Yaamini’s Notebook: Gigas Larvae Day 4

My larvae are 60 microns now!

Today, Laura helped me screen my larvae through 60 micron and 48 micron screens. You can see the data for yourself here, but the majority of my larvae held on a 60 micron screen! Since there was really nothing in the 48 micron screens, I tossed out those samples. While I probably threw some slower-growing larvae down the drain, I also got rid of dead larvae that could possibly disrupt the healthier ones. Laura screened while I counted right away. A few things I learned:

  • It’s much easier to count my larvae on a Sedgewick Rafter cell than in a well-plate
  • Yesterday, I was classifying larvae as dead if they didn’t show any signs of movement (both through the water column and their innards). This is incorrect! Larvae area dead when the shells are empty. I don’t remember seeing any larvae like that yesterday, but I think it’s safe to say that those data points are not high-quality. I shouldn’t use those counts in any analysis.
  • What I thought was a shell deformation was actually the larvae’s…butt?? It’s just a normal D-hinge larvae from a different angle! I realized this when I saw one of those weirdly-angled larvae do a few spings and flips, then start swimming in a way that was recognizable as a D-hinge

I also preserved larvae in the freezer and fed a mix of C.iso and Chagra since those were the most dense. It’s likely that I undercounted algal cells on the hemocytometer since there were so many! I was a little short on time so I also only counted three hemocytometer cells.

Another thing we noticecd when filling buckets is that the water in the heating reservoir had been sitting so long that the 1 micron filter was showing signs of being anoxic. We flushed the line and replenished the header a bit. Laura talked to Stuart, and they suggest draining the header tank tomorrow and flushing fresh seawater through that line so that it sits overnight. I would also have to make sure my water is heating overnight.

For tomorrow:

  • Figure out which AVTECH is which
    • Add to larval tanks and totes strategically
    • Relabel on Dashboard/OA Monitor
  • Relabel HOBO probe tags
  • Finish counting and imaging Plates 2 and 3
  • Flush header and lines
    • Replenish with new seawater
    • Start heating process
  • Feed Joth’s scallops
  • Feed oyster larvae
  • Prepare tripours for Saturday screening
  • Clean up Sealab

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Yaamini’s Notebook: Gigas Larvae Day 3

Counting larvae takes a while

Before I get into that, a quick rundown of other things I did today.

  • Recorded probe numbers in each larval bucket
    • 1: SN6 Amb pH A
    • 2: SN6 Low pH B
    • 3: SN6 Amb B
    • 5: NF6 Amb B
    • 6: NF10 Low pH A
    • 9: SN10 Low pH A
    • 10: SN10 Amb A
    • 11: SN6 Low pH A
    • 12: NF6 Amb A
    • 12: NF10 Amb B
    • 15: NF6 Low pH B
    • 17: NF10 Low pH B
    • 19: SN10 Low B
    • 21: NF6 Low pH A
    • 23: NF10 Amb A
  • Recorded which bucket was in which tote

img_8126

Figure 1. Tote layout. Totes go from 1 to 9, left to right.

  • Did not record morning temperatures
    • HOBOs already in tanks, no need
  • Measured morning food presence
    • Very dense! I think this is because I fed in the afternoon on Tuesday, so the larvae didn’t have a full 24 hours to really feed down

img_8127

Figure 2. Color of larval tanks in the morning, before daily feeding.

  • Steven rigged up the AVTECH in the SeaLab, so I now have 5 probes to work with
  • He also started messing around with the OA system set-up
  • Moved immersion heater probes to inside larval bucket, instead of in tote
  • Counted heaters
    • 8 new from Roberts Lab, 1 not being used right now
    • 2 old heaters (one I ordered, one I took from the lab with the Farenheight controller)
    • Gray heater borrowed from rick
  • Fed larvae

Now to the main event: imaging and counting! I counted all of the larvae in Plate 1 collected yesterday. It took me much longer than I expected, mainly because I was trying to figure out how to work the dissecting scope while also taking pictures for future use. My count data can be found here.

There were lots of larvae that exhibited a normal D-hinge shape, but there were also some that looked like peanuts! I recorded how many of these deformed ones I saw as well.

fig3

fig4

Figures 3-4. Normal D-hinge larvae shape at 40x (top) and 20x (bottom) magnification.

fig5

Figure 5. Weird larvae shape.

I’m storing all of my larvae photos and videos from today here. I’m cataloging each day in a different folder, and labelling which bucket the larvae are from in the image name.

For tomorrow:

  • Screen with 60 and 48 micron screens
  • Sample and count
  • Save samples in -80ºC freezer
  • Finish imaging Plate 2 and 3

Since you made it through this lab post, here are some easy links to larvae videos. Both are from bucket 8:

Video 1

Video 2

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Laura’s Notebook: Oly spawning charts

Here’s a quick visualization of the Olympia oyster spawning data from this spring. Each chart represents a population, and within the charts data is color coded by treatment.

imageimageimageimage

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