Wednesday 11/4/15
Gave a talk to the PSRF monthly meeting in the morning. Afterwards I went with Alice to the dock and brought up the cultch set F2 oysters to the hatchery. As there weren’t very many tile set oysters, I wanted to supplement the experiment with these. I put them in one of the setting tanks outside in static, ambient water.
Thursday 11/5/15 (Experiment Day)
Alice and I built a little “house” for the chiller to protect it from the rain. We set up the chiller in the smaller section of the setting tank in order to cool down the water quicker. The chiller was set to 0degC and turned on at 10am. Water on the other side was at 11degC, a little cooler than ambient due to the air temperature overnight. For the controls, 250+/- 5 cultch set oysters from each population were placed in 180 micron silos hanging in the ambient temperature side of the tank. I put around 350 oysters in silos on the chilled side at 11:30am when the temperature was at 7degC. At 1:00pm the temperature was at 0degC and maintained that temperature all day. At 1:30PM I added some tiles to the control and chilled tanks.
Chilled treatment on the left and controls on the right
Tiles added to treatments. The numbers represent oysters on that tile after sampling for gene expression. F = front of tile, S = side, B= back
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Control
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20 hours 0degC
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3 hours 0degC
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HCA5: 12F, 1S, 13B
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HCA13: 9F, 1S
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HCA11: 10F, 3S
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HCB2: 7F, 2S, 8B
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HCA12: 8F, 1S
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SSA9: 4F, 1S (4 sampled)
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HCA2: 1F, 8B
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HCB5: 1
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NFA6: 2F (4 sampled)
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SSA4: 4F, 2S
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HCB8: 2B
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SSA13: 4F, 1R
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SSA1: 14F, 2S, 1B
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SSB11: 1F
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SSA11: 0 (4 sampled)
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SSA14: 4F, 1S
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NFA4: 14F, 1B
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SSB2: 0F, 2B
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NFA11: 6F, 1B
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NFA7: 3F, 1S
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NFA9: 4F
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NFA12: 3F, 1B
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NFB2: 10F
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NFA15: 11F, 1B
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NFA14: 5F
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NFA2: 26F, 1B
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NFA?: 15F, 1B
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Total HC: 52
Total SS: 19
Total NF: 63
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Total HC: 22
Total SS: 17
Total NF: 41
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Total HC: 13
Total SS: 5
Total NF: 2
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At 4:00pm I sorted out 200 cultch set oysters from the chilled tank and added them to new silos in the control tank labelled with the population name and “3 hr chilled”. In addition to looking for differential mortality, we are interested in looking at temperature-induced gene expression. I put some oysters from each group in small (300 mL) cups with chilled water and then dissected 6 from each population and put whole body tissue in 1.5 mL tubes with RNALater. This took about 45 minutes total but I tried to alternate between populations while dissecting. I took pictures of most dissected oysters with a ruler. At 5:00pm I brought in one tile for each population from the chilled tank and dissected 4 oysters per population, giving 10 samples each. These samples were placed in the fridge overnight. The other ~150 oysters in the chilled tank were left there overnight to be sampled the next day.
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Time
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10:00AM
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– Chiller turned on and set to 0degC.
– Ambient at 11degC
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11:30AM
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– Chilled side: 7degC
– 350 cultch set oysters placed in chilled water
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12:15PM
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– Chilled side: 4degC
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1:00PM
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– Chilled side: 0degC
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1:30PM
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-Added tiles to control and chilled tanks
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4:00PM
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-Sorted out ~200 per population; added to new silo in the control tank
– Dissected 10 samples per pop for gene expression
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Getting My Genes Wet 