Standard Operating Procedure for water sampling on SMART and other Ala Wai projects
Provided by Craig Nelson, June 2018
Three sites on each stream (Mānoa, Pālolo, and Makiki) will performed monthly, one or two more intense efforts especially for tidal & episodic storms;
Manoa – Lyon, Kanewai, Iolani minimum
Palolo – Measures house, Anuenue, St. Louis or Chaminade or USGS 16247000 abandoned site
Makiki – Forest & Wildlife Makiki, Archie Baker Park, Halau Ku Mana, USGS 16238000 King & Kalaukaua
• Cooler prepped, blue ice frozen in advance.
• 500 mL polycarbonate Water collection bottles are acid-leached (2-12h in ~1N HCl), DI rinsed, dried, tape encircled, 1 per site.
• 20 mL amber borosilicate Fluorescent dissolved organic matter fDOM sampling bottles w/ptfe-lined lids are acid washed, DI rinsed, vials combusted/caps dried, assembled, tape encircled, 1/site, cardboard box rack.
• 50 mL polypropylene disposable centrifuge nutrient sampling tubes, 2/site, held in plastic wire rack.
• Sterivex 0.2 um poresize polyethersulfone DNA sampling filters, 1/site, held in ziplock bag.
• 2 mL Flow CytoMetry FCM sampling tubes, amended 80 uL 8% paraformaldehyde (16uL 32%PFA), 2/site.
• Storm and Tidal Supplemental Sampling, multiple samples of above and below per site:
o 25 mm GF/F particulate sampling filters in polypropylene swin-lok housing, aluminum foil packet.
o 25 mm HAWP chlorophyll sampling filters in polypro swin-lok housing, aluminum foil packet.
o 25 ml Kimax clear borosilicate DOC sampling bottles and ptfe-lined lids are acid washed, DI rinsed, vials combusted, dried, assembled, tape encircled, held in cardboard box rack.
o 500 mL polypropylene TSS sampling bottle tape encircled
– Samples (500ml whole water) collected 10 cm subsurface in Water collection bottles after triple-sample rinse,
– Samples in a cooler in the field with “blue ice” until return to the laboratory for processing within 6h collection.
– Sample codes will be continuously numbered throughout project, with site/timestamp records associated.
Field Sample Processing: Equipment is a) Nalgene rig, b) handheld vac pump, c) sharpie, d) tweezers, e) 47mm polyethersulfone 0.2 um filter, f) MoBio Bead Tube, g) nutrient sampling tube, h) fDOM sampling bottle. Filter is installed in rig, 250ml sample added to reservoir, vacuum applied (10 in Hg max), once filtered completely filter is stuffed into bead tube, filtrate is poured into fDOM/nutrient sampling tubes, remainder discarded, samples are frozen.
Lab Sample Processing:
– Duplicate 1ml FCM samples are pipetted out of whole water into FCM sampling tubes, mixed by inversion, labeled side of tube as FCM sample, flash frozen -80C after 0.5-3.0 hours at room temp.
– 45 ml whole water decanted into nutrient sampling tube, labeled side and top with sharpie as “TN/TP” sample and frozen -20C upright in racks.
– 250-500 ml water is pumped through platinum or peroxide cured silicone tubing via peristalsis from bottle after thorough mixing. Flow rates will target 10 minutes per sample or 3L per hour.
– After brief flush first 100ml is pumped through Sterivex DNA sampling filter, measuring volume by collecting filtrate and discarded as waste (cleans the filter of plasticizers).
– Second 100ml is collected sequentially into fDOM (20ml) and dissolved nutrient sampling tubes (45ml). fDOM are labeled on encircled tape with sharpie and refrigerated in the dark. Nutrients are labeled side and top with sharpie as “DIN” sample and frozen -20C upright in racks
– After 250 mL minimum Sterivex are emptied of water (dry filter, some drops OK), labeled on side with sharpie detailing code and volume filtered, and stored frozen -80C in ziplocks until DNA Extraction.
– During storm/tidal sampling, additional 125ml samples from original sample collected through GF/F particulate sampling filters (20 ml filtrate collected as DOC sampling bottles above) and again through HAWP chlorophyll sampling filters. DOC samples will be labeled and acidified to pH 2 with 12N HCl and stored in fridge. Filters will be wrapped in aluminum foil, wrapped in tape and labeled and frozen -80C in ziplocks.
– During storm/tidal sampling, additional 500 mL whole water TSS sampling bottle will be filled and labeled in fridge.
Sample analytical fates:
– FCMs and fDOM processed by Nelson lab on Attune Flow Cytometer and Horiba Aqualog Fluorometer.
– DNA filters processed by Nigro/Alegado/Nelson labs for DNA Extraction and downstream microbial analysis.
– DIN processed by S-LAB and TN/DP processed by S-LAB via in-line oxidation on Seal AA3 SFIA.
– DOC and TDN processed by S-LAB on TOC-L autosampler with TN analyzer.
– GF/F processed by S-LAB on Elemental Analyzer, HAWP processed by S-LAB Turner AU-10 fluorometer.
– TSS processed by Tsang Lab.