Daryle Boyd

Screening for planar chlorobiphenyl congeners in tissues of marine biota by high-performance liquid chromatography with photodiode array detection

A rapid method has been developed to screen for planar chlorobiphenyl (CB) congeners, as well as certain other CBs and DDTs, in tissue samples from marine biota. The analytes were extracted from tissue matrices with 1:1 hexane/pentane (v/v) and interfering compounds were separated from the CBs on a gravity-flow column packed with acidic, basic and neutral silica gel eluted with 1:1 hexane/methylene chloride (v/v). Subsequently, the planar CB congeners were resolved from the DDTs and other CBs by HPLC on Cosmosil PYE analytical columns cooled to 9 degrees C and were measured by an ultraviolet (UV) photodiode array (PDA) detector. Two important advantages of PDA over conventional UV detection were the ability to identify individual analytes by comparing their UV spectra to those of reference standards and the ability to establish the spectral homogeneity (purity) of the analytes by comparing spectra within a peak to the apex spectrum. The HPLC/PDA method was tested with tissue samples from fish, shellfish and marine mammals; concentrations of certain CBs and DDTs in samples determined by screening compared favorably with those in the same samples analyzed by a comprehensive method (e.g., gas chromatography/high resolution mass spectrometry). However, the HPLC/PDA method was about an order of magnitude less sensitive than determinations by comprehensive methods.

Geologically distinct crude oils cause a common cardiotoxicity syndrome in developing zebrafish.

Crude oils from different geological formations vary in composition, yet most crude oils contain a polycyclic aromatic hydrocarbon (PAH) fraction that would be expected to produce cardiotoxic effects in developing fish. To determine whether different crude oils or PAH compositions produce common or distinct effects, we used zebrafish embryos to directly compare two crude oils at different states of weathering. Iranian heavy crude oil (IHCO) spilled in the Yellow Sea following the 2007 Hebei Spirit accident was compared to the intensively studied Alaska North Slope crude oil (ANSCO) using two different exposure methods, water-accommodated fractions containing dispersed oil microdroplets and oiled gravel effluent. Overall, both crude oils produced a largely overlapping suite of defects, marked by the well-known effects of PAH exposure on cardiac function. Specific cardiotoxicity phenotypes were nearly identical between the two oils, including impacts on ventricular contractility and looping of the cardiac chambers. However, with increased weathering, tissue-specific patterns of aryl hydrocarbon receptor (AHR) activation in the heart changed, with myocardial AHR activation evident when alkyl-PAHs dominated the mixture. Our findings suggest that mechanisms of cardiotoxicity may shift from a predominantly AHR-independent mode during early weathering to a multiple pathway or synergistic mode with prolonged weathering and increased proportions of dissolved alkyl-PAHs. Despite continued need for comparisons of crude oils from different sources, the results here indicate that the body of knowledge already acquired from studies of ANSCO is directly relevant to understanding the impacts of other crude oil spills on the early life history stages of fish.

ORGANOCHLORINE CONTAMINANT LEVELS IN ESKIMO HARVESTED BOWHEAD WHALES OF ARCTIC ALASKA

Organochlorine (OC) levels in liver and blubber of 20 bowhead whales (Balaena mysticetus) collected during the Eskimo subsistence harvest at Barrow (Alaska, USA) in 1992 and 1993 are presented. Liver sum DDT (lipid weight) was significantly greater in male whales than in females. Most of the organochlorines measured were at higher levels in longer (older) than in shorter (younger) males. For female bowhead whales, hexachlorobenzene and lipid levels decreased and other OC levels did not change significantly with increasing length. Most organo-chlorine contaminants have low concentrations in tissues of the bowhead whale compared to concentrations in tissues of other cetaceans, especially Odontocetes. Based on allowable daily intakes (ADI) levels established by the Canadian Northern Contaminants Program (Ottawa, Ontario, Canada) “safe” levels of blubber to consume were calculated. Chlordane levels in bowhead whale blubber results in the most restrictive consumption amount (50 g blubber/day). We expect no adverse effects related to these organochlorine contaminants to occur in bowhead whales or in consumers of their tissues. However, investigation of low level chronic exposure effects and a more rigorous assessment of histopathology, biomarkers, and immune status in the bowhead whale would be required to conclude “no effect” with more certainty.

Potent Phototoxicity of Marine Bunker Oil to Translucent Herring Embryos after Prolonged Weathering

Pacific herring embryos (Clupea pallasi) spawned three months following the Cosco Busan bunker oil spill in San Francisco Bay showed high rates of late embryonic mortality in the intertidal zone at oiled sites. Dead embryos developed to the hatching stage (e.g. fully pigmented eyes) before suffering extensive tissue deterioration. In contrast, embryos incubated subtidally at oiled sites showed evidence of sublethal oil exposure (petroleum-induced cardiac toxicity) with very low rates of mortality. These field findings suggested an enhancement of oil toxicity through an interaction between oil and another environmental stressor in the intertidal zone, such as higher levels of sunlight-derived ultraviolet (UV) radiation. We tested this hypothesis by exposing herring embryos to both trace levels of weathered Cosco Busan bunker oil and sunlight, with and without protection from UV radiation. Cosco Busan oil and UV co-exposure were both necessary and sufficient to induce an acutely lethal necrotic syndrome in hatching stage embryos that closely mimicked the condition of dead embryos sampled from oiled sites. Tissue levels of known phototoxic polycyclic aromatic compounds were too low to explain the observed degree of phototoxicity, indicating the presence of other unidentified or unmeasured phototoxic compounds derived from bunker oil. These findings provide a parsimonious explanation for the unexpectedly high losses of intertidal herring spawn following the Cosco Busan spill. The chemical composition and associated toxicity of bunker oils should be more thoroughly evaluated to better understand and anticipate the ecological impacts of vessel-derived spills associated with an expanding global transportation network.

Organic and trace metal contaminants in sediments and English sole tissues from Vancouver Harbour, Canada

As part of a multinational workshop on marine environmental quality, sediments were collected from seven sites in Vancouver Harbour and analyzed for polynuclear aromatic hydrocarbons (PAHs) and organochlorines (OCs), and for trace metals. English sole were collected from five sites, and muscle was analyzed for trace metals and liver for OCs. As expected, sediment PAH and OC concentrations and tissue OC concentrations were higher at sites east of the First Narrows, compared to the outer harbor and reference sites. Sediment PAH concentrations east of the First Narrows were similar to concentrations at moderately contaminated sites in Puget Sound, south of Vancouver Harbour. In contrast, concentrations of OCs in sediments and tissue were low to moderate, even at relatively contaminated sites within Vancouver Harbour. Although several trace metals in sediments were higher than in contaminated sediments from Puget Sound, trace metals measured in fish muscle were lower.

Blood-based biomarkers of selenium and thyroid status indicate possible adverse biological effects of mercury and polychlorinated biphenyls in Southern Beaufort Sea polar bears.

We examined biomarkers of selenium status (whole blood Se; serum Se; glutathione peroxidase activity) and thyroid status (concentrations and ratios of thyroxine, T4; tri-iodothyronine, T3; albumin) in polar bears to assess variations among cohorts, and relationships to circulating concentrations of contaminants. Concentrations of total mercury (Hg) in whole blood were similar among cohorts (prime aged males and females, older animals, ages≥16 years, and young animals, ages 1-5 years; 48.44±35. 81; p=0.253). Concentrations of sum of seven polychlorinated biphenyls (∑PCB7) in whole blood were greater in females (with and without cubs, 26.44±25.82 ng/g ww) and young (26.81±10.67 ng/g ww) compared to males (8.88±5.76 ng/g ww, p<0.001), and significantly related to reduced body condition scores (p<0.001). Concentrations of Se and albumin were significantly greater in males than females (whole blood Se, males, 42.34 pmol/g ww, females, 36.25±6.27 pmol/g ww, p=0.019; albumin, males, 4.34±0.34 g/dl, females, 4.10±0.29 g/dL, p=0.018). Glutathione peroxidase activity ranged from 109.1 to 207.8 mU/mg hemoglobin, but did not differ significantly by sex or age (p>0.08). Thyroid hormones were greater in females (solitary females and females with cubs) compared to males (p<0.001). Biomarkers of Se status and concentrations of T3 were significantly positively related to Hg in all prime aged polar bears (p<0.03). Albumin concentrations were significantly positively related to total TT4, and significantly negatively related to concentrations of ∑PCB7 (p<0.003). Total thyroxine (TT4) was significantly negatively associated with blood concentrations of ∑PCB7 in solitary females (p=0.045). These data suggest that female polar bears were more susceptible to changes in blood-based biomarkers of selenium and thyroid status than males. Further classifications of the physiologic states of polar bears and repeated measures of individuals over time are needed to accurately assess the biological impact of combined toxicant exposures.

Geographic variation of persistent organic pollutant levels in humpback whale (Megaptera novaeangliae) feeding areas of the North Pacific and North Atlantic

Seasonal feeding behavior and high fidelity to feeding areas allow humpback whales (Megaptera novaeangliae) to be used as biological indicators of regional contamination. Biopsy blubber samples from male individuals (n = 67) were collected through SPLASH, a multinational research project, in eight North Pacific feeding grounds. Additional male samples (n = 20) were collected from one North Atlantic feeding ground. Persistent organic pollutants were measured in the samples and used to assess contaminant distribution in the study areas. North Atlantic (Gulf of Maine) whales were more contaminated than North Pacific whales, showing the highest levels of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and chlordanes. The highest dichlorodiphenyltrichloroethane (DDT) levels were detected in whales feeding off southern California, USA. High-latitude regions were characterized by elevated levels of hexachlorocyclohexanes (HCHs) but generally nondetectable concentrations of PBDEs. Age was shown to have a positive relationship with SigmaPCBs, SigmaDDTs, Sigmachlordanes, and total percent lipid. Contaminant levels in humpback whales were comparable to other mysticetes and lower than those found in odontocete cetaceans and pinnipeds. Although these concentrations likely do not represent a significant conservation threat, levels in the Gulf of Maine and southern California may warrant further study.

Relationships Between Anthropogenic Chemical Contaminant Exposure and Associated Changes in Reproductive Parameters in Male English Sole (Parophrys vetulus) Collected from Hylebos Waterway, Puget Sound, Washington

Effects of chemical contaminant exposure on gonadal development in adult male English sole (Parophrys vetulus) from Hylebos Waterway and Colvos Passage, Puget Sound, Washington were investigated. Hylebos Waterway sediment is contaminated with polycyclic aromatic hydrocarbons (PAHs) and organochlorines (OCs), and Colvos Passage, a nearby nonurban area, is minimally contaminated. Fish from Hylebos Waterway had higher concentrations of both PAHs and OCs in tissues than fish from Colvos Passage. Overall, little correlation was observed between PAH exposure and biological parameters, but strong correlations were observed between OCs and the biological parameters. Migration of fish from less contaminated areas into the Hylebos Waterway during the reproductive season might have influenced these results, based on temporal changes in fish age and contaminant concentrations.

Persistent organic pollutants in the endangered Hawaiian monk seal (Monachus schauinslandi) from the main Hawaiian Islands

Little is known about levels or effects of persistent organic pollutants (POPs) in Hawaiian monk seals (HMS) from the main Hawaiian Islands (MHI) subpopulation. This study examined concentrations of a large suite of POPs in blubber and serum of juvenile and adult HMS from the MHI. Adult females have the lowest blubber levels of most POPs, whereas adult males have highest levels. POPs in serum were significantly different in adult males compared with adult females for chlordanes and summed dichlorodiphenyltrichloroethanes (DDTs). Lipid-normalized concentrations of chlordanes, DDTs, polychlorinated biphenyls, and mirex in paired blubber and serum samples were significantly correlated. Contaminant levels from the MHI were at similar or lower levels than those from remote Northwestern Hawaiian Island populations. Determining initial ranges of POPs is an important step towards assessing one of the many potential health threats to this critically endangered species.

Lactational transfer of mercury and polychlorinated biphenyls in polar bears

We examined concentrations of total mercury (tHg, inorganic and methylated forms) and polychlorinated biphenyls (PCBs) in blood and milk from free-ranging Southern Beaufort-Chukchi Sea polar bears (Ursus maritimus) to assess maternal transfer of contaminants during lactation and the potential health risk to nursing young. Concentrations of contaminants in the blood of dependent and juvenile animals (ages 1-5 years) ranged from 35.9 to 52.2 μg kg(-1) ww for tHg and 13.9 to 52.2 μg kg(-1) ww (3255.81-11067.79 μg kg(-1) lw) for ΣPCB(7)s, similar to those of adult females, but greater than adult males. Contaminant concentrations in milk ranged from 5.7 to 71.8 μg tHg kg(-1)ww and 160 to 690 μg ΣPCB(11)s kg(-1) ww (547-5190 μg kg(-1) lw). The daily intake levels for tHg by milk consumption estimated for dependent young were below the tolerable daily intake level (TDIL) of tHg established for adult humans. Although the daily intake levels of PCBs through milk consumption for cubs of the year exceeded the TDIL thresholds, calculated dioxin equivalents for PCBs in milk were below adverse physiological thresholds for aquatic mammals. Relatively high concentrations of non-dioxin like PCBs in polar bear milk and blood could impact endocrine function of Southern Beaufort-Chukchi Sea polar bears, but this is uncertain. Transfer of contaminants during mid to late lactation likely limits bioaccumulation of dietary contaminants in female polar bears during spring. As polar bears respond to changes in their arctic sea ice habitat, the adverse health impacts associated with nutritional stress may be exacerbated by tHg and PCBs exposure, especially in ecologically and toxicologically sensitive polar bear cohorts such as reproductive females and young.