Cellular-Molecular OA response

OBJECTIVE:

TIMELINE:

F0 to F2 summary

Summer 2021

• F0 Bay scallops from CT spawned, exposed to 400 and 800 uatm pCO2 at 4 hours post-fertilization

Summer 2021 to Summer 2022

• F1 Bay scallops exposed to sample pCO2 condition for the totality of their lifespan

Summer 2022

• the two pCO2 cohorts were spawned and F2 Bay scallops were reared under thier matched parental-offspring condition (400 x 400 uatm and 800 x 800 uatm, parental x offspring)

late-Summer 2022

• at settlement stage, the 400 x 400 uatm F2 cohort was split to begin rearing animals under 1200 uatm (severe condition)

late-Summer 2022 to Spring 2023

• three F2 Bay scallop cohorts reared under the low (400 uatm), moderately-elevated (800 uatm), and severely elevated (1200 uatm) pCO2 levels continuously

Reexposure experiment with F2 Adults

March 23rd 2023

• 90 F2 Adult Bay scallop Airradians were tagged, 30 from each of the pCO2 cohort and position back into their treatment tanks

Apri 24th 2023

• Tagged Adults arranged in three tanks at each pCO2 conditions as the following:

  pCO2 level	tank 1	tank 2	tank 3
  400 uatm	#1-10	#11-20	#21-30
  800 uatm	#31-40	#41-50	#51-60
  1200 uatm	#61-70	#71-80	#81-90

April 26th 2023

– pre experiment chemistry full carbonate chem

May 1st 2023

– move animals into the treatments:

NOTE! notice we have 10 tagged animals per tank!

#s 1 - 30

   -    #s 1-10 = 400 x 400 uatm *MATCHED!*

   -	#s 11-20 = 400 x 800 uatm

   -	#s 21-30 = 400 x 1200 uatm

#s 31 - 60

   -	#s 31-40 = 800 x 400 uatm

   -	#s 41-50 = 800 x 800 uatm *MATCHED!*

   -	#s 51-60 = 800 x 1200 uatm

#s 61 - 90

   -	#s 61-70 = 1200 x 400 uatm

   -	#s 71-80 = 1200 x 800 uatm

   -	#s 81-90 = 1200 x 1200 uatm *MATCHED!*

pCO2 level	tank 1	tank 2	tank 3
400 uatm	#s 1-3 (3); 31-34 (4); 61-63 (3)	#s 4-6 (3); 35-37 (3); 64-67 (4)	#s 7-10 (4); 38 -40(3); 68-70 (3)
800 uatm	#s 11-14 (4); 41-43 (3); 71-73 (3)	#s 15-17 (3); 44-47 (4); 74-76 (3)	#s 18-20 (3); 48-50 (3); 77-80 (4)
1200 uatm	#s 21-23 (3); 51-53 (3); 81-84 (4)	#s 24-27 (4); 54-56 (3); 85-87 (3)	#s 28-30 (3); 57-60 (4); 88-90 (3)

May 2nd 2023:

• timepoint 1, destructively sample half of the animals (45 total) paying close attention to acquiring 5 animals per treatment

• full carbonate chemistry for all 9 tanks

NOTE! we will sample half of the animals from each tank, leaving 5 remaining.

Remove the following animals for destructive sampling

pCO2 level	tank 1	tank 2	tank 3
400 uatm	#s 1, 31, 32, 33, 61	#s 4, 35, 64, 65, 66	#s 7, 8, 9, 38, 68
800 uatm	#s 11, 12, 13, 41, 71	#s 15, 44, 45, 46, 74	#s 18, 48, 77, 78, 79
1200 uatm	#s 21, 51, 81, 82, 83	#s 24, 25, 26, 54, 85	#s 28, 57, 58, 59, 88

May 9th 2023:

• mid-point carbonate chemistry for 3 tanks, 1 representative of each treatment

May 16th 2023:

• timepoint 2, destructively sample the remaining animals (45 total)

Remove the following animals for destructive sampling

pCO2 level	tank 1	tank 2	tank 3
400 uatm	#s 2, 3, 34, 62, 63	#s 5, 6, 36, 37, 67	#s 10, 39, 40, 69, 70
800 uatm	#s 14, 42, 43, 72, 73	#s 16, 17, 47, 75, 76	#s 19, 29, 49, 50, 80
1200 uatm	#s 22, 23, 52, 53, 84	#s 27, 55, 56, 86, 87	#s 29, 30, 60, 89, 90