Inconsistencies in Data Requirements of EU Legislation Involving Tests on Animals

European Union (EU) legislation on the protection of animals used for scientific purposes requires that alternative methods must be used instead of animal tests wherever they are available. Unfortunately, this provision is not implemented to its full extent when it comes to risk assessment of chemicals and new products prior to their authorization and placing on the market in the EU. In this study, we screened data requirements of relevant EU law regarding chemicals (REACH), biocides, pesticides, and food safety (Novel Food) and found that data requirements as part of the risk assessment do not always reflect stateof-the-art science and technology. Most of the data requirements we investigated still include testing on animals for many toxicological endpoints, even though more than 40 alternative testing methods accepted at the level of the EU or the OECD are available. This may be due to a multitude of reasons, including a shortage of both manpower to implement existing knowledge and expertise in the field of alternative methods, as well as unclear and misleading statements on the applicability and state of validation of alternative methods. In conclusion, we strongly suggest a homogeneous EU-wide approach for all areas involving risk assessment of substances with the goal of better implementing the 3Rs and complying with Directive 2010/63/EU. This also would streamline data requirements, save costs on various levels, and enhance product safety for consumers.

Treaty on the Functioning of the European Union (TFEU), amended 2009: "In formulating and implementing the Union's agriculture, fisheries, transport, internal market, research and technological development and space policies, the Union and the Member States shall, since animals are sentient beings, pay full regard to the welfare requirements of animals, while respecting the legislative or administrative provisions and customs of the Member States relating in particular to religious rites, cultural traditions and regional heritage."

Common basic principles:
Each experiment has to be essential for a given purpose The number of animals as well as pain, suffering and harm have to be reduced to a minimum Pain, suffering and harm caused to the animals have to be ethically justifiable

Recital 42: […] It is necessary to introduce specific measures in order to increase the use of alternative approaches and to eliminate unnecessary duplication of regulatory testing. […)
Article 4 'Principle of replacement, reduction and refinement' 1. Member States shall ensure that, wherever possible, a scientifically satisfactory method or testing strategy, not entailing the use of live animals, shall be used instead of a procedure.

Article 13 'Choice of methods' 1. […] Member States shall ensure that a procedure is not carried out if another method or testing strategy for obtaining the result sought, not entailing the use of a live animal, is recognised under the legislation of the Union. 2. In choosing between procedures, those which to the greatest extent meet the following requirements shall be selected: (a) use the minimum number of animals; (b) involve animals with the lowest capacity to experience pain, suffering, distress or lasting harm; (c) cause the least pain, suffering, distress or lasting harm;
Directive 2010/63/EU

German Animal Welfare Federation -Animal Welfare Academy
Analysis of data requirements of EU legislation that involves testing on animals: Are accepted alternatives considered/included/referred to?
How are data requirements structured?
Rules for adaptation?
Consistency of legislation/data requirements?

Analysis of EU legislation -what did we look for?
German Animal Welfare Federation -Animal Welfare Academy Some of the accepted alternative methods that are available are not considered in the data requirements we analyzed Structure of data requirements is non-uniform from legislation to legislation Considerable disparities were also identified in wording, terminology and references to waiving criteria and rules for adaptation

Analysis of EU legislation -what did find?
German Animal Welfare Federation -Animal Welfare Academy

German Animal Welfare Federation -Animal Welfare Academy
Lacking or insufficient instructions how testing on animals can be replaced, reduced or refined in BPR (partly) und PPPR (continuously) "Waiving" criteria and rules for adaptation differ from legislation to legislation Structure, terminology and wording of data requirements inconsistent and often confusing

German Animal Welfare Federation -Animal Welfare Academy
To reduce and replace testing on animals for regulatory toxicity testing and to improve consumer protection in the EU: The European Commission has to act immediately to eliminate those animal tests from the data requirements that can be replaced by accepted alternative methods Setting up a central organ or institution that is responsible for the design and update of data requirements Strategy for design, compilation and update of data requirements needs to be revised and harmonized (harmonized structure, harmonized terminology, harmonized wording, "Waiving" criteria and rules for adaptation) Lay down best practice rules for EU acceptance of an AM after its adoption of an alternative method as an OECD TG (do all OECD TGs have to be accepted?Timeline?)

Recommendations
Internationally accepted AMs available since 2009 (OECD, TGs 437 + 438, can partly replace in vivo test) Adaptation to Technical Progress: AMs were updated in TMR (440/2008/EC) via Amending Regulation (EC) No. 1152/2010 in 2010 BUT: Biocidal Products Regulation + Plant Protection Products Regulation still lack an update or inclusion of AMs in respective data requirements PPPR: in vivo test (B.5., TMR) required exclusively (!), no reference to AMs BPR: reference to "Sequential Testing Strategy for Eye Irritation and Corrosion" (appendix to in vivo eye irritation and corrosion test method (B.5.) in TMR) Testing strategy (in appendix to B.5., TMR) "not an integral part of testing method B.5.", was developed by the OECD in 1996, therefore does not mention specific AMs (just mentions "validated and accepted in vitro or ex vivo tests").Example 1: eye irritation German Animal Welfare Federation -Animal Welfare Academy Internationally accepted AM for skin irritation available since 2010 (OECD, TGs 439, can partly or fully replace in vivo test) Adaptation to Technical Progress: AM were updated in TMR (440/2008/EC) via Amending Regulation (EC) No 761/2009 in 2009 BUT: Biocidal Products Regulation + Plant Protection Products Regulation still lack an update or inclusion of AM in respective data requirements PPPR: in vivo test (B.4., TMR) required exclusively (!), no reference to AM BPR: reference to "Sequential Testing Strategy for Skin Irritation and Corrosion" (appendix to in vivo eye irritation and corrosion test method (B.4.) in TMR) Testing strategy (in appendix to B.4., TMR) "not an integral part of testing method B.4.", was developed by the OECD in 1996, accepted AM only mentioned under "References" in description of testing strategy Example 2: skin irritation German Animal Welfare Federation -Animal Welfare Academy No binding procedure for inclusion of newly adopted OECD TGs in the TMR Confusion about status of inclusion of newly accepted AMs because of separate Amending Regulations for Adaptation to Technical Progress Lacking Adaptation to Technical Progress of the TMR: In Vitro Mammalian Cell Micronucleus Test (OECD TG 487) Acute Toxic Class Method for Acute Inhalation Toxicity (OECD TG 436) EOGRTS (OECD TG 443) OECD GD No. 129 on using cytotoxicity tests to estimate starting doses for acute oral systemic toxicity tests Listing of outdated or unnecessary animal test methods for reproductive toxicity and skin sensitization (One generation reproductive toxicity study, Guinea Pig Maximization Test / Buehler-Test) (TMR) Reference to corresponding OECD TG missing (TMR) Issues concerning unclear, inconsistent or confusing structure of the TMR and its register of test methods Results -analysis of data requirements 1 German Animal Welfare Federation -Animal Welfare Academy Lacking reference to accepted alternative methods for skin irritation (BPR, PPPR) Lacking reference to accepted alternative methods for eye irritation (BPR, PPPR) Lacking reference to accepted alternative methods for skin sensitization (PPPR) Lacking reference to accepted alternative methods for reproductive toxicity (PPPR) Requirement of unnecessary 12-month toxicity study in dogs (PPPR) Inconsistencies in terminology, design of data requirements, and rules for adaptation to technical progress (PPPR) Introduction of endpoints that still lack standard testing methods (respiratory sensitization, BPR)

German Animal Welfare Federation -Animal Welfare Academy Animal experiments -legal situation
Directives 86/609/EEC and 2010/63/EU European Convention ETS 123 Animal Welfare Acts of the EU Member States Recitals: […] whereas such harmonization should ensure that the

number of animals used for experimental or other scientific purposes is reduced to a minimum, that such animals are adequately cared for, that no pain, suffering, distress or lasting harm are inflicted unnecessarily and
ensure that, where unavoidable, these shall be kept to the minimum; Whereas, in particular, unnecessary duplication of experiments should be avoided,[…]Article 7 2.An

experiment shall not be performed if another scientifically satisfactory method
of obtaining the result sought, not