6 26 2014 oyster bay repro

Used the modified anesthesia sop with insulation and ice blocks. Collected from the water the lost stack as well as the hhn stack to reshuffle oyster trays. Big things of note, 1 south sound tray was 99% dead. It was a middle tray, had no predators and no fouling. No idea why those animals died but the other two trays in the stack survived. Brooding in all sampled trays seemed to have followed expectations. With no pop with way more or way less brooders than expected. Also collected temp logger info. All trays were kept in ambient seawater during Pretreatment or out plant if not sampled

Lost trays marked with orange ziptie, hhn trays labeled with blue ziptie.

Numbers as follows:

Temps in c

Pretreatment
Initial.       14
45.             14
1.5.            14
2.25.          16

Treatment
Initial.        11
45.              11
1.5.             11
2.25.            9
3.                 8

Recovery
Initial.          13
45.                15
1.5.               16
2.25.             16

Salinity
Pretreatment.         26
Treatment.               68
Recovery.                 24

Trays not used for Brood collection

1S9-12
Live.           1
Dead.         82

1H13-16
Live.          12
Dead.         0

Brood collection

1N1-4. From hhn stack
Brood.       2
Gaping.     72
Dead.         10
Closed.      1

Brooders
#        size.      Sick
1.        25.         W
2.        31.         Grey

(Three previous trays were put out immediately as stack 1. Far left from shore)

1H9-12
Brood.       1
Gaping.      92
Dead.         4
Closed.      1

Brooders
#      size.      Sick
1.      27.          Grey

1S1-4
Brood.       11
Gaping.      85
Dead.          10
Closed.       2

Brooders
#          size.         Sick
1.          29.            W
2.          25.            W
3.          28.            W
4.          30.            W
5.          26.            W
6.          30.            W
7.          25.            W
8.          26.            W
9.          28.            W
10.        31.            W
11.        27.            W

10/11. Were dirty samples, had lots of mud mixed in with larvae.

1N9-12
Brood.          5
Gaping.        84
Dead.            16
Closed.          1

Brooders
#        size.        Sick
1.        31.           W
2.        29.           W
3.        36.           W
4.        28.           W
5.        30.           W

6 25 2014 Manchester Repro Check

Manchester WA

Mid 60's to mid 70s sunny

Participants: L. Christine Savolainen and Jake Heare

Performed the Anesthesia SOP modified with insulation on the treatment tub as well as ice blocks to keep temperatures within range. Found one brooder in the Northern population with minimal larvae and found one brooder from Dabob that had a very strange mix of eggs and what appeared to be semen. This might be evidence for an abortive spawn. also collected all mortalities from trays.

numbers as follow.

Temps in C

Pretreatment
Initial       10
45 min     15
1.5 hr       15

Treatment
Initial       11
45 min     11
1.5 hrs     11
2.25 hr     11

Recovery
Initial        10
45 min      15
1.5 hr        17


Salinity
Pretreatment      27
Treatment          65
Recovery           21


Brood Collection

4S1-4
Brood    0
Gaping     50
Dead      22
Closed    13

4N5-8
Brood        1
Gaping       53
Dead         23
Closed       19

Brooders
#       size      sick
1        25        w

4H5-8
Brood       1
Gaping      52
Dead        7
Closed      33

Brooders
#        size         sick
1        15           W

Harbor Seal watching us work all day. 

6 20 2014 fidalgo repro check

Anacortes wa

Mid 60s cloudy to sunny

Participants: L. Christine savolainen and Jake heare

Followed the anesthesia sop with insulation on the treatment group.  Found one brooder from the southern population. Two animals looked like possible brooders in the northern pop but they closed quickly and I was unable to confirm brooding in them.

Also Brian found the missing stack at oyster bay. They will be worked up next Thursday as they are safely secured to the dock again.

Numbers as follows

Temps in c
Pretreatment.
Initial.       8
45 min.     8
1.5 hrs.     9

Treatment
Initial.           9
45 min.         9
1.5 hrs.         10
2.25 hrs.       10

Recovery
Initial.        7
45 min.      8
1.5 hrs.      8

Salinity in ppt
Pretreatment.      35
Treatment.           65
Recovery.             35

Brood collection
2S1-4
Brood.       1
Gaping.     83
Dead.         0
Closed.      10
Brooders
#       size.     Sick
1.       27.        W

2H9-12
Brood.        0
Gaping.      77
Dead.          0
Closed.       10

2N13-16
Brood.     0
Gaping.    76
Dead.       0
Closed.    34

6 19 2014. Oyster bay repro

Shelton WA

Mid 60s to high 70s

Participants: L. Christine savolainen and Jake heare

Followed the repro sop with insulation and ice blocks to control temp. Also collected all morts from all trays. Surprisingly, the dabob pop had no brooders while the south sound pop had 11 brooders.

Numbers as follow.

Temps in c
Pretreatment
Initial.            16
45 min.          18
1.5 hrs.          18

Treatment
Initial.            11
45 min.          11
1.5 hrs.          11
2.25 hrs.        10

Recovery
Initial.      16
45 min.     20
1.5 hrs.      16

Salinity
Pretreatment.       25
Treatment.            63
Recovery.              25

Brood collection

1H1-4
Brood.    0
Gaping.  70
Closed.    9
Dead.        7

1N5-8
Brood.     3
Gaping.   54
Dead.       8
Closed.      1

Brooders
#             size.              Sick
1.             29.                  W
2.             26.                  W
3.             30.                  W

1S13-16
Brooders       11
Gaping.          80
Dead.              12
Closed.           1

Brooders
#             size.               Sick
1.             27.                  W
2.             22.                  W
3.             26.                  W
4.             25.                  W
5.             26.                  W
6.             28.                  W
7.             31.                  W
8.             33.                  W
9.             22.                  W
10.           21.                  W
11.           29.                  W

Number 11 could not have its Brood collected because it was attached to the tray.

6 18 2014 Manchester Repro Check

Manchester, WA

Hi 50's to mid 60's, mostly cloudy until later in the day

Participants: L. Christine Savolainen and Jake Heare

Performed the Anesthesia SOP modified with insulators and ice blocks for the treatment tub. Found 1 brooder at Manchester. It seems no matter where they are, South Sound oysters just love to spawn. Also collected all mortalities from all trays checked. Failed to take a picture of the single brooder, I absent mindedly put the tray into recovery before taking the pic and then couldn't find the brooder when I did remember.

Numbers as Follow:

Temps in C

Pretreatment
Initial         7
45 min       8
1.5 hr        10

Treatment
Initial        8
45 min      8
1.5 hr       7
2.25 hr      7

Recovery
Initial         8
45 min       8
1.5 hr        10

Salinity in ppt
Pretreatment     30
Treatment         71
Recovery          30

Brood Collection

4H13-16
Brood      0
Gaping     63
Dead       4
Closed     12

4N13-16
Brood       0
Gaping     73
Dead        2
Closed      12

4S5-8
Brood      1
Gaping     55
Dead       10
Closed     28

Brooder
#           size       sick
1           19         w

Temperature Logger Readings from May to Mid June 2014

Last week I pulled temperature data from the logger pendants out in the field using the Shuttle. Everything performed stellarly and I have the temp graphs on the computer. I'll go by site for each temp graph.

Manchester, WA

From the temperature readings it appears that Manchester has stayed below the 12.5 C mark for most of the logging period. Only recently have temperatures crested the 12.5 mark and stayed that way for several days. It may be soon that the animals will begin spawning and brooding (or spawned within the last week or so). 

Oyster Bay, WA

From this data and the amount of brooders we have seen at Oyster Bay, its clear that temps have been above the 12.5 C mark  since early May. This again correlates with expectations and as temperatures steadily increase I believe we may see a slowing of reproduction due to stress demands. 

Fidalgo Bay, WA

Contrary to the observations in the field, it seems that Fidalgo bay has been much warmer since the beginning of June. Though it does look like in early May there was a warm period for several days. As of June 1st temperatures crested the 12.5 C mark and headed into spawning temps. We saw our first brooder approximately 6 days after that and saw a large number of brooders approximately 12 days after temps went above 12.5. Though temps have somewhat decreased recently I believe we will continue to see spawning across the board.

Overall I think that the populations are responding to temperature cues nicely, though it does seem that North sound respond quickly to temperature increases. The south sound animals seem to have nearly synchronous spawning during warm periods which would be of obvious benefit in South Sound where temps continue to increase and only stay within the spawning window for a short amount of time. The dabob animals seem to have reduced response to temperature cues and do not actively spawn unless temps are somewhat stable for a duration of time. 

6 13 2014. Fidalgo repro check

Anacortes wa

Mid 60s partly cloudy lite rain

Participants:  Sean Bennett and Jake Heare

Used the anesthesia sop with insulation on the treatment group. Temps stayed steady due to insulation and weather. Found multiple brooders in only one population. If you can guess which pop, you get a sticker.

Numbers as follow:

Temps in c

Pretreatment
Initial.         7
45 min.       7
1.5 hrs.       10

Treatment
Initial.          6
45 min.        6
1.5 hrs.         6
2.25 hrs.       6

Recovery
Initial.             7
45 min.           8
1.5 hrs.           9

Salinity in ppt
Pretreatment.     27
Treatment.           66
Recovery.             26

2N1-4
Brood.       0
Gaping.     68
Dead.         1
Closed.       27

2S13-16
Brood.     7
Gaping     83
Dead.       1
Closed.     3

Brooders
#           size.              Sick
1.            30.                W
2.             26.                W
3.             26.                W
4.              25.              W
5.              26.               W
6.              28.               W
7.              25.              W

2H5-8
Brood.         0
Gaping.       72
Dead.           0
Closed.        27

Lots of males with semen. Expect more brooders next week.

6 12 2014 oyster bay repro

Shelton wa

Mid 60s partly cloudy to rainy

Participants:  L. Christine savolainen  and Jake heare

Used anesthesia sop with insulation and semi frozen ice blocks. No change in Temps the entire time. Weather definitely helped. Found several more brooders. Some were below the 20 mm mark.

***Update*** I forgot to mention that I also jumped in the water today during lowtide and hunted underneath the docks to find the lost stack of trays. I was unable to find them. I used a 6 ft pole to feel underneath the lower parts of the dock as well as grab items I found. I pulled out half a dozen shell bags and 1 set of oyster growout cages. Our stacks are long gone. It might be possible to search further under the docks with a diver but I couldn't risk my safety going any further under the docks. At this point in time, its probably safe to assume that we will be unable to ever recover the missing trays. 

Numbers as follow:

Temps
Pretreatment
Initial.  15
45 min. 15
1.5 hrs.   15

Treatment
Initial.       13
45 min.      13
1.5 hrs.      13
2.25 hrs.     13

Recovery
Initial.        13
45 min.       13
1.5 hrs.        13

Salinity.
Pretreatment.        26
Treatment.              64
Recovery.               26

Brood collection
1S13-16
Brood.    3
Gaping.   63
Dead.       11
Closed.     6

Brooders.    
#          size.           Sick
1.            27.            W
2.            26.              W
3.            19.             W

1N5-8
Brood.      2
Gaping      48
Dead.        8
Closed.     1

Brooders
#               size.            Sick
1.                32.               Grey
2.                20.              W

1H1-4
Brood.         1
Gaping.       86
Dead.            8
Closed.         1

Brooders
#         size.          Sick
1.         19.             W

Many males with semen in all treatments.

6 11 2014 Manchester Repro

Manchester WA

Temps Mid 60's to High 70s
Sunny

Participants: Joelle Blais and Jake Heare

Used the Anesthesia SOP to check for brooding larvae. I modified this per Steven and Brent's suggestions to temperature control via adding ice blocks to the solution as well as insulate the tub with 2 inch styrofoam insulation with foil top and bottom. Much to my surprise not only did the insulation work, kept water within 1 degree of ambient with out ice blocks, it actually lowered the temps of the treatment when ice blocks were added (2 degrees below ambient). I believe this insulation and ice blocks method will be the way to go from here on out. Even on the warmest days I see it insulating the tubs very well.

Numbers as follow:

Temps in C

Pretreatment

Initial        9
45 min      15
1.5 hrs      19

Treatment

Initial     11
45 min   11
1.5 hrs    10
2.25 hrs   8

Recovery
Initial       11
45 mins    13
1.5 hrs     15


Salinity in ppt

Pretreatment      25
Treatment          65
Recovery           25


Brood Collection

4H1-4
Brood     0
Gapin      63
Dead       2
Closed     31

4S9-12
Brood       0
Gaping      49
Dead         12
Closed       37

4N9-12
Brood      0
Gaping     41
Dead        8
Closed     12

One male in the 4H group had sperm within its shell. I assume these animals are attempting to spawn though water temps seem to be below expectation.

6 6 2014 fidalgo repro check

Followed the anesthesia sop at fidalgo. Attempted to keep the treatment Temps around 11 c where the ambient water was. The direct sun had other plans. Will need way way way more gel bags. even then it might be more than can actually fit in the tub with the samples.

Also to my and hopefully everyone else's surprise. I found a Brooding oyster! If you can guess which population it came from before you look below, you are one smart cookie.

Numbers as follow

Temps
Pretreatment
Initial.      10
45 min.     15
1.5 hrs.      19

Treatment
Initial.         11
45 min.       16
1.5 hrs.        17
2.25 hrs.      19

Recovery
Initial     10
45 min.   15
1.5 hrs.    19

Salinity
Pretreatment.     20
Treatment.           63
Recovery.             21

Brood collection
2S9-12
Brood.      0
Gaping.    53
Dead.        1
Closed.      43

2H13-16
Brood.       0
Gaping.     58
Dead.         0
Closed.       33

2N5-8
Brood.      1
Gaping.    59
Dead.        2
Closed.      36

Brooders
#         size(mm).        Sick
1.          31.                     W

Lots of nukaluka in the water. Some may have been collected with brooding larvae. May muddy up counting.

Picture of brooders follows.

6 5 2014 oyster bay repro check

Shelton, wa

Participants. Katie Jackson and Jake heare.

High 60s to mid 70s sunny

Followed the anesthesia protocol but modified it by adding ice block and bags of Ice into pools. Temperature started at 13 but stabilized at 16 c. Temp increase unavoidable but non lethal.

Numbers as follow

Temps
Pretreatment.   
Initial. 13
45 min.  16
1.5 hrs.   16

Treatment
Initial.    10
45 min    17
1.5 hrs.   16
2.25 hrs.   16

Recovery
Initial  13
45 min. 17
1.5 hrs 17

Salinity
Pretreatment.     26
Treatment.          63
Recovery.            26

Brood collection.
1S13-16
Brood.  2
Gaping. 60
Dead.     12

Brooders
#     size.    Sick
1.     23.        W
2.     29.        W

1H1-4.   
Brood.   2
Gaping 79
Dead.     7

Brooders
#      size.      Sick
1.       25.         W
2.       24.          Grey

1N5-8
Brood.  1
Gaping.  52
Dead    7

Brooders
#      size.      Sick
1.      30.         W

Multiple males with milt in their shells. Possibly spawning at the moment.

6 4 2014 Manchester Repro Check

Manchester WA

Mid 60s to Low 70s

Followed the Anesthesia SOP to check for brooding oysters. Found no brooders. Water temperatures we also quite low. Last week the surface water temp was around 13 C. This week it was closer to 8 C. I think the bay underwent another turn over event which may lead to either a much later spawn timing for the oysters or an abortive spawn which we will not catch. 

Numbers as Follows:

Temps (Celsius)

Pretreatment 

Initial       8

45 min     9 

1.5 hrs.    10

Treatment

Initial         7

45 min     10

1.5 hr       12

2.25 hr      12

Recovery

Initial      8

45 min    9

1.5 hr     9

Salinity (ppt)

Initial          28

Treatment   63

Recovery    28

Brood Check

4S13-16

Brood    0

Gaping  60

Dead     3

4N1-4

Brood     0

Gaping    43

Dead       15

4H9-12

Brood    0

Gaping   55

Dead      11

Also toured the new hatchery at Manchester with Ryan, saw a chipmunk, and the canadian goose on the docks hatched 4 of it 5 eggs. They goslings were adorable.

Brooding Olys in Hatchery

Flow through with bong screen to catch Oly larvae. 

Various flowthroughs for brooding Olys

Oly larvae (invisible)

Larval Tanks and Ryan's Gesticulation

Chipmunk

Baby Goslings!

Clam Bay with glassy surface waters!

5 30 2014 fidalgo repro check

Anacortes wa

Mid 60s to mid 70s

Performed standard reproduction check using the anesthesia sop. Found no brooders. Also surface water Temps at 3 pm were only 9C.

Temps
Pretreatment
Initial 10
45 min. 12
1.5 hrs   16

Treatment
Initial. 9
45 min.  13
1.5 hrs.  16
2.25 hrs.  19

Recovery
Initial.  9
45 min. 13
1.5 hrs.  16

Salinity
Seawater.  30 ppt
Treatment.  65 ppt

Brood
2S5-8
Brood.   0
Gaping.  77
Dead.      1

2N9-12
Brood.   0
Gaping.  46
Dead.     0

2H1-4
Brood.  0
Gaping. 50
Dead.   1

Also forgot to dessicate 2N tray. Placed it straight in treatment from bath. Still got ok reaction from it.

5 29 2014 oyster bay repro check

Shelton wa
Mid 60s

Performed standard repro check from anesthesia sop. Found brooders!

Also took temp, salinity and size of brooding females.

Numbers as follow:

Temps
President treatment bath
Initial.  15.5
45 min.  15.5
1.5 hrs.   16.5

Treatment temp
Initial.  11
45 min. 12
1.5 hrs.  15
2.25 hrs.  16

Recovery bath Temps
Initial.   14
45 min   17
1.5 hrs.   18
2.25 hrs.  18

Salinity in ppt
Pretreatment.  25
Treatment.        70
Recovery.          25

Brooding data
1H1-4
Brood.  1
Gaping 80
Dead.    7
Brooders info
#       size (mm).      Larvae taken
1.        24.                   y

1S13-16
Brood.   5
Gaping  74
Dead.     9
Brooders info
#        size (mm)         larvae taken
1.        36.                       Y
2.         26.                      Y
3.         26.                      Y
4.         25.                      Y
5.         28.                      Y

1N5-8
Brood.   2
Gaping 51
Dead.     6
Brooders info
#            size (mm)         larvae taken
1.            27.                     Y
2.            28.                      Y

Brooding larvae are easy to spot. They appear as pools or swarms of white material around the mouth portion of the visceral mass. If oyster has water still in it they maybe swarming inside of the animal. To confirm their presence use black tip of zip tie to collect small portion for viewing. Small white grainy substance is brooding larvae. Grey or green substance most likely algae or food.

Also hazard note. Small portion of ethanol fell on to some gaping animals when transferring larvae to tube. Washed off immediately but worried that undue mortality may occur. That or drunk oyster party. 

Attempted to get pics of brooding but not good ones. Posted below.

5 28 2014 Manchester Repro Check

Manchester WA

Mid 60s

Started partly cloudy and warm, then rained later in the day while finishing the last tray check.

Took temperatures while the temp probe still worked. Probe failed halfway through the day due to being doused with salt water.

Followed procedure listed in the Reproduction Anesthesia SOP. Modified the procedure at the end by eliminating the time for the final tray to recover due to the torrential rain.

Also took salinity of the water collected as well as the treatment salinity.

Temps as follows:
Seawater for pre treatment in C
Initial:          13.1
45 min:       19.5
1.5 hrs:       23.8

Treatment Temps in C
Initial:          15.3
45 mins:       21.2
1.5 hrs:        23.5

Salinity (taken during initial collection/treatment creation)
Seawater:   26.5 ppt
Treatment:  68.5 ppt

Brooding Conditions as follows:
4S1-4
Brood    0
Gaping   43
Dead      9

4H5-8
Brood    0
Gaping   27
Dead      0

4N5-8
Brood     0
Gaping    54
Dead      15


There were two oysters in 4H5-8 and three in 4N5-8 that looked like candidated for brooding due to darkened gill tissues. Rinsed all 5 thoroughly with seawater in to 52 um screen but collected nothing. Gill tissue color did not change. I assumed these animals were not brooding based on this.

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.