| layout | title | date | categories | tags | projects |
|---|---|---|---|---|---|
post |
E5 Field Sampling Protocol |
2020-01-01 |
Protocols |
in situ coral sampling, coral, physiology |
E5, Putnam Lab |
Original: 220200101
Last Revised: 20201215
Contents
-
- Quart-size plastic bags
- Whirlpaks (2-oz or 4-oz)
- Fishing weights
- Sharpies
- Mesh collection bags
- 1 for each sampling team to hold supplies
- 1 for Turbinaria collection
- 3 to hold phys frags in water from boat
- 1 for shuttling samples between collection and boat
- Underwater camera and color standard
- Dive slate, pencil, and underwater paper
- Map of tagged colonies printed on underwater paper
- Stainless Steel Bone cutters
- Hammer and chisel
- Liquid nitrogen and dry shipper fitted with sleeve/bag to enable removing samples later
- GPS (in plastic bag) - recommend one unit per sampling team
- Dive flag
- Note: dive flag on buoy with anchor/weight that can be towed by snorkeler may be best, and GPS unit (in waterproof case) can be attached to flag above surface
- Extra zipties and cattle tags
- Extra batteries for camera and GPS
-
- Prelabel one quart-sized plastic bag (for physiological fragment) for each colony to be sampled with site, date, species, and tag ID.
- Note: sharpie comes off of plastic bags easily in the water. Consider using bags that have white labeling area, or labeling in multiple places, or using a wax crayon, etc.
- Stack all quart-size bags and pierce hole through upper corner of all bags (above seal so they can still be closed and hold water). Pass a zip-tie through the hole such that bags are held together on a spool. Include a small fishing weight on spool as well to make bags negatively buoyant and easier to handle in the water.
- Allocate specific colonies to be sampled to each sampling team, e.g., the northern half or southern half of colonies for two teams.
- Prelabel one quart-sized plastic bag (for physiological fragment) for each colony to be sampled with site, date, species, and tag ID.
-
- Drive to site and anchor at recommended anchorage coordinates
- Navigate to colony using GPS
- Find and clean off cattle tag/ziptie
- Take a closeup photograph of the tag with number clearly visible, then take a photograph of the whole colony with the color standard visible
- Acropora sampling
- Collect ~3" physiological fragment with apical polyp
- Collect ~1" molecular fragment 1 with no apical poly
- Collect ~1cm molecular fragment 2 with no apical polyp
- Place all fragments in quart size bag
- Recommendation: Use bone-cutters to clip one ~5" intact branch with apical polyp, then clip two smaller fragments from base as molecular fragments. Remainder is physiological fragment
- Pocillopora sampling
- Collect ~3" physiological fragment intact (i.e., only single cut at base)
- Collect ~1" molecular fragment 1 (can be cut from base of larger fragment, or can be a separate branch tip)
- Collect ~1cm molecular fragment 2 (can be cut from base of larger fragment, or can be a separate branch tip)
-
Place all fragments in quart size bag
- Porites sampling
- Collect 2 sq. in. physiological fragment
- Collect 1 sq. in. molecular fragment 1
- Collect <0.5 sq. in. molecular fragment 2
-
Place all fragments in quart size bag - Recommendation: Use hammer and chisel to remove natural lumps from colony. Place chisel at base of lump tangent to colony surface and hammer from the side to dislodge lump.
- When sampling is complete, seal quart bag with seawater inside. Pass off to person shuttling corals back to boat as soon as possible.
- Return samples to boat
- Place molecular fragments inter whirlpaks with no water, roll tightly and seal, and place into liquid nitrogen.
- Seal quart bags with physiological fragments with as much seawater as possible, and place into mesh bag(s) hanging from side of boat.
- Collect 100 large Turbinaria fronds and place in mesh bag. These collections should be spread out across the sampling site. Gloves are recommended. Can be floated in seawater until leaving site, then placed in the boat.
- Drive back to Gump.
- For next steps, see Sample Same Day Processing Protocol
** WATER SAMPLE FOR DIN and DIP**
Modified from Silbiger Lab Protocols - Water Column Nutrient Sampling
Materials
- 50 mL centrifuge tubes
- -20°C freezer
- 0.7 μm 25 mm glass fiber filters (GF/F)
- Flow injection analyzer
- Luer-Lok tip 60 mL syringes
Protocol
- Collect replicate in situ water samples at each site (n=2).
- Carefully, bend over the boat into the water as far down as comfortable (about 50 cm below the surface) holding a closed tube or snorkel/SCUBA to the desired water column collection depth.
- Rinse the tube 3 times by opening it underwater, filling it with seawater, closing it, returning it to the surface and emptying. Avoid going through the surface film with the tube open.
- Fill the tube a final time underwater. Close and bring to the surface.
- Decant to 40 mL and close tightly.
- Filter all samples through a 0.7 μm GF/F using a luer-lok tip 60 mL syringe.
- Place all samples in a -20°C freezer until analysis.
- Submit samples to the University of California, Santa Barbara Marine Science Institute or comporable institution for phosphate, nitrate + nitrite, nitrite, silicate analysis using a flow injection analyzer.
** WATER SAMPLES FOR ZOOPLANKTON BIOMASS AND C:N**