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MARINE SEDIMENT TOXICITY TESTS Richard C. Swartz U.S. Environmental Protection Agency ABSTRACT Sediment toxicity tests have been developed on the basis of virtually all levels of biological organization from sub- cellular through model ecosystems. Rapid, cost-effective techniques based on acute exposures are often used in research and regulatory programs to determine the spatial and temporal distribution of sediment toxicity, and the rela- tive toxicity of individual chemicals and complex wastes spiked into sediment. Sediment toxicity tests are part of several comprehensive methods for generating sediment qual- ity criteria. Major research needs include test methods for chronic exposures, field validation of acute toxicity tests and the geochemical integrity of test materials, the rela- tion between toxicity and the bioavailability/partitioning of contaminants in different sediment phases, models of toxi- cological interactions between sediment contaminants, and sediment wasteload allocation models. INTRODUCTION Marine pollution often results in the chemical contamination of the seabed and detrimental effects on benthic communities. The initial development of sediment toxicity tests in the early 1970s increasing research and regulatory interest in methods of haunch; c deli on (tannin anal R-~nn 1 971 Aim:: -t :;l _ ~ ~ ~ ) _ ~ ~ ~ . ~ ~ an_ r reflected an documenting ~ ~~ _ ~~ , _ ._, ~ ., 1974; Han- son, 1974; Cardwell et al., 1976; Lee and Mariani, 1976~. The Ocean Dumping Regulations promulgated by the U.S. Environmental Protection Agency (EPA) in 1977 included sediment toxicity tests in the evaluation of applications for dredged material disposal permits. Methods for solid-phase bioassays to be used in conjunction with the Ocean Dumping Regulations were published by EPA and the U.S. Army Corps of Engineers (COE) in 1977 (U.S. EPA/COE, 1977~. Research on sediment toxicity tests and their regulatory applications has expanded greatly since 1977. This paper reviews marine sediment toxicity tests with respect to the variety of methods that are available, and their effectiveness, relevance to remedial actions, and applications in research and regu- latory programs. 115 r

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116 SEDIMENT TOXICITY TESTS--A SUMMARY REVIEW Sediment toxicity tests have been applied at virtually all levels of biological organization ranging from inhibition of enzymatic acti- vity at the subcellular level to alterations of the structure and func- tion of macrobenthic assemblages in experimental ecosystems (Table 1~. However, only a few of these methods are commonly used to assess sediment toxicity. The original EPA/COE (1977) solid-phase bioassay simulates a dredged material disposal operation in a 20 liter or larger exposure chamber. A crustacean, infaunal bivalve, and infaunal polychaete must be included among the test species. Twenty individuals of each species are placed on a 3-cm deep layer of clean sediment, allowed to acclimate for 48 hours, and then covered by a 1.5-cm deep layer of test sediment. Controls are covered by a 1.5-cm layer of clean sediment. Five replicates are prepared for the control and each test sediment. The primary response criterion is survival after 10 days relative to controls. This procedure, or modifications of it, has been used by COE to evaluate applications for dredged material disposal permits. The amphipod acute sediment toxicity test is technically well- developed and widely applied, especially on the Pacific coast of the United States. This method evolved from the EPA/COE (1977) bioassay method after early research with the solid-phase test showed that amphi- pods were consistently more sensitive to polluted sediment than other major benthic taxa (Swartz et al., 1979~. The typical experimental design for the amphipod test includes 5 replicates for each sediment treatment. Each replicate consists of 20 individual amphipods placed in a 1 liter beaker containing a 2-cm deep layer of test sediment and 825 ml of overlying water, at a salinity of 28 ppt (parts per thousand) for marine tests. The exposure system is static, aerated, and main- tained at a constant temperature, usually 15C. At the initiation of the test, the amphipods quickly swim to the bottom and burrow com- pletely into the sediment. In the absence of stress, they remain bur- ied during the 10-day exposure period. There are three response cri- teria: mortality after 10 days, ability of survivors to bury in clean sediment, and emergence of amphipods during the exposure. Typical con- trol treatments include clean sediment from the amphipod collection site, sediment with the same particle-size distribution as the test material, carrier control for spiked chemicals (e.g., acetone), and a positive response control based on the effects of a chemical with known amphipod toxicity (e.g., cadmium). The method was originally developed for the phoxocephalid amphipod, Rhepoxynius abronius, but has been used with a variety of other marine, estuarine, and freshwater amphipod genera including Eohaustorius, Corophium, Grandidierella, Ampelisca, Hyalella, and Pontoporeia. Three detailed descriptions of the acute amphipod test are available (EVS Consultants, Inc. and Tetra Tech Inc., 1986; Swartz et al., 1985b; Reish and Lemay, 1988), and Sub- committee E47.03 of the American Society for Testing and Materials (ASTM) is presently adapting the procedure as an ASTM standard method. The literature on the amphipod test includes interlaboratory (Me arns et al., 1986), intermethod (Wil~iams et al. 9 1986), and interspecies

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117 TABLE 1 Biological organization Response criterion Biochemistry Enzyme induction, Microtox Cell Chromosome damage Development Larval abnormalities Physiology Respiration, osmoregulation Behavior Burrowing, feed- ing, sediment and predator avoidance Reproduction Fertilization, fecundity References Lee et al, 1979; Schiewe et al., 1985; Reichert et al., 1985; Varanasi et al., 1985; E.V.S. Consultants, Inc. and Tetra Tech, Inc., 1986; Williams et al., 1986; Tetra Tech, Inc., 1986a; PTI Environmental Services, 1988; Geisy et al., 1988. E.~.S. Consultants, Inc. and Tetra Tech, Inc., 1986; Chapman et al., 1982; Landolt and Kocan, 1984; Landolt et al., 1984; Long and Chapman, 1985; Chapman, 1986 Hoss et al., 1974; Cardwell et al., 1976; E.~.S. Consultants, Inc. and Tetra Tech, Inc., 1986; Williams et al., 1986; Tetra Tech, Inc., 1985, 1986a; PTI Environmental Services, 1988; Long and Chapman, 1985; Chapman and Morgan, 1983; Chapman et al., 1987. Chapman et al., 1982; Long and Chapman, 1985; Chapman, 1986, 1987; Kehoe, 1983; Alden and Butt, 1987. Chapman et al., 1987; Rubinstein, 1979; McGreer, 1979; Pearson et al., 1981, 1984; Olla and Bej da, 1983; Mohlenberg and Kiorboe, 1983; Phelps et al., 1983; Olla et al., 1984, 1988; Oakden et al., 1984a, 1984b; Swartz et al., 1985b, 1986b; Mearns et al., 1986; Clark and Patrick, 1987; Chapman et al., 1983, 1987 ; Nimmo et al., 1982 ; Pathology Fin erosion Hargis et al., 1984 Individual Mortality Population Life cycle, "r" Community Structure, func- tion, recoloniza- Lee and Mariani, 1976; E.V.S. Consultants, Inc. and Tetra Tech, Inc., 1986; Tetra Tech, Inc., 1985, 1986a, 1986b; PTI Environmental Service, 1988; Long and Chapman, 1985; Chapman, 1986; Chapman et al., 1987; Oakden, 1984a; Swartz et al., 1979, 1982, 1984, 1985a, 1985b, 1986a, 1986b; Mearns et al., 1986; Shuba et al., 1978; Tsai et al., 1979; Peddicord, 1980; Tatem, 1980; McLeese and Metcalfe, 1980; McLeese et al., 1982; Alden and Young, 1982; Ott, 1986; Reish and Lemay, 1988; Breteler et al., 1988; DeWitt et al., 1988. Chapman et al., 1987; Chapman and Fink, 1984; Tierjen and Lee 1984 Hansen' 1974; Tagatz and Tobia, 1978; Hansen and Tagatz, 1980; Rubinstein et al., 1980; Elmgren et al., 1980; Grassle et al., 1981; Oviatt et al., 1982, 1984; Perez, 1983; Bauer et al., 1988

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118 (Swartz et al., 1979) comparisons, field validation (Swartz et al., 1982, 1985b, 1986a), field toxicity surveys (Chapman et al., 1982; Tetra Tech, Inc., 1985, 1986b; Swartz et al., 1979, 1985b; Breteler et al., 1988), bioassays of the toxicity of specific chemicals or complex wastes (Reichert et al., 1985; Varanasi et al., 1985; Oakden et al., 1984a, 1984b; Swartz et al., 1986, 1984), and development of sediment quality criteria (Tetra Tech, Inc., 1986a; PTI Environmental Services, 1988; Long and Chapman, 1985; Chapman, 1986; Chapman et al., 1987~. Two other frequently used sediment toxicity tests are based on the development of bivalve larvae and inhibition of bacterial biolumines- cence (M~crotox). These methods lack the direct ecological relevance of the amphipod test, but may be equally or more sensitive to sediment contaminants (Williams et al., 1986~. Descriptions of standard methods are available for both tests (E.V. S . Consultants, Inc . and Tetra Tech, Inc., 1986; Chapman and Morgan, 1983~. Both Pacific oysters (Crassos tree gigas) and blue mussels (Mytilus edulis) are used in the larval test. Response criteria are survival and abnormal shell development of larvae exposed for 48 hours to a suspension of 20 g, wet weight, of sediment in 1 liter of filtered, sterilized, 28 ppt seawater. The bivalve larvae test has been used primarily to document the distribu- tion of sediment toxicity (Cardwell et al., 1976; Williams et al., 1986; Long and Chapman, 1985; Chapman and Morgan, 1983; Chapman et al., 1987; Tetra Tech, Inc., 1985) and to develop sediment quality criteria (Tetra Tech, Inc., 1986a; PTI Environmental Services, 1988~. The Micro- tox technique measures the inhibition of light emission by the lumines- cent bacterium (Photobacterium phosphoreum) exposed for 15 minutes to either organic or saline sediment extracts (E.V.S. Consultants, Inc. and Tetra Tech, Inc., 1986~. Schiewe et al. (1985) demonstrated a sig- nificant relation between the extract concentration causing a 50 per- cent reduction in luminescence and the concentrations of classes of organic chemicals. In comparative studies, the Microtox assay regis- tered a larger proportion of positive responses than lethality tests with Rhepaxynius abronius (Williams et al., 1986) or the freshwater cladoceran, Daphnia magna (Giesy et al., 1988~. Because of uncer- tainty about the bioavailability of extracted chemicals and the irrele- vance of bacterial luminescence to benthic ecosystems, the greater sen- sitivity of the Microtox test may reflect chemical contamination rather than a potential for ecological degradation. Microtox has also been used to examine the distribution of sediment toxicity (Schiewe et al., 1985; Williams et al., 1986; Giesy et al., 1988) and to develop sedi- ment quality criteria (Tetra Tech, Inc., 1986a; PTI Environmental Ser- vices, 1988~. Experimental designs for sediment toxicity tests with bivalve larvae and bacterial luminescence are similar to those des- cribed above for acute amphipod tests. Most of the sediment toxicity tests cited in Table 1 are not routinely used in sediment toxicity surveys or permit application reviews. These include methods to assess the effects of contaminated sediment on complex biological phenomena including predator-prey interactions (Pearson et al., 1981), the intrinsic rate of population growth ("r") (Tietjen and Lee, 1984), recruitment of benthic assem- blages from planktonic eggs and larvae (Hansen and Tagatz, 1980), and

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119 nutrient flux in recovering benthic mesocosms (Oviatt et al., 1984~. These more sophisticated techniques generally have a relatively high- cost in time, expertise, and resources. Their utility lies in evaluat- ing higher level ecological impacts when equivocal results are obtained from the more standard toxicity tests. EFFECTIVENESS OF SEDIMENT TOXICITY TESTS There are some important limitations and advantages of sediment tox- icity tests (Table 2~. Bioavailability and toxicity of sediment contam- inants can be greatly altered by collection, handling, and storage of sediment samples. Freezing and long storage can usually be avoided (U.S. EPA/COE, 1977; E.~.S. Consultants, Inc. and Tetra Tech, Inc., 1986; Swartz et al., 1985b). However, sediment samples are routinely mixed, sieved, or extracted with poorly understood effects on geochem- ical properties. Similarly, chemicals experimentally spiked into sedi- ment in the laboratory may not be bioevailable in the same way as "naturally" contaminated sediment. Research is needed to compare sediment geochemistry in the field with that of sediments used in toxicity tests . TABLE 2 Limitations and Advantages of Sediment Toxicity Tests Limitations Sediment collection, handling and storage may alter bioavailability. Results may reflect test conditions other than chemical toxicity. Route of exposure can be uncertain. Field validation is needed for sediment spiking methods. Few comparisons of methods and species. Few chronic methods. Inherent limitations of lab tests to predict ecological events. Tests applied to field samples can't discriminate effects of individual chemicals. Advantages Provide a direct benthic, biological impact assessment. Legal and scientific precedence; some standard methods . Tests applied to field samples reflect cumulative effects of all contaminants. Tests applied to spiked chemicals provide unequivocal analysis of causal relations. Sediment toxicity tests can be applied to all chemicals of concern. Only method available to examine contaminant interactions. Limited expertise or special equipment is required. Methods are rapid and cost- effective . Toxicity tests are amenable to field validation.

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120 Toxic effects of natural sediment features beyond the tolerance limits of test species can sometimes be confused with contaminant effects. Information such as the salinity (Swartz et al., 1985b) and sediment particle size requirements (Ott, 1986; DeWitt et al., 1988) known for Rhepoxynius abronius should be developed for other test species. The broad tolerance of some benthic taxa to natural sediment features often extends to contaminant effects (Olla et al., 1988), and is not a proper justification of the use of certain pelecypods and polychaetes in sediment toxicity tests. Uncertainty about the route of exposure can obfuscate toxicity results, especially when epibenthic or pelagic organisms are used as test species. Most burrowing species have direct exposures to sediment particles and interstitial water. However, if the primary exposure is through the overlying water, the degree of exposure is determined by the mechanisms controlling transport across the sediment-water inter- face. Relative toxicity may then be determined by factors such as sediment bioturbation, rather than absolute contamination. Although many sediment toxicity tests have been developed, there are no standard chronic methods and few comparisons of species or methods. EPA's Region 10 Office of Puget Sound, is currently comparing the relative sensitivity of 13 acute and chronic test methods. Pre- vious methods comparisons have shown a general concordance of acute tests in identifying the most and least contaminated sediment samples, although concordance is less at intermediate levels of contamination. Different toxicity tests may be particularly sensitive to different kinds of chemicals and, therefore, no single method will necessarily meet all requirements of sediment toxicity surveys (Swartz et al., 1985~. For these reasons, many investigations now employ several test methods (Williams et al., 1986; Chapman et al., 1982, 1987; Long and Chapman, 1985; Tetra Tech, Inc., 1985~. There is an inherent inability of simple, acute laboratory tests to predict or reflect ecological events. For example, in a field valida- tion of the acute amphipod test along the sediment pollution gradient on the Palos Verdes Shelf, off California, there was generally a good correspondence between sediment contamination, toxicity, and benthic community degradation (Swartz et al., 1985, 1986~. However, at one site intermediate between areas of major and minor impacts, there were substantial contamination and biological perturbations, but no acute amphipod toxicity. These simple tests often are not sensitive to the long-term events that effect chronic toxicity and ecological succes- sion. Sediment bioassays determine the cumulative toxicity of all chemi- cals in samples collected from the field. This is a major advantage over other analytical methods because many chemicals and their toxico- logical interactions are unknown or unmeasured. Conversely, this sensi- tivity to cumulative effects makes it impossible to attribute toxicity to specific chemicals on the sole basis of bioassay results on field sediments. Sediment toxicity tests should be part of a comprehensive analysis of sediment quality that also includes chemical, geological, and biological assessments (Swartz et al., 1985~. This is the basic concept of the very effective benthic assessment method often called

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121 the "Sediment Quality Triad" (Long and Chapman, 1985; Chapman, 1986; Chapman et al., 1987; Swartz et al., 1982, 1985, 1986~. Causal relations are unequivocal when toxicity tests are applied to unpolluted sediment spiked with individual chemicals or complex wastes. The spiking method can be applied to any chemical of concern. It also offers the only experimental procedure for examining interactions between sediment contaminants, an important problem that has not yet received much attention (Oakden et al., 1984; Samolloff et al., 1983; Plesha et al., 1988; Swartz et al., 19881. A major advantage of most sediment toxicity tests is that they require limited expertise or equipment and are rapid and cost- effective. Bioassay results are usually available within two weeks of sample collection. Analyses of macrobenthos and chemical samples from the same survey typically require months for completion at much higher costs for equipment and expertise. Toxicity tests can quickly and inexpensively locate "hot spots" where more comprehensive assessments can be focused. APPLICATIONS OF SEDIMENT TOXICITY TESTS Sediment toxicity tests have a variety of applications in research and regulatory programs (Table 3~. They are used principally to deter- mine patterns of toxicity in the field and quantify the toxicity of materials spiked into sediment. Field surveys can examine the distri- bution of toxicity in space, time, or depth in the sediment (Williams et al., 1986; Giesy et al., 1988; Chapman et al., 1982, 1983, 1987; Long and Chapman, 1985; Chapman, 1986; Chapman and Morgan, 1983; Tetra Tech, Inc., 1985, 1986b; Alden and Butt, 1987; Tsai et al., 1979; Swartz et al., 1982, 1985b, 1986a; Alden and Young, 1982; Breteler et al., 1988; Chapman and Fink, 1984~. Relative sediment toxicity is presently used in a variety of impact assessments, disposal permit decisions, and monitoring programs. Wasteload allocation models that combine sediment toxicity distributions with particle/contaminant transport, deposition, and resuspension models are currently being developed. Examination of the vertical distribution of toxicity in a sediment core reflects the historic pattern of contamination in depositional environments. Such data are particularly relevant to remedial investigations that consider capping and "no action" alter- natives. The toxicity of field-collected sediment is also used as a research variable for comparisons with biological effects and geochemi- cal sediment characteristics (Long and Chapman, 1985 ; Chapman, 1986 ; Tetra Tech, Inc., 1985, 1986b; Chapman et al., 1987; Swartz et al., 1982, 1985b, 1986a). The sediment spiking method can be used to determine the toxicity of individual chemicals or complex wastes like sewage effluents, sludges and drilling fluids (Hansen, 1974; Reichert et al ., 1985; Varanasi et al., 1985; Pearson et al., 1981; Olla and Bej da, 1983; Olla et al., 1984, 1988; Oakden et al., 1984a, 1984b; Swartz et al., 1984, 1986b; McLeese and Metcalfe, 1980; McLeese et al., 1982; Ott, 1986; Tagatz and Tobia, 1978; Bauer et al., 1988; Clark and Patrick, 1987;

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122 Hansen and Tagatz , 1980 ; Rubinstein et al ., 1980 ; Elmgren et al ., 1980 ; Gras sle et al., 1981; Oviatt et al., 1982, 1984; Perez, 1983~. Contam- inated sediment can also be mixed into clean sediment to determine an LC50 or other measure of effects (Swartz et al., 1989~. This proce- dure can determine the relative toxicity of field-collected samples that cause 100 percent mortality of test specimens. Layering, rather than spiking, could be used to test the effectiveness of proposed sedi- ment capping materials in experimental designs similar to the original EPA/COE (1977) solid-phase bioassay. TABLE 3 Tests Research and Regulatory Applications of Sediment Toxicity Field sediment Spiked sediment Sediment features Research variable Comprehensive sedi- ment evaluation me thods Regulatory applications Spatial distribution of toxicity Temporal distribution of toxicity Depth distribution of toxicity Dilution--LC50 in clean sediment S ingle chemical LC50 safe concentration sediment quality criterion Multiple chemicals joint action interaction models Complex wastes sewage sludge drilling fluids dredged material Salinity Particle-size distribution Organic carbon concentration Relation to benthic community structure, function, sediment conditions Apparent Effects Threshold Sediment Quality Triad Dredged material permit decisions Environmental impact assessment Wasteload allocation Remedial action alternatives Sediment quality criteria

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123 Sediment spiking provides a toxicological approach to the develop- ment of numerical sediment quality criteria. Safe concentrations of chemicals in sediment can be estimated from dose-response relations by the same rationale used to generate water quality criteria. Sediment toxicity tests are also part of other methods of developing sediment quality criteria, (e.g., Apparent Effects Threshold [Tetra Tech, Inc., 1986a, 1986b; PTI Environmental Services, 1988], Sediment Quality Triad [Chapman, 19863), and are being used to validate criteria based on the equilibrium partitioning method. There is a close agreement between estimates of safe sediment concentrations of fluoranthene, after organic carbon (OC) normalization, based on the methods of equilibrium partitioning--1,330 ~g/g OC (using the chronic lowest observed effect level tU.S. EPA, 19804~; the amphipod Apparent Effects Threshold--891 g/g OC tTetra Tech, Inc., 1986bJ; and fluoranthene sediment toxicity tests--817 ~g/g OC (10-day LC50 for Rhepoxynius abronius; Swartz, unpublished data). Preliminary research based on the toxicological approach indicates that a simple additivity model can predict interactions between sediment contaminants (Swartz et al., 1988~. CONCLUSIONS AND RESEARCH RECOMMENDATIONS Acute sediment toxicity tests are well-developed and have become an integral part of benthic ecosystem impact assessments. There is a broad range of test methods with a variety of biological response criteria. Standard methods have been established for acute toxicity tests that are rapid and cost-effective. They are principally applied to determine spatial/temporal patterns of toxicity, and the relative toxicity of individual chemicals and complex wastes spiked into clean sediment. These methods are used in a variety of regulatory programs including dredged material disposal permits, sediment quality criteria, wasteload allocations, and remedial actions at sites of major sediment contamination. Benthic impact assessments are most effective when toxicity tests are combined with biological, chemical, and geological indicators of sediment degradation. Future research should focus on development of standard methods for chronic sediment toxicity tests and field validation of acute sediment toxicity tests. A toxicological data base should be established for selected chemicals and sensitive infaunal species. Issues concerning the bioavailability of contaminants in different sediment phases, and the toxicological interactions of sediment contaminants must be re- solved as part of the development of sediment quality criteria. Waste- load allocation models should incorporate sediment quality criteria or toxicity predictions with models of the transport, deposition and resus- pension of sediment particles and contaminants.

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124 ACKNOWLEDGMENTS I thank Janet Lamberson and Steve Ferraro for their reviews of this manuscript. Contribution Number N-068, U.S. EPA Environmental Research Laboratory, Narragansett, Rhode Island and Newport, Oregon. REFERENCES Alden, R. W., III, and A. J. Butt. 1987. Statistical classification of the toxicity and polynuclear aromatic hydrocarbon contamination of sediments from a highly industrialized seaport. Environ. Toxicol. Chem. 6:673-684. Alden, R. W., III and R. J. Young, Jr. 1982. Open ocean disposal of materials dredged from a highly industrialized estuary: An eval- uation of potential lethal effects. Arch. Environ. Contam. Toxicol. 11:567-576. Bauer, J. E., R. P. Kerr, M. F. Bautista, C. J. Decker, and D. C. Capone. 1988. Stimulation of microbial activities and polycyclic aromatic hydrocarbon degradation in marine sediments inhabited by Capi tel la capi tata . Mar. Environ. Res. 25:63-84. Breteler, R. J., K. J. Scott, and S. P. Sheperd. 1988. Application of a new sediment toxicity test using marine amphipods, Ampelisca abdi ta, to San Francisco Bay sediments. Submitted to ASTM Twelfth Symposium on Aquatic Toxicology and Hazard Assessment, Sparks, Nevada, April 24-26, 1988. Cardwell, R. D., C. E. Woelke, M. I. Carr, and E. W. Sanborn. 1976. Sediment and elutriate toxicity to oyster larvae. In Proceedings of the Special Conference on Dredging and its environmental effects, P. A. Krenkel, J. Harrison and J. C. Burdick, III, eds., New York: American Society of Civil Engineers. Chapman, P. M. 1987. Oligochaete respiration as a measure of sediment toxicity in Puget Sound, Washington. Hydrobiologia 155:249-258. Chapman, P. M. 1986. Sediment quality criteria from the sediment qual- ity triad: An example. Environ. Toxicol. Chem. 5:957-964. Chapman, P. M., R. N. Dexter, and E. R. Long. 1987. Synoptic measures of sediment cont~mination, toxicity and infaunal community composi- tion (the Sediment Quality Triad) in San Francisco Bay. Mar. Ecol. Prog. Ser. 37:75-96. Chapman, P. M. and R. Fink. 1984. Effects of Puget Sound sediments and their elutriates on the life cycle of Capi tella capi tata . Bull. Environ. Contam. Toxicol. 33:451-459. Chapman, P. M. and J. D. Morgan. 1983. Sediment bioassays with oyster larvae. Bull. Environ. Contam. Toxicol. 31:438-444. Chapman, P. M., D. R. Munday, J. Morgan, R. Fink, R. M. Kocan, M. L. Landolt, and R. N. Dexter. 1983. Survey of Biological Effects of Toxicants upon Puget Sound Biota. II. Tests of Reproductive Impair- ment. Technical Report NOS 102 OMS 1. Rockville, Md.: National Oceanic and Atmospheric Administration. Chapman, P. M., G. A. Vigers, M. A. Farrell, R. N. Dexter, E. A. Quin- lan, R. M. Kocan, and M. Landolt. 1982. Survey of biological

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