National Status and Trends: Mussel Watch Project

Data Acquisition:The sample collection methods used by NOAA and NOAA contractor field crews will be described here.Sampling Objective:Bivalve mollusks are collected for the analysis of trace element and organic chemical constituents. Separate samples from the same site are packaged for organic analyses, trace element analyses, and gonadal index and histopathology analyses. Once on approximately a decadal basis sediment samples are collected from corresponding Mussel Watch bivalve sites. When the core bivalve sites were first established three collections (stations) and subsequent analyses were performed for each site. Likewise three sediment stations were established for each site. Samples from the same sediment site are collected for sediment grain-size analyses.Bivalve Collection Methods Summary Mollusks are collected at marine sites in the months of November-March with each site visited within three weeks days of a prescribed date. In the Great Lakes, collections are made in late August or early September. From 1986 to 1991, six separate composite samples of 20 oysters or 30 mussels were collected at each site. The small size of Dreissena spp. requires that composite samples from the Great Lake sites consisted of up to 100 individuals. Three composites are homogenized for trace element analyses and three for trace organic analyses. That protocol still applies whenever a site is sampled for the first time. Otherwise, since 1992 only two composite samples have been collected; one for organic and one for trace element analysis.The preferred size ranges are 5-8 cm for mussels, 7-10 cm for C. virginica, 2.5-5 cm for O. sandvicensis. Mollusks are not shucked in the field. They are separated when found to be adhering to one another and scrubbed with a nylon or natural fiber brush to remove adhering detritus. Cleaned samples are then packed in ice or dry ice and shipped to the laboratory. Composite samples are prepared by homogenizing the soft parts of 30 mussels or 20 oysters.Sediment Collection Methods Summary:Multiple sediment grabs were collected from each site using a Young-modified Van Veen grab sampler or hand held box corer. Each grab was nominally 440 cm2 in area and up to 10 cm in depth, but only the top two to three centimeter section of a grab was retained for the chemical analyses. Multiple grabs were processed until sufficient sediment were collected. Any large organisms encountered were removed, but otherwise the sample included resident organisms. Sediment samples were separated into two fractions for storage until analysis. One fraction was frozen and used in the chemical analysis reported in this file, and for the total organic carbon and percent moisture parameters reported in the LAB file. Sampling Platform: Samples were collected from gasoline powered boats, 13 to 24 feet in length or directly from shore for intertidal sites. Sampling Equipment :A 1/25 m2, stainless steel Young-modified Van Veen grab sampler or a hand held box corer was used to collect sediments. Intertidal mussels and oysters found in shallow environments were collected by hand. Subtidal mussels and oysters were collected with an oyster dredge or hand held tongs. Zebra mussels in the Great Lakes were collected by hand while snorkeling or with with an epi-benthic dredge towed from a gasoline powered boat. Sample Collection: Calibration: The sampling gear does not require any calibration, although it was inspected daily for wear and damage.Sample Collection: Quality Control: Care was taken to minimize disturbance to the sediment grabs. Grabs that were incomplete, slumped, less than 7 cm in depth, or comprised chiefly of shelly substrates were discarded. The chance of sampling the same location was minimized by repositioning the boat (five meters downstream) after three sampling attempts.Sample Collection: ReferencesO'Connor, T. P. and G. G. Lauenstein (2006) Trends in Chemical Concentrations in Mussels and Oysters Collected along the U.S. Coast: Update to 2003. Marine Environmental Research 62:261-285.Strobel, C.J. 1998. Environmental Monitoring and Assessment Program - Mid-Atlantic Integrated Assessment. Estuaries Component, Field Operations and Safety Manual. U.S. EPA, Office of Research and Development, NHEERL-AED, Narragansett, RI. July 1998.Lauenstein, G. G. and A. Y. Cantillo (1993) Sampling and analytical methods of the National Status and Trends Program National Benthic Surveillance and Mussel Watch Projects 1984-1992: Overview and summary of methods, Volume I NOAA Technical Memorandum NOS ORCA 71, Silver Spring, MD.Kokkinakis, S.A., J.L. Hyland, and A. Robertson. 1994. Carolinian Demonstration Project - 1994 Field Operations Manual. Joint National Status and Trends/Environmental Monitoring and Assessment Program. NOAA/NOS/ORCA, Silver Spring, MD.

Data Preparation and Sample Processing:Sample Processing Objective:Sediment and tissue samples were analyzed for metals, butyltins, PAHs, PCBs, PBDEs and organochlorine-pesticides.Methods Summary:All samples from the Gulf of Mexico Coast collected from 1986 to 1999 were analyzed by scientists from the Geochemical and Environmental Research Group or the Department of Oceanography at Texas A&M University (TAMU) in College Station, TX. For 1986-1988, samples from California and Hawaii were analyzed by Scientific Applications International Corporation in LaJolla, CA. For 1986-1994 all other West Coast samples and all East Coast samples were analyzed by scientists from the Battelle Laboratories in Duxbury, MA and Sequim, WA. For 1994-1999 all samples from all coasts were analyzed by the Geochemical and Environmental Research Group of the Department of Oceanography at Texas A&M University in College Station, TX. Since 2000, all samples have been analyzed by TDI-Brooks in College Station, TX.All laboratories, in all years, have used cold vapor atomic absorption for the analysis of Hg. TDI-Brooks International, Inc who has been performing the analyses since 2000 uses a modified version of EPA method 245.6. At Battelle, analyses for As, Cu, Se, and Zn have been made by X-ray fluorescence and those for Cd, Ni, and Pb by graphite furnace atomic absorption (GFAA). Beginning in 1992, inductively-coupled plasma with mass spectroscopy detection (ICP/MS) was used for Cd, Cr, Ni, and Pb. The analytical instruments used by SAIC and TAMU were GFAA for As, Cd, Ni, Pb, and Se; flame atomic absorption (FAA) for Zn; and both GFAA and FAA for Cu, depending on the concentration present. Since 2000 As and Se have been analyzed by atomic fluorescence spectrometry; As has also been analyzed by ICP-optical emission spectrometry (ICP/OEP). ICP/OEP is also used to determine Cu, Ni and Zn while ICP/MS is used for Cd, Ni and Pb. Digestion procedures prior to elemental analysis varied to some extent among laboratories and years. Prior to 1990, Battelle used concentrated nitric and perchloric acids in aTeflon digestion bomb in a conventional oven. Since 1991, samples have been digested in nitric acid only, and heating has been by microwave irradiation. SAIC used conventional ovens and nitric acid. The Texas A&M Laboratory used nitric and perchloric acids with heating in a conventional oven. From 1991 to 1999, only nitric acid was used. The current analytical laboratory sequentially adds nitric acid and hydrogen peroxide to Teflon bombs to achieve sample dissolution of all but Hg samples.Organic chemical extractions were similar among laboratories. After addition of internal standards and anhydrous sodium sulfate, the tissue was extracted three times with dichloromethane using a tissumizer. After concentration by solvent evaporation and exchange of dichloromethane with hexane, the tissue extract was fractionated by alumina:silica chromatography. The aromatic fraction eluted from the column with 1:1 pentane: dichloromethane was further purified by removing lipids through chromatography on a Sephadex column. Since 1988, this purification has been accomplished via an HPLC procedure. Organic sample extraction since 2000 remains similar to that of previous years but Hydromatrix is used to dry tissue samples and dichloromethane is the only solvent used in accelerated solvent extractor cells. Extracts are purified using alumina/silica gel chromatography followed by high performance liquid chromatography. Purified extracts were chromatographed on 30-m DB-5 fused silica capillary columns. Since 2000, a second column (DB-17HT) has also been used for chlorinated hydrocarbon confirmation. Chlorinated hydrocarbon concentrations were quantified using an electron capture detector (GC-ECD) by all laboratories in all years. For polycyclic aromatic hydrocarbons all analyses by SAIC in 1986-1988 (California and Hawaiian coasts) and those by Battelle in 1986 (East and Northwest Coasts) employed flame ionization detection (GC-FID). Battelle used mass spectrometry detection (GC-MSD) in full-scan mode in 1987. In all other cases the PAH analysis has been by GC-MSD in the selected ion mode (SIM). For butyltin analyses, anhydrous homogenized tissue was extracted with troplone and hexane (East and West Coast samples) or troplone and dichloromethane (Gulf Coast samples). Current methods use either hexane or dichloromethane. The extracts underwent Grignard reactions by addition of hexylmagnesium bromide (Gulf Coast samples) and n-pentyl magnesium bromide (East and West Coast samples). The Gulf Coast method is currently in use for samples from all coasts. Florisil/silica gel or silica alumina column chromatography were used to separate the analytes.Calibration:The analytical instruments were calibrated by standard laboratory procedures including: constructing calibration curves, running blank and spiked quality control samples, and analyzing standard reference materials.Quality Control:Each batch of sediment and tissue samples was accompanied by QC analyses consisting of method blanks, matrix spikes, matrix spike duplicates, and standard reference materials (SRMs). In total, approximately 5% of all analyses were QC analyses. Processing quality was considered acceptable if the following criteria were met: blanks were less than three times the minimum detection limit; accuracy, as determined by analysis of certified reference materials, was within 30% for organic analytes and within 15% for inorganic analytes; and precision, as determined by replicate analyses, was within 30% for organic analytes and within 15% for inorganic analytes. Additional specifications and guidelines are presented in Cantillo and Lauenstein (1998).All laboratories annually participate in annual intercalibration exercise where common samples are analyzed by them, by other laboratories, and by a lead laboratory for the exercise. The lead laboratory for trace element intercalibrations has been the National Research Council of Canada and that for trace organic analysis has been the National Institute of Standards and Technology.

Project related references: Kimbrough, K.L., G.G. Lauenstein, and W.E. Johnson. 2007. Organic Contaminant Analytical Methods of the National Status and Trends Program: Update 2000-2006, NOAA Technical Memoranda, NOS NCCOS 30. 22 Apr. 2008 http://www.ccma.nos.noaa.gov/publications/organicsmethods.pdf. Kimbrough, K.L., and G.G. Lauenstein. 2007. Major and Trace Element Analytical Methods of the National Status and Trends Program: 2000-2006, NOAA Technical Memorandum, NOS NCCOS 29. 22Apr 2008 http://www.ccma.nos.noaa.gov/publications/nsandtmethods.pdf.McDonald, S. J., D. S. Frank, J. A. Ramirez, B. Wang, and J. M. Brooks. 2006. Ancillary Methods of the National Status and Trends Program: Update 2000-2006, NOAA Technical Memorandum, NOS NCCOS 28. 22Apr. 2008 http://coastalscience.noaa.gov/documents/ancillarymethodsnsandt.pdf. O'Connor, T. P. and G. G. Lauenstein (2006) Trends in Chemical Concentrations in Mussels and Oysters Collected along the U.S. Coast: Update to 2003. Marine Environmental Research 62:261-285.Kim, Y., K.A. Ashton-Alcox, and E.N. Powell. 2006. Histological Techniques for Marine Bivalve Molluscs: Update, NOAA Technical Memorandum, NOS NCCOS 27. 22 Apr. 2008 http://www.ccma.nos.noaa.gov/publications/histopathtechmemofinal.pdf. Cantillo, A. Y. and G. G. Lauenstein (1998) Performance-based Quality Assurance - The NOAA National Status and Trends Program Experience. In: U.S. EPA Proceedings of the NWQMC National Conference - Monitoring: Critical Foundations to Protect Our Waters. Washington, DC. pp. III63-III73.Lauenstein, G. G. and A. Y. Cantillo (1998) Analytical Methods of the National Status and Trends Program Mussel Watch Project: 1993 -1997 Update. NOAA Technical Memorandum NOS ORCA 130, Silver Spring, MD.Lauenstein, G. G. and A. Y. Cantillo (1997) Analytical evaluation of laboratories wishing to perform environmental characterization studies. Environ. Toxicol. and Chem. 16(7):1345-1350.Lauenstein, G. G., A. Y. Cantillo, S. Kokkinakis, J. Jobling, and R. Fay (1997) Mussel Watch Project Site Descriptions, through 1997. NOAA Technical Memorandum NOS ORCA 112, Silver Spring, MD.Lauenstein, G. G., A. Y. Cantillo, B. J. Koster, M. M. Schantz, S. F. Stone, R. Zeisler, and S. A. Wise (1996) National Status and Trends Program Specimen Bank: Sampling protocols, analytical methods, results, and archive samples. NOAA Technical Memorandum NOS ORCA 98. Silver Spring, MD.Schantz, M. M., B. A. Benner, Jr., M. J. Hays, W. R. Kelly, R. D. Vocke, Jr., R. Demiralp, R. R. Greenberg, S. B. Schiller, G. G. Lauenstein, and S. A. Wise (1995) Certification of Standard Reference Material (SRM) 1941a, organics in marine sediment. Fresenius J. Anal. Chem. 352:166-173.Cantillo, A. Y., G. G. Lauenstein (1995) Use of reference materials in coastal quality assurance. Fresenius J Anal Chem. 352:152-156.Lauenstein, G. G. and A. Y. Cantillo (1993) Sampling and analytical methods of the National Status and Trends Program National Benthic Surveillance and Mussel Watch Projects 1984-1992: Overview and summary of methods, Volume I NOAA Technical Memorandum NOS ORCA 71, Silver Spring, MD.Lauenstein, G. G. and A. Y. Cantillo (eds.) (1993) Sampling and analytical methods of the National Status and Trends Program National Benthic Surveillance and Mussel Watch Projects 1984-1992: Comprehensive descriptions of complementary measurements, Volume II NOAA Technical Memorandum NOS ORCA 71, Silver Spring, MD.Lauenstein, G. G. and A. Y. Cantillo (eds.) (1993) Sampling and analytical methods of the National Status and Trends Program National Benthic Surveillance and Mussel Watch Projects 1984-1992: Comprehensive descriptions of elemental analytical methods, Volume III NOAA Technical Memorandum NOS ORCA 71, Silver Spring, MD.Lauenstein, G. G. and A. Y. Cantillo (eds.) (1993) Sampling and analytical methods of the National Status and Trends Program National Benthic Surveillance and Mussel Watch Projects 1984-1992: Comprehensive descriptions of trace organic analytical methods, Volume IV NOAA Technical Memorandum NOS ORCA 71, Silver Spring, MD.Cantillo, A. Y. and G. G. Lauenstein (1993) Performance based quality assurance of the NOAA National Status and Trends Project. In: Quality Assurance for analytical laboratories (ed. M. Parkany). Royal Society of Chemistry, Cambridge, England. pp. 34-43.Lauenstein, G. G., M. R. Harmon, and B. Gottholm (1993) National Status and Trends Program for Marine Environmental Quality: Benthic Surveillance and Mussel Watch Projects monitoring sites. NOAA Technical Memorandum NOS ORCA 70, Silver Spring, MD. 353 pp.Cantillo A. Y. and G. G. Lauenstein (1992) NOAA National Status and Trends Program intercomparison exercise results. Marine Technology Society, pp. 216-222.Lauenstein, G. G., and N. Valette-Silver (1991) Evolution of the National Status and Trends Program from 1984 to 1991. Sea Technology, August 1991, pp. 47-52.Shigenaka, G. and G. G. Lauenstein (1988) National Status and Trends Program for Marine Environmental Quality: Benthic Surveillance and Mussel Watch Projects sampling protocols. NOAA Technical Memorandum NOS OMA 40, 30 pp.