alterREACH: Non-animal (alternative) testing methods for REACH

The implementation of REACH challenges our ability to perform rapid, robust and ethically sound ecotoxicological testing. NIVA addresses this challenge in the multi-national and multi-disciplinary research project alterREACH funded by the Norwegian Research Council.


The Registration, Evaluation, Authorisation and Restriction of Chemical substance (REACH) is the new European Community Regulation on chemicals and their safe use.

Estimates indicate that as much as 30,000 single chemicals may be required to be registered with a potentially risk assessment requirement based on substance-specific data for the persistence, bioaccumulation and toxic (PBT) properties for several thousand chemicals.

This introduces a challenge for industry, the regulatory agencies and the toxicological research community to develop and validate alternative high-throughput testing strategies and limit the testing with animals by implementing the reduction, refinement and replacement (the 3Rs) principle in ecotoxicological testing.

This applies in particular to the use of aquatic vertebrates such as fish, which is regulated under the animal research and welfare legislation.

This project seeks to provide theoretical and experimental data to develop and evaluate alternative ecotoxicological test methods for fish in risk assessment and to provide input to risk regulators, industry and academia on the applicability of the different approaches to assess the PBT properties of chemicals.


The Norwegian Institute for Water research (NIVA) is managing a Norwegian Research Council (NRC) funded project “Non-animal (alternative) testing methods for REACH (alterREACH)” that will run from 2010 to 2014.


The aim of the project will be to develop, evaluate and use non-animal (alternative) test methods for performing robust, reproducible and rapid screening of chemicals for their bioaccumulation and toxic potential without excessive use of experimental animals.

These objectives will be achieved through use of in vitro and in vivo experimental systems for studying the bioaccumulation and toxic potency in fish in combination with computational (in silico) assessment of chemical properties.

Work packages

The proposed work will be divided into three work packages (WPs) where data and experiences will be shared between WPs (Fig 1.).

The chemicals chosen for the evaluations of the alternative approaches detailed below (WP1-WP3) will be selected to provide a broad coverage of physico-chemical properties, mode of toxic action and being representative for high-production industrial compounds.


Figure 1. Overview of individual work packages (WP) and their internal relationship.

Workpackage 1 aims to propose a tiered testing strategy for assessing the bioaccumulation and to minimise the need for full fish bioaccumulation studies on the basis of a selection of reference compounds.

Worpackage 2 will focus on assessing the feasibility of alterative methods for toxicity assessment based on a tiered approach of in silico (QSAR) methods, in vitro methods (cell cultures), non-animal tests such as the zebrafish embryo test and in vivo screening methods for risk assessment purposes.

In the proposed work, experiments will be conducted with zebrafish as an experimental model in order to address different toxicity mechanisms ranging from mortality to sub-lethal cellular responses by a selection of the reference compound proposed previously in WP1.

The proposed work aims to take advantage of a combination of classical toxicity endpoint analysis (mortality, histopathology and DNA damage) and mode of action focused biomarker screening by high-throughput microarray-assisted gene analysis to provide links between their acute and chronic toxicity and their toxic mode of action.

The ultimate goal of the project will be to perform a concept evaluation of the chosen approaches for PBT testing in order to determine whether non-animal and alternative testing methods may be applicable to assessing the risk of chemicals within REACH (WP3).

Sist oppdatert 17.09.2015