication rates, for a further 142 countries. This yielded our most conservative estimate of global animal use: 58.3 million animals in 179 countries. However, this figure excludes several uses and forms of animals that are included in the statistics of some countries. With the data available, albeit for only a few countries, we also produced, by extrapolation, a more comprehensive global estimate that includes animals killed for the provision of tissues, animals used to maintain genetically-modified strains, and animals bred for laboratory use but killed as surplus to requirements. For a number of reasons that are explained, this more-comprehensive figure of 115.3 million animals is still likely to be an underestimate.