Today started like many of the previous days, with checking for extruded eggs and counting the larvae in some of the silos from the preliminary experiments. I had a late start at the hatchery because I was picking up a pipette gun from campus.
- 2 new: 112(2), 113(2)
- 1 repeat: 34 (1)
Screened out silos at Day 5 from experiment started 2016-07-22 with OR2T and CA2T larvae. Took pictures under microscope at 10x of all but A84 and A83.
|A86||OR2T||750 in RNALater||Put in clean silo|
|A85||OR2T||750 in RNALater||Put in clean silo|
|B61||CA2T||750 in RNALater||Put in clean silo|
|B59||CA2T||750 in RNALater||Put in clean silo|
|B65||OR2T||750 in RNALater||Put in clean silo|
|B63||OR2T||750 in RNALater||Put in clean silo|
|A84||CA2T||750 in RNALater||Put in clean silo|
|A83||CA2T||750 in RNALater||Put in clean silo|
Then Sean and I started screening out the newly released larvae from the trays and buckets. There did not seem to be enough larvae from all three populations, so I stayed to finish screening out the last couple of buckets and count larvae. While the BC3T oysters were sitting in a bucket waiting to be placed into a clean tray, they started to release A LOT of larvae. Once I finished screening the last of the buckets I realized that we finally had enough from all three populations to start the experiment. Hooray! However, it meant I ended up staying until 10pm setting up by myself.
I had to remove some of the silos from preliminary experiments that were in the system in order to make room for the full experiment. I screened these out and left in tripours overnight to count the next morning.
- B72, A63, A6, B66, B60, B55, A88, B66, B80, A100, A97, B51, A9
There were two family groups for CA and OR that had a lot of larvae, whereas BC had one group with a lot of larvae and one group with ~20,000. I decided to combine the BC groups, then have 2 replicates per treatment for each family group, so 4 silos per population/ treatment for CA and OR and 2 silos per population/treatment for BC. Not ideal, but I wasn’t sure when I would have at all three populations spawning at the same time again. I counted the larvae for each population with 4 subsamples of 0.5 mL, took the average to get larvae/mL and then calculated how many mL to add to each silo to get 15,000 larvae. I dispensed larvae with a pipette gun and 25mL pipettes while plunging constantly to mix, using a new pipette for each family group. When pipetting out replicates, I alternated between treatments to help mitigate possible effects of the plunging/pipetting process (probably a little overkill).
|Family||larvae/mL||mL added||Experiment labels|
|OR1T||152.8||98.2||B20, B18, A72, A73|
|OR5B||153||98||A59, A71, B13, B11|
|CA4B||168||89.3||B19, A58, A57, B06|
|CA2T||290||51.7||A55, A56, B16, B12|
|BC3T+BC2T||232.5||64.5||A53, A54, B35, B45|
I had hoped there would be enough larvae from BC1T, but there was only ~10,000.