CATALOG DOCUMENTATION EMAP-ESTUARIES PROVINCE LEVEL DATABASE CAROLINIAN PROVINCE 1994-1997 SEDIMENT TOXICITY DATA TABLE OF CONTENTS 1. DATA SET IDENTIFICATION 2. INVESTIGATOR INFORMATION 3. DATA SET ABSTRACT 4. OBJECTIVES AND INTRODUCTION 5. DATA ACQUISITION AND PROCESSING METHODS 6. DATA MANIPULATIONS 7. DATA DESCRIPTION 8. GEOGRAPHICAL AND SPATIAL INFORMATION 9. QUALITY CONTROL/QUALITY ASSURANCE 10. DATA ACCESS 11. REFERENCES 12. TABLE OF ACRONYMS 13. PERSONNEL INFORMATION 1. DATA SET IDENTIFICATION 1.1 Title of Catalog Document EMAP-Estuaries Province Level Database Carolinian Province Sediment Toxicity Data 1.2 Authors of the Catalog entry Timothy R. Snoots, Dr. Jeffrey L. Hyland 1.3 Catalog Revision Date November 23, 1999 1.4 Data Set Name CP TOX.DAT ------- 1.5 Task Group Estuari es 1.6 Data set identification codes 7 1.7 Version 001 1.8 Requested Acknowledgment If you plan to publish these data in any way, EPA requires a standard statement for work it has supported: "Although the data described in this article have been funded wholly or in part by the U. S. Environmental Protection Agency through its EMAP-Estuaries Program, it has not been subjected to Agency review, and therefore does not necessarily reflect the views of the Agency and no official endorsement should be inferred." 2. INVESTIGATOR INFORMATION 2.1 Principal Investigator J. Hyland (N0AA/N0S/NCC0S/CCMA) - Carolinian Province Manager A. Ringwood (SCDNR) - Lead P.I. for SC/GA region team C. Hackney (UNC-W) - Lead P.I. for NC region team G. McRae, G. Nelson, J. McKenna, J. Landsberg (FLDEP) - Lead P.I.s for FL region team (depending on year) 2.2 Investigation Participant - Sample Collection Field Sample Collection T. Alphin, S. Bowen, C. Byrum, D. Dye, A. Gospodarek, J. Grace, J. Grimley, C. Hackney, C. Powell, C. Preziosi, H. Riley, S. Roberts, M. Smith, K. Stokesbury, D. Tremain, T. Wheeler (UNC-W); S. Ross (NCNERR); M. Armstrong-Taylor, J. Jones, M. Levinson, P. Powers, A. Ringwood, T. Snoots, G. Steele (SCDNR); L. Balthis, T. Herrlinger, C. Keppler, M. Wert (UC); D. Adams, K. Amendola, D. Cook, C. Harnden, B. Heagey, J. Mckenna, G. Nelson, C. Nowicki, R. Paperno, B. Rosenblatt, M. Wessel (FLDEP); J. Hyland, S. Kokkinakis (N0AA/N0S/0RCA) Field Training and Coordination S. Kokkinakis (N0AA/N0S/0RCA); J. Macauley (EPA-GED); T. Hei tmuller (USGS-GB); D. Keith (EPA-AED) ------- 2.3 Sampling Processing - Principal Investigator Program Management and Coordination J. Hyland, A. Robertson (NOAA/NOS/NCCOS/CCMA); K. Summers (EPA); F. Holland, A. Ringwood (SCDNR); C. Hackney, T. Wheeler (UNC-W); S. Ross (NCNERR); J. Landsberg, J. McKenna, G. McRae, G. Nelson, R. Paperno (FLDEP) Toxicity Testing M. DeLorenzo (CU) ; J. Grimley (UNC-W); J. Jones, C. Keppler, P. Maier, A. Ringwood, R. Van Dolah (SCDNR); P. Ross (Citadel); C. Mueller, J. Scott, G. Thursby (SAIC) QA/QC T. Heitmuller (USGS-GB), S. Kokkinakis (NOAA/NOS/ORCA) Data Management and Statistical Support T. Snoots, F. Holland, R. VanDolah (SCDNR); L. Balthis, T. Herrlinger (UC); J. Rosen, L. Zimmerman (TPMC); S. Rathbun (UGA); M. Adams, L. Harwell (JCWS); V. Engle (EPA-GED); Z. Malaeb (USGS-GB); S. Hale (EPA-AED); K. Summers (EPA); T. Wilson (CU) 3. DATA SET ABSTRACT 3.1 Abstract of the Data set The CP_T0X.DAT data set contains results of up to four different toxicity assays (10-day Ampelisca abdita mortality assay, 10-day Ampelisca verrilli mortality assay, 7-day Mercenaria mercenaria growth assay, and Microtox solid-phase assay) performed on sediment samples from each station sampled in the EMAP Carolinian Province from 1994-1997. The following reports are products of these and other data collected during the 1994-1997 Sampling period in the Carolinian Province. These reports may contain additional information and summary statistics that are not contained in this data set catalog or its respective data sets. We therefore recommend referring to them when using these data. Hyland, J.L., T.J. Herrlinger, T.R. Snoots, A.H. Ringwood, R.F. Van Dolah, C.T. Hackney, G.A. Nelson, J.S. Rosen, and S.A. Kokkinakis. 1996. Environmental quality of estuaries of the Carolinian Province: 1994. Annual statistical summary for the 1994 EMAP-Estuaries Demonstration Project in the Carolinian Province. NOAA Technical Memorandum NOS ORCA 97. NOAA/NOS, Office of Ocean Resources Conservation and Assessment, Silver Spring, MD. 102 p. ------- Hyland, J.L., L. Balthis, C.T. Hackney, G. McRae, A.H. Ringwood, T.R. Snoots, R.F. Van Dolah, and T.L. Wade. 1998. Environmental quality of estuaries of the Carolinian Province: 1995. Annual statistical summary for the 1995 EMAP-Estuaries Demonstration Project in the Carolinian Province. NOAA Technical Memorandum NOS ORCA 123 NOAA/NOS, Office of Ocean Resources Conservation and Assessment, Silver Spring, MD. 143 p. See also: Grimley and Hackney (1996), Ringwood et al. (1997b), Ringwood and Keppler (In Press), Ringwood et al.(1998), Ringwood et al. (1996), Ringwoodet al. (1995), Ringwood et al. (1997a), Science Applications International Corp. (1998a), Science Applications International Corp. (1998b). 3.2 Keywords for the Data Set Sediment toxicity, amphipod, Ampelisca abdita, Ampelisca verrilli, Microtox, Mercenaria mercenaria, EMAP Carolinian Province 4. OBJECTIVES AND INTRODUCTION 4.1 Program Objective EMAP has three primary objectives: 1. To estimate the current status, extent, changes, and trends in indicators of the Nation's ecological resources on a regional basis; 2. To monitor indicators of pollutant exposure and habitat condition, and to seek correlative relationships between human-induced stresses and ecological condition that identify possible causes of adverse effects; and 3. To provide periodic statistical summaries and interpretive reports on ecological status and trends to the EPA Administrator and to the public. 4.2 Data Set Objective The CP_T0X.DAT data set contains results of up to four different toxicity assays (10-day Ampelisca abdita mortality assay, 10-day Ampelisca verrilli mortality assay, 7-day Mercenaria mercenaria growth assay, and Microtox solid-phase assay) performed on sediment samples from each station sampled in the EMAP Carolinian Province from 1994-1997. Where applicable, both assay end results (i.e., % survival, % growth, or EC50) , and statistical test results (P-value for test of control vs. sample results) are reported. This allows a duel criteria to be used in the determination of a sample's toxicity (i.e., test results should be both statistically and biologically significant to declare the overall toxicity result ------- significant). These duel criteria have been applied for each test and results are reported in a coded fashion (1 = significant toxicity, 0 = not toxic). 4.3 Data Set Background Information The standard 10-day sediment bioassay with the marine amphipod Ampelisca abdita (ASTM 1993) has been used to assess sediment toxicity in other EMAP provinces. This assay was also used in the Carolinian Province in 1994, 1995 and 1997 to provide a basis for comparisons among EMAP provinces and between years within the Carolinian Province. However, because Ampelisca abdita proved to be relatively insensitive to sediment contaminants in prior surveys conducted in both the Carolinian and Louisianian Provinces (Hyland et al. 1996, Macauley et al. 1994), an additional amphipod assay with the congeneric species Ampelisca verrilli was included in the 1995 effort. Preliminary testing with A. verrilli and a subset of the 1994 sediment samples indicated that this species was more sensitive to sediment contamination than A. abdita (Ringwood et al. 1995). Furthermore, A. verrilli is a more common member of the infaunal benthos of southeastern estuaries. A third bioassay used to measure potential sediment toxicity at all base sites and selected supplemental sites was the Microtox solid-phase test with the photoluminescent bacterium Vibrio fischeri (formerly Photobacterium phosphoreum). This assay provides a sublethal measure of toxicity based on attenuation of light production by the bacterial cells due to exposure to the sediment sample (Bulich 1979, Ross et al. 1991, Microbics 1992a and b) . Microtox has not been used in other EMAP-E provinces, but its recent application in other coastal assessment programs suggested that it might be a useful tool to consider for the Carolinian Province. Small sample sizes (a 100-mL subsample of the composited surface sediment from each station) and a short processing time (20-min exposures) provide clear logistical advantages. Results of the Carolinian Province 1993 pilot study (Ringwood et al. 1996) and 1994 monitoring demonstration (Hyland et al. 1996) also suggested that this test is more powerful in its ability to discriminate between degraded and reference sites than the amphipod toxicity test. A fourth sediment bioassay used in the 1995-1997 surveys was a 7-d sublethal test of the effects of sediment exposure on growth of juvenile Mercenaria mercenaria (referred to hereafter as "seed clams"). The seed-clam bioassay was developed during the Carolinian Pilot Study (Ringwood et al. 1996, Ringwood and Kepler In Press). Field-validation testing on a subset of the 1994 sediment samples indicated that this bioassay was a more sensitive indicator of sediment contamination than the A. abdita bioassay (Ringwood et al. 1995). There are other practical advantages. For example, newly metamorphosed clams exhibit very rapid growth, thus effects on growth can be detected within a short time frame. Second, because seed clams can be obtained from cultured populations (available ~ 3 months after fertilization), ------- experiments can be conducted with animals of similar size, age, and pre-exposure histories. Third, a relatively small sample volume (500 mL) is required, thus minimizing sampling time and storage needs. Lastly, Mercenaria feed at the sediment-water interface, where maximum contaminant exposure would be expected. Thus the bioassay is representative of a realistic exposure scenario. 4.4 Summary of Data Set Parameters See 4.2 (Data Set Objective) 4.5 Year-Specific Information about Data Toxicity testing in the Carolinian Province from 1994-1997 was performed by the following laboratories: Test Type Ampeli sea Ampeli sea Mercenari a Year abdi ta verri Hi mercenari a Mi crotox 1994 SCDNR,SAIC - - SCDNR 1995 SAIC SCDNR SCDNR SCDNR 1996 - - SCDNR UNC-W 1997 SCDNR,SAIC - SCDNR SCDNR,SAIC Hyphen (-) indicates test was not performed in that year. 5. DATA ACQUISITION AND PROCESSING METHODS 5.1 Data Acqui si ti on 5.1.1 Sampling Objective See section 4.3 (Data Set Background Information) 5.1.2 Sample Collection Method Summary A 1/25 m2, Kynar-coated stainless steel, Young Grab sampler was used to collect sediments. This grab sampled an area of 440 cm2 and a maximum depth of penetration in the sediment of 10 cm. Stainless steel utensils were used to remove the top 2-3 cm of sediment from a grab. The sediment was removed to a stainless steel bowl and placed in a cooler of ice to remain cold, but unfrozen. The grab sampler was rinsed and re-deployed. This procedure was repeated until the volume of sediment required for all contaminant, toxicity, and sediment characteristics analyses had been collected. The sediment was mixed by hand until thoroughly homogenized, and aliquots were placed into pre-cleaned containers (Polyethylene for A. abdita and A. verrilli, and Polypropylene for Microtox and M. mercenaria). In addition, Microtox samples were wrapped in foil immediately to exclude light. All toxicity samples were immediately stored on ice following collection in the field, and then refrigerated at 4 C once back in the lab. ------- 5.1.3 Beginning Sampling Dates 30 June 1994 05 July 1995 09 July 1996 07 July 1997 5.1.4 Ending Sampling Dates 31 August 1994 14 September 1995 19 September 1996 25 August 1997 5.1.5 Platform Samples were collected from various gasoline or diesel powered boats equipped with at least the following equipment: "A" frame boom or davit, winch, LORAN-C or GPS for location, and a depth finder. 5.1.6 Sampling Equipment A 1/25 m2, Kynar-coated stainless steel, Young Grab sampler. This grab sampled an area of 440 cm2 and a maximum depth of penetration in the sediment of 10 cm. 5.1.7 Manufacturer of Sampling Equipment Ted Young Falmouth, MA 5.1.8 Key Variables 5.1.9 Sample Collection Method Calibration The sampling gear does not require any calibration. It required inspection for deformities incurred due to mishandling or impact on rocky substrates. 5.1.10 Sample Collection Quality Control Field technicians were trained to follow Standard Operating Procedures to insure the collection of representative, uncontaminated and high quality samples. QA/QC measures were taken in the field to avoid or reduce contamination and insure the collection of representative samples. These included: use of stainless steel instruments, thorough cleaning of the sampler between grabs, use of pre-cleaned containers for sediment storage and ensuring that engines were shut down when a sample was exposed to the air. A successful grab had relatively level, intact sediment over the entire area of the grab and a sediment depth of 7-10 centimeters. Unacceptable grabs included those: containing no sediments, which were partially filled or had shelly substrates or grossly slumped surfaces. Grabs completely ------- filled to the top, where the sediment was oozing out of the hinged top, were also unacceptable. See: Kokkinakis et al. (1994a) 5.1.11 Sample Collection Method References See: Hyland et al. (1996), Hyland et al. (1998), Kokkinakis et al. (1994b) 5.1.12 Sample Collection Method Deviations None 5.2 Data Preparation and Sample Processing 5.2.1 Sample Processing Objective Determine toxicity of sediment samples using up to four toxicity assays (10-day Ampelisca abdita mortality assay, 10-day Ampelisca verrilli mortality assay, 7-day Mercenaria mercenaria growth assay, and Microtox solid-phase assay). 5.2.2 Sample Processing Methods Summary 5.2.2.1 Field Summary NA 5.2.2.2 Laboratory Summary A. abdita and A. verrilli Testing Procedures followed the general guidelines provided in ASTM Protocol E1367-92 (ASTM 1993) and the EMAP-E Laboratory Methods Manual (U.S. EPA 1994a, 1994b).This is an acute toxicity test which measures the effect of sediment exposure on amphipod survival under static conditions. Approximately 3-3.5 L of surface sediments (composite of upper 2 cm from multiple grabs) were collected for each type of assay from each station and stored in 3.7-L polyethylene jars at 4 C in the dark until testing. Tests were conducted with subsamples of the same sediment on which analyses of contaminants and other sediment characteristics were performed. Wherever possible, sediment samples were tested within 30 days of collection as recommended in the EMAP-E protocol. Amphipods were collected from unpolluted tidal flats. Prior to testing, the animals were acclimated at 20 C for 2-9 days in the case of A. abdita, or for 2-4 days in the case of A. verrilli. During the acclimation period, the amphipods were fed the diatom Phaeodactylum tricornutum. Wherever possible, juvenile amphipods of approximately the same size (usually 3-5 mm in length for A. abdita, and 3-10 mm in length for A. verrilli) ------- were used to initiate the tests. The general health of each batch of amphipods was evaluated by a reference toxicity test (i.e., "positive control"). These tests were run in a dilution series with seawater (no sediment phase) and the reference toxicant sodium dodecyl sulfate (SDS). Tests for both species were run under static conditions in dark and followed the basic methods described by ASTM (1993). The exposure period was 96 h for A. abdita and 24 h for A. verrilli. The shorter exposure period was used for A. verrilli to match previous reference toxicant tests conducted with this species by MRRI. LC50 values were computed for each batch of test animals for comparison against background toxicity data on these same species and reference toxicant. Animals were not used in definitive tests with field samples unless acceptable reference toxicant results were obtained. A test was considered acceptable if the LC50 value was within +/- 2 SD of the mean LC50 based on the preceding 20 (A. abdita) to 22 (A. verrilli) reference toxicant tests. Treatments for the definitive tests with field samples consisted of a single concentration of each sediment sample (100% sediment) and a negative control [i.e., reference sediment], A negative control was run with each batch of field samples. The tests were conducted under static conditions at a temperature of 20 +/- 1 C and salinity range of 26-33ppt for A. abdita and 26-35ppt for A. verrilli. Twenty amphipods were randomly distributed to each of five replicates per each treatment including the control. Amphipods were not fed during the tests. The negative controls provided a basis of comparison for determining statistical differences in survival in the field sediments. In addition, control survival provided a measure of the acceptability of final test results. Test results were considered valid if mean control survival (among the 5 replicates) was >= 85% and survival in any single control chamber was >= 80%. One-liter glass containers with covers were used as test chambers. Each chamber was filled with 200 mL of sediment and 600-800 mL of filtered seawater. The sediment was press-sieved through a 2.0-mm screen to remove ambient fauna prior to placing it in a chamber. All containers were illuminated constantly throughout the 10-day test to inhibit amphipod emergence from the sediment, thus maximizing exposure to the test sediment. Air was supplied using oil-free aerators and glass pipettes inserted into the test chambers. Water tables with recirculating chiller pumps were used to maintain constant temperatures (20 +/- 1 C). Daily recordings were made of temperature and the number of dead vs. living animals. On days two and eight, two of the five ------- replicate chambers for each treatment were selected randomly and measured for salinity, dissolved oxygen, pH, and total ammonia in the overlying water. At the conclusion of a test, the sediment from each chamber was sieved through a 0.5-mm screen to remove amphipods. The number of animals dead, alive, or missing was recorded. Sediments with missing A. abdita were preserved in formalin containing Rose Bengal stain and re-examined under a dissecting microscope to ensure that no living specimens had been missed. Animals still unaccounted for were considered to have died and decomposed in the sediment. Because of their larger size, A. verrilli were much easier to locate with the unaided eye. Thus, if any of these animals were missing after initial examination of the sieved sediment, then they were assumed to have died and decomposed. Differences between survival of Ampelisca abdita in field versus control samples were evaluated by an unpaired heteroscedastic t-test run on untransformed percentage data, under the assumptions of normality and unequal variances. For A. verrilli, differences between field samples and controls were evaluated by either: (i) an unpaired homoscedastic t-test in cases of normal data with equal variances, or (ii) a Mann-Whitney U-test in cases of non-normal data or unequal variances. The A. verrilli comparisons also were performed on untransformed percentage data. For both bioassays, field samples were considered to be significantly toxic if mean survival in comparison to the corresponding negative control was < 80% and statistically different at alpha = 0.05. A variety of quality control procedures were incorporated to assure acceptability of amphipod test results and comparability of the data with other studies. As described above, these provisions included the use of standard ASTM and EMAP protocols, positive controls run with a reference toxicant, negative "performance" controls run with reference sediment, and routine monitoring of water quality variables to identify any departures from optimum tolerance ranges. In addition, during the first year of the program, an inter-laboratory comparison of results using the A. abdita assay was performed by the two participating testing facilities (SAIC and SCDNR/MRRI). Samples from two of the base sites collected in 1994 were tested by each facility. Results were highly comparable: mean survival in field samples relative to controls was 96% for both samples by one lab, and 98 to 100% by the other lab. ------- Microtox Testing Tests were conducted in duplicate following the "large-sample-size" protocol of Microbics Corporation (1992b). Wherever possible, sediment samples were tested within the recommended 10-d holding period. A 7-g aliquot of each sediment sample was used to make a dilution series ranging from 0.01 to 10% sediment in a 2% saline diluent. A reagent solution containing the bacteria was then added to each sediment suspension. After a 20-min incubation period, a column filter was used to separate the liquid phase and bacterial cells from the sediment. Post-exposure light output in each of the filtrates was measured on a Microtox Model 500 Analyzer. A log-linear regression model was used to determine an EC50 - the sediment concentration that reduced light production by 50% relative to a control (nontoxic reagent blank). EC50 values were corrected for percent water content and reported as dry-weight concentrati ons. Assays were run with the reference toxicant phenol with each new batch of bacteria. These tests provided measures of the general quality of the bacterial populations, as well as the ability of the laboratory to produce results consistent with the expected phenol toxicity range (i.e., Microtox EC50 values typically between 13-26 mg/L). Use of the standard Microtox equipment and protocol helped to assure data comparability with results of other Microtox studies. Mercenaria mercenaria Testing Seed clams (~ 1 mm in length) were obtained from Atlantic Clam Farms, Folly Beach, S.C. Replicate subsets were dried and weighed to provide initial weight estimates. On the day before initiation of a test, sediment samples were sieved through a 500-micrometer screen (to remove ambient fauna) and distributed to the test chambers. Approximately 50 mL of sieved sediment were added to each of four replicate 250-mL beakers for each sediment sample. A negative control (same Folly River sediments used as controls in the Ampelisca verrilli assays) was run with each batch of field samples. Filtered seawater (1-micrometer filter), adjusted to 25 ppt with deionized water, was added to each beaker to bring the total volume up to 200 mL. The sediment suspension was allowed to settle overnight and clams (30-50 per replicate) were added the next day (which initiated the test). Tests were conducted for 7 d. All tests were conducted at room temperature (23-25 C) under gentle aeration. Animals were fed three times throughout the test with a phytoplankton mixture consisting of equal volumes of Isochrysis galbana and Chaetocerus gracilis. ------- At the end of the 7-d exposure period, clams were sieved from the sediments, placed in clean seawater, and allowed to depurate for ~ 1 h. Clams were re-captured on a sieve and rinsed briefly with distilled water to remove excess salt. Dead clams were removed and not included in subsequent growth estimates (mortality rates generally were < 10%). The remaining live clams were dried overnight (60-70 C), counted, and weighed on a micro-balance. The pre- and post-exposure measurements were then used to determine growth rates, expressed as ug/clam/d. Effects of sediment exposure on growth rates were evaluated using either a t-test or Mann-Whitney U-test (when assumptions of the parametric test were violated). Samples were considered to be significantly toxic if mean growth rate in comparison to the control was < 80% and statistically different at alpha = 0.05. Each new batch of seed clams was evaluated for suitability and relative sensitivity with a reference toxicant test ("positive control"). These tests were run under static conditions, at room temperature, in a dilution series with 25ppt seawater (no sediment phase) and the reference toxicant cadmium. Treatments within each test consisted of a seawater control and four cadmium concentrations (25, 50, 100, 200 ug/L as CdCl2) . Each treatment was represented by 3-4 replicates. The effective Cd concentration that reduced growth by 50% (EC50) relative to the seawater control was estimated by regression analysis. 5.2.3 Sample Processing Method Calibration See Section 5.2.2.2 (Sample Processing Methods Summary/ Laboratory Summary) 5.2.4 Sample Processing Quality Control See Section 5.2.2.2 (Sample Processing Methods Summary/ Laboratory Summary) 5.2.5 Sample Processing Method Reference See Section 5.2.2.2 (Sample Processing Methods Summary/ Laboratory Summary) 5.2.6 Sample Processing Method Deviations None ------- 6. DATA ANALYSIS AND MANIPULATIONS 6.1 Name of New or Modified Value P_VALUE RESULT SIG P-value for statistical test Test result (relative to control) Toxicity Test Result (1=Hit) 6.2 Data Manipulation Description P VALUE For A. abdita and A. verrilli, P_VALUE is the probability value associated with a statistical test of mean survival of Amphipods in field versus control samples. The test performed was either: (1) an unpaired homoscedastic t-test in cases of normal data with equal variances, or (2) a Mann-Whitney U-test in cases of non-normal data or unequal variances. The A. verrilli comparisons were performed on untransformed percentage data. For M. mercenaria, P_VALUE is the probability value associated with a statistical test of mean growth of M. mercenaria in field versus control samples. Effects of sediment exposure on growth rates were evaluated using either a t-test or Mann-Whitney U-test (when assumptions of the parametric test were violated). For A. abdita and A. verrilli, RESULT is mean survival in field sediment as a percent of mean survival in control sediment (e.g., RESULT = field/control * 100) For Microtox, values reported in RESULT are EC50 values (sediment concentration that reduced light production by 50% relative to a control). These EC50 values have been corrected for percent water content and are reported as percent (dry-weight) sediment concentrations. For M. mercenaria, RESULT is mean clam growth in field sediment as a percent of mean growth in control sediment (e.g., RESULT = field/control * 100). The variable SIG reports a final, coded toxicity result based on a duel criteria of statistical significance (P_VALUE) and biological significance (RESULT). If both the P_VALUE and RESULT for an assay meet the criteria for significance, then the response is considered to be significantly toxic and SIG = 1. If either P_VALUE or RESULT for an assay do not meet the appropriate criteria for significance, then the response is considered not toxic and SIG = 0. Where available data are not adequate to evaluate using this duel criteria approach (e.g., P_VALUE or RESULT data are missing), SIG = (missing). The criteria used to evaluate toxicity responses in each of the RESULT SIG ------- four assays are discussed below. For both A. abdita and A.verrilli assays, field samples were considered to be significantly toxic if mean survival of field samples as a percent of the mean survival for the corresponding negative control (RESULT) was < 80% and statistically different at alpha =0.05 (P_VALUE). For Microtox assays, evaluation criteria were established for two separate silt-clay classes because of the strong inverse relationship between Microtox EC50 values and percent silt-clay content. Sediments with >= 20% silt-clays were classified as being toxic if EC50 values were <= 0.2%; sediments with < 20% silt-clays were classified as being toxic if EC50 values were <= 0.5% (sensu Ringwood et al. 1995). Lower EC50 values in muddier sediments are believed to be caused by physical adsorption of the bacteria to the sediment particles. Ringwood et al. (1995, 1997) demonstrated this effect by conducting Microtox assays in artificial sediment mixtures of pure sand and kaolin clay and evaluating the EC50 values as a function of the finer-particle content. For M. mercenaria assays, samples were considered to be significantly toxic if mean growth rate in field samples as a percent of mean growth rate in the controls (RESULT) was < 80% and statistically different at alpha = 0.05 (P_VALUE). 6.3 Data Manipulation Examples 7. DATA DESCRIPTION 7.1 Description of Parameters Variable Type Format Label DATE P_VALUE QC_C0DE RESULT SIG STA_NAME TEST UNIT Num Num YYMMDD6. 6.4 Char 30. Num Num Char Char 22. Char 10. 8.3 1. 7. Sample collection date (YYMMDD) P-value for statistical test QC Code Test result (relative to control) Toxicity Test Result (1=Hit) Carolinian Province Office Station Name Test Species Test result unit code Note the conventions used in the Format column above: For character (Char) variables, the number given is the maximum width (number of characters) for that variable. For numeric (Num) variables, the format is given in W.D format, where W = maximum width (number of characters) for the number (including all digits and the decimal point), and D = number of digits to the right of the decimal point. ------- 7.1.6 Precision to which values are reported Variable RESULT is reported to 0.001 units, however, values are only valid to: 0.1 units for A. abdita and A. verrilli results, 0.01 units for M. mercenaria results, and 0.001 units for Microtox results. Variable P_VALUE is reported to 0.0001 units, however, values are only valid to: 0.001 units for A. abdita and A. verrilli results, and 0.0001 units for M. mercenaria results. 7.1.7 Minimum Value in Data Set Variable Minimum P_VALUE 0.0000 RESULT -148.690 SIG 0 7.1.8 Maximum Value in Data Set Variable Maximum P_VALUE 1.0000 RESULT 219.420 SIG 1 7.2 Data Record Example 7.2.1 Column Names for Example Records STA_NAME;DATE;TEST;RESULT;UNIT;P_VALUE;SIG;QC_C0DE 7.2.2 Example Data Records CP95121 CP95121 CP95121 CP95121 CP95122 CP95122 CP95122 CP95122 CP95123 CP95123 CP95123 CP95123 950720 950720 950720 950720 950720 950720 950720 950720 950726 950726 950726 950726 Ampeli sea abdi ta;104.200;Survi val_%;0.0790;0;AST-E Ampeli sea verrilli;103.000;Survi val_%;0.2000;0; Mercenaria mercenaria;135.320;Growth_%;.;0;CST-E, CST- Microtox (V. fischeri);0.550;EC50_%;.;0; Ampeli sea abdi ta;100.000;Survi val_%;0.5000;0;AST-E Ampelisca verrilli;97.000;Survival_%;0.3700;0; Mercenaria mercenaria;78.710;Growth_%;0.0286;1;CST-F Microtox (V. fischeri);0.640;EC50_%;.;0; Ampeli sea abdi ta;92.500;Survi val_%;0.0080;0; Ampeli sea verri11i;104.000;Survi val_%;0.3700;0; Mercenaria mercenaria;52.540;Growth_%;0.0024;1;CST-F Microtox (V. fischeri);9.520;EC50_%;.;0; GEOGRAPHIC AND SPATIAL INFORMATION 8.1 Minimum Longitude -81 Degrees, 43.83 Minutes West Longitude ------- 8.2 Maximum Longitude -75 Degrees, 33.82 Minutes West Longitude 8.3 Minimum Latitude 27 Degrees, 12.07 Minutes North Latitude 8.4 Maximum Latitude 36 Degrees, 43.43 Minutes North Latitude 8.5 Name of area or region Coastal distribution of sampling is along the southeastern US from Cape Henry, VA, through St. Lucie Inlet, FL. States represented: Virginia, North Carolina, South Carolina, Georgia, and Florida. 9. QUALITY CONTROL/QUALITY ASSURANCE 9.1 Measurement Quality Objectives See Section 5.2.2.2 (Sample Processing Methods Summary/ Laboratory Summary) 9.2 Quality Assurance/Control Methods See Section 5.2.2.2 (Sample Processing Methods Summary/ Laboratory Summary) 9.3 Quality Assessment Results Unless flagged with one of the following QC codes, all data reported in the CP_T0X.DAT data set met the QA/QC guidelines given above and are acceptable without further qualification. Where necessary, the following QC codes are reported under the variable QC_C0DE and should be considered when interpreting test results. QC Code Description AST-C Fewer than 5 replicates tested AST-E Sample held for longer than 30 days AST-M More than 20 animals inoculated into replicate CST-A Fewer than 4 replicates were tested CST-B Sediment too coarse to sieve through 0.5 mm mesh therefore making it difficult to recover clams. CST-C Sediment held longer than 30 days prior to testing CST-D Folly River control sediment not used. Note that this occurred only once. EXPTNUM 950928, sediment from Breach Inlet used. CST-E Statistical analysis not run because the mean growth rate was >100% of the mean control growth rate ------- CST-G Very high to complete mortality of clams in sample (i.e., sample i s toxi c) CST-H Fewer than 3 replicates were tested (cadmium exposures only) MST-A Samples were processed within 14 days of sampling MST-E Sample held for > 10 days prior to testing MST-F Unable to calculate an EC50 value for this sample due to an insignificant decrease in luminescence or an increase in luminescence (i.e., little or no toxic effects) MST-X Calculated EC50 result was greater than the highest test concentration of 10%. Because the accuracy of an EC50 value above 10% is unknown, EC50 values greater that 10% have been reported as 10.000%. MST-Y Hit/Miss result could not be determined due to missing silt-clay data DATA ACCESS 10.1 Data Access Procedures Data can be downloaded from the WWW site. 10.2 Data Access Restrictions Data can only be accessed from the WWW site. 10.3 Data Access Contact Persons For programmatic/policy matters, contact: Dr. Jeffrey L. Hyland N0AA/N0S National Centers for Coastal Ocean Science Center for Coastal Monitoring and Assessment - Charleston Lab 217 Fort Johnson Rd. (P.O. Box 12559) Charleston, SC 29422-2559 (843)762-5415 (Tel.) (843)762-5110 (FAX) jeff.hyland0noaa.gov (e-mail) For data-related questions, contact: Dr. W. Leonard Balthis N0AA/N0S National Centers for Coastal Ocean Science Center for Coastal Monitoring and Assessment - Charleston Lab 217 Fort Johnson Rd. (P.O. Box 12559) Charleston, SC 29422-2559 (843)762-5652 (Tel.) (843)762-5110 (FAX) len.balthis0noaa.gov (e-mail) ------- Data Librarian, EMAP-Estuaries Melissa M. Hughes OAO Corporation U.S. EPA NHEERL-AED 27 Tarzwell Drive Narragansett, RI 02882-1197 (401) 782-3184 (Tel.) (401) 782-3030 (FAX) hughes.melissa0epa.gov (e-mail) 10.4 Data file Format Delimited ASCII Text 10.5 Information Concerning Anonymous FTP Not accessible 10.6 Information Concerning WWW Data can be downloaded from the WWW. 10.7 EMAP CD-ROM Containing the Data file Data not available on CD-ROM. 11. REFERENCES American Society for Testing and Materials (ASTM). 1993. Guide for conducting 10-day static sediment toxicity tests with marine and estuarine amphipods. ASTM Standard Method E-1367-92, ASTM, Philadelphia, PA. 24p. Bulich, A.A. 1979. Use of luminescent bacteria for determining toxicity in aquatic environments. In: L. L. Marking and R. A. Kimerle (eds.), Aquatic Toxicology, pp. 98-106. ASTM STP 667. American Society for Testing and Materials, Philadelphia, PA Grimley, J. and C.T. Hackney. 1996 Year end summary. EMAP Carolinian Province: North Carolina Estuaries Component. Final year-three report under N0AA Cooperative Agreement No. NA470A0148. University of North Carolina at Wilmington, Center for Marine Research, Wilmington, NC. Hyland, J.L., T.J. Herrlinger, T.R. Snoots, A.H. Ringwood, R.F. Van Dolah, C.T. Hackney, G.A. Nelson, J.S. Rosen, and S.A. Kokkinakis. 1996. Environmental quality of estuaries of the Carolinian Province: 1994. Annual statistical summary for the 1994 EMAP-Estuaries Demonstration Project in the Carolinian Province. N0AA Technical Memorandum N0S 0RCA 97. NOAA/NOS, Office of Ocean Resources Conservation and Assessment, Silver Spring, MD. 102 p. ------- Hyland, J.L., L. Balthis, C.T. Hackney, G. McRae, A.H. Ringwood, T.R. Snoots, R.F. Van Dolah, and T.L. Wade. 1998. Environmental quality of estuaries of the Carolinian Province: 1995. Annual statistical summary for the 1995 EMAP-Estuaries Demonstration Project in the Carolinian Province. NOAA Technical Memorandum NOS ORCA 123 NOAA/NOS, Office of Ocean Resources Conservation and Assessment, Silver Spring, MD. Kokkinakis, S.A., C. Mageau, and A. Robertson. 1994a. Carolinian Demonstration Project - 1994 Quality Assurance Project Plan. Joint National Status and Trends/Environmental Monitoring and Assessment Program. NOAA/NOS/ORCA, Silver Spring, MD. Kokkinakis, S.A., J.L. Hyland, and A. Robertson. 1994b. Carolinian Demonstration Project - 1994 Field Operations Manual. Joint National Status and Trends/Environmental Monitoring and Assessment Program. NOAA/NOS/ORCA, Silver Spring, MD. Macauley, J.M., J.K. Summers, P.T. Heitmuller, V.D. Engle, G.T. Brooks, M. Babikow, and A.M. Adams. 1994. Annual Statistical Summary: EMAP - Estuaries Louisiana Province - 1992. U.S. EPA Office of Research and Development, Environmental Research Laboratory, Gulf Breeze, FL. EPA/620/R-94/002. Microbics Corporation. 1992a. Microtox Manual (5 volume set). Carlsbad, CA. Microbics Corporation. 1992b. Microtox Update Manual, 128 p. Carlsbad, CA. Ringwood, A.H., M.E. DeLorenzo, P.E. Ross, and A.F. Holland. 1997b. Interpretation of Microtox« solid-phase toxicity tests: the effects of sediment composition. Environ. Toxicol. Chem. 16(6):1135-1140. Ringwood, A.H. and C. Keppler. In Press. Seed clam growth: A sediment bioassay developed in the EMAP Carolinian Province, Environ. Monitor. & Assess, Submitted May 1997. Ringwood, A.H., C. Keppler, R.F. Van Dolah, P. Maier, J. Jones, M.E. DeLorenzo. 1998. EMAP Activities in the Carolinian Province by Marine Resources Research Institute in 1997: Results and Summaries. Final Annual Report under NOAA Cooperative Agreement No. NA770A0214. South Carolina Department of Natural Resources, Marine Resources Research Institute, Charleston, SC. Ringwood, A.H., A.F. Holland, R. Kneib, and P. Ross. 1996. EMAP/NS&T pilot studies in the Carolinian Province: Indicator testing and evaluation in southeastern estuaries. Final Report under Grant NA90AA-D-SG790 through S.C. Sea Grant College Program. S.C. Dept. of Natural Resources, Marine Resources Research Institute, Charleston, S.C. NOAA Technical Memorandum NOS ORCA 102. 113 p. ------- Ringwood, A.H., R. Van Dolah, A.F. Holland, and M.G. Delorenzo. 1995. Year one demonstration project studies conducted in the Carolinian Province by Marine Resources Research Institute: Results and summaries. Final Annual Report under NOAA Cooperative Agreement No. NA470A0177. South Carolina Department of Natural Resources, Marine Resources Research Institute, Charleston, S.C Ringwood, A.H., R. Van Dolah, A.F. Holland, M.G. Delorenzo, C. Keppler, P. Maier, J. Jones, and M. Armstrong-Taylor. 1997a. Year two demonstration project studies conducted in the Carolinian Province by Marine Resources Research Institute: Results and summaries. Final Annual Report under NOAA Cooperative Agreement No. NA470A0177. South Carolina Department of Natural Resources, Marine Resources Research Institute, Charleston, SC. Science Applications International Corp. 1998a. Sediment toxicity testing (MAIA 1997), Amphipod test results, final report (21 Jan 1998). Science Applications International Corporation, Narragansett, RI. Science Applications International Corp. 1998b. Sediment toxicity testing (MAIA 1997), Microtox test results, final report (10 Feb 1998). Science Applications International Corporation, Narragansett, RI. Ross, P.E., L.C. Burnett, C. Kermode, and M. Timme. 1991. Miniaturizing a toxicity test battery for screening contaminated sediments. Can. Tec. Rep. Fish. Aquat. Sci., 1774: 331-335. Strobel, C.J., S.J. Benyi, D.J. Keith, H.W. Buffum, and E.A. Petrocelli. 1994. Statistical summary: EMAP - Estuaries Virginian Province - 1992. U.S. EPA Office of Research and Development, Environmental Research Laboratory, Narragansett, RI. EPA/620/R-94/019. Summers, J.K., J.M. Macauley, P.T. Heitmuller, V.D. Engle, A.M. Adams, and G.T. Brooks. 1993. Annual statistical summary: EMAP - Estuaries Louisianian Province - 1991. U.S. EPA Office of Research and Development, Environmental Research Laboratory, Gulf Breeze, FL. EPA/600/R-93/001. U.S. EPA. 1994a. Environmental Monitoring and Assessment Program (EMAP): Laboratory methods manual - estuaries, Volume 1: Biological and physical analyses. Office of Research and Development, Environmental Monitoring and Systems Laboratory, Cincinnati, OH. EPA/620/R-94/xxx. U.S. EPA. 1994b. Methods for assessing the toxicity of sediment- associated contaminants with estuarine and marine amphipods. EPA 600/R-94/025. U.S. EPA Office of Research and Development, Narragansett, R.I. ------- U.S. EPA. 1995. Environmental Monitoring and Assessment Program (EMAP): Laboratory Methods Manual-Estuaries, Volume 1: Biologi and Physical Analyses. U.S. Environmental Protection Agency, Office of Research and Development, Narragansett, RI. EPA/620/R-95/008. TABLE OF ACRONYMS <= Less than or equal to >= Greater than or equal to C Degrees Celsius cm2 Square centimeters CMBAD Coastal Monitoring and Bioeffects Assessment Division CU Clemson University EMAP Environmental Monitoring and Assessment Program EPA U.S. Environmental Protection Agency EPA-AED EPA-Atlantic Ecology Division EPA-GED EPA-Gulf Ecology Division EPA-RTP EPA-Research Triangle Park, NC FLDEP Florida Dept. of Environmental Protection FMRI Florida Marine Research Institute FTP File Transfer Protocol GIS Geographical Information System JCWS Johnson Controls Word Services km2 Square kilometers m2 Square meters mg/L Milligrams per liter mS/cm MilliSiemens per centimeter (equiv. to milliohms/cm) MRRI Marine Resources Research Institute NCNERR North Carolina National Estuarine Research Reserve NCSU North Carolina State University, NC NA Not Applicable ng/g Nanograms per gram NOAA National Oceanic and Atmospheric Administration NOS National Ocean Service ORCA Office of Ocean Resources Conservation and Assessment QA/QC Quality Assurance/Quality Control ppb Parts per billion (equiv. to ng/g) ppm Parts per million (equiv. to ug/g) ppt Parts per thousand SAIC Science Applications International Corporation SCDNR South Carolina Dept. of Natural Resources TOC Total Organic Carbon TAMU/GERG Texas A&M University, Geochemical and Environmental Research Group TPMC Technology Planning and Management Corporation ug/g Micrograms per gram um Micrometers UC University of Charleston, SC UGA University of Georgia, GA UNC-W University of North Carolina - Wilmington, NC USGS-GB US Geological Survey - Gulf Breeze, FL wt. Weight WWW World Wide Web -Internet ------- 13. PERSONNEL INFORMATION Dr. Courtney T. Hackney University of North Carolina - Wilmington, Wilmington, NC Melissa M. Hughes Data Librarian, EMAP-Estuaries 0A0 Corporation U.S. EPA NHEERL-AED 27 Tarzwell Drive Narragansett, RI 02882-1197 (401) 782-3184 (Tel.) (401) 782-3030 (FAX) hughes.melissa0epa.gov (e-mail) For programmatic/policy matters, contact: Dr. Jeffrey L. Hyland NOAA/NOS National Centers for Coastal Ocean Science Center for Coastal Monitoring and Assessment - Charleston Lab 219 Fort Johnson Rd. Charleston, SC 29412-9110 (843)762-8652 (Tel.) (843)762-8700 (FAX) jeff.hyland0noaa.gov (e-mail) For data-related questions, contact: Dr. W. Leonard Balthis NOAA/NOS National Centers for Coastal Ocean Science Center for Coastal Monitoring and Assessment - Charleston Lab 219 Fort Johnson Rd. Charleston, SC 29412-9110 (843)762-8654 (Tel.) (843)762-8700 (FAX) len.balthis0noaa.gov (e-mail) Jan Landsberg Florida Dept. of Environmental Protection Florida Marine Research Institute St. Petersburg, FL James McKenna Florida Dept. of Environmental Protection Florida Marine Research Institute St. Petersburg, FL Dr. Gil McRae Florida Dept. of Environmental Protection Florida Marine Research Institute St. Petersburg, FL Gary A. Nelson Florida Dept. of Environmental Protection Florida Marine Research Institute St. Petersburg, FL ------- Dr. Amy H. Ringwood South Carolina Dept. of Natural Resources, Marine Resources Research Institute Charleston, SC ------- |