Manchester Hardening project final lab sampling plan

manchester-hardening
experimental design
Planning final sampling of Manchester Hardening experiment oysters
Author

Ariana Huffmyer

Published

August 14, 2026

Overview

This post details the experimental design plans for sampling acute stress survival of Manchester Hardening experiment oysters.

Oysters

Oysters were collected from Manchester on Friday, August 15th and transported to UW campus by Mac. They are all contained in one tank in FTR in red seed bags.

There are 20 red mesh bags of oysters labeled with yellow cattle tags numbered with # 1-20. The numbers are written on the back of each cattle tag with a cirle around it. See the images below.


The cattle tag number is crossed out - refer to the back of the tag for the ID number.

Each red bag contains n=20 oysters that correspond to a specific treatment group. Students do not need to worry about the specific treatment group - just track everything by the bag number (1-20). Each bag has a waterproof paper label with the family and treatment information as backup information. I already have this information recorded.

These 20 oysters per group will be used for survival assays.

Sample size:

There are a total of n=20 oysters per bag available for assays. We will use 15-20 oysters from each depending on how the assays and schedule goes.

Survival

Schedule:

  • Aug 17-21: Round 1 of n=4 oysters per bag, n=80 oysters total
  • August 24-28: Round 2 of n=4 oysters per bag, n=80 oysters total
  • August 31-Sept 4: Round 3 of n=4 oysters per bag, n=80 oysters total
  • Sept 7-Sept 11: Round 3 of n=4 oysters per bag, n=80 oysters total

*Note that specific dates can be flexible based on student schedules and what works best for the team.

This will result in n=16 oysters per bag being used for assays, which will leave a few extra in case an oyster is found dead or needs to be replaced.

Materials needed:

  • Numbered plastic cups (n=80 per round)
  • Seawater (Instant Ocean adjusted to ambient levels)
  • Seawater table running for temperature control

Base protocol:

  • Make a datasheet online in a Google Sheet and share it with Ariana.
  • Pre heat the water table such that cups in the water table reach 36°C (may need to have the table heater set at 38-40°C)
  • Obtain n=4 oysters per bag per round (n=80 oysters per round)
  • Place oysters into numbered cups, being sure to record which bag is in represented in each cup (e.g., Bags 1-20).
  • Fill cups with seawater and ensure each oyster is alive.
  • Record which oysters are dead in the morning and evening of each day for 3-5 days until ~80-90% of oysters have died. Follow the standard lab protocols for survival assays.
  • Record all methods used and send to Ariana.
  • Record the temperature in 5 cups in the water table each time mortality is assessed and write down the temperature readings.
  • Send all data to Ariana/add to a Google Sheet.
  • Repeat until we get through ~16 oysters per bag.

Data sheets that will need to be recorded:

All data will need to be recorded for each “round”.

  1. Temperature recorded in n=5 cups at each sampling point
  2. Alive/dead oysters at each time point
  3. The time of the start of the experiment and time of each sampling time point
  4. A metadata sheet that contains the cup number, bag number, round, and any notes