Interlaboratory Comparison of Three Sediment Bioaccumulation Tests.

Standard bioaccumulation tests are commonly conducted using Macoma nasuta (clam), and Alitta virens (polychaete) for marine tests, and Lumbriculus variegatus (an oligochaete) for freshwater tests. Because the interlaboratory variability associated with these tests is unknown, four experienced laboratories conducted standard 28-day bioaccumulation tests with the above species using sediments contaminated with polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). Chemical analysis of tissue samples was performed by a single laboratory. The intralaboratory variance among replicates was relatively low for PCB tissue concentrations, with coefficients of variation (CVs) ranging from 9% to 28% for all laboratories and species, with the exception of one laboratory reporting higher variability for L. variegatus (CV = 51%). Intralaboratory variance for PCB tissue concentrations was higher than interlaboratory variance for A. virens and L. variegatus, and the magnitude of difference (MOD) for laboratory means ranged from 1.4 to 2.0 across species. Intralaboratory variability was also low for lipid content, and lipid normalization of PCB and PAH body residues generally had little impact on variability. In addition to variability across bioassay laboratories, analytical variability was evaluated by different laboratories measuring the concentration of PCBs and total lipids in a subsample of tissue homogenate of sediment-exposed test organisms. Variability associated with tissue analysis was higher than bioassay laboratory variability only in tests with L. variegatus. Statistical differences between samples may be observed due to the low intralaboratory variability; however, the biological significance of these differences may be limited because the MOD is low. Considering the MOD when comparing bioaccumulation across treatments accounts for uncertainty related to inherent variability of the test in the interpretation of statistically significant results. Environ Toxicol Chem 2022;41:1260-1275. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Streamlining Freshwater Bioaccumulation Bioassays: Letting the Worms Do the Hard Work.

When terminating sediment bioaccumulation tests with the oligochaete Lumbriculus variegatus, varying amounts of detrital material are retained along with the recovered worms after sieving, necessitating time-consuming, labor-intensive manual separation of worms from detritus prior to tissue residue analysis. A method to facilitate the worms self-extracting out of the detrital material into a column of gravel was developed, resulting in approximately 90% recovery test organisms (on a mass basis) at test termination. Following exposure to contaminated sediment, polychlorinated biphenyl tissue residues and residual sediment in the gut of self-extracted animals were not significantly different compared to worms recovered by manual separation followed by purging of gut contents. Environ Toxicol Chem 2021;40:1673-1677. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.

Sensitivity of the Marine Calanoid Copepod Pseudodiaptomus pelagicus to Copper, Phenanthrene, and Ammonia.

There are limited acute toxicity test methods for native North American marine species that are considered zooplankton for their entire life cycle. Examples of standardized marine zooplankton methods include mussel, bivalve, and echinoderm development tests that use a relatively short-lived planktonic larval stage, chronic life-cycle toxicity tests using epibenthic copepods, and a 24-h Acartia tonsa copepod test method. The objectives of the present study were to: 1) develop and evaluate a novel, 48-h acute toxicity test method using the marine North American copepod Pseudodiaptomus pelagicus that is planktonic for its entire life cycle, and 2) determine the sensitivity of P. pelagicus relative to commonly tested marine toxicity test species. The average (±1 standard deviation) median lethal concentrations (LC50s) for copper (Cu), phenanthrene, and un-ionized ammonia were 32 ± 15 µg/L, 161 ± 51 µg/L, and 1.08 ± 0.30 mg NH3 /L, respectively. These results placed P. pelagicus on the more sensitive end of Cu and phenanthrene species sensitivity distributions. The copepod was less sensitive to un-ionized ammonia than commonly tested marine species. This finding suggests that the acute P. pelagicus test method will allow a focus on assessing the impacts of persistent contaminants of concern with less confounding impact from naturally occurring ammonia released to the water from sources such as suspended sediments. Environ Toxicol Chem 2019;38:1221-1230. Published 2019 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Effects of sediment amended with Deepwater Horizon incident slick oil on the infaunal amphipod Leptocheirus plumulosus.

Crude oil released from the Deepwater Horizon disaster into the Gulf of Mexico posed potential impacts to infaunal invertebrates inhabiting near shore habitats. The effects of sediment-associated weathered slick oil on the amphipod Leptocheirus plumulosus was assessed using 28-d exposures to total PAH sediment concentrations ranging from 0.3 to 24mg/kg (sum of 50 PAHs or tPAH50). Survival and growth rate were significantly decreased in the 2.6, 11.4 and 24.2mg/kg treatments, but only growth in 5.5mg/kg. Offspring production was dramatically decreased but was variable and significantly different only for 24.2mg/kg. The concentrations associated with 20% decreases relative to reference were 1.05 (95% CI=0-2.89) mg/kg tPAH50 for growth rate and 0.632 (95% CI=0.11-2.15) mg/kg tPAH50 for offspring production. The concentrations of PAHs affecting amphipods are within the range of concentrations measured in marsh areas reportedly impacted by DWH oil after its release.

Evaluation of reduced sediment volume procedures for acute toxicity tests using the estuarine amphipod Leptocheirus plumulosus.

The volume of sediment required to perform a sediment toxicity bioassay is a major driver of the overall cost associated with that bioassay. Sediment volume affects bioassay cost because of sediment collection, transportation, storage, and disposal costs as well as labor costs associated with organism recovery at the conclusion of the exposure. The objective of the current study was to evaluate reduced sediment volume versions of the standard U.S. Environmental Protection Agency (U.S. EPA) 10-d acute Leptocheirus plumulosus method that uses a beaker size of 1,000 ml and 20 organisms. The test design used evaluated the effects of beaker size (250 and 100 ml) and associated sediment volume (75 and 30 ml, respectively) as well as organism loading density (10 and 20 organisms) on test endpoint responsiveness relative to the standard 10-d test method. These comparisons were completed with three different types of contaminated sediments: a field-collected polycyclic aromatic hydrocarbon (PAH)-contaminated sediment, a lead-spiked control sediment, and a control sediment spiked with mineral oil. Assessment criteria included test endpoint sensitivity, endpoint consistency, statistical power, water quality, and logistical assessments. Results indicate that the current U.S. EPA method is preferable to the reduced sediment volume methods we assessed, but that a 250-ml beaker/10 organism experimental design is of comparable utility and may be advantageous when reduced sediment volumes are desirable because of high contaminant (spiking studies) or sediment disposal costs. In addition, the results of the current study provide toxicity reference values for PAHs, lead, and an oil surrogate for petroleum hydrocarbons.

Comparison of methods for evaluating acute and chronic toxicity in marine sediments.

Sublethal test methods are being used with increasing frequency to measure sediment toxicity, but little is known about the relative sensitivity of these tests compared to the more commonly used acute tests. The present study was conducted to compare the sensitivity of several acute and sublethal methods and to investigate their correlations with sediment chemistry and benthic community condition. Six sublethal methods (amphipod: Leptocheirus plumulosus survival, growth, and reproduction; polychaete: Neanthes arenaceodentata survival and growth; benthic copepod: Amphiascus tenuiremis life cycle; seed clam: Mercenaria mercenaria growth; oyster: Crassostrea virginica lysosome destabilization; and sediment-water interface testing with mussel embryos, Mytilus galloprovincialis) and two acute methods (amphipod survival with Eohaustorius estuarius and L. plumulosus) were used to test split sediment samples from stations in California. The test with Amphiascus proved to be the most sensitive sublethal test and the most sensitive overall, identifying 90% of the stations as toxic. The Leptocheirus 10-d test was the most sensitive of the acute tests, identifying 60% of the stations as toxic. In general, the sublethal tests were not more sensitive to sediments than the acute tests, with the sublethal tests finding an average of 35% of the stations to be toxic while the acute found 44%. Of the sublethal tests, only the Amphiascus endpoints and Neanthes growth significantly (p

Comparative and mixture sediment toxicity of trinitrotoluene and its major transformation products to a freshwater midge.

The explosive trinitrotoluene (TNT) is a prevalent contaminant in many military installations worldwide. Limited knowledge of the comparative toxicity of sediment-associated TNT and related compounds contributes to uncertainty when assessing ecological risks in contaminated sites. Trinitrotoluene undergoes transformation when associated with soils and sediments and typically occurs as a mixture dominated by its reduction products. The objective of this study was to comparatively evaluate the single-compound toxicity of TNT and its major transformation products to the freshwater midge Chironomus tentans in 10-day exposures to sediment spiked with TNT, 2-aminodinitrotoluene (2-ADNT), 2,4-diaminonitrotoluene (2,4-DANT), or trinitrobenzene (TNB). In addition, the nature of the toxicological interactions of the latter compounds in a mixture was evaluated. Upon spiking to sediment, TNT and TNB rapidly degraded to reduced products, and disappearance of extractable compounds suggested irreversible binding to sediment particles. The high degree of transformation and reactivity occurring during 10 days at spiking concentrations as high as 4000 micromol/kg dry weight suggests that TNT and related compounds are unlikely to be encountered in fine-grained sediments at contaminated sites. Similar to previous investigations, the high reactivity of the spiked compound hampered determination of accurate toxic concentrations of TNT and related compounds, and of the nature of toxicological interaction of compounds in a mixture in this study. Sediment concentrations associated with decreased survival were similar for all four compounds, with the 10-d median lethal concentrations (LC50s) determined using initial concentrations ranging from 175 (2-ADNT) to 605 (2,4-DANT) micromol/kg dry weight. Sublethal decrease in growth was not observed for any compound. Results from the mixture experiment suggest additive interaction among TNT and related compounds in sediment exposures.

A field test and comparison of acute and chronic sediment toxicity tests with the estuarine amphipod Leptocheirus plumulosus in Chesapeake Bay, USA.

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.