Cephalopods in neuroscience: regulations, research and the 3Rs

Abstract

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.

Camouflage during movement in the European cuttlefish (Sepia officinalis)

“…When experiments ended, all animals were returned to the Gulf of Naples. Animal maintenance and experimentation were in compliance with EU directive 2010/63 on the protection of animals used for scientific purposes, and following the recommendations of the 3Rs (Fiorito et al, 2014).…”

Bem-estar em Invertebrados: um parâmetro ético de responsabilidade científica e social da pesquisa?

“…Contudo, há uma mobilização para que o mesmo princípio seja aplicado para invertebrados visando minimizar a dor e sofrimento em intervenções com diferentes graus de invisibilidade (Crook, 2013). Desta forma, a justificativa e relevância do estudo devem ser fundamentadas em evidências e argumentos lícitos e idôneos; reduzir o número de animais sacrificados; incentivar o desenvolvimento de protocolos e tecnologia de biomarcadores para identificação de baixos graus de BEA e sua imediata mitigação; e buscar a substituição por métodos alternativos (Fiorito et al, 2014). Para Knutsson (2016) as evidências de que os invertebrados podem sentir dor devem ser suficientes para reduzir o sofrimento.…”

“…A senciência cientificamente apoiada nas evidências de estrutura do sistema nervoso, fisiologia, comportamento, evolução, cognição e repertório comportamental, têm sido testadas e paulatinamente identificadas em invertebrados (Crook e Walters, 2011, Fiorito et al, 2014, Knutsson, 2016, contudo apoiadas na crença que quanto mais complexo é o sistema nervoso mais impacto terá um BEA empobrecido (Broom, 2013). No entanto, o benefício-da-dúvida tem sido proposto como princípio ético aliado ao princípio da precaução, uma vez que até o presente momento não há conhecimento científico suficiente para refutar sua senciência, logo a dúvida deve garantir que sejam tratados de maneira a favorecer seu BEA (Elwood, 2011, Oliveira e Goldim, 2014.…”

Octopus arm movements under constrained conditions: adaptation, modification and plasticity of motor primitives

“…The animals were housed individually in 100 l artificial sea water tanks within a closed circulation system and held according to the guidelines for the EU Directive 2010/63/EU for cephalopod welfare (Fiorito et al, 2014). Tanks were enriched with clay-pot dens, gravel, rocks and green algae (Caulerpa prolifera); the temperature of the holding rooms was held constant at about 19°C.…”