RAPID Wilkinson Basin Sampling Protocol

RAPID Wilkinson Basin Sampling Protocol

RAPID Wilkinson Basin Sampling Protocol

UME based trips:2 science people at minimum, follow all the protocols.

UNH based trips: One designated UME person participates and helps where needed. Only net sampling is required at WB-5 and WB-7, same protocols as below. Priority is always WB-7, and if only one cast possible, then live and formalin samples are the priority.LOPC at both stations.

Station Events:Boat knows each stations coordinates by heart.

Record all information on a “Bottle Depth sheet” as a cruise log. Print out several, double-sided so that each station has it’s own page.

WB-5: (located on Jeffreys Ledge) CTD, 2 dual ring casts (2-4% Form, 1 ETOH, no live), LOPC, 5 Niskins (chl-A and TA/DIC)

WB-7: CTD, 5 Niskins (Chl-A and TA/DIC samples), 2 dual ring casts (2 Form, 1 ETOH, 1 Live), LOPC

Protocols:

LOPC: one cast each station at 40 m/min

Dual Ring Net:

Two Casts of 5m from bottom @ 40m/min.

Record start and end flowmeter numbers and depth net went to.

Record which cast was preserved as what.

Rinse nets designated for preserved samples, but not live net

Put labels inside jar and out

Be sure to fill out labels with cast and net number and be sure preservative label is correct to match which cast goes with what.

  1. Cast 1:
  2. One net with COVERED (black tape over holes) live codend to be divided up among live jugs. Rinse jugs first with hose water. Fill jugs with left over Niskin bottle water. No need to use ALL of the live cod-end contents- just add enough for about 100 animals per liter.
  3. One net preserved in 4% Formalin/seawater (rinse sample down with seawater; plop sample into jar with concentrated formalin; top off with seawater to shoulder of jar)
  1. Cast 2:
  2. Switch out covered live codend for a filtering type (not duct taped)
  3. One net preserved in 95% ETOH:
  4. Use the 150um mesh sieve for ETOH
  5. Condense all sample to one edge of sieve with seawater
  6. Use ETOH filled squirt bottle to move sample from sieve to jar, top off with ETOH.
  7. One net for 4% Formalin/seawater
  1. At end of each cruise, check that flowmeters count revolutions and check nets for rips and tears. Replace/repair where needed

CTD/Niskins:

  1. Instrument cast to within 5 meters off bottom
  2. Remove PAR cap (if present)
  3. Attach big weight to bottom of SeaBird cage
  4. Doesn’t have to be slower than 10m/min
  5. R/V Gulf Challenger crew likes to do Niskin Bottles on same wire as CTD. Must fill out Bottle Depth sheet included with this protocol (Dropbox/Wilki RAPID cruises/Meter_Wheel_Cal.xls). Best to download file to laptop to be done in wheelhouse by science member via computer. Trust me, you’ll appreciate that.
  6. Use the Bottle Depth to calculate for the crew running the winch the depths to be stopped at for attaching Niskins to the wire.
  7. Depths for bottle placement: 2m, 10, 20, 40, 120m and deep (just above CTD) where applicable for a total of 6 Niskins.

Water Filtering (to be done after station):

  1. Chl-A: ONLY ON UME TRIPS:
  2. Label centrifuge tubes with: Date, Rep, Filter type (GF, 5um 20um) and Depth
  3. 2m= 2 replicates GF/F; 1 TA/DIC bottle
  4. 10m= 2 reps GF/F; 2 rep 5m, 2 reps 20m
  5. 20m= 2 reps GF/F
  6. 40m= 2 reps GF/F
  7. 120m= 2 TA/DIC bottles
  8. Deep (just above CTD, record depth)= 2TA/DIC bottles

VERY important to follow protocols for TA/DIC water collection. USE GLOVES when handling the mercury preservative.

  1. Wrap centrifuge tubes in foil and put under ice. Put in -80C at GMRI as soon as possible after trip.

TA/DIC: we take care of on UME trips/ on UNH trips they do it:

Follow protocols UNH left us. Grab the kit and bottles from their shed. Clean bottles should be in a dark blue tote box just inside the UNH groups’ storage bay door. Bryan, the Captain, will open the storage bay for you. When done with the trip, full bottles and kit go back in shed and email goes out to Chris Hunt.

2 samples per 2m, 120m and deep- no bubbles in water hose and small bubble to allow expansion during freezing.

Equipment List and Location of Items:

Item / Location of storage / # Needed
CTD / Subject to change / 1
Niskins / UNH storage @ Newcastle / 5
Niskin hanging rack / UNH storage / 1
Weights / UNH (in big basket of bottles and or black plastic tackle box) / 6
Dual ring / UNH / 1
Cod ends / UNH / 1 covered, 2 regular
Filtering Manifold / UNH / 1
Filter bottle (for vacuum) / UNH / 1
Filter pump / UNH / 1
150 um Sieve/squirt bottles / UNH / 1
TC/DIC kit and 3 bottles / UNH / 1
ETOH carboy / GMRI / 1
Buffered Form primed jars / GMRI / 4
Empty ETOH jars / GMRI / 2
Live jugs / GMRI / 7-8
Green cooler / GMRI / 1
LOPC / GMRI / 1
HUGE round weight / UNH / 1
GF/F, 5um and 20um filters, tweezers, foil / GMRI / 1 box each
Large centrifuge tubes / GMRI / 12
ICE / GMRI/gas station / 2 bags
Meter wheel sheet / On computer or printed out

Lab Sorting of Live Sampling #’s:

Stage / Genetics (-80) (individual) / RNA/DNA (-80) (individual) / Fatty Acid (DEWAR)
(3 indiv per tube)
CV / 30 / 30 / 10 vials = 30 animals
Female / 30 / 30 / 10= 30 animals