USER: Any person with Function D who belongs to the project, when the person in

charge of it does not have said Function.

4. ESTABLISHMENT: Indicate the center registered with the competent authority where
the project will be carried out. If it is to be carried out outside a registered
center, this must also be indicated and an exception must also be requested from
the competent authority(18).
5. INTRODUCTION RELEVANCE OF THE PROJECT: It is not only a matter of explaining
what is going to be done with the animals and for what scientific purpose or its
social importance, but it is also necessary to justify why there is no other way to
do it without animals and that the concept of the Three Rs is applied in our
research.

6. PROCEDURE (with animals): Any invasive or non-invasive use of an animal for

experimental or other scientific purposes, with predictable or unpredictable
results, or for educational purposes, that may cause a level of pain, suffering,
distress or lasting damage , equivalent to or greater than that caused by the
introduction of a needle.
This includes any action that, intentionally or accidentally, may cause the birth
or hatching of an animal or the creation and maintenance of a line of genetically
modified animals in conditions, but excludes the slaughter of animals solely for
the use of their organs or tissues.
DESCRIPTION OF THE PROCEDURES: Indicate and describe the chain of actions (actions/
techniques) to be carried out on the live animal during the project. When the
guidelines for a procedure with animals are officially published or validated, the
corresponding copy will be provided.

7. APPLICATION OF THE THREE Rs: It must be justified that in order to achieve the
objectives of the project the Principle of the Three Rs is applied. What is
indicated in this point can / must be the same as that indicated in the Non-
Technical Summary.
a. Replacement: Justifying that the use of animals is essential after an
unsuccessful search for alternative methods and indicating in which databases said
search has been carried out.
b. Reduction: Indicate how the minimum number of animals necessary to obtain the
expected scientific results has been calculated.
c. Refinement: Indicate how all means are used to minimize the suffering of animals
and to improve their welfare.

8. ANESTHESIA AND ANALGESIA: Include protocol data to be used such as: Name of the
drug to be administered, concentration, dose, route and frequency.

9. HUMANITARIAN END POINT: It must be planned how to detect the moment in which an
animal has to withdraw from the experiment (stop it or perform euthanasia) because
its degree of suffering is unacceptable, in addition to the fact that scientific
data could not be obtained. reliable because of it.
According to the legislation, the death of the animal must be avoided as an end
point, so an earlier and bloodless end point must be established to avoid reaching
that level of suffering.
If death is unavoidable, the procedure will be designed in such a way that as few
animals as possible die, the duration and severity of the animal's suffering is
reduced to a minimum, and a painless death is guaranteed, as far as possible.
(Directive 63/2010/EU).
The endpoint criteria must be specific to the procedure to be performed and to the
characteristics of the individuals.

10. EXPERIMENTAL OR OBSERVATION STRATEGY: This refers, on the one hand, to the
method used to monitor the severity of the project and the welfare of the animals.
Normally it will be through a scoring table of the clinical signs or symptoms that
will help us to control the affectation of the animal.
On the other hand, the statistical model used to calculate the number of animals to
be used in it must be indicated prior to the start of the project.

11. REUSE: The reuse of an animal is possible if its level of affectation by the
experiment (severity) has not been the most serious (severe) and the animal has
fully recovered. The final decision will be made by the Center's Designated
Veterinarian.

12. LEVEL OF SEVERITY: It is the degree of affectation of the welfare and health of
the animals due to the project, both during its course and for the rest of the life
of the animal. It must be assigned one of the four legally established levels: No
recovery, Mild, Moderate or Severe. (It is advisable to consult Annex IX of RD
53/2013 where a series of examples of procedures and their estimated level of
severity are given).
NOTE: "Without recovery" is considered only when an animal that has not undergone
any previous procedure during the project is anesthetized and without regaining
consciousness is sacrificed (eg surgery at term).