The toxicity of nitroaromatic (2,4-diaminonitrotoluene [2,4-DANT] and 1,3,5-trinitrobenzene [TNB]) and 14C-labeled cyclonitramine compounds (hexahydro-1,3,5-trinitro-1,3,5-triazine [RDX] and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine [HMX]) to the marine polychaete Neanthes arenaceodentata and the estuarine amphipod Leptocheirus plumulosus following 10- or 28-d exposures to spiked sediments was investigated. Organismal-level effects on survival, growth, and reproduction and cellular-level effects on apoptosis (programmed cell death) were evaluated. Because cyclonitramines have low affinity for sediment, overlying water was not exchanged in the RDX and HMX exposures. Nitroaromatics sorbed strongly to sediment, resulting in near complete resistance to solvent extraction. Cyclonitramines sorbed weakly to sediment, as more 14C-activity was found in the overlying water than in the sediment at exposure termination. No significant decrease in survival or growth was observed with cyclonitramines at initial sediment concentrations as high as 1,000 microg/g. Survival was significantly affected by nitroaromatics at nominal sediment concentrations as low as 200 microg/g, with L. plumulosus being more sensitive than N. arenaceodentata. Growth was significantly decreased at sublethal concentrations of 2,4-DANT for N. arenaceodentata. Reproduction, measured only with L. plumulosus, was significantly decreased only in the highest RDX treatment and also in the lower TNB treatment. However, no decrease was observed in higher concentrations of TNB. Body burden at exposure termination was below detection limit (1 microg/kg) for all compounds. Significant inhibition of apoptosis was not accompanied by significant decreases in growth or reproduction. Because of its critical function in many biological processes. alterations in this endpoint may result in adverse effects on the organism and could be used as an early indicator of toxicity.

A 28-d partial life-cycle test with the estuarine amphipod Leptocheirus plumulosus was developed in response to the need for an assay to mimic chronic exposure to sediment-associated contaminants. To ensure that toxicity tests have environmental relevance, it is essential to evaluate the relationship between laboratory responses and field measures of contamination. Consequently, one objective of the study was to compare the results of the chronic sediment toxicity test with L. plumulosus to gradients of sediment contamination and the in situ benthic community in its native Chesapeake Bay. Chronic tests were conducted by two laboratories, the Army Corps of Engineers Waterways Experiment Station ([WES]; Vicksburg, MS, USA) and the University of Maryland ([UM] College Park, MD, USA) using different feeding regimes, providing the opportunity to evaluate the effect of this variable on response sensitivity. A second objective was to compare the relative sensitivity of acute and chronic tests with L. plumulosus with field-collected sediments. Overall, there was good agreement between the toxicological response of acute and chronic tests with L. plumulosus and field measures of contamination. Survival in the acute test and chronic test conducted by WES was negatively correlated with concentrations of sediment-associated contaminants. Survival in acute exposures was significantly reduced in sediments from 8 of 11 stations. Indigenous L. plumulosus were found only at two of the three stations that did not exhibit acute toxicity. An unexpected finding was the difference in responsiveness of the two chronic tests. Survival in tests conducted by UM and WES was significantly reduced in sediments from 4 and 6 of 11 stations, respectively. No additional sublethal toxicity was detected in the UM chronic test, but the WES test detected reproductive effects at two additional stations. We believe the observed differences were related to the test diet used. Partly as a result of our findings, the recommended diet for the L. plumulosus chronic test was changed in the final methods document.

Abstract-Based on the need for a test to evaluate chronic sublethal toxicity in estuarine sediments, a 28-d sediment bioassay with the estuarine amphipod Leptocheirus plumulosus (Shoemaker) was developed. The test was initiated with animals less than 2 weeks old (i.e., 425-600 m sieved size class). Test endpoints included survival, growth (g dry weight/d), and reproduction (number of neonates/surviving female). Factors with the potential to influence test animal performance (i.e., nontreatment factors) such as artificial sea salts, salinity, food ration, size at test initiation, intraspecific density, sediment grain size, and diet were evaluated. For example, intraspecific densities between 10 and 60 animals/beaker (i.e., 0.18-1.4 animals/cm 2 ) did not affect survival, growth, or reproduction. Similarly, L. plumulosus were tolerant of a wide range of sediment grain sizes with only extremely fine grained (e.g., Ͼ75% clay) or coarse grained (e.g., Ͼ75% sand) material significantly affecting survival, growth, and reproduction. Test performance criteria included control survival (Ͼ80%) and reproduction (production of offspring in all control replicates), and response to a reference toxicant test with cadmium chloride in a control chart format.

A gradient in H. azteca growth was created by reducing food ration . .Tests were initiated with neonates (<48 h old) and the effects of an altered food ration on survival and growth were examined after 10 and 49 days . Growth rates decreased significantly with reduced food ration (10 day growth rates ranged from 1 .2 tg d -1 in the highest feeding regime to 0 .5 tg d -1 in lowest feeding regime) . Survival after 10 days was not affected by ration, ranging from 86 to 96% . A growth rate of 1 .2 jig d -1 at day 10 resulted in mean dry weight (1 .0 mg), survival (62%) and reproduction (9 .3 neonates/female) at day 49 similar to reported values for this species . Growth rates <0 .9 Ag d-1 at day 10 corresponded to significantly reduced reproduction at day 49 (i .e ., < 1 neonate per female) . Time to the onset of amplexus increased with decreasing ration suggesting reduced ration may have delayed reproduction .

Sediment toxicity tests are valuable tools for assessing the potential effects of contaminated sediments in dredged material evaluations because they inherently address complexity (e.g., unknown contaminants, mixtures, bioavailability). Although there is a need to understand the chronic and sublethal impacts of contaminants, it is common to conduct only short-term lethality tests in evaluations of marine sediments. Chronic toxicity methods for marine sediments have been developed but the efficacy of these methods is less documented. In this evaluation of marine sediments collected from the New York/New Jersey (NY/NJ) Harbor, three 10-d acute toxicity test methods (Ampelisca abdita, Leptocheirus plumulosus, Americamysis bahia) and three chronic and sublethal test methods (28-d L. plumulosus, 20- and 28-d Neanthes arenaceodentata) were applied by three testing laboratories. Although the N. arenaceodentata and A. bahia tests did not indicate significant toxicity for the sediments tested in this study, these methods have been reported useful in evaluating other sediments. The 10-d A. abdita, 10-d L. plumulosus and 28-d L. plumulosus tests were comparable between laboratories, indicating 29-43%, 29%, and 43-71% of the tested sediments as potentially toxic. The 28-d L. plumulosus method was the only chronic toxicity test that responded to the test sediments in this study. The 28-d L. plumulosus endpoint magnitudes were related to sediment chemistry and the sublethal endpoints were reduced as much or more than acute lethality endpoints. However, intra-treatment sublethal endpoint variability was greater, compromising detection of statistical significance. In this study, the chronic L. plumulosus test method was less consistent among laboratories relative to acute test methods, identifying potential for toxicity in a similar number (or slightly more) NY/NJ Harbor sediments.

Abstract-The influence of worm age, duration of exposure, and endpoint selection on bioassay sensitivity were evaluated for Neanthes arenaceodentata. Worms were exposed to contaminated sediment collected from Black Rock Harbor (BRH) near Bridgeport, Connecticut, USA. This sediment was diluted with clean control sediment to result in five experimental treatments: 0, 25, 50, 75, and 100% BRH. Three exposure scenarios were employed: (1) a 4-week exposure beginning with newly emerged juveniles (EJ-4w), (2) a 7-week exposure beginning with newly emerged juveniles (EJ-7w), and (3) a 4-week exposure beginning with 3-week-old juveniles (3WO-4w). Six measures of worm size were recorded at the conclusion of each exposure to evaluate differences among measurement endpoints. Survival was significantly reduced at the 25% BRH level for the EJ-7w scenario and at the 100% BRH level for the EJ-4w and 3WO-4w scenarios. Growth was significantly reduced at the 25% BRH level in each exposure scenario. Estimates based on the calculated minimum detectable difference indicated that considerably lower concentrations of BRH (6-10%) should be distinguishable by measuring effects on Neanthes growth. Worm size measured in terms of projected area, dry weight, and ash-free dry weight provided the most sensitive measures of effects. Increasing the length of exposure from 4 to 7 weeks and initiating exposures with emergent juveniles rather than 3-week-old worms increased the sensitivity of the bioassay. The results of this study demonstrate that N. arenaceodentata is sensitive to the presence of sediment-associated contaminants and that test animal age, duration of exposure, and choice of endpoint can have a large effect on the magnitude of the toxic response observed.

Abstract-A 3 ϫ 2 factorial experiment was conducted to evaluate the influence of food ration on observed toxicity in Neanthes arenaceodentata. Worms were fed one of three food rations: 0.25ϫ, 1.0ϫ, and 6.0ϫ. The 1.0ϫ ration was composed of 4 mg TetraMarin/worm/week and 2 mg alfalfa/worm/week. Worms in each food ration were exposed to one of two sediments: a clean control sediment collected from Sequim Bay, Washington, USA (SC) or a contaminated sediment collected from Black Rock Harbor near Bridgeport, Connecticut, USA (BRH) that was diluted with SC such that the final concentration of contaminated sediment was 12%. Survival and growth were monitored every 2 weeks for 8 weeks. Sediment treatment and food ration had strong effects on survival and growth patterns in N. arenaceodentata. Most of the mortality within any given treatment combination occurred during the first 2 weeks of the experiment. This decline in survival was greatest for worms exposed to BRH. Survival was significantly lower for worms in BRH compared to SC at the 0.25ϫ and 1.0ϫ ration levels. Survival was not reduced for worms in BRH compared to SC at the 6.0ϫ ration level. Worm size during the course of the experiment was positively related to ration level. Significantly reduced size in BRH worms was observed at each monitoring period within the 0.25ϫ and 1.0ϫ rations, but only at weeks 6 and 8 for the 6.0ϫ ration level. The percent difference in size between worms exposed to BRH and SC, when those worms were fed the 0.25ϫ or 1.0ϫ rations, ranged between 55 and 66%. The percent difference in size observed between BRH and SC worms fed the 6.0ϫ ration was much smaller (9-17%). The results of this study demonstrate that food ration has a strong influence on observed toxicity in N. arenaceodentata. The importance of food ration effects on toxicity must be considered while designing chronic sublethal bioassays. To ensure the relevance of bioassay results, the food ration used during a chronic bioassay should produce a pattern of growth in control animals that is similar to patterns of growth observed in nature.

Abstract-The influence of exposure source, worm density, and sex on the bioaccumulation and toxicity of 14 C-labeled DDT was evaluated for Neanthes arenaceodentata. Emergent juvenile worms were exposed to DDT-spiked sediment (SED), DDT-spiked supplemental food (TetraMarin; TM), or a combination of both (SEDTM) in 28-d toxicity experiments. The DDT concentrations in the sediment and in the food were equal on an organic carbon basis. Higher body residues were attained under the TM exposure compared to SED, indicating the importance of nonsedimentary organic matter as a source for contaminant uptake. The DDT was more bioavailable when associated with TetraMarin compared to sediment, indicating that factors other than sorption to organic carbon can influence contaminant bioavailability. Body residues as high as 0.5 mol/g wet weight were not associated with decreased survival, suggesting general narcosis as the mode of lethal toxicity of DDT to N. arenaceodentata. Most of the body burden was DDT unmetabolized compound (ϳ70%). Reduced growth, observed under all exposures, was significant at lower concentrations when worms were fed DDT-spiked food (TM and SEDTM). Neanthes arenaceodentata may have detected the presence of DDT in the TetraMarin and decreased their feeding rate, at the cost of reduced growth. The influence of worm density on DDT toxicity and bioaccumulation was examined by exposing one or five worms per beaker (74 and 370 worms/m 2 , respectively) to spiked sediment. Although higher body residues were attained under low density, worms exposed in groups of five were significantly more sensitive to the effects of DDT on growth. Sex also had a large influence on DDT uptake kinetics. In sediment exposures (SED) to a trace concentration, the uptake clearance rate (mg OC /g lipids/h) was two times more efficient and the elimination rate was five times faster in mature males than in mature females.