This entry describes the sampling and respirometry data collection for Acropora hyacinthus in Moorea, French Polynesia at CRIOBE and Gump Research Stations.

Overview

Sampling was conducted by D. Becker and D. Conetta.

Protocols are as described for Montipora capitata in this online notebook: Respirometry Protocol

Sampling Time Points

Samples were collected for two methodologies:

1) Respirometry, including dark and light runs
2) Flash frozen samples

Table of sampling time points is below.

Day	Time of Day	Hours Post Fertilization	Stage	Sampling	Date
0	2200	0	Fertilized Egg	Freeze (CRIOBE)	Nov 9
1	0200	4-5	Cleavage	Freeze (CRIOBE)	Nov 10
1	1000-1200	12-14	Prawn chip	Freeze	Nov 10
2	1000-1200	36-44	Gastrula/Early Swimming	Freeze + Resp	Nov 11
3	1000-1200	72	Larvae	Freeze + Resp	Nov 12
4	1000-1200	96	Larvae	Freeze + Resp	Nov 13
7	1000-1200	144	Larvae	Freeze + Resp	Nov 16

Sampling Details

Respirometry

Deviations from Mcapitata protocol are described here. Respirometry runs were ran with 30 minute dark and 30 minute light runs. These runs were recorded and data stored separately. Runs included one SDR 641 and one SDR 710 plate under an AquaIllumination light set to mean 500 PAR as measured by light measurement across surface of plates.

Photosynthesis light runs were ran first. After 30 min under light, light was turned off and dark respiration run was run for 30 minutes.

Each well contained 6 larvae with 2 blanks per plate per run. One run was conducted each day, which includes both plates for a total of 48 wells per run.

Plates were filled with filtered seawater and sealed with glass coverslips.

Photographs of notebooks can be found here Notebook Photos

Temperature was measured in air and in a well before and at the end of the run. Temperatures were between 25-28C.

In all SDR 641 plates, sensor B6 was broken and excluded from analysis. Wells with bubbles or other problems were excluded from analysis.

Metadata of all runs can be found here Respirometry metadata

All runs were completed between 1000 and 1300 on each sampling day.

Freezing and Sampling

Samples were frozen each day in -40C freezers. Sampling was conducted each day just prior to respiration run start times.

Analysis and Processing

AH will process analysis of respiration and photosynthesis using scripts and protocols as described for Mcapitata. Ahyacinthus respository