Immune priming injection vs. emersion pilot

Update 4/4/2023

  1. 15 diploid and 15 triploid oysters were placed in 50 gL-1 MgCL overnight (18 hours). Here are how many opened:
ploidy time open closed percent
2n 18h 12 3 80%
3n 18h 11 4 73%

  1. poly(I:C) solution was prepared by adding 4 ml of sterile, 0.2 um filtered seawater to a vial w/ 10 mg of lyopholized powder. Solution was incubated at 65C for 10 minutes and allowed to cool to room temp.
  2. 10 diploid and 10 triploid oysters were injected w/ 100 ul of poly(I:C) solution (total of 0.25 mg per animal) in the adductor muscle. Oysters were allowed to sit open for 10 minutes and returned to seawater for recovery.
Oysters injected:
Diploid: D1-3, D5-8, D10-11, D13
Triploid: T3-7, T9-13
  1. 5 diploid and 5 triploid oysters (from MgCl treatment) were returned to seawater to recover; tissue will be sampled as anathesia control.
  2. 10 diploid and 10 triploid oysters (naive) were added to an emersion bath containing 500 ml of seawater contianing 10 ug/ml Poly(I:C), with a bubbler. Lafont et al 2017 found that emersion at 38 ug/ml worked for 24 hrs, so we will see.