Fauna was collected either by hand, hand nets, plankton nets, seines or trawls. Flora was collected either by hand cutting leaves of marsh plants, hand collection of algae , plankton tows or water filtration for particulate organic matter. Stable Isotope Food Web Monitoring Sample Collection and Processing Sediments Collect 5 surface (2cm) cores using 60 ml syringe, place cores in a labeled (date, station, sample type) 4 oz Qorpack jar and in a cooler for transport back to the lab. Freeze if the sample cannot be dried soon. Dry sediments @ 50o C. After the sample is dried, use a dissecting microscope and check sediment for carbonates using a few drops of 10% HCl on a small subsample, if bubbling occurs then a larger subsample should be acidified and redried for isotope analysis. If carbonates exist it is necessary to remove them, as they will interfere with the 13C analysis. Grind a subsample of the sediments (Wig-L-Bug) for stable isotope analysis and place in a clean scint vial. Benthic diatoms Collect benthic diatoms by laying down several (5-7) separate pieces of 210u Nitex on the sediment surface at low tide. Pick a sediment surface that has a golden sheen and still looks fairly wet. If the surface is wet enough the diatoms will migrate up through the Nitex and there will be a fairly pure sample on one side of the Nitex. Use a 500 ml squirt bottle filled with ambient water from the site and gently squirt drops onto the Nitex to get it to lay on the sediment surface without air gaps. After 20 minutes or so, peel the Nitex from the sediment surface and check for an adequate golden sheen on the Nitex, be careful not to contaminate the diatom side with the under side mud or sand. Fold the Nitex (diatom side) carefully in on itself and place in a labeled (date, station, sample type) ziplock bag and put bag in a cooler for transport back to the lab. At the lab unfold the Nitex with the diatom side on the outside, fold it so that the middle of the Nitex forms a corner facing down and carefully using the ambient water squirt bottle, rinse the diatoms off onto an ashed 25 mm GFF filter that has been set up in a 25mm Gelman filter tower and flask (need replicate filters). When the filter/liquid looks green suction it dry and place the filters into separate labeled (date, station, sample type) small petri dishes. Put petri dishes in a freezer if the samples cannot be dried soon. Dry the filters @ 50o C. After the sample is dried, use a dissecting microscope and check the filters for carbonates using a drop of 10% HCl on a portion of the filter, if bubbling occurs then the whole filter should be acidified and re-dried for isotope analysis. If carbonates exist it is necessary to remove them, as they will interfere with the 13C analysis. Nereis Collect Nereis worms by digging with a shovel in the tidal flat sediments; often the peat chunks have more worms. Collect 15 to 20 worms of 2-5 cm lengths and place in a jar with ambient water. Place the jar in a cooler for transport back to the lab. At the lab transfer the worms to another jar and put clean ambient water in the jar, let the worms sit overnight to purge their guts, then count and record the lengths of worms, towel blot and place worms in a labeled (date, station, sample type) glass scint vial and freeze. Dry the worms @ 50o C and then grind the worms (Wig-L-Bug) for stable isotope analysis and place back in the labeled (date, station, sample type) scint vial. Mummichog Using a seine collect 15 to 20 mummichogs, 35-50 mm lengths and place in a labeled (date, station, sample type) ziplock bag and place in cooler for transport back to the lab. At the lab, count and record the lengths of the mummichogs, fillet and save tissue in a labeled (date, station, sample type) glass scint vial and freeze. Dry the mummichogs @ 50o C and then grind the mummichogs (Wig-L-Bug) for stable isotope analysis and place back in the labeled (date, station, sample type) scint vial. Ribbed mussel Collect 15 to 20 ribbed mussels from below the Spartina alterniflora channel bank edge and place in a labeled (date, station, sample type) ziplock bag and place in cooler for transport back to the lab. At the lab, count and record the lengths of the mussels, cut out the abductor muscle and rinse/dip in DI water. Save muscle tissue in a labeled (date, station, sample type) glass scint vial and freeze. Be careful not to get shell fragments mixed in with the abductor muscle. Dry the tissue @ 50o C and then grind the tissue (Wig-L-Bug) for stable isotope analysis and place back in the labeled (date, station, sample type) scint vial. Particulate Organic Matter (POM) Collect between 1 liter and 4 liters (depends on station) of mid ebb channel water into labeled (date, station, sample type) bottles recording date and time of day, place in cooler for transport back to the lab. At the lab, shake the bottles thoroughly and filter water onto ashed, 25 mm GFF filters until clogged (need replicates) and place filters into separate labeled (date, station, sample type) small petri dishes. Put petri dishes in a freezer if the samples cannot be dried soon. Dry the tissue @ 50o C. Blue mussel Collect 15 to 20 blue mussels from the front edge of the channel and place in a labeled (date, station, sample type) ziplock bag and place in cooler for transport back to the lab. At the lab, count and record the lengths of the mussels, cut out the abductor muscle and rinse/dip in DI water. Save muscle tissue in a labeled (date, station, sample type) glass scint vial and freeze. Be careful not to get shell fragments mixed in with the abductor muscle. Dry the tissue @ 50o C and then grind the tissue (Wig-L-Bug) for stable isotope analysis and place back in the labeled (date, station, sample type) scint vial. Zooplankton (pelagic copepods) Collect zooplankton from mid channel at mid ebb tide using 150u plankton net. Pour cod end sample into a labeled (date, station, sample type) wide mouth jar, recording date and time of day, place in cooler for transport back the lab. Don’t let jar overheat, the zooplankton must be kept alive. At the lab, we will conduct a zooplankton light migration technique procedure to purify the sample. Place the jar on a counter and let it settle. When you see a lot of zooplankton in the water column, pour off the water and zooplankton into two glass 250 ml graduated cylinders that have been wrapped with tin foil within 3” of the top. The water height should be about 2 cm above the height of tin foil. Aim a dissecting scope light or flash light at the water surface and let the zooplankton migrate to the light. Set up a vacuum flask for 25 mm filter rig. When there is a dense mass of clean zooplankton at the surface, while avoiding detritus, pipette the zooplankton onto ashed, 25 mm GFF filters (need replicates), squirt filter surface occasionally to migrate zooplankton to the middle of the filter and place filters into separate labeled (date, station, sample type) small petri dishes. Put the petri dishes in a freezer if the samples cannot be dried soon. Dry the filters @ 50o C. Silversides Using a seine collect 45 to 60 silversides, 30 – 70 mm lengths and place in a labeled (date, station, sample type) ziplock bag and place in cooler for transport back to the lab. At the lab separate the silversides into 3 subsamples, then count and record the lengths of the silversides for each subsample, then fillet and save the tissue in three separately labeled (date, station, sample type) glass scint vials and freeze. Dry the silversides @ 50o C and then grind them (Wig-L-Bug) for stable isotope analysis and place back in the labeled (date, station, sample type) scint vial. Mya (softshell clam) Collect 15 to 20 soft shell clams by digging in the tidal flats and place in a labeled (date, station, sample type) ziplock bag and place in cooler for transport back to the lab. At the lab, count and record the lengths of the clams, cut out the abductor muscle, and rinse/dip in DI water. Save muscle tissue in a labeled (date, station, sample type) glass scint vial and freeze. Be careful not to get shell fragments mixed in with the abductor muscle. Dry the tissue @ 50o C and then grind the tissue (Wig-L-Bug) for stable isotope analysis and place back in the labeled (date, station, sample type) scint vial.