ANIMAL TESTING FAQS

COSMETICS

How can safety be ensured without animal testing?

For consumer products, safe existing ingredients are the key. Hundreds of companies have sworn off animal testing, yet still produce new, safe products. They do so by choosing from among thousands of widely available raw ingredients that have been tested in the past or have a history of safe use. In addition, there are many methods and approaches available for assessing safety that do now use animals, and with increasing frequency, new products can be assessed for safe use without any new animal testing. New approaches combine chemistry, computer modelling, cell-based tests, and more complex animal tests if necessary, with improved methods of understanding exposure to identify safe use conditions with confidence. For more, see our AFSA Cosmetics Capacity Building program.

Why do companies still test cosmetics on animals if it’s not legally required?

Some companies believe that “innovation” requires the creation or use of new chemistries, which are invariably subject to new animal testing, either by or on behalf of the cosmetic manufacturer, or more commonly, by the specialty chemical producer itself. And multinational companies that sell cosmetics in countries such as China may be required to conduct new animal testing to meet the requirements of national authorities.

What are the alternatives to animal testing?

Firstly, companies can use ingredients that are already known to be safe, of which there are thousands. These ingredients have been tested in the past and don’t require new testing. This is how so many socially conscious companies have been able to swear off animal testing. Secondly, companies can use non-animal tests where new data need to be generated. More than 40 non-animal tests have been validated for use, and these modern alternatives can offer results that are more relevant to people, often more cheaply and efficiently too. Advanced non-animal tests represent the very latest techniques that science has to offer, replacing outdated animal tests that have been around for many decades and haven’t stood the test of time. For example, there are a number of skin tests available that use human reconstructed skin, such as EpiDerm, as wells as the “3T3 NRU” test for sunlight-induced “phototoxicity”, and the Bovine Cornea Opacity and Permeability test for eye corrosion.

What’s being done to end cosmetics animal testing?

Humane Society International, Humane Society of the United States, and Humane Society Legislative Fund launched the Be Cruelty-Free campaign in 2012—the largest global campaign to end cosmetics animal cruelty. We’re working to:

Prohibit new cosmetic animal testing and trade across key global beauty markets by the end of 2023.
Promote modern science, championing the development of new non-animal tests and training regulators and companies in their use.
Educate consumers, raising awareness about animal testing and how to shop cruelty-free.
Work with companies to help them move away from animal testing, and partnering with companies to lobby for change.

Which countries prohibit animal testing for cosmetics?

Animal testing for cosmetics has been banned throughout the European Union (EU) since 2009, and the sale of cosmetic products or ingredients subject to new animal testing after March 2013 is also illegal. Israel imposed similar bans in 2007 and 2013. To date, 39 major economies have enacted laws prohibiting or restricting cosmetic animal testing and/or trade including, Australia, India, Norway, Israel, Taiwan, Guatemala, New Zealand, South Korea, seven states in Brazil, and three states in the U.S. Today Humane Society International (HSI), The Humane Society of the United States (HSUS) and our partners are driving forward further legislative efforts in Brazil, Canada, Chile, Mexico, the Association of Southeast Asian Nations (ASEAN), Sri Lanka, South Africa, and the United States.

In most other countries, cosmetics animal testing is neither expressly required nor prohibited, and therefore continues to take place at the discretion of cosmetics companies and ingredient suppliers. In a few countries, including China, cosmetics animal testing may still be a legal requirement for some ingredients and finished products.

TOXICOLOGY

What is toxicology?

Toxicology is the study of how chemicals affect people, as well as animals and plants in the environment. Determining the toxicity of a chemical – how harmful it is — can tell us the amount to which humans and the environment can be safely exposed. This is important because we encounter thousands of chemicals every day – from pollutants in the air we breathe to pesticides in the food we eat and ingredients in the products we use and in the medicines we take. For many of these chemicals, exposure at certain levels, or for sustained periods of time, could have harmful effects ranging from mild skin irritation to serious birth defects or cancer.

Toxicologists apply different analytical techniques to determine the potential of a chemical to cause harm. They investigate the physical properties of the chemical, how it might be absorbed into the body, and what happens as it is broken down in the body. Toxicologists also want to know what level of exposure will produce an adverse effect (toxicity), and importantly, to identify the highest exposure level that results in no obvious effect. They also consider the risk – the danger a chemical poses based on the exposure due to how we use it or come across it in daily life. The risk of harm will depend on the type of chemical, the amount of chemical, the route of exposure (whether it was inhaled, eaten, or absorbed through the skin), and the number of times or length of exposure. Different people will have different susceptibilities depending on their age, gender, genetics, if they have any medical conditions, or if they are pregnant.

Toxicologists use information about each chemical to determine safe exposure levels and establish protective measures to help us avoid situations that may lead to harmful effects.

How are animals used in chemical testing?

Historically, animals have been used as substitutes for people to test the toxicity of chemicals. Certain “model” species of animals have also served as stand-ins for other animals found in the environment. To test chemicals that are used in everyday products – everything from personal care products like lotions or shampoos to cleaning products to oils, paint thinners, etc. – common species used are rabbits, rats, guinea pigs, and mice. To test things that are more likely to be dangerous, like pesticides (rat poison, weed killers, insect repellents), dogs are also used. And to test drugs, other species like pigs and monkeys are often used. Primarily rats, mice, and chickens are used to test the safety of food additives, and several different species of birds and fish are used to test the potential environmental effects of chemicals such as food additives, cosmetics, and pesticides. Birds, rats, mice and fish are most commonly used animals in chemical testing. Although the actual numbers of animals used are not available for most countries around the world, based on reporting from those countries where numbers are available, it was estimated that roughly 9.5 million animals were used for chemical safety assessment worldwide in 2014. For more about what animals are used, and in what type of tests, see “How are animals used in chemical testing .”

Why do we need a better approach based on non-animal methods?

In addition to ethical concerns, there are scientific and practical limitations to using animals to assess chemical toxicity. At the same time, improvements in our understanding of biology as well as technical advances in cell culturing, engineering and computer science provide an opportunity to revolutionize both research and chemical safety assessment.

As an example, research for drug discovery and development has relied heavily on the use of preclinical animal models. Nevertheless, more than 90% of compounds entering clinical trials fail to gain regulatory approval, mainly as a result of insufficient efficacy and/or unacceptable toxicity in humans, due in large part to the limited predictive value of preclinical studies (Plenge et al., 2013, in Langley et al. 2017). The lack of translation from animal studies to humans has several causes, including important differences between species, the fact that animal models do not fully mimic human diseases, lack of understanding of underlying mechanisms of diseases, as well as experimental design flaws and bias (Institute of Medicine, 2014; Pound and Bracken 2014).

Overall, there is a growing recognition that, to increase the success rate, a stronger focus on human-relevant data is therefore needed (Langley et al., 2017), moving from a system that relies on animal assays to one that aims to identify and characterize specific mechanisms or pathways that lead to adverse effects in humans, to design assays to measure pathway responses, to develop models that can predict toxicity using the assay data, and to set priorities among chemicals for more comprehensive toxicity testing (NRC, 2007; NASEM, 2017). For more, see “Why do we need a better approach?”

What is a “Pathway-based Approach” to toxicology?

In brief, this approach involves a characterization of the inherent properties of a chemical (or drug – a drug is a chemical with special properties!), and the application of our knowledge of the underlying biology – biological pathways (that’s where the “pathway-based” comes from), combined with an understanding or estimate of exposure, to design a testing strategy that will give exactly the information needed to safely use that chemical (or drug). Sound fantastical? That’s what pretty much everybody thought when the National Academy of Sciences proposed it in 2007 (NRC, 2007), but it turns out that it’s not so far-fetched after all, and enormous progress has been and continues to be made since then. For more on what this is, see our section on What is a “Pathway-based Approach” to toxicology?

PATHWAYS

Adverse Outcome Pathways (AOPs), Mode of Action (MoA), and mechanism of action – what’s the difference?

Adverse Outcome Pathway (AOP)

Conceptually, an AOP can be viewed as a sequence of events commencing with initial interactions of a stressor with a biomolecule in a target cell or tissue (i.e., molecular initiating event), progressing through a dependent series of intermediate events and culminating with an adverse outcome. AOPs are typically represented sequentially, moving from one key event to another, as compensatory mechanisms and feedback loops are overcome (OECD No. 184, 2017). AOPs describe biological pathways that can be perturbed, but are stressor agnostic (not specific to a particular chemical or other stressor). AOPs include the entire cascade of biology, all the way to potential adverse outcomes at the individual or population level.

Mode of action (MOA)

Mode of action is defined by WHO as “A biologically plausible sequence of KEs leading to an observed effect supported by robust experimental observations and mechanistic data. A mode of action describes key cytological and biochemical events – that is, those that are both measurable and necessary to the observed effect – in a logical framework.” (OECD No. 184, 2017).

While conceptually similar, a critical difference is that a MoA is specific to a chemical and includes chemical related key events (KEs) such as metabolism and focus on the upstream KE events. MOA can result in either adversity or beneficial outcomes. MoA analysis for species concordance additionally takes into account chemical-specific absorption, distribution, metabolism and elimination. It is anticipated, then, that increasingly, AOPs will be developed that contribute to chemical-specific MoA analysis (as one application) and vice versa (Bal-Price, et al., 2017).

Mechanism of action

Mechanism of action for toxicity is the detailed molecular description of key events in the induction of cancer or other health endpoints. Mechanism of action represents a more detailed understanding and description of events than is meant by mode of action or is generally considered in AOPs (OECD No. 184, 2017).

What are Integrated Approaches for Testing and Assessment (IATA)?

Integrated Approaches for Testing and Assessment (IATA)

IATA are pragmatic, hypothesis-based approaches for chemical hazard characterization that rely on an integrated analysis of existing information coupled with the generation of new information using testing strategies. IATA follow an iterative approach to answer a defined question in a specific regulatory context, taking into account the acceptable level of uncertainty associated with the decision context. There is a range of IATA – from more flexible, non-formalized judgment-based approaches (e.g., grouping and read-across) to more structured, prescriptive, rule-based approaches [e.g., integrated testing strategy (ITS), defined approach (DA)]. IATA can include a combination of methods and can be informed by integrating results from one or many methodological approaches [(Q)SAR, read-across, in chemico, in vitro, ex vivo, in vivo] or omic technologies (e.g., toxicogenomics). Ideally, an IATA should, wherever possible, be mechanistically informed, where the AOP concept can be applied as a framework to develop IATA (OECD No. 260, 2016). IATA defined approaches based on AOPs are under development for the prediction of the skin sensitization potential of chemicals (OECD No. 256, 2016).

What are New Approach Methodologies (NAMs)?

According to the U.S. EPA’s “Strategic plan to promote the development and implementation of

alternative test methods within the TSCA program” of 2018, NAMs refer to any technology, methodology, approach, or combination thereof that can be used to provide information on chemical hazard and risk assessment that avoids the use of intact animals. In 2019, U.S. EPA also released an updated list of NAMs that do not require new vertebrate animal testing. Updates on this list are expected to be released at least once a year.

In the ECHA’s workshop proceedings of 2016, NAMs are reported to include in silico approaches, in chemico and in vitro assays, as well as the inclusion of information from the exposure of chemicals in the context of hazard assessment. They also include a variety of new testing tools, such as “high-throughput screening” and “high-content methods” e.g., genomics, proteomics, metabolomics; as well as some “conventional” methods that aim to improve understanding of toxic effects, either through improving toxicokinetic or toxicodynamic knowledge for substances.

How are AOPs useful?

A variety of potential uses have been described for AOPs; the extent to which decisions can be supported by a given AOP depends on the level of uncertainty and quantitative understanding of the KERs (OECD No. 184, 2017):

by identifying and describing the KEs, AOPs describe the rationale for the use of particular methods and also by identifying potentially more predictive methods for development. In this way, AOPs provide curated and transparent support for the development and regulatory use of New Approach Methodologies (NAMs).
AOPs can also be used as a basis for developing an IATA or an ITS, and for assisting in the identification of the nature and the extent of new data that should be generated (Bal-Price and Meek, 2017).
They can also be used for further development and application of alternative approaches, such as read-across, where categories are first formed and data gaps filled within the category, leading to potential refinement, reduction and/or replacement of conventional in vivo testing (OECD No. 184, 2017).
AOPs can also be used to contribute to a number of regulatory contexts, including but not limited to: (1) priority setting for further testing, (2) hazard identification, (3) classification and labelling, and (4) risk assessment (OECD No. 184, 2017).
AOPs can also serve as the starting point for MOA analysis for specific chemicals, incorporating consideration of chemical space and ADME (OECD No. 184, 2017).
The development of many AOPs will form networks of biological pathways, thus ensuring a more realistic representation of a biological disease/process.
Safety assessment of chemicals could benefit from the development of quantitative understanding and description of the AOPs by means of Systems Toxicology.

What is the AFSA’s strategy to support the development of the AOP program?
AFSA’s strategy focuses on three pillars: i) development & dissemination of AOP training resources, ii) support to the OECD AOP Program, and iii) promotion of the paradigm shift in health research.

In particular, we plan to i) give AOPs lecture series across selected European universities, industries, and SMEs, alongside global conferences and/or workshops, ii) organize nation-wide AOPs training programs in other countries than Europe and US, and iii) engage with health scientists and funding bodies in dedicated workshops in the biomedical area. We are also actively contributing in the ongoing AOPs & IATA case studies projects at the OECD level.

Each of these activities are taken forward by several members of the R&T HSI team globally, with the support of the AFSA Tox21 partners.

If you’re interested in AOP training programs, or in giving your contribution, please contact us.

VACCINES

If you are so convinced that animals are obsolete and represent the past, why are they still used?

Vaccines are a powerful instrument of public health, administered to healthy people: their safety is paramount, and as such, authorities are extremely cautious with change, because they fear it might endanger public safety. Switching from animals to new methods requires much information, trust in current manufacturing process, and the willingness to move forward even if it means time and resources need be invested.
Also, test methods able to simply replace animal tests are practically impossible to create and validate: animal methods are usually so unprecise, varying and subject for their results from a variety of external conditions that no single alternative method can replicate them.
Unfortunately, regulatory authorities have always required that tests intended to substitute animal tests be able to perfectly mimic them, something we have come to understand as impossible.

We are now, finally, at a crossroads, where it has become widely understood and accepted that alternative methods do not replace, but “substitute” animal methods, that is: alternative methods guarantee efficacy and safety of vaccines in methods that are unrelatable to animal methods.

This key realization will help propel the transition forward.

And the more we remove legacy animal methods, the more authorities – everywhere – will be convinced of all that is possible, the more the transition will accelerate.

Why should a third party be needed to promote a transition to alternative methods? Aren’t WHO/OIE or other international standardization bodies enough to secure change?

WHO/OIE and other international bodies like the International Councils for Harmonization of Technical Requirements for Pharmaceuticals for Human and Veterinary (ICH and VICH) are doing much work to harmonize the regulatory landscape worldwide. But the speed at which they must move, and the opportunities they have to freely engage the stakeholders are limited by their institutional role and its duties, as is their ability to reach out to smaller manufacturers and regulators of developing countries.

AFSA proposes to take on itself a different, complementary role: that of a facilitator, directly engaging stakeholders from all over the world, creating venues for discussion and information exchange (in so doing also disseminating WHO/OIE’s and ICH/VICH’s work), offering opportunities to smaller countries in the developing world to voice their doubts, concerns, fears, and ideas, and helping getting them in contact with experts and regulatory authorities that are steps forward in acceptance of alternative methods.

As a third party supported by already active stakeholders, AFSA can be of service without generating obligations, be proactive without its actions taking an immediate political meaning, and make proposals that others are freer to embrace due to their neutrality.

AFSA is also willing to tackle a key unmet need in this sector, that is the creation of a vaccine-specific database, an open resource for all the interested parties to locate regulations, ongoing and past projects and their results, access points and responsible for projects, be informed of opportunities for collaboration, and directly see what is the state of regulatory acceptance of alternative methods in other Countries/Regions.

Such a resource must be independent to be credible, and this is yet another reason for AFSA’s involvement as an active and reliable co-player.

What is the strategy AFSA has in mind to pursue change in the vaccine field?

AFSA believes, first and foremost, in inclusion: bringing all the involved stakeholders to the table so that through cooperation trust can be built, ideas be floated, and commitments be undertaken to seed the sows of change.

The strategy sees regulatory harmonization at its core, because the more national regulations converge, the easier it will be to standardize on alternative methods and for less advanced Countries to be convinced of the feasibility, and desirability of replacing animal methods.

To this end, AFSA is ready to discuss and work with all the relevant stakeholders, striving to promote the very concept of harmonization and its practical implementation.

Key to our efforts is also the belief in the power of information and its free flow: the easier for information to be diffused and retrieved, the higher the chances that alternative methods can end up on the agendas of regulatory authorities, and the better the possibilities that conscious efforts are invested to transition away from animal methods.

The database mentioned in the previous FAQ point is an example of this deep trust in the power of information and the conviction of the importance of its unhindered circulation.

Lastly, AFSA believes that example can work as a powerful instrument to motivate others to change.

Sharing information on the progress of certain areas/regions is fundamental, but key will also be achieving success in the elimination of a series of old, extremely deeply-rooted animal tests, like the General Safety Test (GST) and the Rabbit Pyrogen Test (RPT).
Those tests are the staple of practically every Pharmacopoeia (but for the most advanced): securing their worldwide deletion would not only make life easier for manufacturers and regulators while also sparing countless animals, but it would also be a demonstration that change for the better is truly possible, and that if tests of titanic import as those are removed, then regulators everywhere can see their confidence in alternative methods bolstered.

AFSA is profusing its efforts to make worldwide elimination of such tests a reality.