HTL-PIE-YearlyIsotopeSurvey.02 Linda Deegan

Ecosystems Center MBL 7 MBL St Woods Hole MA 02543 USA

Plum Island Ecosystems LTER Program

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7485 pie_im@mbl.edu http://ecosystems.mbl.edu/PIE/ Charles Hopkinson additional investigator Hap Garritt additional investigator 2004 Stable isotopes of primary producers will be compared to stable isotopes of functional groups of organisms at primarily three sites within the estuary that have different dominant sources of organic matter. The three sites are: Lower (IBYC, SO-3, mouth of Plum Island Sound, 2-3 km upstream of the mouth of the estuary), Middle (OTL, PR-10.5, upper Sound, lower Parker, 8-11 km upstream of the mouth of the estuary) and Upper (P2, PR-22, upper Parker, above Middle Rd Bridge (22 km upstream of the mouth odf the estuary). The Lower site is dominated by marine phytoplankton, the Middle site is dominated by a mixture of salt marsh and phytoplankton and the Upper site is dominated by oligohaline phytoplankton and fresh marsh. Ten functional groups will be sampled at each site (Surface sediment, benthic diatoms, Nereis, mummichog, ribbed mussels, POM, blue mussels, pelagic copepods (Acartia), silversides and soft shell clams (Mya). Marsh, benthic algae and phytoplankton inputs or benthic vs. pelagic pathways will be evident in the isotopic signals of these functional groups. Samples will be collected between the middle and end of August to reflect a growing season using recently produced OM. Samples of 15 – 20 individuals will be pooled for analysis. Silversides will have 3 different pooled samples for determination of variance. PIE LTER organic matter population dynamics Massachusetts stable isotope nitrogen carbon food web trophic fish invertebrates organic particulate zooplankton algae Acceptance and utilization of PIE-LTER data requires that: The Principal Investigator be sent a notice stating reasons for acquiring any data and a description of the publication intentions. The Principal Investigator of the data set be sent a copy of the report or manuscript prior to submission and be adequately cited in any resultant publications A copy of any resultant publications should be sent to: Principal Investigator Ecosystems Center Marine Biological Laboratory Woods Hole, MA 02543 http://ecosystems.mbl.edu/PIE/data/HTL/HTL-PIE-YearlyIsotopeSurvey.html P2 -70.91732887 -70.91732887 42.74988851 42.74988851 P5 -70.87290193 -70.87290193 42.75506932 42.75506932 Old Town Landing -70.8365313 -70.8365313 42.76236849 42.76236849 Nelson Island -70.8210343 -70.8210343 42.74691643 42.74691643 IBYC -70.79657702 -70.79657702 42.70899476 42.70899476 1999-08-23 2008-09-24 Study conducted annually, mid August to early September Version 1, July 16, 2007, Data for 2005 and 2006 added, metadata updates using June 2007, Laundre Excel template and macro Version 2, Dec. 17, 2009, added 2007/2008 data Data Manager

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7485 pie_im@mbl.edu http://ecosystems.mbl.edu/PIE/ Plum Island Ecosystems LTER

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7485 pie_im@mbl.edu http://ecosystems.mbl.edu/PIE/ Functional Groups Collected Sediments Pool top 2cm from 5, 60 ml syringe cores, put in 4 oz Qorpack jar and dry @ 50o C Benthic diatoms Nitex migration (Craig/Jeff technique), at low tide lay down 210u Nitex on sediment surface, wait 20 min for diatoms to migrate, rinse diatoms onto GFF at lab, dry @ 50o C Nereis Using shovels, dig up15-20, 2-5 cm lengths, place in jar with ambient water to purge guts for 24 hr, dry @ 50o C Mummichog Using seine, collect 15-20, 35-50 mm lengths, fillet and save flesh tissue (tail end), dry @ 50o C Ribbed mussel Pool 15-20, below S. alt. on main stem, abductor muscle, dry @ 50o C POM mid ebb, clogged GFF, dry @ 50o C Blue mussel Pool 15-20, on front edge of main stem, abductor muscle, dry @ 50o C Pelagic copepod mid ebb, Plankton net, 150 u, light migration, GFF, dry @ 50o C Silverside Seine, pool 15-20, flesh tissue (tail end) like mummichogs, dry @ 50o C Mya (softshell clam) Pool 15-20 from tidal flats, abductor muscle, dry @ 50o C 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. Field Gear and Field lab supplies Jon boat, 25 HP Honda 150 u plankton net with cod end Cooler with ice Dry cooler Field notebook Pencils Sharpies (fine and large) Label tape 3 buckets thermometer Box of 1 gallon ziplock bags Box of 4 oz Qorpack jars, minimum of 6 jars 2, 60 ml syringe cores 5, 6” x 6” squares of 210u Nitex 500 ml squirt bottle with fine nozzle shovel small seine 4, one liter bottles for POM 4, one liter wide mouth Nalgene bottles for zooplankton tows box of ashed 25 mm GFF filters filter forceps small petri dishes for 25 mm filters hand vacuum pump/flask and Gelman tower set up for 25 mm filters dissecting kit with scalpels and tweezers flat of glass scint vials with caps tin foil 2, 250 ml glass graduated cylinders 2, dissecting scope lights or flash lights (with batteries) Anne Giblin

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7488 agiblin@mbl.edu Lead PI Hap Garritt

The Ecosystems Center Marine Biological Lab 7 MBL St Woods Hole MA 02543 USA

(508) 289 7485 pie_im@mbl.edu Information Manager The Plum Island Ecosystems (PIE) LTER is an integrated research, education and outreach program whose goal is to develop a predictive understanding of the long-term response of watershed and estuarine ecosystems at the land-sea interface to changes in climate, land use and sea level. The principal study site is the Plum Island Sound estuary, its coupled Parker, Rowley and Ipswich River watersheds and the adjacent coastal ocean, the Gulf of Maine. The PIE LTER focuses on how several aspects of global change influence organic matter and inorganic nutrient biogeochemistry and estuarine foodwebs. The inputs of organic matter and nutrients from land, ocean and marshes interact with the external drivers (climate, land use, river discharge, sea level) to dictate the extent and degree of nutrient and organic matter processing and determine the spatial patterns of estuarine productivity and trophic structure. This material is based upon work supported by the National Science Foundation under Grants OCE-9726921, OCE-0423565. Any opinions, findings, conclusions, or recommendations expressed in the material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. The Plum Island Ecosystems (PIE) LTER site, is located in northeastern Massachusetts, 42.72 N, 70.85 W. The PIE LTER lies at the interface of a thinly soiled, formerly glaciated New England land mass and the highly productive Gulf of Maine. Three watersheds comprise the estuarine drainage basin: Parker (155 km2), Rowley (26 km2) and Ipswich (404 km2). The Ipswich River watershed is highly urbanized with Boston "bedroom" communities encroaching in the headwater region while the Parker is less urbanized and retains a higher proportion of forest. -71.2198 -70.7568 42.8276 42.4965 0 100 meter uid=PIE,o=lter,dc=ecoinformatics,dc=org all public read HTL-PIE-YearlyIsotopeSurvey.dat Stable isotope (carbon and nitrogen) data for functional groups in the Plum Island Sound Estuary. HTL-PIE-YearlyIsotopeSurvey.dat 1 column , http://ecosystems.mbl.edu/PIE/data/HTL/data/HTL-PIE-YearlyIsotopeSurvey.dat Sample # Sample # Sample number identifier Sample number identifier Date Date Date of sampling dd-mmm-yyyy Site Site Name of sampling site Name of sampling site Kilometer Kilometer Two letter location-kilometer name relative to the mouth of the estuary, mouth is 0 kilometers Two letter location-kilometer name relative to the mouth of the estuary, mouth is 0 kilometers Organisms Organisms Names of organisms sampled Names of organisms sampled 13C 13C delta 13C, isotope ratio 13C/12C partsPerThousand real 15N 15N delta 15N, isotope ratio 15N/14N partsPerThousand real ratio of two quantities as parts per thousand (1:1000)