Did a brood check today at manchester. Found no brooders.
Numbers as follows.
4S5-8
Brood. 0
Gaping. 76
Dead. 3
4H13-16
Brood. 0
Gaping. 16
Dead. 2
4N13-16
Brood. 0
Gaping. 40
Dead. 3
Enjoy this picture of an eagle.
Wednesday, May 21, 2014
Tuesday, May 20, 2014
Capsized boat at Fidalgo Marina
Apparently a boat capsized roughly 500 feet from the samples at Fidalgo Bay. Depending on what leaked into the water, the size of the wake and other disturbances caused by rescue efforts. This might cause some issues with the animals. You can watch a time lapse video of the recovery effort here.
Monday, May 19, 2014
5-14 to 15-16 2014 Reproduction Work Up
5-14-2014
Manchester WA
Participants: Brent Vadopalas and Jake Heare
We retrieved the next set of trays in the dosing sequence and allowed each tray to dessicate for 45 minutes. Trays were then treated in a 10 gallon bath of 50% sea water to 50% freshwater mixed with 7 lbs of epsom salt. The tray was treated for 45 minutes, at which point it was removed and gaping animals were examined for signs of brooding. After examination the trays were then placed into a recovery tub with 100% sea water until the last tray examined had be in the recovery tub for 45 minutes. After each treatment the treatment water was replaced with fresh treatment to reduce temperature flux. Then trays were rebuilt into a stack and hung off the dock.
We counted the number of gaping animals and brooders for each tray.
They are as follows:
4H1-4
brooders 0
Gaping 25
Closed 72
% Open 25.8%
% Open 25.8%
4S9-12
Brooders 0
Gaping 43
Closed 55
% Open 43.9%
% Open 43.9%
4N9-12
Brooders 0
Gaping 31
Closed 28
% Open 52.5%
% Open 52.5%
5-15-2014
Oyster Bay WA
Participants: Katie Jackson and Jake Heare
We followed a procedure similar to that at Manchester. The difference being that the treatment water was not replaced after each treatment. We also counted the dead in each tray.
1H1-4
Brooders 0
Gaping 49
Dead 7
Closed 33
% Open 59.8%
% Open 59.8%
1N5-8
Brooders 0
Gaping 46
Dead 7
Closed 14
% Open 76.7%
% Open 76.7%
1S13-16
Brooders 0
Gaping 59
Dead 8
Closed 26
% Open 69.4%
% Open 69.4%
5-16-2014
Fidalgo WA
Participants: Steven Roberts and Jake Heare
Same procedure as at Oyster Bay.
2N1-4
Brood 0
Open 53
Dead 0
Closed 46
% Open 53.5%
% Open 53.5%
2S13-16
Brood 0
Gaping 55
Dead 0
Closed 39
% Open 58.5%
% Open 58.5%
2H5-8
Brood 0
Gaping 48
Dead 0
Closed 52
% Open 48%
Closed 52
% Open 48%
Monday, May 12, 2014
DNA Extraction Attempt 2 5-12-14
Attempting to extract DNA from 5-10 mm whole oysters minus the shell. This is needed so that sequencing can occur in the future with completed extractions. Unlike the last attempt I will be switching out 95% EtOH for Isopropanol to get better extraction results.
Protocol is as follows:
Incubation for 24 hours at room temp:
Isolation:
Protocol is as follows:
Incubation for 24 hours at room temp:
| 1 | ml | DNAzol |
| 2.35 | ul | Proteinase K (100ug/ml) |
| 1 | whole | oyster 5-10 mm |
| X5 | Reactions |
- Pipette 1 ul DNAzol and 2.35 ul Proteinase K in tube containing oyster
- Homogenize each oyster with disposable pestle.
- Centrifuge briefly
- Incubate overnight at ROOM TEMP.
Isolation:
| 500 | ul | Isopropanol |
| 1 | ml | 75% EtOH |
| 1 | ml | 75% EtOH |
| 200 | ul | Sterile H2O |
- Centrifuge @ 10,000 g for 10 minutes.
- Collect and Transfer Supernant; Discard pellet
- Add 500 ul Isopropanol
- Mix through inversion
- Incubate for 1 minute at ROOM TEMP.
- Spin down DNA precipitate
- Remove supernant
- Wash with 1 ml 75% EtOH
- Spin down
- Remove supernant
- Wash with 1 ml 75% EtOH
- Spin down until solid pellet
- Elute with 200 ul Sterile H2O
- Quant.
Friday, May 9, 2014
Spawning Oly Predictions
After looking through the new temp data I pulled off the temp loggers yesterday I notice a few trends that have lead to some predictions as to which site will spawn first.
Oyster Bay
Temps are trending upward to the 12.5 C mark that the animals require to begin spawning. It seems that they will have hit that temperature this week and will possibly begin spawning next week. The forecast for next week is also encouraging with the daily ambient air temps to be 75 or higher. This will bring these animals to fruitition. The only problem being that it will be during a spring tide which is not good for spawning.
If they don't spawn next week it will occur during the following neap tide during the second to last week in May.
Manchester
While these animals have experienced a warming trend, it seems as in recent weeks there has been a stalling of the warm temps. Water temperatures need to hit the 12.5 C mark and maintain them for several days to iniate spawning. It looks like it might be a slow start at Manchester or even a possible abortive spawn if the temps don't stay high enough long enough. I expect these guys to spawn in the last week of May, first week in June.
Fidalgo
Temps here are colder than elsewhere but are experiencing consistent increases throughout time. It will probably be the last week of May when they spawn.
Thursday, May 8, 2014
5-8-2014 Anesthetization Issues
Dr. Roberts recently asked me why I was getting such a low response rate from the treatments. I was seeing between 5 and 50% of the animals become anesthetized. Monitoring the situation I have a few ideas on why. Which I stated in a response to him in a message which is copied below.
I think it was a combination of the warm ambient air temps that caused an increase in treatment temperature which led to animals not opening in the treatment.
I also checked my treatment calculations and the epsom salt concentration was about half. I calculated it fo 5 gallons instead of the full ten. I'll just double the amount of epsom salt to make it the proper concentration. Hopefully it will increase response from the animals.
Brent has also suggested doing sub samples instead of dosing the whole tray to limit exposure by all the animals and reduce the amount of epsom salt we are using. I think its better to dose the whole tray because I can't guarantee that the percentage of animals that open will stay the same. On top of this variable percentage, there will only be 10-20% of the animals brooding at any given time. If we don't treat enough animals we will be more likely to miss spawning effort.
Brent also had some useful suggestions about previous work with the treatments as well as some recalculations of the treatment solution to increase treatment concentration.
Katie consistently saw about 30% response (without dessicating them first. I don't have her data on dessication, but response rate was higher). If we stick with the 5% threshold to see brooders and a 30% response, you'd need to treat 67 oysters to = 1. If the response with dessication is closer to 60%, you can decrease the subsample by half.
It sounds like the oysters may have been too warm to respond--important to bring a thermometer, and probably some ice packs to control treatment temp.
85g/L = ~7 lbs of MgSO4 for 10 gallons.
It can be argued that we only treat half the tray at any given time to limit downstream effects from the treatment which may affect spawning including abortive spawns or gonadal regression. The problem with this is that too see any signs of spawning we would be looking for 1-2 animals in any group to be brooding. I don't think this is a reliable way to determine if the populations are spawning. We would be better served in treating the whole tray to avoid mixups and hopefully have a higher number of brooding animals to strongly suggest that they animals have spawned.
I think it was a combination of the warm ambient air temps that caused an increase in treatment temperature which led to animals not opening in the treatment.
I also checked my treatment calculations and the epsom salt concentration was about half. I calculated it fo 5 gallons instead of the full ten. I'll just double the amount of epsom salt to make it the proper concentration. Hopefully it will increase response from the animals.
Brent has also suggested doing sub samples instead of dosing the whole tray to limit exposure by all the animals and reduce the amount of epsom salt we are using. I think its better to dose the whole tray because I can't guarantee that the percentage of animals that open will stay the same. On top of this variable percentage, there will only be 10-20% of the animals brooding at any given time. If we don't treat enough animals we will be more likely to miss spawning effort.
Brent also had some useful suggestions about previous work with the treatments as well as some recalculations of the treatment solution to increase treatment concentration.
Katie consistently saw about 30% response (without dessicating them first. I don't have her data on dessication, but response rate was higher). If we stick with the 5% threshold to see brooders and a 30% response, you'd need to treat 67 oysters to = 1. If the response with dessication is closer to 60%, you can decrease the subsample by half.
It sounds like the oysters may have been too warm to respond--important to bring a thermometer, and probably some ice packs to control treatment temp.
85g/L = ~7 lbs of MgSO4 for 10 gallons.
It can be argued that we only treat half the tray at any given time to limit downstream effects from the treatment which may affect spawning including abortive spawns or gonadal regression. The problem with this is that too see any signs of spawning we would be looking for 1-2 animals in any group to be brooding. I don't think this is a reliable way to determine if the populations are spawning. We would be better served in treating the whole tray to avoid mixups and hopefully have a higher number of brooding animals to strongly suggest that they animals have spawned.
Wednesday, May 7, 2014
Fidalgo Bay 5-2-14
5-2-14
Anacortes WA
Fidalgo Bay
Start Time: 9:30 AM
Finish Time: 5:00 PM
Temps: Mid 60s to Mid 70s
Participants: Jake Heare
I arrived earlier in the morning. I pulled up the hanging devices and broke them down into individual trays. I collected dead, counted live, and photographed trays for size/growth for later analysis. Upon completing these metrics the trays were reorganized into stacks containing one tray from each population at a random level in stack (top, middle, bottom). 3 stacks were then rebuilt and hung off the docks.
The 4th stack was treated with an epsom salt mixture to anesthetize the animals for brooding larvae inspection. One tray was dosed at a time in a 10 gallon mixture of 50/50 sea water/freshwater with 6-8 cups of epsom salt in them (roughly 75-80 g/L). Gaping oysters in each tray were individually inspected for brooding larvae. Once inspection was completed, trays were then transferred to a tub filled with 10-15 gallons of fresh seawater to recover from treatment for 30 minutes to 1 hour depending on recovery rate. In the first tray we had 25 animals open up, in the second we had 40 animals open, in the third there were 30 animals. None of these animals showed any signs of brooding larvae.
Anacortes WA
Fidalgo Bay
Start Time: 9:30 AM
Finish Time: 5:00 PM
Temps: Mid 60s to Mid 70s
Participants: Jake Heare
I arrived earlier in the morning. I pulled up the hanging devices and broke them down into individual trays. I collected dead, counted live, and photographed trays for size/growth for later analysis. Upon completing these metrics the trays were reorganized into stacks containing one tray from each population at a random level in stack (top, middle, bottom). 3 stacks were then rebuilt and hung off the docks.
The 4th stack was treated with an epsom salt mixture to anesthetize the animals for brooding larvae inspection. One tray was dosed at a time in a 10 gallon mixture of 50/50 sea water/freshwater with 6-8 cups of epsom salt in them (roughly 75-80 g/L). Gaping oysters in each tray were individually inspected for brooding larvae. Once inspection was completed, trays were then transferred to a tub filled with 10-15 gallons of fresh seawater to recover from treatment for 30 minutes to 1 hour depending on recovery rate. In the first tray we had 25 animals open up, in the second we had 40 animals open, in the third there were 30 animals. None of these animals showed any signs of brooding larvae.
 |
| Temperatures logged at Fidalgo Bay from Mar 1 to May 2, 2014. Temps logged every 15 minutes. |
| Fidalgo | 2N1-4 | 2N5-8 | 2N9-12 | 2N13-16 | Total |
| Live | 99 | 100 | 96 | 110 | 405 |
| 2H1-4 | 2H5-8 | 2H9-12 | 2H13-16 | Total | |
| 91 | 100 | 83 | 94 | 368 | |
| 2S1-4 | 2S5-8 | 2S9-12 | 2S13-16 | Total | |
| 92 | 104 | 93 | 94 | 383 |
 |
| Calm Day at Fidalgo |
 |
| Sea Otter hangin out by the dock |
 |
| Sea Otter Hunting for Prey |
 |
| Sea Otter after capturing large red rock crab |
 |
| Sea Otter Eating Large Eel. |
Oyster Bay 5-1-14
5-1-14
Shelton WA
Oyster Bay/Totten Inlet
Start Time: 9:30 AM
Finish Time: 5:30 PM
Temps: Mid 60s to Mid 80s
Participants: Jake Heare
I arrived earlier in the morning. I pulled up the hanging devices and broke them down into individual trays. I collected dead, counted live, and photographed trays for size/growth for later analysis. Upon completing these metrics the trays were reorganized into stacks containing one tray from each population at a random level in stack (top, middle, bottom). Since one stack is missing we were unable to completely reorganize the trays as such. One hanging tray has 2 Hood Canal Pops and 1 North Sound pop tray. 2 stacks were then rebuilt and hung off the docks.
The 3rd stack was treated with an epsom salt mixture to anesthetize the animals for brooding larvae inspection. One tray was dosed at a time in a 10 gallon mixture of 50/50 sea water/freshwater with 6-8 cups of epsom salt in them (roughly 75-80 g/L). Gaping oysters in each tray were individually inspected for brooding larvae. Once inspection was completed, trays were then transferred to a tub filled with 10-15 gallons of fresh seawater to recover from treatment for 30 minutes to 1 hour depending on recovery rate. In the first tray we had 25 animals open, in the second we had 3, in the third we had 4. None of these animals showed signs of brooding larvae.
Shelton WA
Oyster Bay/Totten Inlet
Start Time: 9:30 AM
Finish Time: 5:30 PM
Temps: Mid 60s to Mid 80s
Participants: Jake Heare
I arrived earlier in the morning. I pulled up the hanging devices and broke them down into individual trays. I collected dead, counted live, and photographed trays for size/growth for later analysis. Upon completing these metrics the trays were reorganized into stacks containing one tray from each population at a random level in stack (top, middle, bottom). Since one stack is missing we were unable to completely reorganize the trays as such. One hanging tray has 2 Hood Canal Pops and 1 North Sound pop tray. 2 stacks were then rebuilt and hung off the docks.
The 3rd stack was treated with an epsom salt mixture to anesthetize the animals for brooding larvae inspection. One tray was dosed at a time in a 10 gallon mixture of 50/50 sea water/freshwater with 6-8 cups of epsom salt in them (roughly 75-80 g/L). Gaping oysters in each tray were individually inspected for brooding larvae. Once inspection was completed, trays were then transferred to a tub filled with 10-15 gallons of fresh seawater to recover from treatment for 30 minutes to 1 hour depending on recovery rate. In the first tray we had 25 animals open, in the second we had 3, in the third we had 4. None of these animals showed signs of brooding larvae.
 |
| Temperatures logged at Oyster bay from Feb 28th to May 1st, 2014. Temps logged every 15 minutes. |
| Oyster Bay | 1N1-4 | 1N5-8 | 1N9-12 | 1N13-16 | Total |
| Live | 84 | 67 | 100 | 251 | |
| 1H1-4 | 1H5-8 | 1H9-12 | 1H13-16 | Total | |
| 89 | 101 | 96 | 14 | 300 | |
| 1S1-4 | 1S5-8 | 1S9-12 | 1S13-16 | Total | |
| 74 | 93 | 167 |
 |
| Treatment set up at Oyster Bay |
 |
| Artsy photo of science |
 |
| Gaping Oyster |
 |
| Jack trying to help out |
 |
| Jack decided to drink some epsom salt water behind my back. He made this face when I caught him. |
Manchester 4-30-14
4-30-14
Manchester WA
Clam Bay
Start Time: 10:00 AM
Finish Time: 6:00 PM
Temps: Mid 60s to Mid 80s
Participants: Sammi Brombacher and Jake Heare
We arrived earlier in the morning. We pulled up the hanging devices and broke them down into individual trays. We collected dead, counted live, and photographed trays for size/growth for later analysis. Upon completing these metrics the trays were reorganized into stacks containing one tray from each population at a random level in stack (top, middle, bottom). 3 stacks were then rebuilt and hung off the docks.
The 4th stack was treated with an epsom salt mixture to anesthetize the animals for brooding larvae inspection. One tray was dosed at a time in a 10 gallon mixture of 50/50 sea water/freshwater with 6-8 cups of epsom salt in them (roughly 75-80 g/L). Gaping oysters in each tray were individually inspected for brooding larvae. Once inspection was completed, trays were then transferred to a tub filled with 10-15 gallons of fresh seawater to recover from treatment for 30 minutes to 1 hour depending on recovery rate. In the first tray we had 30 animals open, in the second tray we had 20, in the 3rd tray we had 45. None of these animals had any signs of brooding larvae.
For More info see the oly wiki
Manchester WA
Clam Bay
Start Time: 10:00 AM
Finish Time: 6:00 PM
Temps: Mid 60s to Mid 80s
Participants: Sammi Brombacher and Jake Heare
We arrived earlier in the morning. We pulled up the hanging devices and broke them down into individual trays. We collected dead, counted live, and photographed trays for size/growth for later analysis. Upon completing these metrics the trays were reorganized into stacks containing one tray from each population at a random level in stack (top, middle, bottom). 3 stacks were then rebuilt and hung off the docks.
The 4th stack was treated with an epsom salt mixture to anesthetize the animals for brooding larvae inspection. One tray was dosed at a time in a 10 gallon mixture of 50/50 sea water/freshwater with 6-8 cups of epsom salt in them (roughly 75-80 g/L). Gaping oysters in each tray were individually inspected for brooding larvae. Once inspection was completed, trays were then transferred to a tub filled with 10-15 gallons of fresh seawater to recover from treatment for 30 minutes to 1 hour depending on recovery rate. In the first tray we had 30 animals open, in the second tray we had 20, in the 3rd tray we had 45. None of these animals had any signs of brooding larvae.
 |
| Temps logged for Manchester from Feb 28th to April 30th, 2014. Temps logged every 15 minutes. |
| Manchester | 4N1-4 | 4N5-8 | 4N9-12 | 4N13-16 | Total |
| Live | 90 | 97 | 59 | 91 | 337 |
| 4H1-4 | 4H5-8 | 4H9-12 | 4H13-16 | Total | |
| 97 | 85 | 89 | 84 | 355 | |
| 4S1-4 | 4S5-8 | 4S9-12 | 4S13-16 | Total | |
| 92 | 91 | 98 | 94 | 375 |
 |
| Tray being treated with epsom salt |
 |
| Oysters being treated with Epsom Salt |
 |
| 1 tray being treated, 1 tray being examined, 1 tray recovering |
 |
| Gaping oyster |
 |
| Sammi processing Trays. |
 |
| Ferry on the Sound. |
Dabob Final Sampling 4-18-14
4-18-2014
Dabob WA
Tarboo Creek
Start Time: 11:30 AM
Finish Time: 2:00 PM
Temp: Low to Mid 60's
Participants: Sammi Brombacher and Jake Heare
We reached the site at roughly 10 AM on Friday morning. There was a slight breeze, misty rain, and overall warm temperatures. We walked out to the site at 11:30 when the tide was low enough to reach the trays. We collected all the trays and placed them on a plastic sled and then dragged them back to the truck. This took approximately 2 hours and some swear words. The trays were then driven back to Seattle and cold stored at 12C. Trays stayed here for 14 hours before being processed the following day by Jake Heare.
 |
| Logged Temps for Dabob from Jan 30th to April 17th 2014. Temps logged every 15 minutes. |
Dabob
|
3H1-4
|
3H5-8
|
3H9-12
|
3H13-16
|
Total
|
Live
|
2
|
16
|
32
|
19
|
69
|
Dead
|
1
|
1
|
1
|
0
|
3
|
3N1-4
|
3N5-8
|
3N9-12
|
3N13-16
|
Total
| |
Live
|
0
|
16
|
0
|
17
|
33
|
Dead
|
0
|
7
|
2
|
11
|
20
|
3S1-4
|
3S5-8
|
3S9-12
|
3S13-16
|
Total
| |
Live
|
14
|
19
|
11
|
15
|
59
|
Dead
|
12
|
6
|
8
|
6
|
32
|
For more info see the Oly Wiki
Tuesday, May 6, 2014
Inaugural Post
This is the first post in what hopes to be a long running record of the scientific work performed by Jake Heare. This should include wet lab, computer science, and field work posts with informational videos and links to supportive information.
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