Darrin J. Greenstein

Analysis, occurrence, and toxic potential of pyrethroids, and fipronil in sediments from an urban estuary

Eight pyrethroids and fipronil and its three major degradates were analyzed in urban estuarine sediments that exhibited a range of toxic effects to an amphipod test species. Sediments from Ballona Creek, an urban estuary in Southern California (USA), collected during three dry season events were analyzed by gas chromatography with electron capture and negative chemical ionization mass spectrometric detection (GC-ECD and GC-NCI-MS). The two detection methods were in agreement for intermediate levels of pyrethroid contamination (10-50 ng/g dry wt) but deviated for both low and high concentrations (< 5 and > 50 ng/g). Sediments contained total pyrethroids as high as 473 ng/g with permethrin, bifenthrin, and cypermethrin as the most abundant compounds. In contrast, fipronil and its desulfinyl, sulfide, and sulfone degradates were detected at much lower levels (<or= 0.18-16 ng/g). Toxic units estimated for these compounds revealed that bifenthrin and cypermethrin were likely contributors to the mortality observed in tests with the estuarine amphipod Eohaustorius estuarius. Although fipronil was not a likely contributor to the observed mortality, the concentrations detected may be of concern for more sensitive crustacean species. Furthermore, the spatial pattern of pyrethroid contamination and potential toxicity was highly correlated with fine-grained substrate, which shifted to downstream stations within a three-month period during the dry season.

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 <or= 0.05) correlated with sediment chemical concentrations. Poor correspondence occurred between the toxicity endpoints and the indicators of benthic community condition. Differences in test characteristics such as mode of exposure, species-specific contaminant sensitivity, changes in contaminant bioavailability, and influence of noncontaminant stressors on the benthos may have been responsible for variation in response among the tests and low correspondence with benthic community condition. The influence of these factors cannot be easily predicted, underscoring the need to use multiple toxicity methods, in combination with other lines of evidence, to provide an accurate and confident assessment of sediment toxicity.

Pyrethroids in Southern California coastal sediments

Little is known about pyrethroid fate and effects in estuarine and marine environments. In the present study, the extent and magnitude of pyrethroids in coastal embayments of the Southern California Bight (SCB), USA, were assessed. Using a stratified probabilistic design, 155 sediment samples were collected from four embayment habitats (estuaries, marinas, open bays, and ports) and analyzed for eight common-use pyrethroids. Total pyrethroid concentrations ranged from less than 0.5 to 230 µg/kg dry weight (area-weighted mean concentration=5.1 ± 3.1 µg/kg) and were detected in 35% of the total SCB embayment area. Estuaries and marinas had the greatest areal extent of detectable concentrations (up to 65%) and the greatest area-weighted mean concentrations (22.1 ± 26.5 µg/kg). Sites with the greatest pyrethroid concentrations were located near sources of runoff from urban watersheds. Bifenthrin and cyfluthrin were detected in 32 and 15% of all samples, respectively, whereas the other six pyrethroids were detected in ≤ 5% of samples. Permethrin and bifenthrin had the highest concentrations at 132 and 65 µg/kg. Toxic units estimated for the marine amphipod Eohaustorius estuarius ranged from 0 to 5.8, exceeding unity in 9 and 32% of the total and estuary habitat areas, respectively, and were not correlated with mortality, suggesting that other factors (e.g., co-occurring contaminants, reduced bioavailability) may affect the predictive capability using a single test species.

Concentrations of methyl- tert-butyl ether (MTBE) in inputs and receiving waters of Southern California

The occurrence and concentration of the fuel additive methyl-tert-butyl ether (MTBE) were measured in dry weather runoff, municipal wastewater and industrial effluents, and coastal receiving waters in southern California. Combined, refineries and sewage treatment plants release approximately 214 kg day(-1) of MTBE into the marine environment, with Santa Monica Bay receiving most (98%) of this discharge. Dry weather urban runoff was analysed for samples collected from 25 streams and rivers, and accounted for less than 0.5% of the mass of MTBE discharged to coastal waters. Receiving water samples were collected from 23 stations in Santa Monica Bay, Los Angeles Harbour and Mission Bay or San Diego Bay. MTBE was detected at low concentrations near effluent discharges, however there was no evidence of baywide MTBE contamination related to these outfalls. Marinas and areas used intensively for recreational boating had the highest average MTBE concentration (8.8 microg l(-1)). Surface water contamination was most widespread in San Diego Bay and Mission Bay, areas with no refinery or sewage treatment plant inputs.

Municipal wastewater contamination in the Southern California Bight. Part II. Cytosolic distribution of contaminants and biochemical effects in fish livers

Abstract The objective of this study was to examine the cytosolic distribution of metals and oxygenated organic metabolites (MTBs), and biochemical effects, in livers of fish collected from both highly contaminated and less contaminated southern California coastal sites. Cytosolic extracts were separated by Sephadex G-75 column chromatography into high molecular weight (> 20 000 daltons) enzyme-containing (ENZ) pools, medium molecular weight (3000–20000 daltons) metallothionein- or metallothionein-like-containing (MT) pools, and low molecular weight ( Concentrations of Cd, Cu and Zn were frequently lower in cytosolic pools of longspine combfish, yellowchin sculpin, and California tonguefish from highly contaminated Palos Verdes (PV) relative to those from less contaminated Santa Monica Bay (SMB) despite much higher concentrations of these metals in sediments at PV. Patterns of cytosolic metal distribution differed more between metals than between species or sampling locations. Most Cd, Cu and Zn occurred in the MT pools of these three species, with the exception of Zn in California tonguefish which occurred predominately in the ENZ pool. In all three species, ENZ-Cu showed positive slopes when regressed against total cytosolic Cu, while ENZ-Cd showed no significant slopes when regressed against total cytosolic Cd. Patterns for Zn were the least consistent among species, with higher ENZ-Zn slopes occurring in fish livers with lower cytosolic Zn concentrations. The largest portion of DDT and PCB oxygenated MTBs occurred in GSH pools of scorpionfish livers from PV or less contaminated Cortes Bank (CB). Concentrations of MTBs in ENZ- and MT-pools of CB scorpionfish livers showed positive slopes when regressed against total cytosolic- and GSH-MTBs. Positive slopes for regressions of ENZ-Cu, -Zn and -MTBs against total cytosolic concentrations are consistent with the model of an equilibrium-dependent exchange of these among cytosolic pools. The lower metal concentrations, higher glutathione concentrations, and higher catalase activities found in fish from PV relative to those from SMB are in accordance with effects known to result from exposure to organic contaminants.

Genomic and phenotypic response of hornyhead turbot exposed to municipal wastewater effluents

Laboratory tests with marine flatfish were conducted to investigate associations among gene expression, higher biological responses and wastewater effluent exposure. In the present study, male hornyhead turbot (Pleuronichthys verticalis) were exposed to environmentally realistic (0.5%) and higher (5%) concentrations of chemically enhanced advanced-primary (PL) and full-secondary treated (HTP) effluents from two southern California wastewater treatment plants (WWTP). Hepatic gene expression was examined using a custom low-density microarray. Alterations in gene expression (vs. controls) were observed in fish exposed to both effluent types. Fish exposed to 0.5% PL effluent showed changes in genes involved in the metabolism of xenobiotics, steroids, and lipids, among other processes. Fish exposed to 5% PL effluent showed expression changes in genes involved in carbohydrate metabolism, stress responses, xenobiotic metabolism, and steroid synthesis, among others. Exposure to 5% HTP effluent changed the expression of genes involved in lipid, glutathione and xenobiotic metabolism, as well as immune responses. Although no concentration-dependent patterns of response to effluent exposure were found, significant Spearman correlations were observed between the expression of 22 genes and molecular and/or higher biological responses. These results indicate that microarray gene expression data correspond to higher biological responses and should be incorporated in studies assessing fish health after exposure to complex environmental mixtures.

Using Gene Expression to Assess the Status of Fish from Anthropogenically Influenced Estuarine Wetlands

The diverse mixture of contaminants frequently present in estuaries complicates their assessment by routine chemical or biological analyses. We investigated the use of gene expression to assess contaminant exposure and the condition of southern California estuarine fish. Liver gene expression, plasma estradiol concentrations, and gonad histopathology were used to study biological condition in longjaw mudsuckers (Gillichthys mirabilis). Metals, legacy organochlorine pesticides, PCBs, and contaminants of emerging concern were detected in sediments and whole fish. Overall gene expression patterns were characteristic to each of four sites investigated in this study. Differentially expressed genes belonged to several functional categories including xenobiotic metabolism, detoxification, disease, and stress responses. In general, plasma estradiol concentrations were similar among fish from all areas. Some fish gonads had pathologic changes (e.g., infection, inflammation) that could indicate weakened immune systems and chronic stress. The differential expression of some genes involved in stress responses correlated with the prevalence of histologic gonad lesions. This study indicates that gene expression is a promising tool for assessing the biological condition of fish exposed to environmental contaminants.

Gene expression of fathead minnows (Pimephales promelas) exposed to two types of treated municipal wastewater effluents.

Contaminants of emerging concern (CECs) in treated municipal effluents have the potential to adversely impact exposed organisms prompting elevated public concern. Using transcriptomic tools, we investigated changes in gene expression and cellular pathways in the liver of male fathead minnows (Pimephales promelas) exposed to 5% concentrations of full secondary-treated (HTP) or advanced primary-treated (PL) municipal wastewater effluents containing CECs. Gene expression changes were associated with apical end points (plasma vitellogenin and changes in secondary sexual characteristics). Of 32 effluent CECs analyzed, 28 were detected including pharmaceuticals, personal care products, hormones, and industrial compounds. Exposure to both effluents produced significantly higher levels of plasma VTG and changes in secondary sexual characteristics (e.g., ovipositor development). Transcript patterns differed between effluents, with <10% agreement in the detected response (e.g., altered production of transcripts involved in xenobiotic detoxification, oxidative stress, and apoptosis were observed following exposure to both effluents). Exposure to PL effluent caused changes in transcription of genes involved in metabolic pathways (e.g., lipid transport and steroid metabolism). Exposure to HTP effluent affected transcripts involved in signaling pathways (e.g., focal adhesion assembly and extracellular matrix). The results suggest a potential association between some transcriptomic changes and physiological responses following effluent exposure. This study identified responses in pathways not previously implicated in exposure to complex chemical mixtures containing CECs, which are consistent with effluent exposure (e.g., oxidative stress) in addition to other pathway responses specific to the effluent type.

Exposure of scorpionfish (Scorpaena guttata) to cadmium: biochemical effects of chronic exposure

Abstract Scorpionfish (Scorpaena guttata) were exposed to sublethal levels of cadmium in seawater (10 and 20 mg/l) for 4 wk. Measurements of enzyme activities were conducted in order to examine the relationship between toxic effects and the subcellular distribution of Cd, Cu and Zn. Liver, kidney, intestine and gill tissue were analyzed for the activity of the enzymes alkaline phosphatase, succinate dehydrogenase, glyceraldehyde phosphate dehydrogenase and CuZn-superoxide dismutase (SOD). Exposure to Cd resulted in the inhibition of only CuZn-SOD activity in the intestine. The activity of SOD was reduced by sixfold in intestines at the 20-mg Cd/l exposure level. Intestine SOD activity was also correlated positively with the concentration of Cu in the high-molecular-weight fraction of the cytosol (ENZ). The sulfhydryl-containing enzymes glyceraldehyde phosphate dehydrogenase and succinate dehydrogenase were not affected by Cd exposure, suggesting that the amounts of Cd found in the ENZ pool after the exposures examined here were insufficient to have a direct impact on enzyme activities. The mechanism of reduction of SOD activity in these fish appeared to be related instead to Cd-induced alterations of Cu and/or Zn metabolism.

Toxicity assessment of sediment cores from Santa Monica Bay, California

During the summer of 1997, sediment core samples were taken at 25 stations in Santa Monica Bay. Toxicity testing was performed on 4-cm sections of the entire length of each core using purple sea urchin fertilization and amphipod survival tests. The sea urchin test identified sections as being toxic at six stations, all located near current or former Hyperion Treatment Plant (HTP) wastewater outfall locations. The amphipod test identified sections from 17 stations as having toxic sediments. The stations having toxic sediments were scattered throughout the bay and toxicity was identified at numerous core depths. Spatial and temporal patterns indicated that toxicity was most strongly associated with the historical disposal of municipal wastewater sludge. Many of the sections toxic to the amphipods did not have chemical levels expected to cause toxicity and were in locations where a source of toxicity was not apparent.