#Heat / Mechanical Stress Post Mortem
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Last Wednesday (4/22/15) I ran both the heat and mechanical stress treatments on all three populations of Oly oysters. The experiment had a few kinks but otherwise worked well. 

First animals were held in the cold room for 2 weeks before the experiment. Feeding occurred regularly but not during the same time of day. Water changes occurred once a week. 

On the day of the experiment animals were transferred to mesh bags. The plan called for 24 per population (8 1hr, 8 24hr, 8 mort). But I accidentally placed 22 in each bag because it was the number in the previous workplans. Now there are 8 1hr, 8 24hr, and 6 morts. Per population.

Heat treatment required that 500 ml of salt water be preheated for the animals prior to the experiment. Animals were transferred to the beakers while still in the mesh bags. The bags caused a few of the animals to be partially out of the water. Also the water was not aerated during the treatment. Animals were sampled according to the schedule (1 hr post end of treatment) which worked smoothly by having the treatments staggered by 1 hour. 

The mechanical stress also went smoothly due to the staggering of the treatments. The biggest issue was that the dewar ran out of Liquid Nitrogen before I was able to sample the Dabob and Oyster Bay. The samples were placed on ice and transferred to the -80 as soon as possible. 

The following day (4/23/15) I collected fresh liquid nitrogen immediately prior to sampling. I began collecting samples at 2 pm and worked continuously through the afternoon and evening. I started with the heat shock groups (N,H,S in order), then the Mechanical Stress (N,H,S in order) and finally collected the controls (N,H,S in order). The liquid nitrogen ran out prior to collecting the controls, which were placed on ice and transferred to the -80 as quickly as possible. 

Both days took approximately 14 hours to complete treatments and sampling. 

Things to improve:

- Larger dewar needed
- More dissection scissors and forceps (broke a pair while working)
- More -80 freezer space.
- Run heat and mechanical treatments a day apart to reduce sampling fatigue
- Increase speed on centrifuge (no mortalities from mech stress)
- Digital thermometer in heat treatment
- Account for thermal mass of oysters when heat treating
- Larger volume of prewarmed water (1 L or more) to offset thermal mass
- Create bulletted reference sheet with relevant information
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