Laura’s Notebook: Manchester, 5/31 to 6/4

From where I stand, these days: img_8324

And check out these Olympia oyster larvae horsing around

May 31st

Spent the AM moving buckets around: K & HL broodstock moved to small table against the wall, and all larval buckets to top shelf of the center table in the hopes that this would improve survival. New arrangement:

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Katie helped today, and spent her morning collecting new larvae and counting. New larvae – note the very large number from HL-10 Ambient! Likely from 2 individuals, according to the ~200k larvae/oyster estimates from the literature: b4c101c4-8091-4827-8568-12e9d0b288e5

Sent Katie home with the excess larvae and 15 adult oysters from each K treatment group (5 per replicate). While getting her oysters, Katie counted live/dead in each K group:

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Spoke to Katie about potential summer projects – see her notebook post for details.

Drilled and tapped into the air line for the larval buckets, enough for a total of 32 buckets. This provides for aeration during the double-bucket challenge, and once I separate >180um and <180um.

June 1st

Arrival inspection:

  • Larvae possibly present in:
    • NF-10 low B
    • SN-10 amb B
    • SN-6 low A
    • K-6 amb
    • K-10 amb
  • Currently spawning (foamy):
    • NF-6 amb A
    • SN-6 low B
  • New larval setup looks good- no issues overnight.
  • Changed all 3 in-line filters
  • Screened larvae in double-bucket system, sampled all buckets (1 rep per bucket) and counted live/dead. Reminder: 5-gal is the first bucket, which should catch all dead larvae. 2-gal bucket is the second, which should only house live larvae.

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Counts with “?” had lots of larvae which I didn’t have time to count; I fixed with lugols and will count tomorrow. Some buckets were more successful than others. Notice, the 3 buckets with the largest %live in the first, 5-gal bucket are all low pH groups, indicating they weren’t as strong swimmers or liked to hang around the bottom of the bucket. I saved the 5-gal bucket contents from K-6 Low and SN-10 Low, since there were so many, and put them in a separate 2gal bucket. I’ll check them early next week and determine if I should incorporate them into the stock.

Other observation: the HL group looks great- very active, jet black, and very little mortality. The other groups, as a whole, did not look as good. In speaking with Rick, he said that oxygen levels had been through the roof late last week, and that it likely crashed over the weekend (not confirmed with data yet). Is it possible that the HL groups are healthier because they can withstand more extreme changes in oxygen?

Collected new larvae; I only sampled 15k from SN-10 ambient B, and set up a double-bucket challenge on the rest. 0c7c53dd-d919-46c6-98ff-24dcc2728808

June 2nd

@ UW

Determined setting tank dimension: 26.75”W x 81.5”L x 20”D = 25.24 cubic ft = ~190 gal = ~715 L Purchased 2x 800 watt immersion heaters on Amazon for the setting tank. Also created a Skyline project with the new .blib file, re-uploading my converted Lumos files

June 3rd

Back @ Manchester today:

Arrival inspection

  • Larvae possibly present in:
    • K-6 Amb
    • K-10 Amb
    • NF-10 low B
    • SN-6 amb B
    • NF-10 amb A
    • SN-10 amb A
    • SN-6 low A
    • NF-6 low A
    • NF-6 amb B
  • Spawning (foamy) in:
    • SN-6 amb B
  • Fed with ~1/2 CGW & 1/2 Tetraselmos (sp?)
  • Imaged lots of well plates to catch up- halfway through imaging I decided to stop taking photos of each group prior to imaging the corresponding wells. Instead, I took a photo of the plate & date, and can refer back to the plate map when naming my image files.
  • Collected new larvae – lots today!

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  • This is my sampling setup: img_8327
  • Counted the 2-bucket challenge group. I wanted to set up another 2-bucket challenge today, but space is limited, and this data could be collected 1x/week when I screen.
  • Counts for SN-10 amb B after 48 hrs

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  • Inspected the setting tank screens that Ryan pointed me towards. Discovered they are all 450um, which is too big for the initial setting (I need <224um). I can still use these for post-set oysters with the tank upwelling.
  • Set up a small setting tank in my space, using a plastic tub and 180um screen silos. One bucket can fit 8. I think I can squeeze another bucket in there, too, so I can do all my setting here (which is great, since I can easily control the feeding and temperature). I got water flowing before I left, so I can see how things look after 24 & 48hrs.

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June 4th

Arrival inspection:

  • Larvae preset in
    • K-10 LOW
      • NF-10 Low B
      • NF-10 Amb A
      • SN-10 Amb A
  • Probes located in:
    • T1 – setting tank
    • T2 – gigas
    • T3 – Header
    • T4 – Larval tank, East manifold #3
    • D1 – broodstock, West manifold #8
    • D2 – Setting tank
    • D3 – broodstock, East manifold #8
  • Fed:
    • small table: header still 1/2 left, did not top off
    • middle table: 1/2 Tet and 1/2 Tiso, as no diatoms ready to go
  • Set up 100L tank, filled with FSW from heated line (~14degC) and placed immersion heater set point to max T (34degC) for Yaamini’s gigas heat shock tomorrow. Set @ 10:45am, after ~2.5hrs had reached 33degC.
  • Collected larvae:
    • Started a new bucket (#2) for SN-10 amb A group
    • Fixed well plates with lugols, did not image image
  • Prepped another downwelling setting tub for a total capacity of 16x 180um silos
  • Placed 16 upwelling 450um silos & corresponding manifold pieces in upwelling tank, filled ~2/3 with freshwater and added 250mL vortex to achieve 1 mL vortex/mL freshwater, left to clean overnight
  • Screened through shell hash pile to make microcultch – screened through 2400um, 1000um, and kept what held on 450um. Rinsed thoroughly with freshwater. Took home to sanitize in oven.
  • Pump sounds very weird again

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