News

Published: 05 August 2025

New scientific study published

A new scientific publication, co-authored by Doctors Against Animal Experiments (DAAE) and the European Coalition to End Animal Experiments (ECEAE), shows that despite the availability of modern, animal-free methods, mice continue to suffer for Botox tests in Europe. The study was conducted as part of the SAFE consortium (Safety Assessment Through Animal Free Evolution) in the Netherlands, which aims to accelerate the transition to human-relevant, animal-free science.

Botulinum neurotoxin (BoNT), known under the brand name Botox, is used for both aesthetic and medical purposes. However, all production units (batches) are usually tested using the so-called LD50 test, in which different concentrations of the BoNT neurotoxin are injected into the abdominal cavity of groups of mice with in order to determine the dose at which half of the animals die. These tests are carried out at several production stages. The new study, co-authored by Dr Dilyana Filipova, scientific officer at DAAE, and Dr Katy Taylor, ECEAE staff member, shows that more than 200,000 mice across Europe still die in agony every year in cruel Botox experiments - often while fully conscious (1). Nowadays, instead of the LD50 test there are validated, human cell-based test methods that do not involve animal testing.

DAAE and ECEAE fight for an end to Botox animal testing

DAAE launched the campaign against Botox animal testing back in 2007, which was expanded across Europe two years later with the ECEAE (2).

As part of an international petition against Botox animal testing, the ECEAE collected over 164,000 signatures from all over Europe and handed them over to the European Medicines Agency (EMA) in 2023 - a clear signal from the public that animal testing for Botox must be ended (3).

Some progress in Germany and many challenges

The current study analyses publicly available data from the UK, Ireland and Germany. It is striking that in Germany the LD50 test for BoNT products has presumably been largely discontinued. At least there have been no more corresponding entries in the animaltetinfo.de database since 2021. However, due to the lack of transparency, it is not possible to determine with certainty whether all such tests have actually been replaced or merely outsourced to other countries. Nevertheless, it is clear that cruel LD50 tests are still being carried out in countries such as the UK and Ireland.

Despite the existence of recognised cell-based test methods, LD50 tests continue to be used for e.g. reference and validation tests as well as regulatory requirements. The implementation of non-animal methods is inconsistent and slow, which emphasises the urgency of regulatory reform.

Demands and outlook

The study clearly advocates the deletion of the outdated LD50 test from the European Pharmacopoeia and calls for the mandatory introduction of human-relevant, animal-free cell test procedures as the legal standard. Standardised regulatory requirements and improved support for manufacturers during the transition are also necessary.

For DAAE and, one thing is certain: animal testing for Botox is superfluous and must be stopped urgently ended. We firmly continue to advocate that Germany and the whole of Europe consistently implement animal-free test methods and abolish the LD50 test once and for all.

References

1. Watkins J.C. et al. Botulinum neurotoxin: Tracking the transition from lethal dose to in vitro models. NAM Journal 2025; doi: 10.1016/j.namjnl.2025.100040:100040
2. Taylor K. et al. Botulinum toxin testing on animals is still a Europe-wide issue. ALTEX - Alternatives to animal experimentation 2019; 36(1):81–90
3. 165,000 signatures against botox animal testing. Doctors Against Animal Experiments, press release, 20.04.2023