Development and application of whole-sediment toxicity test using immobilized freshwater microalgae Pseudokirchneriella subcapitata

A method for a whole-sediment toxicity test using alginate immobilized microalgae Pseudokirchneriella subcapitata was developed using spiked sediments and applied to contaminated field sediment samples. For method development, a growth inhibition test (72 h) with algal beads was conducted for the sediments spiked with Cu or diuron. The method was validated by determining dose-response relationships for Cu and diuron in both fine-grained and coarse-grained sediments. The results of a spiked sediment toxicity test suggested that sediment particle size distribution (clay content) had a significant effect on the growth of P. subcapitata. The developed method using immobilized microalgae P. subcapitata beads was applied successfully in the toxicity test and toxicity identification evaluation (TIE) for the four field sediment samples. After a series of extractions with 0.01 M CaCl(2) solution, acetone, and dichloromethane, the extracted sediment, which was shown to be nontoxic to algae, was used as the control and diluent for the same sediment in the whole-sediment toxicity test. The results showed that all investigated field sediment samples were found to be toxic to the immobilized algae P. subcapitata, with their median effective concentration (EC50) values ranging from 41.4 to 79.0% after 72 h exposure. In the whole sediment TIE, growth of P. subcapitata was improved to varying degrees after adding zeolite, resin, or activated charcoal, suggesting different contributions to toxicity from ammonia, metals, and organic contaminants in the tested sediments.     

Establishing cause-effect relationships in hydrocarbon-contaminated sediments using a sublethal response of the benthic marine alga, Entomoneis cf punctulata

A sublethal whole-sediment toxicity test that uses flow cytometry to measure inhibition of esterase activity in the marine microalga Entomoneis cf punctulata was applied to the assessment of hydrocarbon-contaminated sediments and toxicity identification and evaluation (TIE). Concentration-response relationships were developed, and a 20% effect concentration for total polycyclic aromatic hydrocarbons (PAHs) of 60 mg/kg normalized to 1% total organic carbon was calculated. Relationships between toxic effects and sediment organic carbon concentrations, organic carbon forms (e.g., black carbon), and sediment particle size indicated that further normalization of hydrocarbon concentrations to sediment particle size may improve concentration-response relationships. The algal toxicity test was applied as a rapid whole-sediment TIE procedure that involved the addition to sediment of powdered coconut charcoal (PCC), a hydrophobic, carbon-based material that strongly adsorbs PAHs and decreases the pore-water exposure pathway. Sediments with PCC concentrations of up to 15% (w/w) provided acceptable responses in control sediments. For six sediments with total PAH concentrations of 1,060, 4,060, 5,120, 9,150, 9,900, and 15,900 mg/kg, inhibition of E. cf punctulata esterase activity (% of control) was 75, 97, 94, 93, 100, and 97%, respectively. Following a 15% PCC amendment to these sediments, inhibition of esterase activity was 0, 1, 11, 69, 32, and 68%, respectively, indicating a decrease in toxicity in all sediments. Because the alga E. cf punctulata is exposed to toxicants via both pore water and overlying water, the reduction in toxicity achieved by 15% PCC additions can be related to the efficient removal of dissolved hydrocarbons released from sediment particles. The sediment-PCC manipulations coupled with algal whole-sediment toxicity tests provided an effective and rapid TIE method to determine whether hydrocarbon contaminants are responsible for toxicity in sediments.     

An in situ toxicity identification evaluation method Part I: Laboratory validation

Identification of individual chemical groups is critical in evaluating sediment quality and fractionating these groups of chemicals in a mixture is important to determine the primary chemicals causing toxicity. The in situ toxicity identification evaluation (iTIE) is a novel method that was developed to fractionate chemicals in contaminated sediments and waters and assess toxicity organisms. The study objectives were to verify that the iTIE can help identify contaminant chemical classes and improve the toxicity assessment process; and compare the iTIE and U.S. Environmental Protection Agency's (U.S. EPA) toxicity identification evaluation (TIE) methods. The iTIE exposure chamber is powered by a portable air pump that suctions pore water, via a Venturi system, through selective sorption materials. After passing through the sorptive materials, the pore water passes into an exposure chamber containing Daphnia magna. The chemical sorption materials included Ambersorb 563 for nonpolar organic chemical adsorption, Chelex for metals chelation, and multiple zeolite types for ammonia adsorption. The laboratory studies were performed using water and sediments spiked with ammonia, cadmium, and fluoranthene. The laboratory validation of the iTIE approach showed that different classes of compounds readily could be separated via the resin treatments, resulting in significant differences in concentrations and thus exposures to in situ exposed organisms. Ammonia, cadmium, and fluoranthene were significantly removed by zeolite, Chelex, and Ambersorb, respectively. Although there was some cross-adsorption to the other nontarget resins, it was limited and allowed for treatment differences to be detected. Survival in the treatment resins exposed to the target compounds was as high as control survivals. A 24-h exposure period appeared optimal, allowing for replacement of initial culture water with pore waters, while longer exposures occasionally allowed for breakthrough of contaminants. The iTIE was more sensitive than the U.S. EPA TIE method, in that it detected toxicity more readily due to the greater loss of contaminant concentrations in the TIE manipulation process.     

Causes of Sediment Toxicity to <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> in San Francisco Bay,California

Since the San Francisco Regional Monitoring Program (RMP) sampling began, elutriate samples prepared with sediment from the Grizzly Bay monitoring station have been consistently toxic to bivalve larvae (Mytilus galloprovincialis). An investigation into the cause of toxicity was initiated with a Phase I Toxicity Identification Evaluation (TIE) using bivalve embryos. TIE results and chemical analyses of elutriate samples suggested that divalent metals were responsible for the observed toxicity. Following the initial characterization of trace metals as toxicants, additional TIEs were performed on elutriates prepared from three additional Grizzly Bay samples collected between 1997 and 2001. Additional TIEs included ethylenediamine tetraacetic acid (EDTA) treatments in a sediment-water interface (SWI) exposure system, and the use of a cation exchange column with serial elution of sample fractions with hydrochloric acid of increasing normality. EDTA significantly reduced toxicity in overlying water in the SWI system. The cation exchange column reduced both toxicity and concentrations of trace metals, and serial elution of the column added back both toxicity and specific metals contained in individual acid fractions. Chemical analyses of three elutriate samples demonstrated copper concentrations were within the range toxic to bivalves. Results of Phase I TIEs, additional Phase II treatments, SWI exposures, and metals analyses indicate the potential for metal toxicity in sediments from this estuarine site. When combined with the results of standard TIE methods, a solid-phase cation extraction and elution approach identified copper as the most probable cause of toxicity.     

Toxicity of uranium, molybdenum, nickel, and arsenic to Hyalella azteca and Chironomus dilutus in water-only and spiked-sediment toxicity tests

A series of laboratory spiked-sediment toxicity tests with the amphipod Hyalella azteca and the midge Chironomus dilutus were undertaken to determine acute and chronic toxicity thresholds for uranium (U), molybdenum (Mo), nickel (Ni), and arsenic (As) based on both whole-sediment (total) and pore water exposure concentrations. Water-only toxicity data were also generated from separate experiments to determine the toxicities of these metals/metalloids under our test conditions and to help evaluate the hypothesis that pore water metal concentrations are better correlated with sediment toxicity to benthic organisms than whole-sediment metal concentrations. The relative toxicity of the four elements tested differed depending on which test species was used and whether whole-sediment or pore water metal concentrations were correlated with effects. Based on measured whole-sediment concentrations, Ni and As were the two most acutely toxic elements to H. azteca with 10-d LC50s of 521 and 532 mg/kg d.w., respectively. Measured pore water concentrations indicated that U and Ni were the two most acutely toxic elements, with 10-d LC50s to H. azteca of 2.15 and 2.05 mg/L, respectively. Based on pore water metal concentrations, the no-observed-effect concentrations (NOECs) for growth were (H. azteca and C. dilutus, respectively) 0.67 and 0.21 mg/L for U, <0.37 and 0.60 mg/L for Ni, and 16.43 and <0.42 mg/L for As. Pore-water lowest-observed-effect concentrations (LOECs) for growth were (H. azteca and C. dilutus, respectively) 2.99 and 0.48 mg/L for U, 0.37 and 2.33 mg/L for Ni, and 58.99 and 0.42 mg/L for As. For U and Ni, results from 96-h water-only acute toxicity tests correlated well with pore water metal concentrations in acutely toxic metal-spiked sediment. This was not true for As where metalloid concentrations in overlying water (diffusion from sediment) may have contributed to toxicity. The lowest whole-sediment LOEC reported here for As was 6.6- and 4-fold higher than the Canadian Council of Ministers of the Environment interim sediment quality guideline and the Canadian Nuclear Safety Commission (CNSC) lowest effect level (LEL), respectively. The lowest whole-sediment LOECs reported here for Ni, U and Mo were 4-, 17.5-, and >260-fold higher, respectively, than the CNSC LELs for these metals/metalloids. Data on pore water metal concentrations in toxic sediment would be a useful addition to future Guidelines documents.     

Biological and Chemical Characterization of Metal Bioavailability in Sediments from Lake Roosevelt,Columbia River,Washington, USA

We studied the bioavailability and toxicity of copper, zinc, arsenic, cadmium, and lead in sediments from Lake Roosevelt (LR), a reservoir on the Columbia River in Washington, USA that receives inputs of metals from an upstream smelter facility. We characterized chronic sediment toxicity, metal bioaccumulation, and metal concentrations in sediment and pore water from eight study sites: one site upstream in the Columbia River, six sites in the reservoir, and a reference site in an uncontaminated tributary. Total recoverable metal concentrations in LR sediments generally decreased from upstream to downstream in the study area, but sediments from two sites in the reservoir had metal concentrations much lower than adjacent reservoir sites and similar to the reference site, apparently due to erosion of uncontaminated bank soils. Concentrations of acid-volatile sulfide in LR sediments were too low to provide strong controls on metal bioavailability, and selective sediment extractions indicated that metals in most LR sediments were primarily associated with iron and manganese oxides. Oligochaetes (Lumbriculus variegatus) accumulated greatest concentrations of copper from the river sediment, and greatest concentrations of arsenic, cadmium, and lead from reservoir sediments. Chronic toxic effects on amphipods (Hyalella azteca; reduced survival) and midge larvae (Chironomus dilutus; reduced growth) in whole-sediment exposures were generally consistent with predictions of metal toxicity based on empirical and equilibrium partitioning-based sediment quality guidelines. Elevated metal concentrations in pore waters of some LR sediments suggested that metals released from iron and manganese oxides under anoxic conditions contributed to metal bioaccumulation and toxicity. Results of both chemical and biological assays indicate that metals in sediments from both riverine and reservoir habitats of Lake Roosevelt are available to benthic invertebrates. These findings will be used as part of an ongoing ecological risk assessment to determine remedial actions for contaminated sediments in Lake Roosevelt.     

Toxicity Identification Evaluation of Five Metals Performed with Two Organisms (Daphnia magna and Lactuca sativa)

When trying to identify the main toxicants in effluents, natural waters, sediments, soil leachates, and leachates from products, the Toxicity Identification Evaluation (TIE) procedure has proven useful. To enhance the use of this procedure for soil, sewage, and sediment samples, we wanted to evaluate this TIE procedure, regarding metal toxicity, for the 96-h root elongation test performed with Lactuca sativa (lettuce) seeds. We also wanted to evaluate the effect of TIE treatment on the toxicity of Mn and Fe to Daphnia magna. Bioassays were performed with Daphnia magna (48-h immobility) and lettuce seeds (96-h root elongation) to determine the effect concentrations for both organisms of Ag, Cu, Fe, Mn, and Zn. The TIE was then performed at the determined Daphnia 48-h EC₈₄ and Lactuca 96-h EC₅₀ for each metal. Our results showed that the order of the metal toxicity was Ag>Cu>Zn>Fe>Mn, for Daphnia and Ag = Zn = Fe = Cu > Mn for lettuce seeds. We also found that toxicity of the metals for Daphnia magna was reduced according to the prevailing knowledge regarding Cu, Zn, and Ag. However, the toxicity of Ag and Cu for Daphnia was also reduced by filtration through a C18 resin. Toxicity of Mn and Fe was reduced by filtration through a CM resin and increase of pH. For lettuce seeds, toxicity of the metals was reduced by the same treatments as for Daphnia magna with the exception of EDTA addition, which did not affect Cu toxicity to lettuce seeds. No effects were found for filtration through a C18 resin. We suggest that the TIE procedure using lettuce seeds can be used in toxicity identification of metals. However, the effects of pH manipulations were often stronger with lettuce and should be interpreted with care.     

Development of a whole-sediment toxicity test using a benthic marine microalga

An acute whole-sediment toxicity test with a benthic marine microalga was developed and optimized using flow cytometry to distinguish algae (based on their chlorophyll a autofluorescence) from sediment particles. Of seven benthic marine algae screened, the diatom Entomoneis cf punctulata was most suitable because of its tolerance of a wide range of water and sediment physicochemical parameters, including salinity, pH, ammonia, and sulfide. A whole-sediment and water-only toxicity test based on inhibition of esterase activity in this species was developed. Enzyme activity rather than growth was used as the test endpoint, as nutrient release from sediments has previously been found to stimulate algal growth, potentially masking contaminant toxicity. The sensitivity of the bioassay to a range of metals (copper, zinc, cadmium, lead, arsenic, manganese) and phenol in water-only exposures was compared to the standard 72-h growth rate inhibition test. The esterase enzyme inhibition test was sensitive to copper, with a 3-h inhibitory concentration to cause a 50% (IC50) reduction in a fluorescein diacetate fluorescence value of 97 +/- 39 microg Cu/L. A concentration-dependent response was also observed in the presence of sediment particles (copper tailings), with and without dilution, using a control clean sediment. The primary route of exposure to copper was via pore water rather than by direct contact with tailings particles. This is the first whole-sediment bioassay developed with a benthic alga suitable for sediment quality assessment in marine/estuarine systems, and its advantages and limitations are discussed.     

Identifying the causes of oil sands coke leachate toxicity to aquatic invertebrates

A previous study found that coke leachates (CL) collected from oil sands field sites were acutely toxic to Ceriodaphnia dubia; however, the cause of toxicity was not known. Therefore, the purpose of this study was to generate CL in the laboratory to evaluate the toxicity response of C. dubia and perform chronic toxicity identification evaluation (TIE) tests to identify the causes of CL toxicity. Coke was subjected to a 15-d batch leaching process at pH 5.5 and 9.5. Leachates were filtered on day 15 and used for chemical and toxicological characterization. The 7-d median lethal concentration (LC50) was 6.3 and 28.7% (v/v) for pH 5.5 and 9.5 CLs, respectively. Trace element characterization of the CLs showed Ni and V levels to be well above their respective 7-d LC50s for C. dubia. Addition of ethylenediaminetetraacetic acid significantly (p?≤?0.05) improved survival and reproduction in pH 5.5 CL, but not in pH 9.5 CL. Cationic and anionic resins removed toxicity of pH 5.5 CL only. Conversely, the toxicity of pH 9.5 CL was completely removed with an anion resin alone, suggesting that the pH 9.5 CL contained metals that formed oxyanions. Toxicity reappeared when Ni and V were added back to anion resin-treated CLs. The TIE results combined with the trace element chemistry suggest that both Ni and V are the cause of toxicity in pH 5.5 CL, whereas V appears to be the primary cause of toxicity in pH 9.5 CL. Environmental monitoring and risk assessments should therefore focus on the fate and toxicity of metals, especially Ni and V, in coke-amended oil sands reclamation landscapes.     

Determining stressor presence in streams receiving urban and agricultural runoff: development of a benthic in situ toxicity identification evaluation method

Determining toxicity in streams during storm-water runoff can be highly problematic because of the fluctuating exposures of a multitude of stressors and the difficulty of linking these dynamic exposures with biological effects. An underlying problem with assessing storm-water quality is determining if toxicity exists and then which contaminant is causing the toxicity. The goal of this research is to provide an alternative to standard toxicity testing methods by incorporating an in situ toxicity identification evaluation (TIE) approach. A benthic in situ TIE bioassay (BiTIE) was developed for separating key chemical classes of stressors in streams during both low- and high-flow events to help discern between point and nonpoint sources of pollution. This BiTIE method allows for chemical class fractionation through the use of resins, and these resins are relatively specific for removing nonpolar organics (Dowex Optipore), ammonia (zeolite), and polywool (control). Three indigenous aquatic insects, a mayfly (Isonychia spp.), a caddisfly (Hydropsyche spp.), and a water beetle (Psephenus herricki), were placed in BiTIE chambers that were filled with natural substrates. Acute 96-h exposures were conducted at Honey Creek, New Carlisle, Ohio, USA (reference site), and Little Beavercreek, Beavercreek, Ohio, USA (impaired site). At both sites, significant (p < 0.025) stressor responses were observed using multiple species with polywool or no resin (control) treatments exhibiting < 80% survival and resin treatments with >80% survival. The BiTIE method showed stressor-response relationships in both runoff and base flow events during 96-h exposures. The method appears useful for discerning stressors with indigenous species in situ.     

Toxicity Identification Evaluation of Lake Orta (Northern Italy) Sediments Using the Microtox System

Pore waters extracted by centrifugation from Lake Orta (Northern Italy) sediments were studied with a modified Toxicity Identification Evaluation (TIE) procedure using the Microtox bacterial luminescence toxicity test system. The most toxic pore water samples were from stations near a rayon factory, known as a source of copper and ammonium discharges. The TIE manipulations used were filtration, EDTA chelation, and C18 solid-phase resin adsorption. The most effective treatments to remove toxicity were the EDTA and C18, indicating that both metals and nonpolar organic compounds contribute to the observed toxicity.     

Whole-sediment toxicity identification evaluation tools for pyrethroid insecticides: I. Piperonyl butoxide addition

Piperonyl butoxide (PBO) is a synergist used in some pyrethroid and pyrethrin pesticide products and has been used in toxicity identification evaluations (TIEs) of water samples to indicate organophosphate or pyrethroid-related toxicity. Methods were developed and validated for use of PBO as a TIE tool in whole-sediment testing to help establish if pyrethroids are the cause of toxicity observed in field-collected sediments. Pyrethroid toxicity was increased slightly more than twofold in 10-d sediment toxicity tests with Hyalella azteca exposed to 25 microg/L of PBO in the overlying water. This concentration was found to be effective for sediment TIE use, but it is well below that used in previous water and pore-water TIEs with PBO. The effect of PBO on the toxicity of several nonpyrethroids also was tested. Toxicity of the organophosphate chlorpyrifos was reduced by PBO, and the compound had no effect on toxicity of cadmium, DDT, or fluoranthene. Mixtures of the pyrethroid bifenthrin and chlorpyrifos were tested to determine the ability of PBO addition to identify pyrethroid toxicity when organophosphates were present in a sample. The PBO-induced increase in pyrethroid toxicity was not seen when chlorpyrifos was present at or above equitoxic concentrations with the pyrethroid. In the vast majority of field samples, however, the presence of chlorpyrifos does not interfere with use of PBO to identify pyrethroid toxicity. Eleven field sediments or soils containing pyrethroids and/or chlorpyrifos were used to validate the method. Characterization of the causative agent as determined by PBO addition was consistent with confirmation by chemical analysis and comparison to known toxicity thresholds in 10 of the 11 sediments.     

The behavior and identification of toxic metals in complex mixtures: Examples from effluent and sediment pore water toxicity identification evaluations

Toxicity caused by heavy metals in environmental samples can be assessed by performing a suite of toxicity identification evaluation (TIE) methods. The behavior of metals during TIEs can vary greatly according to sample matrix. Some approaches and precautions in using TIE to identify metal toxicants in a sample are discussed, using case studies from three effluent and one sediment TIEs. These approaches include responses of metals that erroneously suggest the presence of other toxicants, the bioavailability of metals retained by glass-fiber filtration, and cautionary steps in Phase III to avoid dilution water effects on sample toxicity.Mention of trade names does not constitute endorsement by the U.S. Environmental Protection Agency.     

Whole sediment toxicity identification evaluation tools for pyrethroid insecticides: III. Temperature manipulation

Since the toxicity of pyrethroid insecticides is known to increase at low temperatures, the use of temperature manipulation was explored as a whole-sediment toxicity identification evaluation (TIE) tool to help identify sediment samples in which pyrethroid insecticides are responsible for observed toxicity. The amphipod Hyalella azteca is commonly used for toxicity testing of sediments at a 23 degrees C test temperature. However, a temperature reduction to 18 degrees C doubled the toxicity of pyrethroids, and a further reduction to 13 degrees C tripled their toxicity. A similar response, though less dramatic, was found for 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT), and dissimilar temperature responses were seen for cadmium and the insecticide chlorpyrifos. Tests with field-collected sediments containing pyrethroids and/or chlorpyrifos showed the expected thermal dependency in nearly all instances. The inverse relationship between temperature and toxicity provides a simple approach to help establish when pyrethroids are the principal toxicant in a sediment sample that could be used as a supplemental tool in concert with chemical analysis or other TIE manipulations. The phenomenon appears to be, in part, a consequence of a reduced ability to biotransform the toxic parent compound at cooler temperatures. The strong dependence of pyrethroid toxicity on temperature has important ramifications for predicting their environmental effects, and the standard test temperature of 23 degrees C dramatically underestimates risk to resident fauna during the cooler months.     

Toxicity of Sediment Cores Collected from the Ashtabula River in Northeastern Ohio,USA, to the Amphipod <Emphasis Type="Italic">Hyalella azteca</Emphasis>

This study was conducted to support a Natural Resource Damage Assessment and Restoration project associated with the Ashtabula River in Ohio. The objective of the study was to evaluate the chemistry and toxicity of 50 sediment samples obtained from five cores collected from the Ashtabula River (10 samples/core, with each 10-cm-diameter core collected to a total depth of about 150 cm). Effects of chemicals of potential concern (COPCs) measured in the sediment samples were evaluated by measuring whole-sediment chemistry and whole-sediment toxicity in the sediment samples (including polycyclic aromatic hydrocarbons [PAHs], polychlorinated biphenyls [PCBs], organochlorine pesticides, and metals). Effects on the amphipod Hyalella azteca at the end of a 28-day sediment toxicity test were determined by comparing survival or length of amphipods in individual sediment samples in the cores to the range of responses of amphipods exposed to selected reference sediments that were also collected from the cores. Mean survival or length of amphipods was below the lower limit of the reference envelope in 56% of the sediment samples. Concentrations of total PCBs alone in some samples or concentrations of total PAHs alone in other samples were likely high enough to have caused the reduced survival or length of amphipods (i.e., concentrations of PAHs or PCBs exceeded mechanistically based and empirically based sediment quality guidelines). While elevated concentrations of ammonia in pore water may have contributed to the reduced length of amphipods, it is unlikely that the reduced length was caused solely by elevated ammonia (i.e., concentrations of ammonia were not significantly correlated with the concentrations of PCBs or PAHs and concentrations of ammonia were elevated both in the reference sediments and in the test sediments). Results of this study show that PAHs, PCBs, and ammonia are the primary COPCs that are likely causing or substantially contributing to the toxicity to sediment-dwelling organisms. An erratum to this article can be found at     

A Site-Specific Evaluation of Mercury Toxicity in Sediment

A site-specific evaluation of mercury toxicity was conducted for sediments of the Calcasieu River estuary (Louisiana, USA). Ten-day whole-sediment toxicity tests assessed survival and growth (dry weight) of the amphipods Hyalella azteca and Leptocheirus plumulosus under estuarine conditions (10 ppt salinity). A total of 32 sediment samples were tested for toxicity, including 14 undiluted site sediment samples and 6 sediment dilution series. All sediment samples were analyzed for total mercury and numerous other chemical parameters, including acid volatile sulfide (AVS) and simultaneously extracted metals (SEM). No toxicity attributable to mercury was observed, indicating that a site-specific threshold for total mercury toxicity to amphipods exceeds 4.1 mg/kg dry weight. Site-specific factors that may limit mercury bioavailability and toxicity include relatively high sulfide levels. Additionally, the chemical extractability of mercury in site sediments is low, as indicated by SEM mercury analyses for three sediment samples containing a range of total mercury concentrations. Received: 17 November 1998/Accepted: 7 June 1999     

Toxicity of sediments and pore water from Brunswick Estuary,Georgia

A chlor-alkali plant in Brunswick, Georgia, USA, discharged >2 kg mercury/d into a tributary of the Turtle River-Brunswick Estuary from 1966 to 1971. Mercury concentrations in sediments collected in 1989 along the tributary near the chlor-alkali plant ranged from 1 to 27 μg/g (dry weight), with the highest concentrations found in surface (0–8 cm) sediments of subtidal zones in the vicinity of the discharge site. Toxicity screening in 1990 using Microtox^® bioassays on pore water extracted on site from sediments collected at six stations distributed along the tributary indicated that pore water was highly toxic near the plant discharge. Ten-day toxicity tests on pore water from subsequent sediment samples collected near the plant discharge confirmed high toxicity to Hyalella azteca, and feeding activity was significantly reduced in whole-sediment tests. In addition to mercury in the sediments, other metals (chromium, lead, and zinc) exceeded 50 μg/g, and polychlorobiphenyl (PCB) concentrations ranged from 67 to 95 μg/g. On a molar basis, acid-volatile sulfide concentrations (20–45 μmol/g) in the sediments exceeded the metal concentrations. Because acid-volatile sulfides bind with cationic metals and form metal sulfides, which are generally not bioavailable, toxicities shown by these sediments were attributed to the high concentrations of PCBs and possibly methylmercury.     

The Effects of Sieving and Spatial Variability of Estuarine Sediment Toxicity Samples on Sediment Chemistry

In 1998, we conducted a field-validation study of the chronic 28-day whole-sediment toxicity test with Leptocheirus plumulosus in Baltimore Harbor, MD, an area where this amphipod is indigenous. This study included an evaluation of the effect of sieving on sediment chemical concentrations and the use of field replicates, or separate grabs from the same site, which provided an estimation of within-site chemical and toxicologic variability. Six stations in Baltimore Harbor, MD, were included in this evaluation. Chemical analysis of two separate unsieved field replicates from the six sites indicated that, overall, the chemical concentrations of replicates within each site were similar, especially for metals. Organic contaminants particularly total PCBs, had the highest variability between replicates. Chemical variability did not appear to be related to differences in organic carbon content or grain size or to variability in toxicologic end points. Results supported the use of composite samples in sediment toxicity tests. In addition, in most cases, sieving had little effect on sediment chemistry. For the metals and trace elements, only selenium showed a substantial change after sieving, with some samples increasing after sieving and others decreasing. Concentrations of acid-volatile sulfide (AVS) increased 194.6% at one station after sieving, although in most other cases, AVS and simultaneously extracted metals remained relatively unchanged. As expected, concentrations of organics generally decreased after sieving, but in the majority of cases this decrease was small (i.e., coefficient of variation 25%). Total benzene hexachloride and total chlordanes had the greatest changes, whereas polychlorinated biphenyl concentrations decreased at only two stations after sieving. Concentrations of polyaromatic hydrocarbons showed little change after sieving. These changes in sediment chemistry due to sieving must be viewed in the larger context of the potentially confounding effects that indigenous organisms may have on the interpretation of test results from whole-sediment toxicity tests.     

Assessment of supercritical fluid extraction use in whole sediment toxicity identification evaluations

Supercritical fluid extraction (SFE) with pure CO(2) was assessed as a confirmatory tool in phase III of whole sediment toxicity identification evaluations (TIEs). The SFE procedure was assessed on two reference sediments and three contaminated sediments by using a combination of toxicological and chemical measurements to quantify effectiveness. Sediment toxicity pre- and post-SFE treatment was quantified with a marine amphipod (Ampelisca abdita) and mysid (Americamysis bahia), and nonionic organic contaminants (NOCs) polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) were measured in sediments, overlying waters, and interstitial waters. In general, use of SFE with the reference sediments was successful, with survival averaging 91% in post-SFE treatments. Substantial toxicity reductions and contaminant removal from sediments and water samples generated from extracted sediments of up to 99% in two of the contaminated sediments demonstrated SFE effectiveness. Furthermore, toxicological responses for these SFE-treated sediments showed comparable results to those from the same sediments treated with the powdered coconut charcoal addition manipulation. These data demonstrated the utility of SFE in phase III of a whole sediment TIE. Conversely, in one of the contaminated sediments, the SFE treatments had no effect on sediment toxicity, whereas sediment concentrations of PCBs and PAHs were reduced. We propose that, for some sediments, the SFE treatment may result in the release of otherwise nonbioavailable cationic metals that subsequently cause toxicity to test organisms. Overall, SFE treatment was found to be effective for reducing the toxicity and concentrations of NOCs in some contaminated sediments. However, these studies suggest that SFE treatment may enhance toxicity with some sediments, indicating that care must be taken when applying SFE and interpreting the results.     

Use of Zeolite for Removing Ammonia and Ammonia-Caused Toxicity in Marine Toxicity Identification Evaluations

Ammonia occurs in marine waters including effluents, receiving waters, and sediment interstitial waters. At sufficiently high concentrations, ammonia can be toxic to aquatic species. Toxicity identification evaluation (TIE) methods provide researchers with tools for identifying aquatic toxicants. For identifying ammonia toxicity, there are several possible methods including pH alteration and volatilization, Ulva lactuca addition, microbial degradation, and zeolite addition. Zeolite addition has been used successfully in freshwater systems to decrease ammonia concentrations and toxicity for several decades. However, zeolite in marine systems has been used less because ions in the seawater interfere with zeolites ability to adsorb ammonia. The objective of this study was to develop a zeolite method for removing ammonia from marine waters. To accomplish this objective, we performed a series of zeolite slurry and column chromatography studies to determine uptake rate and capacity and to evaluate the effects of salinity and pH on ammonia removal. We also assessed the interaction of zeolite with several toxic metals. Success of the methods was also evaluated by measuring toxicity to two marine species: the mysid Americamysis bahia and the amphipod Ampelisca abdita. Column chromatography proved to be effective at removing a wide range of ammonia concentrations under several experimental conditions. Conversely, the slurry method was inconsistent and variable in its overall performance in removing ammonia and cannot be recommended. The metals copper, lead, and zinc were removed by zeolite in both the slurry and column treatments. The zeolite column was successful in removing ammonia toxicity for both the mysid and the amphipod, whereas the slurry was less effective. This study demonstrated that zeolite column chromatography is a useful tool for conducting marine water TIEs to decrease ammonia concentrations and characterize toxicity.