Overview

One of the latest major developments in the quest for greater use of new approach methodologies (NAMs), which are broadly defined as alternatives to animal testing for studying environmental and human health hazards, is the EU’s publication of its “Roadmap Towards Phasing Out Animal Testing for Chemical Safety Assessments.” Adopted by the European Commission (EC) on June 1, 2026, the roadmap follows a string of NAMs announcements, including the US FDA’s “Roadmap to Reducing Animal Testing in Preclinical Safety Studies,” published in April of last year, and the UK government’s “Replace Animals in Science: A Strategy to Support the Development, Validation and Uptake of Alternative Methods” policy paper, published in November 2025. Also released last year was Canada’s “Strategy to Replace, Reduce or Refine Vertebrate Animal Testing under the Canadian Environmental Protection Act, 1999 (CEPA).”

As is commonly a starting point for the greater use of NAMs, the EU Roadmap is not only focused on eliminating animal testing altogether but also reducing the volume of animal tests themselves, the number of animals used in tests, and mitigating animal distress during tests. These goals are in line with the “Replacement, Reduction and Refinement,” or 3Rs, approach.

The EC’s Roadmap is a plan to guide actions related to existing and proposed EU legislation, scientific bodies, resources, and regulations. One goal specifically defined by the Roadmap is the identification of regulatory needs. Other topics highlighted are the need for validation, qualification, standardization, and R&D of non-animal approaches, as well as knowledge sharing, progress monitoring, and engagement with stakeholders. The report opts for the use of the word “non-animal approaches” rather than NAMs.

The Roadmap sets out recommendations for short- and long-term actions to phase out of animal testing for assessing chemical safety, dividing them among three pillars: “pathways for making change happen,” “aligning institutional efforts and resources,” and “a robust organizational framework for working together in Europe and beyond.” In addition, specific types of tests for assessing human health and/or environmental safety are classified according to the goal of replacement, reduction, or refinement.

The Tests

The individual tests listed in the Roadmap as replacements for animal testing fall into one of three categories, computational methods, in-vitro assays, or in silico approaches, though tests from more than one category may be combined. The EC’s EU Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM) defines in-vitro tests as methods that “use tissues, reconstructed tissues, whole cells, or parts of cells.” Employing a more inclusive definition, the EU Scientific Committees define in vitro as “an artificial environment outside a living organism or body.”

The EURL ECVAM describes computational methods, which encompass in silico, as “mathematical approaches such as Quantitative Structure-Activity Relationship (QSAR) modelling and Physiologically Based Kinetic and Dynamic (PBK/D) modelling.” The description continues, “QSAR models predict biological or toxicological properties on the basis of the physicochemical and structural properties of chemicals,” and “physiologically based kinetic (PBK) models are mathematical descriptions of how chemicals distribute in humans or other animal species.”

A Working Paper published in conjunction with the Roadmap provides further details on the NAMS available and under development for environmental safety assessment and/or human health safety assessment. The Working Paper discusses in depth the latest scientific advancements, needs, and recommendations in each of these categories of tests.

Specific tests for environmental safety assessments using animal tests that could be replaced in the short term, according to the paper, are fish acute toxicity, aquatic and terrestrial bioaccumulation, fish chronic toxicity, endocrine disruption, and bird and mammal tests. The EURL ECVAM has already validated methods for several of these tests.

For human health safety assessment tests done on animals, the areas of opportunity described in the paper are tests for acute toxicity, genotoxicity, carcinogenicity, repeated dose toxicity, developmental neurotoxicity, developmental and reproductive toxicity, and endocrine disruption—human health. As with the environmental safety assessments, the EURL ECVAM has already validated methods for several of these tests.

Having validated replacement methods for animal testing since 1995, the EURL ECVAM lists validated methods spanning a wide range of NAMs, including those for skin irritation, skin corrosion, skin sensitization, and phototoxicity. The lab has validated a smaller set of methods for partial replacement and reduction of tests for eye irritation, skin sensitization, genotoxicity, carcinogenicity, and acute toxicity.

The EURL ECVAM’s Tracking System for Alternative Methods Towards Regulatory Acceptance (TSAR) database shows 36 tests that have been adopted and published by the EURL ECVAM originating from various organizations worldwide. Skin sensitization tests account for the highest percentage (17%), followed by pyrogenicity (14%) (see pie chart).

*Exceeds 100% due to rounding

Awareness

A survey conducted in late 2024 and early 2025 by the EC regarding NAMs, referred to as non-animal approaches in the survey, was used to inform the creation of the Roadmap, providing insight into the current use of NAMs. The survey was comprised of 193 stakeholders. Most respondents were EU or Member State authorities/agencies (29%), companies/industry associations (27%), or institutes/universities/commission partners (22%).

For human health hazard assessments, skin corrosion/irritation and skin sensitization are the non-animal approaches of which the highest percentage of respondents–over 60% each (more than one answer allowed)–were aware and believed the tests could be readily used to replace an animal-based method. Asked about non-validated tests that they were aware of, considered scientifically mature, and can use already, the top two choices were neurotoxicity and acute toxicity.

The survey also sought to determine which tests respondents did not see as ready to be used. The top two percentages of respondents chose reproductivity toxicity and repeated dose toxicity as non-animal approaches they were aware of but felt could not be used for the foreseeable future. The methods about which the highest percentage of respondents by far did not know about were aspiration hazards and respiratory sensitization.

For the same question, but for environmental hazard endpoints, the highest number of respondents chose aquatic/acute/short-time studies and bioaccumulation as validated methods that exist and can be used. The non-animal approach chosen as non-validated but scientifically mature and ready to be used was aquatic chronic/long-term studies. The two methods most chosen as those for which only animal tests can be used for the foreseeable future were aquatic chronic/long-term studies and avian toxicity.

Of the 40% of respondents whose organizations were developing non-animal test methods, the most common tests under development were in-vitro (using cells or other biological materials outside of a living organism) and integrated approaches to testing and assessment (combining multiple methods, including read-across and tiered testing). The EC’s Joint Research Center defines read-across as using “existing data on chemical analogues.”

For most respondents, the top two challenges to greater use of non-animal test methods were scientific barriers/lack of methodologies and regulatory acceptance, at over 80% each (multiple answers possible). Asked about the cost comparison between animal-based tests and non-animal tests, the highest percentage of respondents stated the cost varies greatly by hazard end-point.

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