Larval Care – Pacific oyster larvae at PSRF Manchester

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping.

Here’s a pic:

Oyster food!

Larval Care – Pacific oyster larvae at PSRF Manchester

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping.

Here’s a pic:

Collecting & cleaning larvae on nylon screen.

Larval Care – Pacific oyster larvae at PSRF Manchester

It’s Saturday. Yep, Saturday…

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping.

Here’s a pic:

Saturday morning at the hatchery.

 

Larval Care – Pacific oyster larvae at PSRF Manchester

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping.

Prepared a spreadsheet for Dan to use to calculate necessary quantities of algae (two species) to achieve target concentrations:

Google Sheet: AlgaeFeedingCalcs

Here’re some pics of the day:

Larval Care – Pacific oyster larvae at PSRF Manchester

Continued helping on Dan’s Pacific oyster familial crosses project at the Puget Sound Restoration Fund (PSRF) hatchery at Manchester Research Station in Manchester, WA. Pacific oyster (Crassostrea gigas) families were crossed based on earlier genotyping. Here’re some pics of the day:

Morphometrics – Olympia Oyster OA Larvae Completed

Finished measuring Ostrea lurida larvae using NIS-Elements BR (10x objective and 10x nosepiece setting in software). That’s 2,620 larvae measured!

NOTE: Robyn previously measured 33 tubes worth of larvae, but I’m not certain where that data was saved.

All measurements by me can be found here (Google Sheet): FHL_Oly_larvae_measurements

Measured 30 larvae from each tube of the following samples:

  Box Label Color Tube Label Notes
70 FHL Oly OA x DO Box 2 etOH Friedman Purple 4.3.15 U FB HL.L-1
71 FHL Oly OA x DO Box 2 etOH Friedman Purple 4.3.15 V FB HL.L.2
72 FHL Oly OA x DO Box 2 etOH Friedman Purple 4.3.15 W FB HH.L.1
73 FHL Oly OA x DO Box 2 etOH Friedman Purple 4.3.15 X FB HH.L.2
74 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 Y SS LLH3 Many transluscent shells
75 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 Z SS LL.H.4 Many transluscent shells
76 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 AA SS LHH3 Mostly transluscent shells
77 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 BB SS LHH4
78 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 CC SS LL.L.3
79 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 DD SS LL.L.4
80 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 EE SS LH.L.3
81 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 FF SS LH.L.4
82 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 SS LLL-1 4.3.E
83 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 SS LLL2 4.3.F
84 FHL Oly OA x DO Box 2 etOH Friedman White 4.3.15 SS LHL-1 4.3.G
85 FHL Oly OA x DO Box 2 etOH Friedman Pink 4.4.15 FB release Low CO2 103A
86 FHL Oly OA x DO Box 2 etOH Friedman White SS Low CO2 4.4.15 release 103A All shells partially transluscent.
87 FHL Oly OA x DO Box 2 etOH Friedman Blue SS High CO2 4/5-6/15 release 103B All shells mostly transluscent.
88 FHL Oly OA x DO Box 2 etOH Friedman Pink 44/5-6/15 release FB LCO2 FB 103A

 

Example images from notes above:

 

Sample #74

 

 

Sample #75

 

 

Sample #77

 

 

Sample #85

 

 

Sample #86

 

 

Sample #87

Morphometrics – Olympia Oyster OA Larvae Continued

Continued measurements of larvae samples from yesterday. Measured length (hinge to growing edge) and width of 30 oysters from each tube using NIS-Elements BR (10x objective and 10x nosepiece setting in software).

The measurement data can be found here (Google Sheet): FHL_Oly_larvae_measurements

Here’s the list of tubes that were measured today and their labels:

  Box Label Color Tube Label Notes
14 2015 – DO x pH Olys FHL Friedman White AM 103B-SS LH-H-2 3.31.15
15 2015 – DO x pH Olys FHL Friedman White AL 103B-SSLH-H-1 3.31.15
16 2015 – DO x pH Olys FHL Friedman White AK 3.31.15 103A-SS LL-H-2
17 2015 – DO x pH Olys FHL Friedman White AJ 3.31.15 108A-SS LL-H-1
18 2015 – DO x pH Olys FHL Friedman Blue AG 108A-SS HL-L-2 4.1.15
19 2015 – DO x pH Olys FHL Friedman White 3.30.15 SS-103A LL-H-3 Day 3 3.30.15
20 2015 – DO x pH Olys FHL Friedman White 3.30.15 Day 3 SS-103A LLH-4 3.30.15
21 2015 – DO x pH Olys FHL Friedman White 3.30.15 SS-103B Day 3 LH-H-3 3.30.15
22 2015 – DO x pH Olys FHL Friedman White 3.30.15 Day 3 SS 103B LHH-4 Day 3
23 2015 – DO x pH Olys FHL Friedman White 3.30.15 Day 3 SS-108A LL-L-3 3.30.15
24 2015 – DO x pH Olys FHL Friedman White 3.30.15 Day 3 SS 108A LLL-4 Day 3
25 2015 – DO x pH Olys FHL Friedman White 3.30.15 SS 108B Day 3 LHL-3 3.30.15
26 2015 – DO x pH Olys FHL Friedman White 3.30.15 Day 3 SS 108B LH-L-4 3.30.15 Day 3
27 2015 – DO x pH Olys FHL Friedman Blue AH 108B-SS HH-L-1 4.1.15
28 2015 – DO x pH Olys FHL Friedman White 3.29.15 Day 5-103A LL-H1 SS-D5
29 2015 – DO x pH Olys FHL Friedman White 3.29.15 Day 5 103A LL-H-2 D5 SS
30 2015 – DO x pH Olys FHL Friedman White 3.29.15 Day 5 103B LH-H-1 SS D5
31 2015 – DO x pH Olys FHL Friedman White 3.29.15 Day 5103B LH-H-2 D5 SS
32 2015 – DO x pH Olys FHL Friedman White 3.29.15 Day 5 108A LL-L-1 D5-SS
33 2015 – DO x pH Olys FHL Friedman White 3.29.15 Day 5 108A LL-L-2 -SS D5

 

Morphometrics – Olympia Oyster OA Larvae

Olympia oyster larvae were measured using NIS Elements-BR (Nikon) imaging software to measure larvae length (hinge to growing edge) and width (10x objective and 10x nosepiece setting on software). Below is a screenshot of what the software, larvae, and measurement controls look like:

Measured 30 individuals from each tube.

The following samples were measured (these are the tube labels; currently not certain what shorthand labelling scheme should be used):

  Box Label Color Tube Label Notes
1 2015 – DO x pH Olys FHL Friedman White/Blue Released: 3.25.15 103B-SS Olys High CO2 3-25-15
2 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 103A HLH-1 3.28.15 3/25/release
3 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 103A HL-H-2 3.28.15 3.25 release
4 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 103B HH-H-1 3.28.15 3/25 release
5 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 103B HH-H-2 3.27.15 release
6 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 108A HL-L-1 3.27.15 3/25 release
7 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 108A HL-L2 3.27.15 3.25 release
8 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 108B HH-L-1 3.27.15 3/25 release
9 2015 – DO x pH Olys FHL Friedman Blue SS Day 3 HH-L-2 3.27.15 3.25 release
10 2015 – DO x pH Olys FHL Friedman Blue AI 108B-SS HH-L-2 4.1.15
11 2015 – DO x pH Olys FHL Friedman White Ap 108B-SS LH-L-1 3.31.15
12 2015 – DO x pH Olys FHL Friedman White AN 108A-SS LL-L-1 3.31.15
13 2015 – DO x pH Olys FHL Friedman White AO 103A-SS LL-L-2 3.31.15

Measurement data is here (Google Sheet): FHL_Oly_larvae_measurements

Histology – Fixed Oyster Larvae Embedded in Agar

Oyster larvae were received 5/19/2014 in Davidson’s fixative. There were three different groups (A, B, and C), with each group consisting of 5 different tubes. Confirmed with Hamdi that the tubes in each group were just replicate sample collections from the same exact population of oyster larvae.

Made a 4% agar solution in distilled H2O and cooled to 56C.

Transferred 1mL of larvae in Davidson’s fixative from a single tube in each group to their own separate weigh boats. Weigh boats were angled to help pool the larvae solution. Mixed the larvae with an equal volume of 4% agar solution. Allowed mix to solidify. Transferred to the appropriate cassettes (labelled A, B, and C) with a sponge and stored in freshly made Davidson’s fixative solution.

Here’s an image of the interior box lid that explains what the A, B, and C group labelling means:

RNA Isolation – Olympia Oyster Larvae from May 2012

Isolated RNA from 11 Olympia (O. lurida) oyster larvae samples, from 4 different pCO2 treatments using TriReagent (MRC) according to the manufacturer’s protocol. RNA was resuspended in 100uL of 0.1% DEPC-H2O, spec’d on the Roberts Lab NanoDrop 1000, and stored @ -80C.

Results:

RNA looks good. Will prepare for submission for Illumina HiSeq sequencing. Sample pooling info is here.