Reproduction Survey Spring 2014 Standard Operating Procedures

SOP Anesthesia Procedure for Brood Check/Collection

1 stack consisting of 1 tray from each population is pulled out of the water where they are held for anesthesia. The stack will be different each week until all 4 stacks have been checked and then the process will repeat starting with the stack that was checked first.

Two of the trays are placed individual in roughly 10 gallons of ambient sea water in 2 twenty gallon pools.

The third tray remains outside side of the pool for approximately 45 minutes to deprive the oysters of oxygen.

Once the 45 minute mark is reached the tray is then placed in 10 gallons of a 50/50 seawater/85 g/L epsom salt freshwater mixture. These animals are then allowed 45 minutes to open up inside the treatment. Once open the animals are anesthetized by the epsom salts and remain gaping until flushed with sea water.

While the first tray is being treated the second tray is being dessicated. This repeats with the second tray treatment and the third tray dessication.

The animals are then visually inspected for signs of larvae including a grayish mass in the gills or on the lower deep cup of the shell. If anything suspect is seen, these animal is then flushed with seawater into a 52 um screen to capture any possible larvae. If larvae are present, they are washed off the screen into a 50 ml falcon with 75% Ethanol solution for preservation.

After all gaping animals are inspected, a quick count of all dead, gaping, and brooding animals is performed. Then the tray is placed in a pool of fresh ambient sea water to help the animals recover and avoid dessication until the process is completed for all trays being observed.

Once the final tray has been surveyed. The trays are then randomly organized back into a stack, secured to the hanging rope, and then returned back to their holding area.

Any brood collected will be brought back to the lab and have their solution switched out with 95% Ethanol.

5 24 2014 fidalgo repro

Anacortes, WA

Temps mid 50 to 60
Light rain

Participants L. Christine savolainen and Jake heare

Checked the third stack for reproductive activities using Anesthesia SOP. Also checked the first stack to see if there were any mortality events. The first stack was completely fine with minimal mortalities.

The third stack had no signs of brooding.

Numbers for pops as follows

2H9-12
Brood. 0
Gaping. 24
Dead 0

2N13-16
Brood. 0
Gaping 53
Dead 0

2S1-4
Brood. 0
Gaping. 77
Dead. 0

#priorities

5 23 2014 oyster bay emergency sampling

Oyster bay, WA

Mid 60s 70s

Participants Steven Roberts, Brent vadopalas, and Jake heare

We came back to oyster bay today to check if mortality occurred across the board.

Upon inspection of the three remaining stacks. There was no indication that the mortality event was spread to the other trays. It seems that a mix of extended exposure, high temperatures, and the treatment caused a mass mortality event. Dabob was the first group to be treated and thus did not experience the same duration of high heat and low oxygen that the north and south sound trays did. We decided to same from the affected trays anyway.  We collected 8 from the north sound tray, 4 from the south sound tray, and 10 from the dabob tray. We also individually bagged the shells from the dead oyster in hopes of using them for future data efforts.

On a side note, there appeared to be 2 dabob oysters that had recently spawned as evidenced by the post gonadal canals. Since they showed no signs of brooding it is assumed they spawned as male and may spawn again in the near future as female.

We also did a semi brood check on the other full set of trays and found no brooders.

Shells were placed on dry ice, tissue samples stored in rna later. Will be transferred to cold storage in the near future.

Will continue repro check at fidalgo tomorrow.

Enjoy this selfie of Brent and I in the car as I write reports.

5 22 2014 oyster bay repro

Oyster bay, WA

High mid 70s.

Participants Katie Jackson and Jake heare.

Tested for reproductive active today. Very bad news.

There has been a huge mortality event at oyster bay. Numbers for brooding, gaping, dead, and alive but not open are as follows.

1n13-16
Brood. 0
Gaping. 3
Dead 93
Alive 5

1s5-8
Brood 0
Gaping 2
Dead 68
Alive 4

1h5-8
Brood 0
Gaping 47
Dead  51
Alive. 9

Clearly dabob has survived whatever mortality event much better than north and south sounds populations. We will be sampling this site tomorrow to ensure fresh samples.

5 21 2014 Manchester repro

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.

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.

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%

4S9-12

Brooders 0

Gaping  43

Closed  55
% Open  43.9%

4N9-12

Brooders 0

Gaping 31

Closed  28
% 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%

1N5-8

Brooders 0

Gaping    46

Dead      7

Closed  14
% Open  76.7%

1S13-16

Brooders 0

Gaping     59

Dead        8

Closed    26
% 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%

2S13-16

Brood 0

Gaping  55

Dead   0

Closed 39
% Open  58.5%

2H5-8

Brood 0

Gaping 48

Dead  0
Closed  52
% Open   48%