Solé, J.
Arnedo, M. A.
Barat, J.
Enguídanos, A.
Pujade-Villar, J.
Franch, M.
Cadiach, O.
Aguilar, F.
García, G.
Pascual, R. http://orcid.org/0000-0001-9937-9644
Funding for this research was provided by:
Generalitat de Catalunya (2014SGR1604)
Diputació de Tarragona (8004330008-2016-0005574, 8004330008-2017-0001118)
Article History
Received: 30 January 2018
Accepted: 24 September 2018
First Online: 3 October 2018
Compliance with ethical standards
:
: The work does not involve any conflict of interest.
: To obtain the data presented in the manuscript it has been necessary to involve animals. These have been used either for measuring morphometric characters or for molecular analysis of DNA. I declare that, despite invertebrates not being included in the current legislation on the protection of animals in scientific research, we have followed the regulations established in the <i>Directive 2010</i>/<i>63</i>/<i>EU of the European Parliament and of the Council, of 22 September 2010, on the protection of animals used for scientific purposes</i>, as well as in the <i>Real Decreto 53</i>/<i>2013, de 1 de febrero, por el que se establecen las normas básicas aplicables para la protección de los animales utilizados en experimentación y otros fines científicos, incluyendo la docencia</i>. That means that we have followed, when possible, the ‘Three Rs’ rules enshrined in these documents: to <b>refine</b> the procedures so that the degree of suffering is kept to a minimum. To <b>reduce</b> the number of animals used in research to the minimum required for meaningful results. To <b>replace</b> the use of live animals by non-animal alternatives. The application of these rules in our particular research has entailed the following: (1) To comply with the first ‘R’ (refinement): (a) Biometric measurements have been carried out by means of an innovative procedure based on a calibrated imaging system, avoiding any manipulation which could lead to stress and suffering. This procedure involves only a slight anaesthesia followed by several photographs and the releasing of the individual in the location where it was collected and (b) The individuals used for molecular analyses have been euthanized after anaesthesia, because although the procedure can be performed on a separate member of the animal we have considered that this could cause stress and suffering when the mutilated individual was released in the environment. (2) To comply with the second ‘R’ (reduction): the number of euthanized animals has been the minimum to achieve significant results through its DNA analysis. In fact, for some populations the number of individuals used has been even smaller because we were not able to collect more specimens. (3) It has not been possible to comply with the third ‘R’ (replacement) as a consequence of the nature of the research.