Jennifer P. Voorhees

Characterization of the metabolic actions of crude versus dispersed oil in salmon smolts via NMR-based metabolomics.

With maritime transport of crude oil from Alaska to California, there is significant potential for a catastrophic spill which could impact migrating salmon. Therefore, this study compared the lethal and sublethal metabolic actions of the water-accommodated fraction (WAF) and the chemically enhanced WAF (CEWAF, via Corexit 9500) of Prudhoe Bay crude oil in smolts of Chinook salmon (Onchorhyncus tshawytscha). After 96-h exposure to the CEWAF, the resulting LC50 was some 20 times higher (i.e., less toxic) than that of the WAF. Muscle and liver samples from surviving fish were collected and low-molecular weight metabolites were analyzed using one-dimensional (1)H and projections of two-dimensional (1)H J-resolved NMR. Principal component analysis (PCA), employed to analyze NMR spectra and identify most variance from the samples, revealed age-related metabolic changes in the fish within the replicated studies, but few consistent metabolic effects from the treatments. However, ANOVA results demonstrated that the dose-response metabolite patterns are both metabolite- and organ-dependent. In general, exposure to either WAF or CEWAF resulted in an increase of amino acids (i.e., valine, glutamine and glutamate) and a decrease of both organic osmolytes (i.e., glycerophosphorylcholine) and energetic substrates (i.e., succinate). The simultaneous increase of formate and decrease of glycerophosphorylcholine in the liver, or the decrease of glycerophosphorylcholine in muscle, may serve as sensitive sublethal biomarkers for WAF or CEWAF exposures, respectively. In conclusion, dispersant treatment significantly decreased the lethal potency of crude oil to salmon smolts, and the NMR-based metabolomics approach provided a sensitive means to characterize the sublethal metabolic actions.

Evaluation of methods to determine causes of sediment toxicity in San Diego Bay, California, USA.

Regulation of waterbodies impaired due to sediment toxicity may require development of Total Maximum Daily Load (TMDL) allocations to reduce chemicals of concern. A key step in this process is the identification of chemicals responsible for toxicity, and sediment toxicity identification evaluation procedures (TIEs) are the primary tools used to accomplish this. Several sites in San Diego Bay (CA, USA) are listed as impaired due to sediment toxicity associated with organic chemicals and metals, and due to degraded benthic macroinvertebrate communities. Sediment was collected from one of these sites, at the confluence of Switzer Creek in San Diego Harbor. The sediment was subjected to selected whole-sediment TIE treatments to evaluate the efficacy of these procedures for identifying the causes of toxicity at Switzer Creek. Toxicity was assessed using the estuarine amphipod Eohaustorius estuarius. The results indicated that toxicity of San Diego Bay sediment was likely partly due to mixtures of pyrethroid pesticides. These experiments showed that the effectiveness of the individual TIE procedures varied by treatment. Variability was mainly due to inconsistency between results of samples subjected to various Phase II TIE procedures, including chemical analyses of samples subjected to high-pressure liquid chromatography and direct analyses of acetone extractions of carbonaceous resin. The procedures require further refinement to ensure maximum sorption and complete elution and detection of sorbed chemicals. Despite these inconsistencies, the results indicate the utility of these procedures for identifying chemicals of concern in this system.

Pyrethroid and organophosphate pesticide‐associated toxicity in two coastal watersheds (California, USA)

Portions of the Santa Maria River and Oso Flaco Creek watersheds in central California, USA, are listed as impaired under section 303(d) of the Clean Water Act and require development of total maximum daily load (TMDL) allocations. These listings are for general pesticide contamination, but are largely based on historic monitoring of sediment and fish tissue samples that showed contamination by organochlorine pesticides. Recent studies have shown that toxicity in these watersheds is caused by organophosphate pesticides (water and sediment) and pyrethroid pesticides (sediment). The present study was designed to provide information on the temporal and spatial variability of toxicity associated with these pesticides to better inform the TMDL process. Ten stations were sampled in four study areas, one with urban influences, and the remaining in agriculture production areas. Water toxicity was assessed with the water flea Ceriodaphnia dubia, and sediment toxicity was assessed with the amphipod Hyalella azteca. Stations in the lower Santa Maria River had the highest incidence of toxicity, followed by stations influenced by urban inputs. Toxicity identification evaluations and chemical analysis demonstrated that the majority of the observed water toxicity was attributed to organophosphate pesticides, particularly chlorpyrifos, and that sediment toxicity was caused by mixtures of pyrethroid pesticides. The results demonstrate that both agriculture and urban land uses are contributing toxic concentrations of these pesticides to adjacent watersheds, and regional water quality regulators are now using this information to develop management objectives.

Evaluation of phase II toxicity identification evaluation methods for freshwater whole sediment and interstitial water.

Phase I whole sediment toxicity identification evaluation (TIE) methods have been developed to characterize the cause of toxicity as organic chemicals, metals, or ammonia. In Phase II identification treatments, resins added to whole sediment to reduce toxicity caused by metals and organics can be separated and eluted much like solid-phase extraction (SPE) columns are eluted for interstitial water. In this study, formulated reference sediments spiked with toxic concentrations of copper, fluoranthene, and nonylphenol were subjected to whole sediment and interstitial water TIE treatments to evaluate Phase I and II TIE procedures for identifying the cause of toxicity to Hyalella azteca. Phase I TIE treatments consisted of adding adsorbent resins to whole sediment, and using SPE columns to remove spiked chemicals from interstitial water. Phase II treatments consisted of eluting resins and SPE columns and the preparation and testing of eluates for toxicity and chemistry. Whole sediment resins and SPE columns significantly reduced toxicity, and the eluates from all treatments contained toxic concentrations of the spiked chemical except for interstitial water fluoranthene. Toxic unit analysis based on median lethal concentrations (LC50s) allowed for the comparison of chemical concentrations among treatments, and demonstrated that the bioavailability of some chemicals was reduced in some samples and treatments. The concentration of fluoranthene in the resin eluate closely approximated the original interstitial water concentration, but the resin eluate concentrations of copper and nonylphenol were much higher than the original interstitial water concentrations. Phase II whole sediment TIE treatments provided complementary lines of evidence to the interstitial water TIE results.

NMR-based characterization of the acute metabolic effects of weathered crude and dispersed oil in spawning topsmelt and their embryos

Oil spill responders require information on the relative toxicity of dispersed and un-dispersed oil in order to make informed decisions regarding the use of chemical dispersants during spill events. Toxicity of the water-accommodated fraction (WAF) and the chemically-enhanced WAF (CEWAF; via the dispersant Corexit 9500) of weathered Prudhoe Bay crude oil was investigated using adult and embryonic topsmelt; topsmelt are an ecologically important atherinid in California bays and estuaries and an important indicator species. Following 96-h exposures, metabolite profiles were measured using 1D (1)H nuclear magnetic resonance (NMR) spectroscopy and compared via principal component analysis. Similar metabolic profiles were obtained between WAF- and CEWAF-exposed adults and embryos. Although metabolic changes, for the adults lacked significance, significant increasing and decreasing metabolic changes were observed for embryos directly exposed. Furthermore, no mortality was observed for embryos, exposed to WAF and normal development occurred, whereas CEWAF exposed embryos lead to mortality and cardiovascular abnormalities. Observed toxicological information, specifically for developing fish, can aide resource managers in the relative risk of treating oil spills with dispersant.

Bioswales reduce contaminants associated with toxicity in urban storm water

Contamination and toxicity associated with urban storm water runoff are a growing concern because of the potential impacts on receiving systems. California water regulators are mandating implementation of green infrastructure as part of new urban development projects to treat storm water and increase infiltration. Parking lot bioswales are low impact development practices that promote filtering of runoff through plants and soil. Studies have demonstrated that bioswales reduce concentrations of suspended sediments, metals, and hydrocarbons. There have been no published studies evaluating how well these structures treat current-use pesticides, and studies have largely ignored whether bioswales reduce toxicity in surface water. Three storms were monitored at 3 commercial and residential sites, and reductions of contaminants and associated toxicity were quantified. Toxicity testing showed that the majority of untreated storm water samples were toxic to amphipods (Hyalella azteca) and midges (Chironomus dilutus), and toxicity was reduced by the bioswales. No samples were toxic to daphnids (Ceriodaphnia dubia) or fish (Pimephales promelas). Contaminants were significantly reduced by the bioswales, including suspended solids (81% reduction), metals (81% reduction), hydrocarbons (82% reduction), and pyrethroid pesticides (74% reduction). The single exception was the phenypyrazole pesticide fipronil, which showed inconsistent treatment. The results demonstrate these systems effectively treat contaminated storm water associated with surface water toxicity but suggest that modifications of their construction may be required to treat some contaminant classes. Environ Toxicol Chem 2016;35:3124-3134.

Hypersalinity Toxicity Thresholds for Nine California Ocean Plan Toxicity Test Protocols

Currently, several desalination facilities have been proposed to operate or are actually operating in California. These facilities’ use of reverse osmosis (RO) may discharge hypersaline reject brine into the marine environment. The risks, if any, this brine would pose to coastal receiving waters are unknown. To test the toxicity of hypersaline brine in the absence of any additional toxic constituents, we prepared brine and tested it with the seven toxicity test organisms listed in the 2009 California Ocean Plan. The most sensitive protocols were the marine larval development tests, whereas the most tolerant to increased salinities were the euryhaline topsmelt, mysid shrimp, and giant kelp tests. Reject brines from the Monterey Bay Aquarium’s RO desalination facility were also tested with three species. The effects of the aquarium’s brine effluent on topsmelt, mussels, and giant kelp were consistent with those observed in the salinity tolerance experiments. This information will be used by regulators to establish receiving water limitations for hypersaline discharges.

Monitoring the aquatic toxicity of mosquito vector control spray pesticides to freshwater receiving waters

Pesticides are applied to state and local waterways in California to control insects such as mosquitoes, which are known to serve as a vector for West Nile Virus infection of humans. The California State Water Resources Control Board adopted a National Pollutant Discharge Elimination System General Permit to address the discharge to waters of the United States of pesticides resulting from adult and larval mosquito control. Because pesticides used in spray activities have the potential to cause toxicity to nontarget organisms in receiving waters, the current study was designed to determine whether toxicity testing provides additional, useful environmental risk information beyond chemical analysis in monitoring spray pesticide applications. Monitoring included a combination of aquatic toxicity tests and chemical analyses of receiving waters from agricultural, urban, and wetland habitats. The active ingredients monitored included the organophosphate pesticides malathion and naled, the pyrethroid pesticides etofenprox, permethrin, and sumithrin, pyrethrins, and piperonyl butoxide (PBO). Approximately 15% of the postapplication water samples were significantly toxic. Toxicity of half of these samples was attributed to the naled breakdown product dichlorvos. Toxicity of 2 other water samples likely occurred when PBO synergized the effects of pyrethroid pesticides that were likely present in the receiving system. Four of 43 postapplication sediment samples were significantly more toxic than their corresponding pre-application samples, but none of the observed toxicity was attributed to the application events. These results indicate that many of the spray pesticides used for adult mosquito control do not pose significant acute toxicity risk to invertebrates in receiving systems. In the case of naled in water, analysis of only the active ingredient underestimated potential impacts to the receiving system, because toxicity was attributed to the breakdown product, dichlorvos. Toxicity testing can provide useful risk information about unidentified, unmeasured toxicants or mixtures of toxicants. In this case, toxicity testing provided information that could lead to the inclusion of dichlorvos monitoring as a permit requirement.

An integrated vegetated ditch system reduces chlorpyrifos loading in agricultural runoff

Agricultural runoff containing toxic concentrations of the organophosphate pesticide chlorpyrifos has led to impaired water body listings and total maximum daily load restrictions in Californias central coast watersheds. Chlorpyrifos use is now tightly regulated by the Central Coast Regional Water Quality Control Board. This study evaluated treatments designed to reduce chlorpyrifos in agricultural runoff. Initial trials evaluated the efficacy of 3 different drainage ditch installations individually: compost filters, granulated activated carbon (GAC) filters, and native grasses in a vegetated ditch. Treatments were compared to bare ditch controls, and experiments were conducted with simulated runoff spiked with chlorpyrifos at a 1.9 L/s flow rate. Chlorpyrifos concentrations and toxicity to Ceriodaphnia dubia were measured at the input and output of the system. Input concentrations of chlorpyrifos ranged from 858 ng/L to 2840 ng/L. Carbon filters and vegetation provided the greatest load reduction of chlorpyrifos (99% and 90%, respectively). Toxicity was completely removed in only one of the carbon filter trials. A second set of trials evaluated an integrated approach combining all 3 treatments. Three trials were conducted each at 3.2 L/s and 6.3 L/s flow rates at input concentrations ranging from 282 ng/L to 973 ng/L. Chlorpyrifos loadings were reduced by an average of 98% at the low flow rate and 94% at the high flow rate. Final chlorpyrifos concentrations ranged from nondetect (<50 ng/L) to 82 ng/L. Toxicity to C. dubia was eliminated in 3 of 6 integrated trials. Modeling of the ditch and its components informed design alterations that are intended to eventually remove up to 100% of pesticides and sediment. Future work includes investigating the adsorption capacity of GAC, costs associated with GAC disposal, and real-world field trials to further reduce model uncertainties and confirm design optimization. Trials with more water-soluble pesticides such as neonicotinoids are also recommended. Integr Environ Assess Manag 2017;13:423-430.

Relative toxicity of bifenthrin to Hyalella azteca in 10 day versus 28 day exposures.

Many watersheds in the Central Valley region of California are listed as impaired due to pyrethroid-associated sediment toxicity. The Central Valley Regional Water Quality Control Board is developing numeric sediment quality criteria for pyrethroids, beginning with bifenthrin. Criteria are being developed using existing data, along with data from 10 d and 28 d toxicity tests with Hyalella azteca conducted as part of the current study. A single range-finder and 2 definitive tests were conducted for each test duration. Median lethal concentrations (LC50s), as well as LC20s and inhibition concentrations (IC20s) were calculated based on measured whole sediment bifenthrin concentrations and interstitial water concentrations. Sediment LC50s were also corrected for organic C content. Average LC50s were not significantly different in 10 d versus 28 d tests with H. azteca: 9.1 and 9.6 ng/g bifenthrin for 10 d and 28 d tests, respectively. Average LC20 values were also similar with concentrations at 7.1 and 7.0 for 10 d and 28 d tests, respectively. Bifenthrin inhibition concentrations (IC20s) based on amphipod growth were variable, particularly in the 28 d tests, where a clear dose-response relationship was observed in only 1 of the definitive experiments. Average amphipod growth IC20s were 3.9 and 9.0 ng/g for 10 d and 28 d tests, respectively. Amphipod growth calculated as biomass resulted in IC20s of 4.1 and 6.3 ng/g for the 10 d and 28 d tests, respectively. Lack of a clear growth effect in the longer term test may be related to the lack of food adjustment to account for amphipod mortality in whole sediment exposures. The average C-corrected LC50s were 1.03 and 1.09 μg/g OC for the 10 d and 28 d tests, respectively. Interstitial water LC50s were determined as the measured dissolved concentration of bifenthrin relative to interstitial water dissolved organic carbon. The average LC50s for dissolved interstitial water bifenthrin were 4.23 and 4.28 ng/L for the 10 d and 28 d tests, respectively. In addition, a set of 10 d and 28 d tests were conducted at 15 °C to assess the relative toxicity of bifenthrin at a lower temperature than the standard 23 °C test temperature. These results showed that bifenthrin was more toxic at the lower temperature, with LC50s of 5.1 and 3.4 ng/g bifenthrin in 10 d and 28 d tests, respectively. Amphipod growth at 15 °C after a 28 d exposure resulted in the lowest effect concentration of all experiments conducted (IC20 = 0.61 ng/g). This article discusses how bifenthrin dose-response data from 10 d and 28 d exposures inform development of sediment quality criteria for this pesticide for California Central Valley watersheds.