An in situ toxicity identification evaluation method Part II: Field validation

When sediments are found to be toxic usually there is a mixture of chemicals present. Often it is important to establish which chemicals contribute to the toxicity. Establishing causality can be difficult and often requires fractionation with subsequent toxicity testing. The sample collection and manipulation process can alter chemical bioavailability and toxicity. An in situ toxicity identification evaluation (iTIE) chamber is described that was placed in sediments and fractionated pore-water chemicals into nonpolar chemicals, metals, and ammonia-type groups. This method was field tested and compared to the laboratory-based, U.S. Environmental Protection Agency (U.S. EPA) toxicity identification evaluation (TIE) method. Field studies were performed at three sites contaminated primarily with polycyclic aromatic hydrocarbons (PAHs) (Little Scioto River, OH, USA), polychlorinated biphenyls (PCBs) (Dicks Creek, OH, USA), and chlorobenzenes (Sebasticook River, ME, USA). Both the iTIE and the U.S. EPA TIE methods used Daphnia magna in 24-h exposures. Although the iTIE and TIE were conducted on sediments from the same location, there was significantly more toxicity observed in the iTIE testing. The dominant chemical classes were separated by the iTIE method and revealed which fractions contributed to toxicity. The loss of toxicity in the TIE approach did not allow for subsequent fractionation and stressor identification. Advantages of the iTIE over the TIE method were greater sensitivity and ability to detect causative toxic chemical fractions; lack of sediment collection and subsequent manipulation; and, thus, reduction in potential artifacts, more realistic exposure with slow, continual pore-water renewal in situ, ability to evaluate pore waters in sandy or rocky substrates where pore waters are difficult to collect, and a quicker phase I evaluation. Limitations of the iTIE method as compared to the TIE methods were extensive pretest assembly process, fewer phase I fractionation possibilities, and restriction to shallow waters. The results of these studies suggest that the iTIE method provides a more accurate and sensitive evaluation of pore water toxicity than the laboratory TIE method.     

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.     

Metal Content and Toxicity of Produced Formation Water (PFW): Study of the Possible Effects of the Discharge on Marine Environment

A preliminary chemical and ecotoxicological assessment was performed on the produced formation water (PFW) and superficial sediment around a gas platform (Fratello Cluster), located in the Adriatic Sea (Italy), in order to evaluate the effects of PFW discharged from the installation. The ecotoxicological bioassays, with the marine bacterium Vibrio fischeri and the sea urchin Paracentrotus lividus, were associated with chemical data to estimate the possible effects on living organisms. PFW collected on the platform was toxic, but no significant effect was recorded on marine sediment. Only the sediment station nearest to the discharge point showed higher values of some contaminants (zinc and arsenic) in comparison to other sites and only some stations showed low toxicity.     

Causes of water toxicity to Hyalella azteca in the New River, California, USA

The New River (CA, USA) was created in 1905 to 1907 when the Colorado River washed out diversionary works and flowed into the Salton Basin, creating the Salton Sea. Approximately 70% of the river's current flow is agricultural wastewater from the Imperial Valley. The river is contaminated with pesticides, industrial organic chemicals, metals, nutrients, bacteria, and silt. Monitoring for the State of California Surface Water Ambient Monitoring Program has indicated persistent water column toxicity to the epibenthic amphipod Hyalella azteca. Four toxicity identification evaluations (TIEs), along with chemical analyses, were performed, and the results indicated multiple and varying causes of toxicity. The first two TIEs characterized the causes of toxicity as a combination of metals and organics, but only the second sample contained enough total copper to contribute to toxicity. The third TIE used an emerging method for characterizing and identifying toxicity caused by pyrethroid pesticides. This TIE characterized organics as the cause of toxicity, and a carboxylesterase enzyme treatment further identified the cause of toxicity as pyrethroids. The final TIE used the enzyme and Phase II procedures to identify cypermethrin as the cause of toxicity. The TIE results demonstrate the evolving causes of toxicity in the New River and should assist regulators with implementing the total maximum daily load process for pesticides, particularly pyrethroids. Further research will determine if pyrethroids and other New River contaminants are having an impact on the Salton Sea.     

Evaluation and use of sediment toxicity reference sites for statistical comparisons in regional assessments

Sediment reference sites were used to establish toxicity standards against which to compare results from sites investigated in San Francisco Bay (California, USA) monitoring programs. The reference sites were selected on the basis of low concentrations of anthropogenic chemicals, distance from active contaminant sources, location in representative hydrographic areas of the Bay, and physical features characteristic of depositional areas (e.g., fine grain size and medium total organic carbon [TOC]). Five field-replicated sites in San Francisco Bay were evaluated over three seasons. Samples from each site were tested with nine toxicity test protocols and were analyzed for sediment grain size and concentrations of trace metals, trace organics, ammonia, hydrogen sulfide, and TOC. The candidate sites were found to have relatively low concentrations of measured chemicals and generally exhibited low toxicity. Toxicity data from the reference sites were then used to calculate numerical tolerance limits to be used as threshold values to determine which test sites had significantly higher toxicity than reference sites. Tolerance limits are presented for four standard test protocols, including solid-phase sediment tests with the amphipods Ampelisca abdita and Eohaustorius estuarius and sea urchin Strongylocentrotus purpuratus embryo/larval development tests in pore water and at the sediment-water interface (SWI). Tolerance limits delineating the lowest 10th percentile (0.10 quantile) of the reference site data distribution were 71% of the control response for Ampelisca, 70% for Eohaustorius, 94% for sea urchin embryos in pore water, and 87% for sea urchins embryos exposed at the SWI. The tolerance limits are discussed in terms of the critical values governing their calculation and the management implications arising from their use in determining elevated toxicity relative to reference conditions.     

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.     

Evidence of pesticide impacts in the Santa Maria River watershed, California, USA

The Santa Maria River provides significant freshwater and coastal habitat in a semiarid region of central California, USA. We conducted a water and sediment quality assessment consisting of chemical analyses, toxicity tests, toxicity identification evaluations, and macroinvertebrate bioassessments of samples from six stations collected during four surveys conducted between July 2002 and May 2003. Santa Maria River water samples collected downstream of Orcutt Creek (Santa Maria, Santa Barbara County, CA, USA), which conveys agriculture drain water, were acutely toxic to cladocera (Ceriodaphnia dubia), as were samples from Orcutt Creek. Toxicity identification evaluations (TIEs) suggested that toxicity to C. dubia in Orcutt Creek and the Santa Maria River was due to chlorpyrifos. Sediments from these two stations also were acutely toxic to the amphipod Hyalella azteca, a resident invertebrate. The TIEs conducted on sediment suggested that toxicity to amphipods, in part, was due to organophosphate pesticides. Concentrations of chlorpyrifos in pore water sometimes exceeded the 10-d median lethal concentration for H. azteca. Additional TIE and chemical evidence suggested sediment toxicity also partly could be due to pyrethroid pesticides. Relative to an upstream reference station, macroinvertebrate community structure was impacted in Orcutt Creek and in the Santa Maria River downstream of the Creek input. This study suggests that pesticide pollution likely is the cause of ecological damage in the Santa Maria River.     

A large-scale categorization of sites in San Francisco Bay, USA, based on the sediment quality triad, toxicity identification evaluations, and gradient studies

Sediment quality was assessed in San Francisco Bay, California, USA, using a two-tiered approach in which 111 sites were initially screened for sediment toxicity. Sites exhibiting toxicity were then resampled and analyzed for chemical contamination, recurrent toxicity, and, in some cases, benthic community impacts. Resulting data were compared with newly derived threshold values for each of the metrics in a triad-based weight-of-evidence evaluation. Sediment toxicity test results were compared with tolerance limits derived from reference site data, benthic community data were compared with threshold values for a relative benthic index based on the presence and abundance of pollution-tolerant and -sensitive taxa, and concentrations of chemicals and chemical mixtures were compared with sediment quality guideline-based thresholds. A total of 57 sites exceeded threshold values for at least one metric, and each site was categorized based on triad inferences. Nine sites were found to exhibit recurrent sediment toxicity associated with elevated contaminant concentrations, conditions that met program criteria for regulatory attention. Benthic community impacts were also observed at three of these sites, providing triad evidence of pollution-induced degradation. Multi- and univariate correlations indicated that chemical mixtures, heavy metals, chlordanes, and other organic compounds were associated with measured biological impacts in the Bay. Toxicity identification evaluations indicated that metals were responsible for pore-water toxicity to sea urchin larvae at two sites. Gradient studies indicated that the toxicity tests and benthic community metrics employed in the study predictably tracked concentrations of chemical mixtures in Bay sediments.     

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 of Parking Lot Runoff After Application of Simulated Rainfall

Stormwater runoff is an important source of toxic substances to the marine environment, but the effects of antecedent dry period, rainfall intensity, and duration on the toxicity of runoff are not well understood. In this study, simulated rainfall was applied to parking lots to examine the toxicity of runoff while controlling for antecedent period, intensity, and duration of rainfall. Parking areas were divided into high and low use and maintained and unmaintained treatments. The parking stalls were cleaned by pressure washing at time zero. Simulated rainfall was then applied to subplots of the parking lots so that antecedent periods of 1, 2, and 3 months were achieved, and all of the runoff was collected for analysis. On a separate parking lot, rainfall was applied at a variety of intensities and durations after a 3-month antecedent period. Runoff samples were tested for toxicity using the purple sea urchin fertilization test. Every runoff sample tested was found to be toxic. Mean toxicity for the sea urchin fertilization test ranged from 2.0 to 12.1 acute toxic units. The toxicity increased rapidly during the first month but then decreased approximately to precleaning levels and remained there. No difference in toxicity was found between the different levels of use or maintenance treatments. The intensity and duration of rainfall were inversely related to degree of toxicity. For all intensities tested, toxicity was always greatest in the first sampling time interval. Dissolved zinc was most likely the primary cause of toxicity based on toxicant characterization of selected runoff samples.     

Integrative ecotoxicological assessment of sediment in Portmán Bay (southeast Spain)

Portmán Bay, southeast Spain, contains the most seriously metal-contaminated sediments of the Mediterranean Sea. From 1958 to 1991, approximately 50 million tons of mine tailings were dumped into the bay, completely filling up the bay and dispersing over an extensive area of the continental platform and continental slope. The objective of our study was to characterize the nature and extent of metal contamination and the responses of natural communities to it and to assess the toxicity of the sediment deposits 10 years after mining had ceased. We studied the physical and chemical characteristics of the sediments and toxicity (of the porewater and sediment–water interface) using two sea urchin species (Arbacia lixula and Paracentrotus lividus). Metal bioavailability and patterns of macroinvertebrate community composition along the contamination gradient were also studied. Univariate and multivariate analyses showed positive correlation between the sediment metal concentrations associated to the all biological effects (sea urchins toxicity tests and benthic indices). The effects of sediment contamination on the benthic community structure are visible among sampling stations.     

Is the 1:4 elutriation ratio reliable? Ecotoxicological comparison of four different sediment: water proportions

Methodological research was carried out to evaluate the discriminatory capability of three toxicity bioassays toward different elutriation ratios (1:4, 1:20, 1:50, and 1:200 sediment:water ratios). Samples from six sampling stations of the Lagoon of Venice have been investigated. The toxicity bioassay results (sea urchin Paracentrotus lividus Lmk sperm cell and embryo toxicity bioassays and bivalve mollusk Crassostrea gigas Thunberg embryo toxicity bioassays) have shown that elutriates generated from the widely used 1:4 ratio were less toxic than those from intermediate ratios (1:20 and 1:50).     

Spatial and temporal variability in receiving water toxicity near an oil effluent discharge site

The distribution of point-source pollutants in the field can vary in both time and space. This study examined spatial and temporal patterns of toxicity from a produced water (an oil production effluent) discharge plume using a sea urchin fertilization toxicity test. Specifically, the sensitivity and response patterns of sea urchin gametes exposed to receiving waters sampled along a 1 Km transect near an active produced water outfall were tested. Fertilization success varied significantly with proximity to the outfall, with reduced fertilization found closer to the outfall. Although toxicity in receiving water samples, based on fertilization success, was variable in time—perhaps responding to variation in the quantity or make-up of produced water discharges–the general spatial pattern of toxicity along the transect remained relatively constant. The discharge plume was well established in the westerly direction throughout the experimental period. Toxicity data from samples of effluent and receiving waters, which were collected simultaneously, were used to determine the effective plume concentrations of produced water at seven sampling stations along a 1 km transect down-field from an active outfall. Strong evidence that field toxicity was directly attributable to the presence of produced water was provided by sampling the discharge plume during a period while the produced water discharge was not operating. During this period, no toxicity was found at any of the field sites.     

Bauxite manufacturing residues from Gardanne (France) and Portovesme (Italy) exert different patterns of pollution and toxicity to sea urchin embryos

This study was designed to investigate the composition and toxicity of solid residues from bauxite manufacturing plants. Soil and dust samples were collected in the proximity of two bauxite plants (Gardanne, France, and Portovesme, Italy). Samples were analyzed for their content of some selected inorganic contaminants by means of inductively coupled plasma optical emission spectroscopy (ICP-OES) either following acid digestion procedures or by seawater release of soluble components. Toxicity was tested by sea urchin bioassays to evaluate a set of toxicity endpoints including acute embryotoxicity, developmental defects, changes in sperm fertilization success, transmissible damage from sperm to the offspring, and cytogenetic abnormalities. Inorganic analysis showed two distinct sets of inorganic contaminants in Gardanne versus Portovesme, including Al, Cr, Cu, Fe, Mn, Pb, Ti, and Zn; sample composition (seawater-soluble contaminants) and toxicity showed a noteworthy association. The most severe toxicity to embryogenesis and to sperm fertilization success was exerted by some Portovesme samples (0.03-0.5% w/v), with a significant association between toxicity and dose-related seawater release of Zn, Pb, and Mn. Seawater extraction of a toxic dust sample (G20) from the Gardanne factory showed increasing seawater release of Al, Fe, and Mn; the G20 sample, at the level of 0.5%, affected both developing sea urchin embryos and sperm (offspring quality). Soil samples around the Gardanne factory showed the highest frequency of toxic soil sites eastward from the factory. The present data point to solid deposition from bauxite plants as a potential subject of environmental health concern. The results suggest that extraction methods for evaluating the toxicity of complex mixtures should be based on the environmental availability of mixture components. The differences in sample toxicity among the tested sites, however, suggest possible site-to-site variability in geochemical and/or technological parameters.     

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.     

Echinoderms as Bioindicators,Bioassays, and Impact Assessment Tools of Sediment-Associated Metals and PCBs in the North Sea

The study assessed the occurrence, possible toxicity, and impact of sediment-associated metals and PCBs in the coastal zone of the southern North Sea using echinoderms as representatives of the macrobenthos. Metals and PCBs were analyzed in the sediments and in the body compartments of the starfish Asterias rubens from 11 stations. The general toxicity of sediment-associated contaminants was assessed by bioassays using embryonic and larval developments of both A. rubens and the sea urchin Psammechinus miliaris. The impact of contamination was assessed by measuring cellular immune responses of A. rubens collected in the same stations.Contamination of the starfish by metals and PCBs closely reflected that of the sediments. However, bioaccumulation was element-specific for metals and depended on the chlorination pattern for PCBs. The sediment-associated contaminants appeared to be toxic in both the A. rubens and P. miliaris developmental assays. Moreover, metals were shown to affect the immune responses of starfishes living in contaminated stations. The most significant effects on biological responses were recorded in the plumes of the Scheldt/Rhine/North Sea Canal and the Elbe/Weser Rivers.     

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.     

Toxicological profiling of sediments using in vitro bioassays, with emphasis on endocrine disruption

In vitro bioassays are valuable tools for screening environmental samples for the presence of bioactive (e.g., endocrine-disrupting) compounds. They can be used to direct chemical analysis of active compounds in toxicity identification and evaluation (TIE) approaches. In the present study, five in vitro bioassays were used to profile toxic potencies in sediments, with emphasis on endocrine disruption. Nonpolar total and acid-treated stable extracts of sediments from 15 locations in the Rhine Meuse estuary area in The Netherlands were assessed. Dioxin-like and estrogenic activities (using dioxin-responsive chemical-activated luciferase gene expression [DR-CALUX] and estrogen-responsive chemical-activated luciferase gene expression [ER-CALUX] assays) as well as genotoxicity (UMU test) and nonspecific toxic potency (Vibrio fischeri assay) were observed in sediment extracts. For the first time, to our knowledge, in vitro displacement of thyroid hormone thyroxine (T4) from the thyroid hormone transport protein thransthyretin by sediment extracts was observed, indicating the presence of compounds potentially able to disrupt T4 plasma transport processes. Antiestrogenic activity was also observed in sediment. The present study showed the occurrence of endocrine-disrupting potencies in sediments from the Dutch delta and the suitability of the ER- and DR-CALUX bioassays to direct endocrine-disruption TIE studies.     

Toxicity ranking of estuarine sediments on the basis of Sparus aurata biomarkers

Sparus aurata biomarkers were used to rank sediments from the Sado River estuary (Portugal) according to their toxicity. Initially, the activities of liver ethoxyresorufin-O-deethylase, liver and gill glutathione S-transferases, muscle lactate dehydrogenase, and brain acetylcholinesterase were tested in a laboratory bioassay with the reference compound benzo[a]pyrene. Enzymatic activities were determined in different tissues of fish exposed for 48, 96, or 240 h to three concentrations of benzo[a]pyrene (25, 50, and 100 microg/L). Induction of liver ethoxyresorufin-O-deethylase was observed at all the exposure periods and concentrations, suggesting a continuous response of this system to toxicant exposure. Induction of liver glutathione S-transferases activity was only observed after 240 h of exposure, whereas gill glutathione S-transferases activity was significantly inhibited at all the exposure periods, suggesting a direct or indirect effect of the toxicant on these enzymes. Inhibition of lactate dehydrogenases activity was only observed after 96 h of exposure to 25 microg/L of benzo[a]pyrene. No significant effects were observed on acetylcholinesterase activity, suggesting that cholinergic function of S. aurata is not affected by benzo[a]pyrene. In a second phase, fish were exposed for 240 h to sediments collected at five sites of the Sado River estuary, and the same biomarkers were analyzed. For all the enzymes assayed, significant differences among sites were found. In this study, the battery of biomarkers used allowed to discrimination among sites with different types of contamination, levels of contamination, or both, after multivariate data analysis. Discrimination of sites was similar to the ranking provided by a more complex and parallel study (including chemical analysis of sediments, macrobenthic community analysis, amphipod mortality toxicity tests, and sea urchin abnormality embryo assays), suggesting its suitability to evaluate the toxicity of estuarine sediments.     

Toxicity of bauxite manufacturing by-products in sea urchin embryos.

By-products from a bauxite manufacturing plant located in Seydi?ehir, Turkey, were investigated for their composition and any toxicity to sea urchin embryogenesis. Samples from three other bauxite plants located in France, Greece, and Italy were simultaneously tested for toxicity in sea urchin embryos. Samples included sludge and solid residues in the plant and sediment and water columns from two holding ponds (red sludge or cryolite residues). Samples were analyzed for their inorganic content by inductively coupled plasma optical emission spectroscopy (ICP-OES). Analyses were carried out either following strong acid extraction or after release of soluble components from seawater-suspended pellets. Toxicity was tested by sea urchin bioassays, to evaluate the following endpoints: (a) acute and/or developmental toxicity, (b) changes in fertilization success, and (c) transmissible damage from sperm to offspring. The results revealed the following: (1) inorganic analysis, following strong acid extraction, showed a prevalence of Al and Fe; (2) seawater release of soluble contaminants was confined to Fe and Mn, whereas Al levels were not changed by suspending increasing sample amounts in seawater; (3) the most severe toxicity to sea urchin embryos was exerted by a 2% water column from the red sludge holding pond and by soil and sludge collected near the plant reactor; (4) sludge supernatant was the most toxic sample to sperm and offspring. The data showed a prevailing association of free Fe (and possibly Mn) levels with Seydi?ehir sample toxicity. The water column of the red sludge holding pond showed both excess levels of free Al and high pH, thus suggesting a combined effect. The differences in sample toxicity in the Seydi?ehir plant compared with other bauxite manufacturing plants suggest a possible variable toxicity as related to bauxite ore composition and/or manufacturing processes.