Comparison of Two Total Mercury Screening and Assessment Methods in Fishes: Biopsy Punch and Dried Muscle Samples.

Rapid and effective quantification of total mercury concentrations ([THg]) in fish muscle is an important part of ongoing monitoring to provide reliable and near real-time public health guidance. Methods for quantifying THg in fish muscle frequently require the use of large sample mass and numerous preparation steps. Wet (aka fresh weight) biopsy punch samples of fish muscle have been used to quantify THg directly, without drying and homogenization. Both methods have advantages and disadvantages. We compare the use of fresh weight biopsy punches for quantifying THg to using larger, dried homogenized samples. The [THg] determination for the two sampling methods was EPA method 7473. Three separate biopsy punch samples and a large muscle sample were taken from each fish and analyzed on a Direct Mercury Analyzer. There were no statistical differences between mean log transformed wet weight [THg] from biopsy punches and homogenized muscle across all samples or within individual species. Similarly, across the range of [THg] (7.5-612.7 ng/g ww), linear regression of [THg] from biopsy punch and homogenized muscle samples was not different from a 1:1 linear relationship. Linear regression statistics of [THg] with fish fork length produced similar results for both biopsy punch and homogenized muscle samples. However, the coefficient of variation among biopsy punch replicates for individual fish was frequently above the acceptable threshold of 15%. We recommend biopsy punches be used as an effective tool for broad-scale rapid monitoring of fish resources for Hg, while homogenized muscle samples be used for fine-scale ecological and health questions.

Metals in the stomach contents and brain, gonad, kidney, and liver tissues of subsistence-harvested northern sea otters (Enhydra lutris kenyoni) from Icy Strait, Alaska.

Southeast Alaska sea otters (Enhydra lutris) have had a rapid rise in their population. As they feed primarily on sessile prey, they are excellent sentinels for examining metals contamination. Objectives of this study on sea otters were to determine: (1) concentrations of metals in different tissues; (2) whether metals biomagnify from stomach contents (i.e., the prey) to other tissues; (3) whether selenium and mercury concentrations indicate an overall health benefit or risk; and (4) if metals concentrations in tissues vary with body size. Brain, kidney, gonad, liver, and stomach contents were collected from freshly harvested sea otters in Icy Strait, Alaska, and analyzed for arsenic (As), cadmium (Cd), copper (Cu), lead (Pb), total mercury (THg), and selenium (Se). Metals concentrations varied significantly, and some were biomagnified, with livers and kidneys harboring the highest concentrations. Lead and arsenic appeared to be readily excreted. This study represents baseline metals concentrations to assist in monitoring the health of sea otters.

Subsistence fish consumption in rural Alaska: Using regional monitoring data to evaluate risk and bioavailability of dietary methylmercury.

On average, Alaskans in rural communities consume over three times the Federally recommended maximum weekly fish ingestion rate (IR), the overwhelming majority of which is Pacific salmon. Results of statewide monitoring efforts consistently show that Pacific salmon from Alaska have low concentrations of mercury, yet concerns regarding dietary exposure to methylmercury (MeHg) and other aquatic contaminants continue to contribute to declining subsistence fish consumption rates in rural communities. Therefore, the goal of the present study was to use statewide biomonitoring datasets and regional fish IRs to quantitatively evaluate potential risk from dietary MeHg exposure via subsistence consumption of salmon from Alaska. Hazard Indices (HIs) did not exceed 1 for any of the groups evaluated, indicating negligible risk for the average Alaskan subsistence consumer. Selenium health benefit values (HBVSe) of various fish species from AK were also calculated, with positive results for all commonly consumed subsistence species. Additionally, mercury concentrations in the hair of Alaskan women were evaluated as a proxy for dietary MeHg exposure. Results reveal that Alaskan women of childbearing age have substantially lower hair Hg concentrations than their counterparts in other large-scale biomonitoring studies, despite similar fish IRs. Collectively, results of the present study suggest that MeHg in Pacific salmon does not pose an unacceptable hazard for the average subsistence consumer in Alaska.

Sodium perchlorate induces non-alcoholic fatty liver disease in developing stickleback.

Perchlorate is a pervasive, water-soluble contaminant that competitively inhibits the sodium/iodide symporter, reducing the available iodide for thyroid hormone synthesis. Insufficient iodide uptake can lead to hypothyroidism and metabolic syndromes. Because metabolism, obesity and non-alcoholic fatty liver disease (NAFLD) are tightly linked, we hypothesized that perchlorate would act as an obesogen and cause NAFLD via accumulation of lipids in liver of developing threespine stickleback (Gasterosteus aculeatus). We performed an upshift/downshift exposure regime (clean water to perchlorate treated water or perchlorate treated water to clean water) on stickleback embryos at two concentrations (30 mg/L and 100 mg/L) plus the control (0 mg/L) over the course of 305 days. Adult stickleback were euthanized, H&E stained and analyzed for liver morphology. Specifically, we counted the number of lipid droplets, and measured the area of each droplet and the total lipid area of a representative section of liver. We found that perchlorate treated fish had more and larger lipid droplets, and a larger percentage of lipid in their liver than control fish. These data indicate that perchlorate causes NAFLD and hepatic steatosis in stickleback at concentrations commonly found at contaminated sites. These data also indicate the potential of perchlorate to act as an obesogen. Future studies should investigate the obesogenic capacity of perchlorate by examining organ specific lipid accumulation and whether perchlorate induces these effects at concentrations commonly found in drinking water. Work is also needed to determine the mechanisms by which perchlorate induces lipid accumulation.

Developmental timing of sodium perchlorate exposure alters angiogenesis, thyroid follicle proliferation and sexual maturation in stickleback.

Perchlorate, a common aquatic contaminant, is well known to disrupt homeostasis of the hypothalamus-pituitary-thyroid axis. This study utilizes the threespine stickleback (Gasterosteus aculeatus) fish to determine if perchlorate exposure during certain windows of development has morphological effects on thyroid and gonads. Fish were moved from untreated water to perchlorate-contaminated water (30 and 100mg/L) starting at 0, 3, 7, 14, 21, 42, 154 and 305 days post fertilization until approximately one year old. A reciprocal treatment (fish in contaminated water switched to untreated water) was conducted on the same schedule. Perchlorate exposure increased angiogenesis and follicle proliferation in thyroid tissue, delayed gonadal maturity, and skewed sex ratios toward males; effects depended on concentration and timing of exposure. This study demonstrates that perchlorate exposure beginning during the first 42 days of development has profound effects on stickleback reproductive and thyroid tissues, and by implication can impact population dynamics. Long-term exposure studies that assess contaminant effects at various stages of development provide novel information to characterize risk to aquatic organisms, to facilitate management of resources, and to determine sensitive developmental windows for further study of underlying mechanisms.

Developmental timing of perchlorate exposure alters threespine stickleback dermal bone.

Adequate levels of thyroid hormone are critical during development and metamorphosis, and for maintaining metabolic homeostasis. Perchlorate, a common contaminant of water sources, inhibits thyroid function in vertebrates. We utilized threespine stickleback (Gasterosteus aculeatus) to determine if timing of perchlorate exposure during development impacts adult dermal skeletal phenotypes. Fish were exposed to water contaminated with perchlorate (30mg/L or 100mg/L) beginning at 0, 3, 7, 14, 21, 42, 154 or 305days post fertilization until sexual maturity at 1year of age. A reciprocal treatment moved stickleback from contaminated to clean water on the same schedule providing for different stages of initial exposure and different treatment durations. Perchlorate exposure caused concentration-dependent significant differences in growth for some bony traits. Continuous exposure initiated within the first 21days post fertilization had the greatest effects on skeletal traits. Exposure to perchlorate at this early stage can result in small traits or abnormal skeletal morphology of adult fish which could affect predator avoidance and survival.

Perchlorate trophic transfer increases tissue concentrations above ambient water exposure alone in a predatory fish.

This study examined effects of varying concentrations of the environmental contaminant perchlorate in northern pike (Esox lucius) based on exposure in water and/or from prey (threespine stickleback, Gasterosteus aculeatus). Routes of exposure to pike were through contaminated water at 0, 10, or 100 mg/L perchlorate for 49 d and/or through feeding, 1 per day over 14 d, sticklebacks that were previously maintained in water at 0, 10, or 100 mg/L perchlorate. Both water and food significantly contributed to pike tissue concentrations of perchlorate as compared to controls, but, as expected for a water-soluble contaminant, perchlorate did not biomagnify from prey to predatory fish. Pike gastrointestinal tissue retained significantly more perchlorate than other tissues combined. Route of exposure and concentration of perchlorate in various media are important to consider in risk assessment when evaluating uptake and tissue concentration of perchlorate because significantly higher tissue concentrations may result from combined prey and water exposures than from prey or water exposures alone in a concentration-dependent manner.

Partial reproductive isolation of a recently derived resident-freshwater population of threespine stickleback (Gasterosteus aculeatus) from its putative anadromous ancestor.

We used no-choice mating trials to test for assortative mating between a newly derived resident-freshwater population (8-22 generations since founding) of threespine stickleback (Gasterosteus aculeatus) in Loberg Lake, Alaska and its putative anadromous ancestor as well as a morphologically convergent but distantly related resident-freshwater population. Partial reproductive isolation has evolved between the Loberg Lake population and its ancestor within a remarkably short time period. However, Loberg stickleback readily mate with morphologically similar, but distantly related resident-freshwater stickleback. Partial premating isolation is asymmetrical; anadromous females and smaller resident-freshwater males from Loberg Lake readily mate, but the anadromous males and smaller Loberg females do not. Our results indicate that premating isolation can begin to evolve in allopatry within a few generations after isolation as a correlated effect of evolution of reduced body size.