Temporal trends in aluminum smelter-derived polycyclic aromatic hydrocarbons in outmigrant juvenile Chinook salmon from Kitimat, British Columbia, Canada.

Aluminum smelter-derived polycyclic aromatic hydrocarbons (PAHs) in outmigrant juvenile Chinook salmon were evaluated in Kitimat Arm, British Columbia, Canada from 2000 to 2004, and in 2015. Decades of continual smelter operations by Rio Tinto resulted in PAH contamination of marine sediments at levels associated with adverse effects in juvenile salmon. Recently, smelter operations have undergone process changes to reduce PAH input to the environment. The PAH concentrations in juvenile Chinook salmon observed in 2000 to 2004, at sites nearest the smelter were comparable to salmon in other urban areas where reduced disease resistance was observed; the levels were lower in 2015 than 2000-2004 suggesting that the recent process changes were effective. Further, these data establish a benchmark for assessing any future changes affecting PAH input and the potential risks to the receiving environment.

Polycyclic aromatic hydrocarbons and risk to threatened and endangered Chinook salmon in the Lower Columbia River estuary.

Polycyclic aromatic hydrocarbons (PAHs), derived from oil and fuel combustion, are ubiquitous nonpoint source pollutants that can have a number of detrimental effects on fish and wildlife. In this study, we monitored PAH exposure in outmigrant juvenile Chinook salmon from the Lower Columbia River to evaluate the risk that these contaminants might pose to the health and recovery of threatened and endangered salmonids. Juvenile Chinook salmon (Oncorhynchus tshawytscha) were collected by beach seine from five sites in the Lower Columbia River from Bonneville Dam to the mouth of the estuary (Warrendale, the Willamette-Columbia Confluence, Columbia City, Beaver Army Terminal, and Point Adams) and from a site in the Lower Willamette near downtown Portland (Morrison Street Bridge). Sediment samples were also collected at the same sites. Concentrations of PAHs in sediment samples were relatively low at all sites with average total PAH concentrations <1000 ng/g dry weight (wt.). However, we found PAHs in stomach contents of salmon from all sites at concentrations ranging from <100 to >10,000 ng/g wet wt. Metabolites of low and high molecular-weight PAHs were also detected in bile of salmon from all sites; for metabolites fluorescing at phenanthrene (PHN) wavelengths, concentrations ranged from 1.1 to 6.0 µg/mg bile protein. Levels of PAHs in stomach contents and PAH metabolites in bile were highest in salmon from the Morrison Street Bridge site in Portland and the Willamette-Columbia Confluence, Columbia City, and Beaver Army Terminal sites. Mean PAH concentrations measured in some stomach content samples from the Columbia City, Beaver Army Terminal, and Morrison Street Bridge sites were near the threshold concentration (approximately 7200-7600 ng/g wet wt.) associated with variability and immune dysfunction in juvenile salmonids (Meador et al., Can J Fish Aquat Sci 63:2364-2376, 2006; Bravo et al., Environ Toxicol Chem 30:704-714, 2011). Mean levels of biliary fluorescent aromatic compounds (FACs)-PHN in juvenile Chinook collected at the Morrison Street Bridge site in Portland, at the Confluence and Columbia City sites, and at the Beaver Army Terminal site were at or above a threshold effect concentration of 2 µg/mg protein for FACs-PHN linked to growth impairment, altered energetics, and reproductive effects (Meador et al., Environ Toxicol Chem 27(4):845-853, 2008). These findings suggest that PAHs in the food chain are a potential source of injury to juvenile salmon in the Lower Columbia and Lower Willamette rivers.

Development of an enzyme-linked immunosorbent assay for quantifying vitellogenin in Pacific salmon and assessment of field exposure to environmental estrogens.

A competitive enzyme-linked immunosorbent assay was developed to quantitate vitellogenin (VTG) in plasma and serum of coho (Oncorhynchus kisutch) and chinook (O. tshawytscha) salmon. The working range of the assay was 9 to 313 ng/ml (80-20% binding), with 50% binding at 54 ng/ml. The intra-assay and interassay variations at approximately 50% binding were 8.1% (n = 9) and 9.0% (n = 9), respectively. Dilution curves of plasma or serum from coho and chinook females and estrogen-treated males were parallel to the purified coho VTG standard curve. Male plasma samples could be assayed at a minimum dilution of 1:40 (chinook) or 1:75 (coho) without assay interference because of high sample concentration, whereas minimum acceptable dilutions of male serum samples were 1:200 (chinook) or 1:600 (coho). Identification of proper techniques for preserving VTG integrity in plasma and serum samples showed that VTG from both species was robust; both sample types required no protease inhibitor despite subjection to two freeze-thaw cycles. To test its applicability, this assay was used to measure VTG in out-migrating juvenile chinook that were collected from urban and nonurban areas in Puget Sound, Washington, USA. Results showed a small but significant plasma VTG elevation at two urban sites, suggesting that these juveniles may be exposed to environmental estrogens at an early life stage. Also, wild fish tended to have higher plasma VTG levels than hatchery fish collected in the field. Elevation of mean VTG levels was similar to that previously reported in male English sole from the same area, where both males and females exhibited alterations in timing of spawning.

Xenoestrogen exposure and effects in English sole (Parophrys vetulus) from Puget Sound, WA.

Vitellogenin, a yolk protein produced in the liver of oviparous animals in response to estrogens, normally occurs only in sexually mature females with developing eggs. However, males can synthesize vitellogenin when exposed to environmental estrogens, making the abnormal production of vitellogenin in male animals a useful biomarker for xenoestrogen exposure. In 1997-2001, as part of the Washington State's Puget Sound Assessment and Monitoring Program, we surveyed English sole from a number of sites for evidence of xenoestrogen exposure, using vitellogenin production in males as an indicator. Significant levels of vitellogenin were found in male fish from several urban sites, with especially high numbers of fish affected in Elliott Bay, along the Seattle Waterfront. Intersex fish were rare, comprising only two fish out of more than 2900 examined. Other ovarian and testicular lesions, including oocyte atresia, were also observed, but their prevalence did not appear to be related to xenoestrogen exposure. However, at the Elliott Bay sites where abnormal vitellogenin production was observed in male sole, the timing of spawning in both male and female English sole appeared altered. Sources of xenoestrogens and types of xenoestrogens present in Elliott Bay are poorly documented, but the compounds are likely associated with industrial discharges, surface runoff, and combined sewer outfalls.

The relationships of biochemical endpoints to histopathology and population metrics in feral flatfish species collected near the municipal wastewater outfall of Orange County, California, USA.

In July 2000, 330 individuals of three flatfish species were collected from reference locations and nine sites surrounding the outfall of the Orange County (CA, USA) Sanitation District (OCSD) municipal wastewater discharge. Species included hornyhead turbot (Pleuronichthys verticalis), English sole (Pleuronectes vetulus), and bigmouth sole (Hippoglossina stomata). Livers of sampled animals were examined for the expression of vitellogenin (in males), cytochrome P450 1A (CYP1A), and DNA damage (comet assay). Biliary fluorescent aromatic compounds (FACs) were also measured. Comparisons with tissue and sediment contaminant residues, liver histopathology, and population estimates were assessed to determine whether relationships exist between levels of biological organization. The CYP1A results indicated exposure to planar aromatic hydrocarbons at one nearshore site. Regression analysis of all English sole CYP1A to total sediment polychlorinated biphenyls (PCBs) (r2 = 0.97; p < 0.05) indicated a significant correlation. Widespread exposure to estrogens was observed at all sites without correlation to the abundance of the three species examined. Fluorescent aromatic compounds demonstrated no correlation to CYP1A expression. Overall, histopathology and biochemical endpoints did not indicate significant adverse effects in fish at the OCSD outfall.