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 .