Ministerial Protocol

Annex VI

In Italy, in agreement with the 2010/63/UE European Directive, the Legislative Decree D.Ig. 26/2014 has been emanated for the regulating the procedures on all animals used for scientific purposes.

Moreover, according to the new Formational Decree of 05/08/2021, all persons who have to work on the animal models will necessarily have to receive an adequate training for the specific functions.

As early as the late 1950s, Russel and Butch published a book entitled “The principles of humane experimental technique” in which they introduced the principles of the 3Rs (Replacement, Reduction, Refinement).


Reduction refers to the need for decreasing the number of animals used in an experiment. This can be achieved with the choice of suitable procedures for a given experiment, allowing for the reduction of the variability and consequently the number of animals used necessary for a statistically significant result


Refinement refers to any possible reduction in the severity of painful procedures, towards a better understanding of the biological needs of an animal.


In order to obtain the approval of a project, the Italian legislation requires the approval of several documents, including Annex VI, which describes in a very detailed and justified manner the number of animals and the procedures intended to be carried out on them.

Our OPBA with the Wellness Manager constantly interacts with researchers, the Ministry of Health and the Evaluator Agency (ISS), to prepare the Annex VI and submit all documents related to the approval of ministerial projects involving in vivo testing.

Our OPBA has a high rate of success for approved projects, which in 2022 was equal to 100% (16 new projects and 12 amendments).