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.

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.

Atmospheric deposition of current-use and historic-use pesticides in snow at national parks in the western United States.

The United States (U.S.) National Park Service has initiated research on the atmospheric deposition and fate of semi-volatile organic compounds in its alpine, sub-Arctic, and Arctic ecosystems in the Western U.S. Results for the analysis of pesticides in seasonal snowpack samples collected in spring 2003 from seven national parks are presented herein. From a target analyte list of 47 pesticides and degradation products, the most frequently detected current-use pesticides were dacthal, chlorpyrifos, endosulfan, and gamma-hexachlorocyclohexane, whereas the mostfrequently detected historic-use pesticides were dieldrin, alpha-hexachlorocyclohexane, chlordane, and hexachlorobenzene. Correlation analysis with latitude, temperature, elevation, particulate matter, and two indicators of regional pesticide use reveal that regional current and historic agricultural practices are largely responsible for the distribution of pesticides in the national parks in this study. Pesticide deposition in the Alaskan parks is attributed to long-range transport because there are no significant regional pesticide sources. The percentage of total pesticide concentration due to regional transport (%RT) was calculated for the other parks. %RT was highest at parks with higher regional cropland intensity and for pesticides with lower vapor pressures and shorter half-lives in air.

Trace analysis of semivolatile organic compounds in large volume samples of snow, lake water, and groundwater.

An analytical method was developed for the trace analysis of a wide range of semivolatile organic compounds (SOCs) in 50-L high-elevation snow and lake water samples. The method was validated for 75 SOCs from seven different chemical classes (polycyclic aromatic hydrocarbons, organochlorine pesticides, amides, triazines, polychlorinated biphenyls, thiocarbamates, and phosphorothioates) that covered a wide range of physical-chemical properties including 7 orders of magnitude of octanol-water partition coefficient (log K(ow) = 1.4-8.3). The SOCs were extracted using a hydrophobically and hydrophilically modified divinylbenzene solid-phase extraction device (modified Speedisk). The average analyte recovery from 50 L of reverse osmosis water, using the modified Speedisk, was 99% with an average relative standard deviation of 4.8%. Snow samples were collected from the field, melted, and extracted using the modified Speedisk and a poly(tetrafluoroethylene) remote sample adapter in the laboratory. Lake water was sampled, filtered, and extracted in situ using an Infiltrex 100 fitted with a 1-microm glass fiber filter to trap particulate matter and the modified Speedisk to trap dissolved SOCs. The extracts were analyzed by gas chromatographic mass spectrometry with electron impact ionization and electron capture negative ionization using isotope dilution and selective ion monitoring. Estimated method detection limits for snow and lake water ranged from 0.2 to 125 pg/L and 0.5-400 pg/L, respectively. U.S. historic and current-use pesticides were identified and quantified in snow and lake water samples collected from Rocky Mountain National Park, CO. The application of the analytical method to the analysis of SOCs in large-volume groundwater samples is also shown.

Quantitative analysis of 39 polybrominated diphenyl ethers by isotope dilution GC/low-resolution MS.

A GC/low-resolution MS method for the quantitative isotope dilution analysis of 39 mono- to heptabrominated diphenyl ethers was developed. The effects of two different ionization sources, electron impact (EI) and electron capture negative ionization (ECNI), and the effects of their parameters on production of high-mass fragment ions [M - xH - yBr](-) specific to PBDEs were investigated. Electron energy, emission current, source temperature, ECNI system pressure, and choice of ECNI reagent gases were optimized. Previously unidentified enhancement of PBDE high-mass fragment ion [M - xH - yBr](-) abundance was achieved. Electron energy had the largest impact on PBDE high-mass fragment ion abundance for both the ECNI and EI sources. By monitoring high-mass fragment ions of PBDEs under optimized ECNI source conditions, quantitative isotope dilution analysis of 39 PBDEs was conducted using nine (13)C(12) labeled PBDEs on a low-resolution MS with low picogram to femtogram instrument detection limits.

The effects of temperature and artificial rain on the metabolism of American kestrels (Falco sparverius).

The effect of rainfall on the metabolism of birds is poorly understood. We measured the metabolism as rate of oxygen consumption (VO2) of four male and four female American kestrels (Falco sparverius) using open-circuit respirometry. We measured VO2 during the spring at ambient temperatures (Ta) of 5, 10, 15, and 25 degrees C in air without rainfall and with simulated rainfall of 2.5 (low rainfall) and 6.1 cm h(-1) (high rainfall). Kestrel metabolism was significantly higher when exposed to the two rainfall levels compared to no rainfall. However, kestrel metabolism was not significantly different at the two rainfall levels. Body temperature (Tb) was significantly lower under high rainfall compared to low rainfall. In addition, under both rainfall levels Tb decreased with decreasing Ta. Calculated thermal conductance was significantly higher in kestrels exposed to rain compared to no rainfall. Kestrels may use sleeking behavior at high rainfall levels to decrease water penetration of the plumage. Daily energy expenditure (DEE) of kestrels exposed to rain may increase markedly, and kestrel energetics may be further exacerbated by wind that often accompanies natural rainstorms.