Mar 06, 2025

Public workspaceProtocol for anesthesia and euthanasia in Octopus maya

DOI
dx.doi.org/10.17504/protocols.io.6qpvr928pvmk/v1
  • Daisy Pineda-Suazo1,
  • Wendy Escobedo-Hinojosa2,
  • Pedro Gallardo1,
  • Carlos Rosas1
  • 1Unidad Multidisciplinaria de Docencia e Investigación. Facultad de Ciencias, Universidad Nacional Autónoma de México. Puerto de abrigo s/n, Sisal, Yucatán, México. C. P. 97356;
  • 2Unidad de Química en Sisal. Facultad de Química, Universidad Nacional Autónoma de México, Puerto de abrigo s/n, Sisal, Yucatán, México. C. P. 97356
Daisy Pineda-Suazo
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DOI: dx.doi.org/10.17504/protocols.io.6qpvr928pvmk/v1
Protocol Citation: Daisy Pineda-Suazo, Wendy Escobedo-Hinojosa, Pedro Gallardo, Carlos Rosas 2025. Protocol for anesthesia and euthanasia in Octopus maya. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr928pvmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 20, 2025
Last Modified: March 06, 2025
Protocol Integer ID: 118834
Keywords: Octopus maya, Anesthesia, Euthanasia, Feeding, Gastric secretion stimulation
Abstract
This protocol details the procedure of anesthesia and euthanasia in Octopus maya.
Materials
Materials and equipment

  • Euthanasia container
  • Seawater (prepared at 15°C and natural salinity levels)

Feeding and stimulation materials

  • Fresh crabs (Callinectes spp.) for feeding.
  • Bags for holding dead crabs (Callinectes spp.) during gastric secretion stimulation. Ensure the bags are designed to allow contact but prevent ingestion.
Feeding protocol for validation
Feeding protocol for validation
20m
20m

Note
Gastric juice for the assay is obtained after gastric secretion stimulation, while gastric juice post-digestion (chyme), obtained from animals fed with fresh crab (Callinectes spp.), is used as the standard.
Feed three octopuses ad libitum with fresh crab (Callinectes spp.) for Duration00:20:00 to stimulate gastric juice production.

20m
Fasting protocol for assay
Fasting protocol for assay
12h 20m
12h 20m
Fasting protocol:

Fast three octopuses for Duration12:00:00 to ensure complete digestion of prior meals.

12h
Gastric secretion stimulation:


Place bags containing dead crabs (Callinectes spp.) into the tank with the octopuses.
Ensure the design of the bags allows the animals to contact the prey without ingesting them.
Leave the octopuses in contact with the bags for Duration00:20:00 to stimulate gastric secretion.
20m
Anesthesia protocol
Anesthesia protocol
5m
5m
Prepare seawater at a natural salinity level and chill it to Temperature15 °C .

Place the octopus in the chilled seawater for Duration00:05:00 to induce hypothermia, which acts as a mild anesthetic.

5m
Monitor the octopus for signs of reduced activity and responsiveness, ensuring it is sufficiently anesthetized before proceeding.
Euthanasia protocol
Euthanasia protocol
Gently remove the anesthetized octopus from the chilled seawater.
Using sterile dissection tools, perform euthanasia by cutting between the eyes into the brain. This method ensures immediate cessation of brain activity, adhering to ethical standards for cephalopod handling.

Note
Notes and Precautions:
  • Maintain the seawater at Temperature15 °C throughout the anesthesia process to ensure effective hypothermia.
  • Sterilize all dissection tools prior to use to prevent contamination of subsequent samples.
  • Handle the octopus gently to minimize stress during the procedure.
  • Follow all ethical guidelines and local regulations regarding cephalopod euthanasia.