NGI - Methods

Protocol for sampling for NGI Marsh project:

In each of the proposed locations, we will visit 1 CONTROL and 1 OIL IMPACTED marsh location.

At each location we will sample 3 quadrats (1m2), located no more than 5 meters apart; i.e. 10meters of shoreline.

All samples will be collected in DUPLICATE where possible, ONLY sample “A” will be analyzed by Micromethods, with sample B as a backup.

Go to CONTROL Site first! Do IMPACTED site last, to avoid potential contamination of tools and samples.

At the site:

• 1. Get GPS coordinates of location, record. Get photos from boat of the shoreline.

  2. Run 10meter transect along shore parallel to Spartina, place Q1 at 0m, Q2 at 5m and Q3 at 10m approx to capture “typical” marsh conditions.

  3. Place first 1m2 quadrat in Spartina alterniflora zone (no more and 3m = 10ft from open water). Get photos of the quadrat location, close-up of plants, sediments.

  4. Record GPS location of center of quadrat plus/minus accuracy in feet. Get air temperature and record. Record the time of collection on datasheet

  5. Cut 20-30 Spartina alterniflora stems near base, cut into pieces that will fit in 1000ml glass jar, divide sample equally between 2 jars (duplicate A and B). Record number of stems harvested. Place labels on jars.

  6. Using gardening trowel, collect sediments to a depth of no more than 3-4” (8-12cm) and fill two 250ml glass jars about 2/3 full (duplicate A and B). Preference for shallow sediments with oil, over deeper sediments where possible. Place labels on jars.

  7. Dipnet Fundulus grandis and Palaemonetes near low tide and collect a minimum of 50 shrimp and 10+ fish from the location. You will ONLY have one sample at CONTROL and one at IMPACT site. Estimate meters of shoreline sampled and record on datasheet. The sample from each site needs to be in one labeled ziplock bag (fish and shrimp together from one collection) and maintained on ice until returned to Mark Peterson's lab for processing.

  8. Repeat steps 2 – 5 on two more quadrats placed no more than 5 meters apart.

  9. Return all jars and bags to coolers and place on ice. Record the jar/bag numbers on datasheet.

  10. Clean all hardware with acetone. First, put on CLEAN gloves. Rinse in water. Get clean paper towel, squirt with acetone, wipe down plant cutters and trowel. Repeat twice more with fresh paper towels. Give a final rinse with tap water. Discard all gloves, paper towels. Want to make sure we minimize any possible contamination of samples between sites.

Next, go to OIL IMPACT site and get 3 more quadrats worth of samples. Make sure to clean gear well after this site!

Before leaving a site make sure all samples are on ice in cooler(s). All jar/bag numbers have been recorded for each site on the datasheet, along with GPS coordinates.

On return to GCRL, fill coolers with fresh ice, store inside overnight. Transfer fish/shrimp samples in their ziploc bags to Peterson lab prior to freezing. Download pictures from camera to desktop computer. Separate images by location and quadrat within location.

ANIMAL TISSUE PROCESSING:

Fish will be processed in Mark Peterson's lab by removing the head, opening up the alimentary track and removing the liver from F. grandis. These will be pooled for a min weight of 1-5grams. Shrimp will also be beheaded and the carapace removed and the hepato-pancreas/organs removed and these be pooled (min weight 1-5grams).

In addition, pool the muscle tissues of the fishes as one sample, and the meaty tail of the shrimps as a second sample. Each sample (fish liver, shrimp hepato-pancreas, fish muscle, shrimp tails) will then be frozen at -20C in 7ml glass scintillation tubes until analysis by Micromethods. Remember to have duplicate A and B samples in Epindorff tubes where possible.

Fill out Chain of Custody form, indicating Test Needed as ORO (Oil Range Organics) and PAH (Poly Aromatic Hydrocarbons). Then arrange to drop off samples in jars (sediments and plant tissues) at Micromethods.