Mar 06, 2025

Public workspaceChyme sampling from the digestive tract of Octopus maya

DOI
dx.doi.org/10.17504/protocols.io.n92ldrj87g5b/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.n92ldrj87g5b/v1
Protocol Citation: Daisy Pineda-Suazo, Wendy Escobedo-Hinojosa, Pedro Gallardo, Carlos Rosas 2025. Chyme sampling from the digestive tract of Octopus maya. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ldrj87g5b/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: 118835
Keywords: Octopus maya, Extraction of enzymes , Chyme
Abstract
This protocol details the chyme sampling from the digestive tract of Octopus maya.
Materials
Materials and equipment

  • Surgical tools: scissors and clamps
  • Centrifuge capable of reaching Centrifigation13370 x g
  • Tissue homogenizer (e.g., ULTRA-TURRAX IKA T18 basic)
  • Liquid nitrogen
  • Cryogenic storage containers (Temperature-40 °C freezer)
  • Eppendorf tubes (1.5 mL or 2 mL)
  • Protective equipment: gloves and lab coat
Preparation
Preparation
Ensure all tools and equipment are clean and sterilized before starting.
Prepare the workspace with necessary materials, ensuring easy access to surgical instruments, centrifuge, liquid nitrogen, and Eppendorf tubes.

Use proper personal protective equipment (PPE) throughout the procedure.
Dissection of the Digestive Tract
Dissection of the Digestive Tract
Carefully dissect the digestive tract of Octopus maya, ensuring the integrity of each section.
Identify and separate the following sections:

  • Anterior salivary glands (BM)
  • Crop (Cr)
  • Stomach (St)
  • Caecum (Ca)
  • Digestive Gland (DG)
Use surgical clamps to isolate and prevent mixing of digestive contents between sections.
Sampling of chyme
Sampling of chyme
5m
5m
Open each section carefully to access the chyme.
Transfer the chyme into labeled Eppendorf tubes.
Centrifuge the chyme samples at Centrifigation13370 x g, 00:05:00 to separate solids from the supernatant.

5m
Centrifigation
Collect the supernatant and immediately freeze it in liquid nitrogen.
Store the frozen samples at Temperature-40 °C until analysis.
Temperature
Extraction of enzymes from the buccal mass (BM)
Extraction of enzymes from the buccal mass (BM)
5m
5m
Dissect the anterior salivary glands.
Liquefy and homogenize the glands using a tissue homogenizer (e.g., ULTRA-TURRAX IKA T18 basic) at an appropriate speed for uniform homogenization.
Transfer the homogenized sample into an Eppendorf tube.
Centrifuge the sample at Centrifigation13370 x g, 00:05:00 to obtain a clear supernatant.
5m
Centrifigation
Freeze the supernatant in liquid nitrogen immediately after centrifugation.
Store the sample at Temperature-40 °C until further analysis.

Note
Notes and precautions:

  • Ensure the surgical clamps are properly placed to prevent cross-contamination of digestive contents.
  • Work quickly when handling biological samples to preserve enzyme activity.
  • Properly label all tubes with the section of the digestive tract and the sample type for accurate identification.
  • Use liquid nitrogen with caution and follow appropriate safety protocols to avoid burns or injuries.

Storage:

Samples should remain frozen at Temperature-40 °C to preserve enzyme and metabolite integrity until the time of analysis.

Temperature