engleski

Procedure of hemolymph sampling in Octopus vulgaris

Cephalopods (i.e. octopus, squid, cuttlefish and nautilus) are class of exclusively marine animals belonging to the phylum Mollusca, containing about 700 known living species. Unlike other molluscs, cephalopods have a functionally closed circulation system, sophisticated nervous system with well- developed sense organs and large, highly differentiated brain, and are thought to be the most intelligent invertebrates. Live cephalopods have been used in studies of neurobiology, physiology, immunology and environmental toxicology and consequently have been involved in scientific procedures where there was need to immobilize, anesthetize and euthanize them. However, only just recently, cephalopods became the only invertebrate animals included in the Directive 2010/63/EU on the protection of animals used for scientific purposes. In cephalopod research community this regulation opened a lot of questions about objective criteria for identification and assessment of pain, suffering, distress and lasting harm in cephalopods and revealed the lack of protocols for cephalopod surgery, anaesthesia, analgesia and humane euthanasia. In this work, we present a procedure of Octopus vulgaris anesthesia, using combination of magnesium chloride and ethanol solution, and procedure of hemolymph sampling. Hemocytes, cephalopod blood cells, could be used for in vitro investigations, such as cytotoxicity assays, gene transcription expression patterns under conditions of thermal stress, stress- induced neuroendocrine and immune changes, thus gaining valuable information without invasive experiments on animals. This research was funded by COST Action FA1301: 'A network for improvement of cephalopod welfare and husbandry in research, aquaculture and fisheries (CephsInAction)'.

cephalopods, anesthesia, hemocytes, welfare impact

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