A wide variety of substances have been used to anaesthetise invertebrates, but many are not anaesthetics and merely incapacitate animals rather than preventing pain. In essence, the role of an ideal general anaesthetic is to act as a muscle relaxant, an analgesic, an anaesthetic, and an amnesic. To achieve all these properties with a single substance is difficult, and various adjuvants usually need to be administered, resulting in a cocktail of drugs. In a clinical setting, the vast majority of patients are unaware of surgery being carried out and have no memory of it, so they can claim to have felt no pain, but this is much more difficult to demonstrate in invertebrates. Here, we show that 1% MgCl_2, a muscle relaxant_, is a useful adjuvant for the clinical anaesthetic isoflurane on Octopus vulgaris when applied alone in seawater for 10 min before the clinical anaesthetic. After this, full anaesthesia can be achieved in 5 min using 1% isoflurane insufflated into the saline still containing MgCl₂. Full recovery takes place rapidly in about 10 to 15 min. The depth of anaesthesia was monitored using changes in respiratory rate, chromatophore pattern, and withdrawal movements of the arms and siphon. This methodology reduces stress on the animal and minimises the quantity of anaesthetic used.

A wide variety of substances have been used to anaesthetise invertebrates, but many are not anaesthetics and merely incapacitate animals, rather than preventing pain. Much is now known about the mode of action of modern clinical and veterinary anaesthetics because of their use on human beings and other vertebrates. In essence, the role of an ideal general anaesthetic is to act as a muscle relaxant, an analgesic, an anaesthetic and an amnesic. To achieve all these properties with a single substance is difficult and various adjuvants usually need to be administered, resulting in a cocktail of drugs. In a clinical setting, the vast majority of patients are unaware of surgery being carried out and have no memory of it, so they can claim to have felt no pain, but this is much more difficult to demonstrate in invertebrates. Here we show that 1% MgCl2, a muscle relaxant, is a useful adjuvant for the clinical anaesthetic isoflurane on Octopus vulgaris when applied for 10 minutes before the clinical anaesthetic. After this, full anaesthesia can be achieved in 5 minutes using only 1% isoflurane and from which full recovery takes place rapidly in about 10 to 15 minutes. This reduces stress on the animal and minimises the quantity of anaesthetic used.

The common octopus (Octopus vulgaris), among coleoid cephalopods, has evolved the most complex nervous system and sophisticated behaviors. Historically, O. vulgaris was a key animal model for neurophysiology research, and today, it is studied for its genomic innovations. However, unlike other models, there is no octopus farming for research, so specimens must be collected from the wild. This study assessed the impact of fishing on octopuses used in research, considering those caught using artisanal pots in the ‘Regno di Nettuno’ Marine Protected Area, Ischia (NA). To evaluate fishing stress, we identified morphological stress indicators such as chromatophore pattern and posture, and three potential molecular markers, estrogen receptor (ER), catalase (CAT), and heat shock protein (HSP70). We measured the percentage of stress signals shown by fished specimens and analyzed their differential gene expression. The transcriptional levels of octopuses caught using traps were compared to control specimens acclimated in captivity. Results indicated fluctuations in gene expression due to fishing stress. These findings suggest that an acclimation period after the stress event of fishing is crucial for ensuring the welfare of octopuses used in research, thus enhancing the quality of physiological and ethological studies.