Picchietti, Simona http://orcid.org/0000-0002-6116-213X
Buonocore, Francesco
Guerra, Laura
Belardinelli, Maria Cristina
De Wolf, Tania
Couto, Ana
Fausto, Anna Maria
Saraceni, Paolo Roberto
Miccoli, Andrea
Scapigliati, Giuseppe
Article History
Received: 4 June 2020
Accepted: 10 November 2020
First Online: 12 January 2021
Compliance with ethical standard
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: The authors declare that they have no conflict of interest.
: Animal manipulation during the diet experiments complied with the guidelines of the European Union Directive (2010/63/EU) and Portuguese legislation (DL113/2013) for animal treatments. The Organism for Animal Welfare of CIIMAR and the Portuguese Directorate-General for Food and Veterinary approved all the protocols performed in the present study (approval 0421/000/000/2017 DGAV). Fish handling was performed under ethylene glycol monophenyl ether anesthesia (0.3 mL/L; ref.: 8.07291.2500, Merck, Whitehouse Station, USA), and discomfort, stress, and pain to the experimental animals was avoided, as much as possible, along the experiment. For sampling, fish were euthanized by anesthetic overdose (ethylene glycol monophenyl ether; ref.: 8.07291.2500, Merck, Whitehouse Station, USA).For the experiments performed in Italy, ethical approval by ethics committee was not required, as per the Explanatory note of the Italian Ministry of Health’s Directorate-General for Animal Health and Veterinary Medicinal Products (DGSAF) of 26 July 2017. Organs and tissues were dissected out following temporary immobilization with 1 mg/ml tricaine methane sulfonate (MS-222) (Sigma) in aquarium water, buffered with 0.6 g/L sodium bicarbonate. Anesthesia was followed by exsanguination or decapitation. All efforts were made to minimize suffering. The study was carried out in strict accordance to Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes and on National Legislative Decree 26 of 4 March 2014 “Attuazione della Direttiva 2010/63/UE sulla protezione degli animali utilizzati a fini scientifici.”