Matt, Brent, Nate (from hatchery in Lilliwaup?), and Josh present
Biopsies of animals from treatment and control groups (tanks B1, B2, B4, and B5)
Grabbed 5 animals from each tank and labeled with pencil and marker:
Used 15mL falcon tube “speculum” to hold pedal gap open and sampled gonad with 23G 1.5” needle on a 5mL syringe.
Gave each animal shellfish tag
pictures of gonad biopsies:
- biopsies for tank 1 animals cross-contaminated on the slide before I could take pictures so no tank 1 animals are pictured below
Because all animals seemed mostly immature, we decided to biopsy animals from Tank 6
Biopsies of animals from tank B6
Biopsied all animals from October harvest and found 5 ripe females and 4 ripe males.
Images of ripe females:
Decided to strip spawn these and not the animals from the control and treatment tanks
Strip spawning animals from tank B6
Followed the protocol outlined here: https://shellytrigg.github.io/225th-post/
- Pooled all eggs together in one 4L beaker
- Rinsed in 4 batches so to not clog the screens
- Did 2 KCl priming baths at a time to save time
- Rinsed KCl off and transferred back into beaker and Josh counted eggs
- 15 M eggs
- Divided eggs up in 4 equal volumes and added sperm from each male
- Transferred fertilized eggs to LRT
- SIDE EXPERIMENT: Did a fertilization with leftover eggs from gonads WITHOUT KCl. Let fertilization sit in container until d-hinge count a few days later.
End of the day notes
At the end of the day, tank 2 had a volitional spawn. Matt and Josh saved 4M eggs.
Noticed a couple ports in manifold in the silo set up in tank B3 were clogged. I took pictures of animals in these silos, they look alive and ok. I flushed the manifold out with fresh water, reconnected, and made sure each port had flow before I left.