Kim A. Anderson

Impact of the deepwater horizon oil spill on bioavailable polycyclic aromatic hydrocarbons in Gulf of Mexico coastal waters.

An estimated 4.1 million barrels of oil and 2.1 million gallons of dispersants were released into the Gulf of Mexico during the Deepwater Horizon oil spill. There is a continued need for information about the impacts and long-term effects of the disaster on the Gulf of Mexico. The objectives of this study were to assess bioavailable polycyclic aromatic hydrocarbons (PAHs) in the coastal waters of four Gulf Coast states that were impacted by the spill. For over a year, beginning in May 2010, passive sampling devices were used to monitor the bioavailable concentration of PAHs. Prior to shoreline oiling, baseline data were obtained at all the study sites, allowing for direct before and after comparisons of PAH contamination. Significant increases in bioavailable PAHs were seen following the oil spill, however, preoiling levels were observed at all sites by March 2011. A return to elevated PAH concentrations, accompanied by a chemical fingerprint similar to that observed while the site was being impacted by the spill, was observed in Alabama in summer 2011. Chemical forensic modeling demonstrated that elevated PAH concentrations are associated with distinctive chemical profiles.

Neurodevelopmental low-dose bisphenol A exposure leads to early life-stage hyperactivity and learning deficits in adult zebrafish

Developmental bisphenol A (BPA) exposure has been implicated in adverse behavior and learning deficits. The mode of action underlying these effects is unclear. The objectives of this study were to identify whether low-dose, developmental BPA exposure affects larval zebrafish locomotor behavior and whether learning deficits occur in adults exposed during development. Two control compounds, 17β-estradiol (an estrogen receptor ligand) and GSK4716 (a synthetic estrogen-related receptor gamma ligand), were included. Larval toxicity assays were used to determine appropriate BPA, 17β-estradiol, and GSK4716 concentrations for behavior testing. BPA tissue uptake was analyzed using HPLC and lower doses were extrapolated using a linear regression analysis. Larval behavior tests were conducted using a ViewPoint Zebrabox. Adult learning tests were conducted using a custom-built T-maze. BPA exposure to <30μM was non-teratogenic. Neurodevelopmental BPA exposure to 0.01, 0.1, or 1μM led to larval hyperactivity or learning deficits in adult zebrafish. Exposure to 0.1μM 17β-estradiol or GSK4716 also led to larval hyperactivity. This study demonstrates the efficacy of using the zebrafish model for studying the neurobehavioral effects of low-dose developmental BPA exposure.

Comparative developmental toxicity of environmentally relevant oxygenated PAHs

Oxygenated polycyclic aromatic hydrocarbons (OPAHs) are byproducts of combustion and photo-oxidation of parent PAHs. OPAHs are widely present in the environment and pose an unknown hazard to human health. The developing zebrafish was used to evaluate a structurally diverse set of 38 OPAHs for malformation induction, gene expression changes and mitochondrial function. Zebrafish embryos were exposed from 6 to 120h post fertilization (hpf) to a dilution series of 38 different OPAHs and evaluated for 22 developmental endpoints. AHR activation was determined via CYP1A immunohistochemistry. Phenanthrenequinone (9,10-PHEQ), 1,9-benz-10-anthrone (BEZO), xanthone (XAN), benz(a)anthracene-7,12-dione (7,12-B[a]AQ), and 9,10-anthraquinone (9,10-ANTQ) were evaluated for transcriptional responses at 48hpf, prior to the onset of malformations. qRT-PCR was conducted for a number of oxidative stress genes, including the glutathione transferase(gst), glutathione peroxidase(gpx), and superoxide dismutase(sod) families. Bioenergetics was assayed to measure in vivo oxidative stress and mitochondrial function in 26hpf embryos exposed to OPAHs. Hierarchical clustering of the structure-activity outcomes indicated that the most toxic of the OPAHs contained adjacent diones on 6-carbon moieties or terminal, para-diones on multi-ring structures. 5-carbon moieties with adjacent diones were among the least toxic OPAHs while the toxicity of multi-ring structures with more centralized para-diones varied considerably. 9,10-PHEQ, BEZO, 7,12-B[a]AQ, and XAN exposures increased expression of several oxidative stress related genes and decreased oxygen consumption rate (OCR), a measurement of mitochondrial respiration. Comprehensive in vivo characterization of 38 structurally diverse OPAHs indicated differential AHR dependency and a prominent role for oxidative stress in the toxicity mechanisms.

Developmental Exposure to Polychlorinated Biphenyls Interferes with Experience-Dependent Dendritic Plasticity and Ryanodine Receptor Expression in Weanling Rats

Background Neurodevelopmental disorders are associated with altered patterns of neuronal connectivity. A critical determinant of neuronal connectivity is the dendritic morphology of individual neurons, which is shaped by experience. The identification of environmental exposures that interfere with dendritic growth and plasticity may, therefore, provide insight into environmental risk factors for neurodevelopmental disorders. Objective We tested the hypothesis that polychlorinated biphenyls (PCBs) alter dendritic growth and/or plasticity by promoting the activity of ryanodine receptors (RyRs). Methods and Results The Morris water maze was used to induce experience-dependent neural plasticity in weanling rats exposed to either vehicle or Aroclor 1254 (A1254) in the maternal diet throughout gestation and lactation. Developmental A1254 exposure promoted dendritic growth in cerebellar Purkinje cells and neocortical pyramidal neurons among untrained animals but attenuated or reversed experience-dependent dendritic growth among maze-trained littermates. These structural changes coincided with subtle deficits in spatial learning and memory, increased [3H]-ryanodine binding sites and RyR expression in the cerebellum of untrained animals, and inhibition of training-induced RyR upregulation. A congener with potent RyR activity, PCB95, but not a congener with negligible RyR activity, PCB66, promoted dendritic growth in primary cortical neuron cultures and this effect was blocked by pharmacologic antagonism of RyR activity. Conclusions Developmental exposure to PCBs interferes with normal patterns of dendritic growth and plasticity, and these effects may be linked to changes in RyR expression and function. These findings identify PCBs as candidate environmental risk factors for neurodevelopmental disorders, especially in children with heritable deficits in calcium signaling.

Determination of parent and substituted polycyclic aromatic hydrocarbons in high-fat salmon using a modified QuEChERS extraction, dispersive SPE and GC-MS

A fast and easy modified QuEChERS (quick, easy, cheap, rugged and safe) extraction method has been developed and validated for determination of 33 parent and substituted polycyclic aromatic hydrocarbons (PAHs) in high-fat smoked salmon that greatly enhances analyte recovery compared to traditional QuEChERS procedures. Sample processing includes extraction of PAHs into a solution of ethyl acetate, acetone and isooctane followed by cleanup with dispersive SPE and analysis by GC-MS in SIM mode. Method performance was assessed in spike recovery experiments (500 μg/g wet weight) in three commercially available smoked salmon with 3-11% fat. Recoveries of some 2-, 3- and 5-ring PAHs were improved 50-200% over traditional methods, while average recovery across all PAHs was improved 67%. Method precision was good with replicate extractions typically yielding relative standard deviations <10%, and detection limits were in the low ng/g range. With this method, a single analyst could extract and clean up ≥60 samples for PAH analysis in an 8 h work day.

Ketone and quinone-substituted polycyclic aromatic hydrocarbons in mussel tissue, sediment, urban dust, and diesel particulate matrices.

Polycyclic aromatic hydrocarbons (PAHs) substituted with a ketone or quinone functionality (OPAHs) may be important environmental contaminants. The OPAHs from environmental samples have demonstrated toxicity and may be more harmful than PAHs. Knowledge gaps concerning the occurrence of OPAHs in the total environment arise from analytical difficulties, as well as limited standards and methodologies. An optimized method was developed to quantify five ketone and four quinone OPAHs from matrices ranging from biological tissue to diesel particulates. Five National Institute of Standards and Technology Standard Reference Materials (SRMs) were analyzed. This is the first report of OPAH quantitation in SRM 2977 (mussel tissue), SRM 1944 (New York/New Jersey, USA waterway sediment), SRM 1975 (diesel extract), and SRM 1650b (diesel particulate matter) and among the few to report concentrations from SRM 1649 (urban dust). Furthermore, this is one of the first reports of OPAHs in biological tissue. Σ₉OPAHs were 374 ± 59 mg/kg (mussel tissue), 5.4 ± 0.5 mg/kg (sediment), 16.9 ± 1.6 mg/kg (urban dust), 33.4 ± 0.4 mg/kg (diesel extract), and 150 ± 43 mg/kg (diesel particulate matter). In all SRMs, the levels of OPAHs were similar to or exceeded levels of PAHs. Of the OPAHs tested, the most frequently occurring in the environmental matrices were 9-fluorenone, 9,10-anthraquinone, benzofluorenone, and 7,12-benz[a]anthracenequinone.

Trace element concentration in tree-rings biomonitoring centuries of environmental change.

Inductively coupled plasma mass spectrometry (ICP-MS) was used to examine trace element concentration in tree-rings over three and half centuries to assess macro-trends of environmental change. Tree-rings of a 350+ year old mammoth ponderosa pine (Pinus ponderosa) were analyzed for element concentration and evaluated versus local and global historical events. The ponderosa pine was located 100 miles south of the Canada/USA border and 180 miles east of the Pacific Ocean, and grew near apple orchards, a public road, and Swakane Creek in western Washington, USA. The elements tested did not all display the same time versus concentration patterns. Copper and chromium displayed cyclic concentration patterns over the last 350+ years, which appear to be associated with local events. Strontium, barium, zinc and cadmium were found to be relatively constant between the mid 1600s and the early 1800s. Strontium, barium, zinc, and cadmium then increased beginning in the early 1800s for approximately 50 years then decreased to present day 2000. Significantly, similar changes seen in Ca, Mg, and Zn in other studies have been attributed to acid rain, whereas, in our study area there is no history of anthropogenic acid rain. Most importantly, our data goes back to the mid-1600s several hundred years further back than most other studies of this nature. This additional time data provides for a better context of trend data not previously available.

Silicone Wristbands as Personal Passive Samplers

Active-sampling approaches are commonly used for personal monitoring, but are limited by energy usage and data that may not represent an individual’s exposure or bioavailable concentrations. Current passive techniques often involve extensive preparation, or are developed for only a small number of targeted compounds. In this work, we present a novel application for measuring bioavailable exposure with silicone wristbands as personal passive samplers. Laboratory methodology affecting precleaning, infusion, and extraction were developed from commercially available silicone, and chromatographic background interference was reduced after solvent cleanup with good extraction efficiency (>96%). After finalizing laboratory methods, 49 compounds were sequestered during an ambient deployment which encompassed a diverse set of compounds including polycyclic aromatic hydrocarbons (PAHs), consumer products, personal care products, pesticides, phthalates, and other industrial compounds ranging in log Kow from −0.07 (caffeine) to 9.49 (tris(2-ethylhexyl) phosphate). In two hot asphalt occupational settings, silicone personal samplers sequestered 25 PAHs during 8- and 40-h exposures, as well as 2 oxygenated-PAHs (benzofluorenone and fluorenone) suggesting temporal sensitivity over a single work day or week (p < 0.05, power =0.85). Additionally, the amount of PAH sequestered differed between worksites (p < 0.05, power = 0.99), suggesting spatial sensitivity using this novel application.

Structurally distinct polycyclic aromatic hydrocarbons induce differential transcriptional responses in developing zebrafish.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the environment as components of fossil fuels and by-products of combustion. These multi-ring chemicals differentially activate the aryl hydrocarbon receptor (AHR) in a structurally dependent manner, and induce toxicity via both AHR-dependent and -independent mechanisms. PAH exposure is known to induce developmental malformations in zebrafish embryos, and recent studies have shown cardiac toxicity induced by compounds with low AHR affinity. Unraveling the potentially diverse molecular mechanisms of PAH toxicity is essential for understanding the hazard posed by complex PAH mixtures present in the environment. We analyzed transcriptional responses to PAH exposure in zebrafish embryos exposed to benz(a)anthracene (BAA), dibenzothiophene (DBT) and pyrene (PYR) at concentrations that induced developmental malformations by 120 h post-fertilization (hpf). Whole genome microarray analysis of mRNA expression at 24 and 48 hpf identified genes that were differentially regulated over time and in response to the three PAH structures. PAH body burdens were analyzed at both time points using GC-MS, and demonstrated differences in PAH uptake into the embryos. This was important for discerning dose-related differences from those that represented unique molecular mechanisms. While BAA misregulated the least number of transcripts, it caused strong induction of cyp1a and other genes known to be downstream of the AHR, which were not induced by the other two PAHs. Analysis of functional roles of misregulated genes and their predicted regulatory transcription factors also distinguished the BAA response from regulatory networks disrupted by DBT and PYR exposure. These results indicate that systems approaches can be used to classify the toxicity of PAHs based on the networks perturbed following exposure, and may provide a path for unraveling the toxicity of complex PAH mixtures.

DGT estimates cadmium accumulation in wheat and potato from phosphate fertilizer applications

Cadmium is a common impurity in phosphatic fertilizers and may contribute to soil Cd accumulation. Changes in total and bioavailable Cd burdens to agricultural soils and the potential for plant Cd accumulation resulting from fertilizer input was investigated. Three year field studies were conducted using three dose levels of cadmium-rich, commercial, phosphate fertilizers applied at four agricultural sites. Labile Cd concentrations, measured using the passive sampling device Diffusive Gradients in Thin Films (Cd(DGT)), increased with increasing fertilizer application rates. Cd also accumulated in the edible portion of wheat and potato crops grown at the sites, and showed strong positive dose response with fertilizer treatment. Regression models were calculated for each site, year, and for individual crops. Model comparisons indicated that soil physical and chemical parameters in addition to soil Cd fractions, were important determinants of Cd(DGT). Significant factors contributing to Cd(DGT) concentrations were Cd from fertilizer input (Cd(fertilizer)), pH, cation exchange capacity (CEC), and total recoverable Cd (Cd(total)). Important factors used to determine Cd concentrations in wheat grain (Cd(wheat)) and in potato (Cd(potato)) were as follows: Cd(wheat):Cd(fertilizer), and Cd(DGT); and Cd(potato):Cd(fertilizer), Cd(DGT), % O.M. The effective concentration, C(E), calculated from DGT did not correlate well with Cd(wheat) or with Cd(potato). Direct measurements of Cd(DGT) correlated better with Cd found in edible plant tissue. The modeling approach presented in this study helps to estimate Cd accumulation in plant tissue over multiple years and in distinct agricultural soil systems.