Grace’s Notebook: Geoduck Egg Development – 3 days later

Today we took a look under the scope of the eggs in the silos from Friday’s egg development trial (Notebook post).

Looking at the eggs

I didn’t really look at the ones that were fertilized.

Benoit and I looked at eggs from the three silos that were not fertilized (although I did see polar bodies, cleavage).

Treatment group (all not fertilized) D-hinge? Notes
H50 yes don’t look great; lots of ciliates; some trochophores
C maybe lots of ciliates; remnants of trochophores; potential D-hinges, but hard to know because a lot of things could have been eaten
50 no some trochophores; lots of ciliates

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Grace’s Notebook: Day 1 of Trial 2 of Geoduck Stripspawn experiment – KCl dose + duration

Today was Day 1 of the second trial of geoduck stripspawn. We created an experimental design that takes into account both KCl dosage, and duration of the dose exposure. Overall things went pretty well (details in post), and on Wednesday, I’ll check for D-hinge development.

Google doc with plan

(written in prep for the day, things changed and will be noted in this post)

8:30am – Pt Whitney

Started at Pt Whitney, biopsy punching gonad tissue and checking for sex and ripeness.

I started out getting two or three males, and used a new biopsy punch with each geoduck. I placed the biopsied males in the right tank and grabbed new geoduck to biopsy from the left tank.

I then found two females, and then I found one that looked like a hermaphrodite, which was pretty cool! I’ve never seen that before:

img

Then I kept looking and finally found another female, though it may not have been as ripe as the other two. I left a little after 9:45am.

10:15am – Taylor Hatchery

I started out with cleaning the work area, cleaning the 20 20um screened mini-silos, gathering materials. Benoit, Michelle, and Molly helped by getting a filtered saltwater line available for me to use in the work area.

Once I had the filtered saltwater, I started setting up the tripours with the different mM KCl dosages, as well as some next to them full of filtered saltwater. While I was doing this, Benoit was setting up buckets in the bucket room with airstones and water in which the fertilized eggs will grow out in at the end of the dosing and fertilizing.

Setting up the dosages:

I filled the tripours up to 800ml (any more than that and the water will spill over once the 20um silo is placed in). When making the dosed saltwater, I made them as though they’d be filled to 1000ml so that the math would be easier.

I made them in a 3L pitcher, so I did them in sets of two. Example: to make the 20mM, I placed 20mL 2M KCl and 1960ml filtered saltwater. I then dispersed it into two tripours, up to 800ml. I did that again, and then moved on to the next dose.

KCl mM 2 M KCl stock to add (ml) ml saltwater to add
20 10 980
50 25 975
60 30 970
80 40 960

Tripour treatments:
img

Check female geoduck ploidy

Before dissecting the females and stripping the eggs, I measured the shells:

female number length width ratio diploid? (yes if ratio < 1.62)
1 127 73 1.739 no
2 121 71 1.704 no
3 124 80 1.55 yes

Benoit ran the ctenidia samples from all three females on the flow (? is that what it’s called) to check for ploidy. All three had the same ploidy, but it isn’t known if they are all three diploid, or all three triploid. Benoit believes they are likely all three diploid.

Stripspawn females

I dry stripped some eggs from all three directly into the screened silo sitting in tripour 19 (50mM with no hydration), and timed for 20 mins.

During those 20 mins, I stripped eggs from the three females into some saltwater. Benoit helped get a lot of eggs out. One female was much more ripe than the other two. Of the remaining two, one female was not very ripe at all unfortunately.

Benoit helped with screening and cleaning the eggs, and counted them using the coulter counter. The first counts showed that there was about 4 million eggs in the 3L pitcher.

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Kaitlyn’s notebook: ANOVA on heath stack juveniles

I preformed an ANOVA on the lengths of the juveniles in the heath stacks at Pt.Whitney. I did a post-hoc test (Tukey HSD) to determine significance between he treatments.

length-treatment

length-treatment

homogenization

With blocking the anova gives a p=0.028 for treatment and p=0.245 for tray.

Here is the Tukey HSD p-values on the anova with blocking:

Tukey multiple comparisons of means
95% family-wise confidence level

AE-AA 0.54267725
EA-AA 0.25261761
EE-AA 0.72476397
EA-AE 0.91813748
EE-AE 0.10032636
EE-EA 0.03504769

H0_T-H0_B 0.9999666
H1_B-H0_B 0.9999930
H1_T-H0_B 0.9303406
H2_B-H0_B 0.9999448
H2_T-H0_B 0.9970459
H3_B-H0_B 0.9994060
H3_T-H0_B 0.9988241
H1_B-H0_T 0.9980676
H1_T-H0_T 0.8107845
H2_B-H0_T 0.9963141
H2_T-H0_T 0.9703042
H3_B-H0_T 0.9875827
H3_T-H0_T 0.9999998
H1_T-H1_B 0.9588824
H2_B-H1_B 1.0000000
H2_T-H1_B 0.9996848
H3_B-H1_B 0.9999842
H3_T-H1_B 0.9708530
H2_B-H1_T 0.9898375
H2_T-H1_T 0.9991863
H3_B-H1_T 0.9962770
H3_T-H1_T 0.4563048
H2_T-H2_B 0.9999849
H3_B-H2_B 0.9999999
H3_T-H2_B 0.9641553
H3_B-H2_T 0.9999998
H3_T-H2_T 0.8273796
H3_T-H3_B 0.9074722

Tank Treatment
H0_T EA
H0_B EE
H1_T AE
H1_B AA
H2_T EA
H2_B AA
H3_T AE
H3_B EE