Tag Archives: manchester

Oyster Sampling – Olympia Oyster OA Populations at Manchester

I helped Katherine Silliman with her oyster sampling today from her ocean acidification experiment with Olympia oysters (Ostrea lurida) at the Kenneth K. Chew Center for Shellfish Research & Restoration, which is housed at the NOAA Northwest Fisheries Science Center at Manchester in a partnership with the Puget Sound Restoration Fund (PSRF). We sampled the following tissues and stored in 1mL RNAlater:

  • adductor muscle (A)
  • ctenidia (C)
  • mantle (M)

When there was sufficient ctenidia tissue, an additional sample was stored in 75% ethanol for potential microbial analysis.

Tissue was collected from two oysters from each of the following oyster populations:

  • British Columbia (BC)
  • California (CA)
  • Oregon (OR)

Oysters were sampled from each of the following tanks:

  • 1A
  • 2A
  • 3A
  • 4A
  • 1B
  • 2B
  • 3B
  • 4B

Tubes were labeled in the following fashion:

  1. Population & Tank (e.g. OR3B)
  2. Tag#
  3. Tissue

If no tag was present on the oyster, the oyster was assigned a number (beginning at 150 and increased sequentially) and photographed with a ruler for future measurement. White colored tags were written with the number followed by the letter ‘W’ (e.g. 78W) – no tag color info was recorded for other tag colors.

Additionally, gonad developmental stage was roughly assessed: ripe, kinda ripe, or not ripe.

All info was recorded by Katherine in her notepad. All samples were retained by Katherine (not sure where she stored them).

Utensils were flame sterilized between oysters and gloves/work surfaces were washed with a 10% bleach solution between oysters.

 

Here are a few pics from the day:

 

 

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Bringing Ocean Acidification System online

I was out at Manchester yesterday to help Laura out with getting things going.

tldr- Water is flowing, rising temperature seems to be an issue. This could be attributed to air temp and/or pumps.

When we arrived water temperature was up to 19C after about a week. We decided to drain down system refill, calibrate Durafets, and monitor system over next few days with respect to temperature and pH.

Here is the system draining..
[youtube https://www.youtube.com/watch?v=CtDfMPkK69E?rel=0&showinfo=0]

 

As the system drained we calibrated with NBS buffers (7-4-10). In actuality I think they were only calibrated at 7 and 4. Need to confirm calibration system with Honeywells.

Probes are designated pink, blue, green, and yellow. Two in treatment tanks and two in each of experimental systems. As we placed in 7 buffer they initially read as follows

pink – 6.6

blue – 6.98

green – 6.82

yellow – 6.89

After all were calibrated we went through buffers and just read.

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Here are more #s

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Tour time (if you listen closely you can hear a narration)
[youtube https://www.youtube.com/watch?v=K5xkpAAVSuI?rel=0&showinfo=0]

 

As the system started to refill with ambient water (10c) this is how the pH probes read.

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This is without any C02 input. We then “sample calibrated” experimental system to read same pH

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At the end of the day pH was set to 7.5 in treatment tank and we will monitor to see how temperature and pH holds (assuming it can adjust with high flow rates). For more on this day check out Laura’s post.

I will leave you with an inside look at treatment tanks. Note that the first tank in the video (Tank #2) has less water coming in from the head tank as compared to Tank #1.

[youtube https://www.youtube.com/watch?v=ctiNrf2DKEU?rel=0&showinfo=0]

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Larval Care – Pacific oyster larvae at PSRF Manchester

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping.

Prepared a spreadsheet for Dan to use to calculate necessary quantities of algae (two species) to achieve target concentrations:

Google Sheet: AlgaeFeedingCalcs

Here’re some pics of the day:

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Larval Care – Pacific oyster larvae at PSRF Manchester

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping. Here’re some pics of the day:

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No code required

Today Sam and I took care of the daily maintenance of F2 Olympia oysters grown at the Ken Chew Shellfish Hatchery at Manchester, WA. In short, larvae are being described from Fidalgo (North: NF), Oyster Bay (South: SS), and Hood Canal (HC) broodstock. Though I had done most aspects before, never without Katherine, thus she provided instructions.

When I arrived the algae tank was empty and pump was off.
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The system was flushed with freshwater (with Sam soaked with said water) along with bleach. The algae tank was filled. The first set of tanks that were processed were the larger larvae (+160um).
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Using 100um screen larvae were collected, brought up in 800ml and 0.5ml (x3) taken for counts.
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The remaining larvae were placed back into cleaned systems.

Next, the smaller larvae tanks were processed, using 160 over 100 um screens, with larger ones moving over to +160um tanks (above). For this counts were done for both size classes and DNA samples taken for 160 size class. The smaller larvae placed back in the same tanks.
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For DNA samples, 4mls of larvae from 800ml beaker were taken, rinsed with ethanol and placed in 1.5ml centrifuge tube with 1ml of RNAlater (This was also done with fresh larvae- below).
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The last systems tackled were the “new”, fresh larvae from collectors..
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For this, larvae were captured using 200um / 100um screen, with larvae moved to larger tanks, counts done, and samples collected for DNA. As expected very little larvae, they were brought up in 400ml, 1ml used for counting, and 8ml used for DNA samples.
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Sam counted the larvae, probably still very wet (Sam and the larvae).
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And here are the counts!
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