OA oyster data

Carolyn and Brent went to Taylor and fertilized eggs for the experiment. 21 females and 20 males were used in the fertilization. 80,000 eggs were aliquoted to each jar for transport (each jar would then be emptied into the larger larval chambers at NWFSC) and the eggs were fertilized in the jars with pooled sperm. We began putting the fertilized eggs into the chambers containing treatment water around 5 pm (treatment pCO2= 280 ppm, 400 ppm 1000 ppm). The system is being kept at 14C. Fertilized eggs in the larval chambers were left static overnight so that the larvae could hatch.

Began cleaning and sampling jars around 8:30 am. Each jar was completely emptied into a 20 um mesh sieve and then rinsed out with the appropriate treatment water. A new, clean jar/larval chamber was filled with new water from the appropriate treatment and if that jar was not scheduled to be sampled, all larvae were rinsed into the new jar. If the jar was scheduled to be sampled (jars 5 and 6 for May 10), then a clean jar was prepped while larvae were washed into a 50 mL Falcon tube, which was then filled to 45 mL. The tube was gently mixed ~5x, 3 aliquots of a fixed volume were taken and put into a welled plate (1 well) and then these two steps were repeated so that for each jar sampled there were 2 wells of a fixed volume to count. For shelf 1, the volume taken was 45 uL x3, for shelves 3 and 6 the volume was 25 uL x3 (135 uL yielded too many larvae). After the aliquots were taken from the Falcon tube, it was poured into the new, clean jar and replaced in the appropriate box. At the end of sampling all the jars, they were plugged into the system so that the water would flow through.

Counting of larvae began ~10 am. First, live/dead were determined on the inverted microscope at 40x. Then the larvae were dropped with 100% EtOH and total counts were taken per well on the inverted scope at 4x. Count data are here

Here's how we started the experiment:

System       Treatment
1                  1000
2                  400 +/- 100
3                  280
4                 280 +/- 100
5                  1000 +/- 100
6                  400

We ended up stopping the dynamic pH control and finished with:

System       Treatment
1                  1000
4                 280
6                  400

Thanks,
Mike