The use of performance-based outcomes in achieving better global harmonisation in the care and use of animals in research

Abstract

Animal research remains a crucial component in our efforts to enhance both human and animal health. To better ensure research reproducibility and welfare of animals in research, harmonisation must be improved globally. This review explores the current state of harmonisation in animal research and the associated challenges, along with the roles of existing animal welfare principles, legislative guidelines, codes of practice, international and national organizations, and professional bodies in facilitating harmonisation. We discuss the current obstacles to harmonisation and suggest a performance-based solution. Examples of attainable performance-based outcomes are examined, encompassing areas such as ethical review and oversight, animal housing, environment, training, veterinary care, experimental design and reporting, and openness/transparency in research. In conclusion, the establishment of harmonised performance standards and the promotion of performance-based outcomes hold the potential to significantly improve global animal welfare in research and contribute to scientific advancement.

Introduction

There is no doubt that the use of animals in research has played a pivotal role in advancing biomedical science and remains essential for developing new disease treatments. The impact of the benefits of the use of animals in research cannot be underestimated, as evidenced by numerous Nobel Prize achievements, the development of COVID vaccines, and breakthroughs in diabetes and cancer research among others.^{1 –3}

Despite efforts to replace animal use with alternatives such as tissues, cells and computer modelling, as well as advancements in Artificial Intelligence and organ-on-a-chip technology, research animals continue to be a cornerstone of scientific progress.^4,5 Given the importance of research animals, it is incumbent upon scientists not only to reduce their use as much as possible but also to refine the way animals are utilised. Deficiencies in experimental design and differences in housing conditions are well documented to impact research reproducibility and robustness, ⁶ and with the ever growing global nature of research with greater public scrutiny, a more comprehensive method of harmonisation of animal use is still needed. ⁷

Current state of harmonisation

Welfare principles

For many years, numerous principles, guidelines and legislations have been developed in order to improve the welfare of research animals. The 3Rs principles developed by Russell and Burch in 1959 ⁸ are now indeed truly global, with references made to the 3Rs in codes of practice and legislation in all parts of the world. The five freedoms (from hunger and thirst; from discomfort; from pain, injury or disease; to express normal behaviour; and from fear and distress) first developed in 1965 ⁹ for the Farm Animal Welfare Council are used to a lesser degree than the 3Rs. For example, current legislation in the European Union refers explicitly to the 3Rs, but not to the five freedoms. ¹⁰ However, they may still be used as a metric during harms versus benefit analysis during protocol review by many institutions.

Legislation, guidelines and codes

A number of legislative documents are available globally. ¹¹ Legal documents largely focus on engineering standards often based on prescribed and measurable parameters, for example, cage space, cage change frequency, humidity or temperature ranges and air change frequency. There is, however, a number of differences between these standards, as well as the level of enforcement in each jurisdiction making harmonisation difficult. As opposed to legislation, other guidelines^{12 –16} focus more on common performance standards (e.g. evaluation of the sanitation practices), which can be adopted regardless of the legislation in place.

International and national organisations

Global organisations have also played a part in improving harmonisation. The Council for International Organizations of Medical Sciences (CIOMS) and the International Council for Laboratory Animal Science (ICLAS) published the International Guiding Principles for Biomedical Research Involving Animals (2012), ¹⁷ which provides a strong basis and outline, including expectations for the 3Rs, the need for standardisation to ensure scientific validity and reproducibility; the need for veterinary medical care; welfare and environmental and housing conditions; minimising pain and distress; humane endpoints; adequate training, and the need for euthanasia appropriate to the species. Also, the World Organization for Animal Health describes similar expectations in its Terrestrial Code. ¹⁸

National organisations, albeit with international reach, such as the UK’s NC3Rs and Norway’s Norecopa, have promoted concepts such as the ARRIVE and PREPARE Guidelines.^19,20 The ARRIVE Guidelines focus on improvements in experimental design and reporting. Scientific publications often lack sufficient information and detail as to how the study was designed, performed or data analysed, thus limiting the possibility for adequate scrutiny, or reproducibility. With questions being asked over reproducibility of animal research ⁶ the ARRIVE Guidelines serve as an important resource to guide researchers to consider a robust experimental design and provide the necessary information in publications. The PREPARE Guidelines focus on study formulation such as having a clear hypothesis and literature review; interaction between the scientist and animal care personnel, including necessary infrastructure, training and safety; and quality control, including characteristics to ensure standardisation, housing conditions, refined procedures and humane euthanasia. ²⁰

Both the ARRIVE and PREPARE Guidelines are complementary to improving harmonisation, in experimental planning and design, conduct and reporting, and, if followed, represent a significant improvement in harmonisation of information and practices leading to published research.

There has also been a recent drive to improve openness and transparency in research with the Concordat on Openness on Animal Research being agreed in the UK and being launched in Oceania by Understanding Animal Research; ²¹ as well as several other transparency agreements in European countries promoted by the European Animal Research Association and in the United States an Animal Research Openness initiative has also been launched.^22,23 So far, over 500 institutions worldwide have agreed to openness and transparency, so such agreements contribute to harmonisation since they encourage institutions to be open and report on how and why animals are used, and thus indirectly contribute to better practices.^{21 –23}

Professional organisations

Professional organisations have long been contributing indirectly to harmonisation by providing standardised training of animal care staff, as is the case with the Institute of Animal Technology and the American Association of Laboratory Animal Science (AALAS), with certified training for animal technicians.^7,24,25

Regional organisations such as Federation of European Laboratory Animal Science Associations (FELASA) regularly release guidance documents on various areas of animal research that include animal health surveillance, training and education, that are often adopted globally. ²⁶ The Asian Federation of Laboratory Animal Science Associations, ²⁷ Australian and New Zealand Laboratory Animal Association, ²⁸ the Federación de Sociedades Sudamericana de Ciencia en Animales de Laboratorio ²⁹ and Federación de Sociedades y Asociaciones Hispanas de América del Norte, Centro América y el Caribe de la Ciencia de los Animales de Laboratorio ³⁰ are also increasing their reach and membership in Asia, Oceania, Latin and Central American associations respectively, whereas ICLAS continues to promote ethical use of animals worldwide.

AALAS and FELASA established years ago a liaison body that has set up several joint working groups who have published recommendations on several topics, in an attempt to harmonise practices between the United States and Europe which could also have impact in the rest of the world. ²⁶ The American Veterinary Medical Association (AVMA) have played a significant role in harmonising euthanasia techniques, with their Guidelines for the Euthanasia of animals (2020) being widely adopted globally. ³¹

Veterinary care for laboratory animals is also enhanced through the work of regional collages of laboratory animal medicine (e.g. in Europe, America, Japan, South Korea and India), who all subscribe to follow the International Association of Colleges of Laboratory Animal Medicine, with the goal to promote the welfare and responsible use of laboratory animals through the certification of veterinary specialists.^7,32,33

One organisation that is uniquely positioned to contribute to harmonisation is AAALAC International (AAALAC). With over 1100 accredited institutions in over 50 countries, AAALAC applies standards from national legislation and codes as well as the three Primary Standards: the Guide for the Care and Use of Laboratory Animals, the Guide for the Care and Use of Agricultural Animals in Research and Teaching, and the European Convention ETS 123. ³⁴ Combining these standards with Positions Statements, FAQs and Reference Resources, AAALAC is able to adapt relatively quickly to emerging trends in new knowledge and best practices. Many of the guidelines and codes released by the International and National organisations described above (e.g. CIOMS/ICLAS Guiding Principles, FELASA health monitoring, AVMA Euthanasia Guidelines, etc.) are indeed included as Reference Resources by AAALAC’s Council on Accreditation, and thus become part of the evaluation of accredited programmes, thus directly impacting and improving global harmonisation.^7,34

Harmonisation versus standardisation

There is a tendency to use both terms as synonymous in presentations and dialogues at meetings in our field, when discussing having common standards and, even, legislation. However, in the authors’ opinion, they should not be considered as such. While standardisation would mean to follow exactly the same procedure for a particular activity, harmonisation would mean to agree on the same outcome of that activity, regardless of the procedure (performance-based approach). For example, we cannot aim to have all animals of a certain species housed all around the world with the same type of bedding and the same environmental enrichment items (standardisation), but we can promote whatever (there may be several options) bedding and enrichment items are appropriate for that species (harmonisation). The same idea on a more global level has already been presented some time ago by others. ³⁵

Current challenges and weaknesses

Despite all the harmonisation efforts and progress described above, further harmonisation has been challenging due to differences in legislation, guidelines, cultural norms and history. The experience obtained by AAALAC through the evaluation of animal care and use programmes globally allows us to say that they vary greatly between countries and regions, as does the enforcement of guidelines and legislation. International and national organisations that produce guidance documents often rely on voluntary adherence but otherwise are powerless to insist on full implementation and thus better harmonisation.^{33,36 –38}

It is possible to improve compliance with these principles and guidelines, but incentives need to be increased. For example, if granting bodies, funding agencies and publications were to insist on adherence to a set of principles, then improvement in harmonisation in these principles could potentially occur rapidly. But this approach needs commitment from such organisations to fully implement to be effective.

Greater harmonisation (as long as it is implemented with consideration given to performance standards that have been proven effective) would certainly lead to improved animal welfare, facilitate data sharing and collaboration between scientists, and improved science in general, and build public trust in the scientific process. However, given the diverse range of legislations and standards available, how could a unified set of principles be developed? Is there a way to combine all the schemes based on welfare principles, guidelines, principles and professional recommendations together?

Performance-based outcomes

One possible approach to overcome these differences is the use of a performance-based system that harmonises minimum expectations for the care and use of research animals. ³⁵ Utilising outcome-based goals or expected results rather than the process used, a performance-based approach may be a method to achieve an appropriate level of animal care when engineering standards differ, and thus achieve a more harmonised result.^33,35,37

It is hoped that when used correctly, performance expectations can ensure humane treatment and justified use of animals, improve research data quality by promoting best practices, and harmonise animal care and use practices globally.

Examples of achievable performance-based outcomes

Harmonisation in ethical review and oversight

Broad-based protocol review

The composition of Institutional Animal Care and Use Committees (IACUCs) or Animal Ethics Committees (AECs) varies globally, as does the level of review and post-approval monitoring. For example, members from a non-scientific background and public members referred to in the ILAR Guide are not considered in the European Directive.^10,12 Better harmonisation can be achieved by ensuring broad representations of differing views within the committee. It is common to have IACUC/AEC representation from a veterinarian knowledgeable about the species used and a scientist familiar with research design and implementation, but have community views been considered? Engaged and committed lay/non-scientist members and external members can have a significant impact on protecting animal needs as well as institutional reputation by conveying societal views and concerns. The performance outcome in this case is to ensure a balanced and representative review of animal care and use protocols.

Considerations of factors that directly impact animal welfare

Details of what is included during protocol review may also vary, for example in the US it is typical to address occupational health and safety issues whereas this is not common practice in Europe, but there are some aspects that directly impact on animal welfare that should always be considered. Essential aspects include personnel training and competency, harm–benefit analysis, experimental design (powering, randomization, blinding), housing and behavioural management needs, procedure details, analgesic and anaesthesia use, use of non-pharmaceutical grade compounds, humane endpoints and details of euthanasia technique. Insisting that all these aspects are reviewed better ensures a harmonised and unified approach to ethical review. A good example of harmonization, within the European Union, is the consensus document on Project Evaluation and Retrospective Assessment. ³⁶

The entire programme must have oversight

IACUCs/AECs should not limit their activity to protocol review, but also oversee entire animal care programmes, ensuring compliance with facility standards, protocols, personnel training and welfare expectations.^7,37 While some jurisdictions assign this oversight to regulatory bodies, establishing a minimum level of oversight, facility inspections and post-approval compliance and monitoring should be an international minimum expectation.

Harmonisation in animal housing, environment, training and veterinary care

Animal housing

Standards for space requirements are often defined in legislation based on engineering standards.^{12 –16} Whilst these space requirements differ regionally and form a basis for adequate space, the performance outcome is to ensure that the wellbeing and behavioural needs of the animals are met. On a performance basis, if the behavioural needs for space, social housing, complexity and enrichment are met, then the animals’ welfare can be better assured. Institutions should regularly review behavioural management of animals to ensure that the wellbeing needs of animals are always met.

Animal environment

Air changes, temperature and humidity standards are also often quoted as engineering standards,^{12 –16} but on a performance basis, the goal of these standards is to ensure an environment that is appropriate for the species concerned as well as safe for personnel. An evaluation or review of animal health, environmental monitoring and safety risk assessments should be considered from a performance basis to ensure an appropriate environment.

Training and competence

Personnel training and competence in animal care and use programmes can have a significant impact on animal welfare and research robustness and reproducibility. Not all jurisdictions mandate training or competency assessments for researchers or animal workers, as, for example, in many European countries. At a minimum, there should be some form of education and training, and competency-based assessment for all personnel engaged in animal care and use. Institutions should develop training, education or competency programmes designed to meet their specific needs.

Veterinary care

Within laboratory animal science there are often established engineering standards to define humane endpoints (e.g. tumour size or weight loss), anaesthetic/analgesic regime, or clinical treatments for laboratory animal species. However, due to the diverse range and changing nature of research, it is often needed for the veterinarian to exercise professional judgement on what action/treatment to take, for example, when an analgesic may confound the research data, or when to remove the animal from the study. As qualification and education standards and curriculum vary significantly globally, for a performance-based approach to work it is vital that veterinary staff are appropriately experienced and qualified and receive continual education in the species used. ⁷ It is also vital that veterinarians have the institutional support and authority to administer the programme under their professional judgement so that animal welfare needs can be balanced with research outcomes.

Harmonisation in experimental design and reporting and openness

Experimental design

Without robust experimental design, reproducibility and reliability of animal research will be severely compromised. ³⁸ Poorly designed animal experiments lacking in statistical power, blinding, randomisation and appropriate controls undermine scientific progress, whereas inadequate reporting of methods, housing conditions, genetics, disease status et cetera make true evaluation of the study impossible. Ideally, scientific journals and granting/funding agencies should mandate appropriate planning and reporting of animal experiments using established systems such as the ARRIVE ¹⁹ and PREPARE ²⁰ Guidelines, as the current voluntary inclusion of much of the needed information is not effective. In the absence of incentivising by granting bodies and scientific journals, IACUC/AEC should ensure that these experimental design and reporting standards are met.

Openness and transparency

Whilst the use of animals in research remains critical to scientific progress, it is also vital that we improve our openness and transparency as to how and why we do animal research.^{21 –23} Only by having open dialogue and scrutiny by the public will we push harder to raise standards of animal care and use and ensure continued public trust.

Conclusions

The use of performance-based outcomes is a promising method to achieve better harmonisation in the care and use of animals in research. By focusing on outcomes rather than the methodology, performance metrics can provide a flexible framework for improving humane care of animals used in science and medicine. Such an approach could lead to improved animal welfare, ensuring that they are humanely treated and that there is improved research quality that promotes better experimental design, leading to more reproducible and reliable research data, and increased global harmonisation by bridging the gap between national and international standards. Very importantly, this approach could also facilitate improvements in regions lacking specific legislation on the protection and/or enforcement of standards, of animals used in research, where the same performance outcomes could be achieved regardless of the legal framework.

Whilst significant challenges remain to implementing performance-based outcomes, such as the development of clear and measurable parameters, the potential benefits to both animal welfare and research are significant. International organisations such as AAALAC and ICLAS, as well as national/regional organisations and associations, could play a significant role in developing these metrics, and thus shape the future of global animal research practices.

Some specific steps that may be considered include: development of clear performance standards on aspects that directly impact welfare, for example, pain management, humane endpoints, behavioural management and veterinary care; training and competency expectations for animal care staff and researchers; and promotion of performance-based outcomes such as the ARRIVE, PREPARE and openness agreements in funding applications and scientific publications.

By following such steps, significant improvements could be made in global research animal welfare as well as the scientific and medical advancement.