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:
- 1A-1E
- 2A-2E
- 4A-4E
- 5A-5E
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
| Tank | Treatment | Written Label | Tag | Sex |
|---|---|---|---|---|
| 1 | pH 7.2 | 1A | 019 | NA |
| 1 | pH 7.2 | 1B | 018 | NA |
| 1 | pH 7.2 | 1C | 013 | male |
| 1 | pH 7.2 | 1D | 012 | male |
| 1 | pH 7.2 | 1E | 009 | NA |
| 2 | ambient | 2A | 031 | male |
| 2 | ambient | 2B | 028 | NA |
| 2 | ambient | 2C | 027 | NA |
| 2 | ambient | 2D | 022 | NA |
| 2 | ambient | 2E | 025 | female |
| 4 | ambient | 4A | 057 | NA |
| 4 | ambient | 4B | 040 | NA |
| 4 | ambient | 4C | 059 | NA |
| 4 | ambient | 4D | 044 | NA |
| 4 | ambient | 4E | 058 | NA |
| 5 | pH 7.2 | 5A | 097 | male |
| 5 | pH 7.2 | 5B | 085 | female |
| 5 | pH 7.2 | 5C | 093 | female |
| 5 | pH 7.2 | 5D | 088 | female |
| 5 | pH 7.2 | 5E | 090 | male |
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
| Animal Label | Image |
|---|---|
| 2A | |
| 2B | |
| 2C | |
| 2D | |
| 2E | |
| 4A | |
| 4B | |
| 4C | |
| 4D | |
| 4E | |
| 5A | |
| 5B | |
| 5C | |
| 5D | |
| 5E |
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://shellywanamaker.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
- made KCl as follows:
- 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.