Cephalopods in neuroscience: regulations, research and the 3Rs

Graziano Fiorito, Andrea Affuso, David B. Anderson, Jennifer Basil, Laure Bonnaud, Giovanni Botta, Alison Cole, Livia D’Angelo, Paolo De Girolamo, Ngaire Dennison, Ludovic Dickel, Anna Di Cosmo, Carlo Di Cristo, Camino Gestal, Rute Fonseca, Frank Grasso, Tore Kristiansen, Michael Kuba, Fulvio Maffucci, Arianna Manciocco, Felix Christopher Mark, Daniela Melillo, Daniel Osorio, Anna Palumbo, Kerry Perkins, Giovanna Ponte, Marcello Raspa, Nadav Shashar, Jane Smith, David Smith, António Sykes, Roger Villanueva, Nathan Tublitz, Letizia Zullo, Paul Andrews

Cephalopods have been utilised in neuroscience research for more than 100 years particularly because of their phenotypic plasticity, complex and centralised nervous system, tractability for studies of learning and cellular mechanisms of memory (e.g. long-term potentiation) and anatomical features facilitating physiological studies (e.g. squid giant axon and synapse). On 1 January 2013, research using any of the about 700 extant species of "live cephalopods" became regulated within the European Union by Directive 2010/63/EU on the "Protection of Animals used for Scientific Purposes", giving cephalopods the same EU legal protection as previously afforded only to vertebrates. The Directive has a number of implications, particularly for neuroscience research. These include: (1) projects will need justification, authorisation from local competent authorities, and be subject to review including a harm-benefit assessment and adherence to the 3Rs principles (Replacement, Refinement and Reduction). (2) To support project evaluation and compliance with the new EU law, guidelines specific to cephalopods will need to be developed, covering capture, transport, handling, housing, care, maintenance, health monitoring, humane anaesthesia, analgesia and euthanasia. (3) Objective criteria need to be developed to identify signs of pain, suffering, distress and lasting harm particularly in the context of their induction by an experimental procedure. Despite diversity of views existing on some of these topics, this paper reviews the above topics and describes the approaches being taken by the cephalopod research community (represented by the authorship) to produce "guidelines" and the potential contribution of neuroscience research to cephalopod welfare.