Paul E. Drevnick

Toxicity of dietary methylmercury to fish: Derivation of ecologically meaningful threshold concentrations

Threshold concentrations associated with adverse effects of dietary exposure to methylmercury (MeHg) were derived from published results of laboratory studies on a variety of fish species. Adverse effects related to mortality were uncommon, whereas adverse effects related to growth occurred only at dietary MeHg concentrations exceeding 2.5 µg g(-1) wet weight. Adverse effects on behavior of fish had a wide range of effective dietary concentrations, but generally occurred above 0.5 µg g(-1) wet weight. In contrast, effects on reproduction and other subclinical endpoints occurred at dietary concentrations that were much lower (<0.2 µg g(-1) wet wt). Field studies generally lack information on dietary MeHg exposure, yet available data indicate that comparable adverse effects have been observed in wild fish in environments corresponding to high and low MeHg contamination of food webs and are in agreement with the threshold concentrations derived here from laboratory studies. These thresholds indicate that while differences in species sensitivity to MeHg exposure appear considerable, chronic dietary exposure to low concentrations of MeHg may have significant adverse effects on wild fish populations but remain little studied compared to concentrations in mammals or birds.

Gene Expression Changes Related to Endocrine Function and Decline in Reproduction in Fathead Minnow (Pimephales promelas) after Dietary Methylmercury Exposure

Background Methylmercury (MeHg) is a known neurotoxic agent, but the mechanisms by which MeHg may act on reproductive pathways are relatively unknown. Several studies have indicated potential changes in hormone levels as well as declines in vertebrates with increasing dietary MeHg exposure. Objectives The purpose of this study was to identify alterations in gene expression associated with MeHg exposure, specifically those associated with previously observed changes in reproduction and reproductive biomarkers. Fathead minnows, Pimephales promelas, were fed one of three diets that were similar to documented concentrations of MeHg in the diets of wild invertivorous and piscivorous fish. We used a commercial macroarray in conjunction with quantitative polymerase chain reaction to examine gene expression in fish in relation to exposure to these environmentally relevant doses of MeHg. Results Expression of genes commonly associated with endocrine disruption was altered with Hg exposure. Specifically, we observed a marked up-regulation in vitellogenin mRNA in individual Hg-exposed males and a significant decline in vitellogenin gene expression in female fish with increasing Hg concentrations. Other genes identified by the macroarray experiment included those associated with egg fertilization and development, sugar metabolism, apoptosis, and electron transport. We also observed differences in expression patterns between male and female fish not related to genes specifically associated with reproduction, indicating a potential physiological difference in the reaction of males and females to MeHg. Conclusion Gene expression data may provide insight into the mechanisms by which MeHg affects reproduction in fish and indicate how MeHg differs in its effect from other heavy metals and endocrine-disrupting compounds.

Atmospheric Hg Emissions from Preindustrial Gold and Silver Extraction in the Americas: A Reevaluation from Lake-Sediment Archives

Human activities over the last several centuries have transferred vast quantities of mercury (Hg) from deep geologic stores to actively cycling earth-surface reservoirs, increasing atmospheric Hg deposition worldwide. Understanding the magnitude and fate of these releases is critical to predicting how rates of atmospheric Hg deposition will respond to future emission reductions. The most recently compiled global inventories of integrated (all-time) anthropogenic Hg releases are dominated by atmospheric emissions from preindustrial gold/silver mining in the Americas. However, the geophysical evidence for such large early emissions is equivocal, because most reconstructions of past Hg-deposition have been based on lake-sediment records that cover only the industrial period (1850-present). Here we evaluate historical changes in atmospheric Hg deposition over the last millennium from a suite of lake-sediment cores collected from remote regions of the globe. Along with recent measurements of Hg in the deep ocean, these archives indicate that atmospheric Hg emissions from early mining were modest as compared to more recent industrial-era emissions. Although large quantities of Hg were used to extract New World gold and silver beginning in the 16th century, a reevaluation of historical metallurgical methods indicates that most of the Hg employed was not volatilized, but rather was immobilized in mining waste.

Mercury toxicity in livers of northern pike (Esox lucius) from Isle Royale, USA

Many laboratory studies have documented that mercury can be toxic to fish, but it is largely unknown if mercury is toxic to fish in their natural environments. The objective of our study was to investigate the toxic effects of mercury on northern pike (Esox lucius) at Isle Royale, Michigan. In 124 northern pike from eight inland lakes, concentrations of total mercury in skin-on fillets ranged from 0.069 to 0.622 microg/g wet mass (wet wt). Concentrations of total mercury in livers increased exponentially compared with concentrations in fillets, to a maximum of 3.1 microg/g wet wt. Methylmercury constituted a majority of the mercury in livers with total mercury concentrations <0.5 microg/g wet wt, but declined to 28-51% of the mercury in livers with total mercury concentrations >0.5 microg/g wet wt. Liver color (absorbance at 400 nm) varied among northern pike and was positively related to liver total mercury concentration. The pigment causing variation in liver color was identified as lipofuscin, which results from lipid peroxidation of membranous organelles. An analysis of covariance revealed lipofuscin accumulation was primarily associated with mercury exposure, and this association obscured any normal accumulation from aging. We also documented decreased lipid reserves in livers and poor condition factors of northern pike with high liver total mercury concentrations. Our results suggest (i) northern pike at Isle Royale are experiencing toxicity at concentrations of total mercury common for northern pike and other piscivorous fish elsewhere in North America and (ii) liver color may be useful for indicating mercury exposure and effects in northern pike at Isle Royale and possibly other aquatic ecosystems and other fish species.

Increase in mercury in Pacific yellowfin tuna

Mercury is a toxic trace metal that can accumulate to levels that threaten human and environmental health. Models and empirical data suggest that humans are responsible for a great deal of the mercury actively cycling in the environment at present. Thus, one might predict that the concentration of mercury in fish should have increased dramatically since the Industrial Revolution. Evidence in support of this hypothesis has been hard to find, however, and some studies have suggested that analyses of fish show no change in mercury concentration. By compiling and re-analyzing published reports on yellowfin tuna (Thunnus albacares) caught near Hawaii (USA) over the past half century, the authors found that the concentration of mercury in these fish currently is increasing at a rate of at least 3.8% per year. This rate of increase is consistent with a model of anthropogenic forcing on the mercury cycle in the North Pacific Ocean and suggests that fish mercury concentrations are keeping pace with current loading increases to the ocean. Future increases in mercury in yellowfin tuna and other fishes can be avoided by reductions in atmospheric mercury emissions from point sources.

Laser Ablation ICP-MS Co-Localization of Mercury and Immune Response in Fish

Mercury (Hg) contamination is a global issue with implications for both ecosystem and human health. In this study, we use a new approach to link Hg exposure to health effects in spotted gar (Lepisosteus oculatus) from Caddo Lake (TX/LA). Previous field studies have reported elevated incidences of macrophage centers in liver, kidney, and spleen of fish with high concentrations of Hg. Macrophage centers are aggregates of specialized white blood cells that form as an immune response to tissue damage, and are considered a general biomarker of contaminant toxicity. We found elevated incidences of macrophage centers in liver of spotted gar and used a new technology for ecotoxicology studies, laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS), to colocalize aggregates and Hg deposits within the tissue architecture. We conclude that Hg compromises the health of spotted gar in our study and, perhaps, other fish exposed to elevated concentrations of Hg.

Evidence of impaired health in yellow perch (Perca flavescens) from a biological mercury hotspot in northeastern north America

Few studies have investigated the effects of mercury (Hg) on wild fish from remote areas, even though these fish can have high total Hg concentrations. In Kejimkujik National Park and National Historic Site (KNPNHS), Nova Scotia, Canada, concentrations of total Hg in many yellow perch (Perca flavescens) currently exceed the estimated threshold level for adverse effects in fish (0.2 µg Hg g(-1) (wet wt), whole body). To determine whether Hg exposure is adversely affecting the general health of these fish, the authors collected male and female perch in the fall of 2009 and 2010 from 12 lakes within KNPNHS. The health endpoints condition, liver somatic index (LSI), and macrophage aggregates (MAs; indicators of oxidative stress and tissue damage) in the liver, kidney, and spleen were examined, and in female perch were compared between lakes and related to Hg concentrations measured in the muscle and liver tissue. No negative relationships between fish condition or LSI and Hg were found. However, within the liver, kidney, and spleen tissues of females, the relative area occupied by MAs was positively related to both muscle and liver Hg concentrations, indicating the health of these perch was adversely affected at the cellular level. These findings raise concerns for the health of these perch as well as for other wild fish populations known to have similarly elevated Hg concentrations.

DETERMINATION OF MERCURY SPECIATION IN FISH TISSUE WITH A DIRECT MERCURY ANALYZER

Knowledge of Hg speciation in tissue is valuable for assessing potential toxicological effects in fish. Direct Hg analyzers, which use thermal decomposition and atomic absorption spectrometry, have recently gained popularity for determining organic Hg after procedural solvent extraction from some environmental media, although quantitative recovery from lipid-rich materials, such as fish liver, has been problematic. The authors developed a new method by which organic Hg in fish liver and muscle is estimated by the difference between direct measurements of inorganic Hg in an acid extract and total Hg in whole tissue. The method was validated by analysis of a certified reference material (DOLT-4 dogfish liver) and naturally contaminated fish tissues with comparison to an established Hg speciation method (gas chromatography cold vapor atomic fluorescence spectrometry). Recovery of organic Hg from DOLT-4, estimated by difference, averaged 99 ± 5% of the mean certified value for methylmercury. In most liver samples and all muscle samples, estimates of organic Hg from the proposed method were indiscernible from direct speciation measurements of methylmercury (99% ± 6%). Estimation of organic Hg by the difference between total Hg and inorganic Hg was less accurate in liver samples with a high percentage of inorganic Hg (90%). This was because of the increased uncertainty that results from estimating a third value (i.e., organic Hg) by using the difference between two large concentrations (inorganic and total Hg). The proposed method is a useful tool for examining the speciation of Hg in fish muscle and liver, and by extension, potentially other tissues and environmental media.

Reproductive health of yellow perch (Perca flavescens) from a biological mercury hotspot in Nova Scotia, Canada.

Methylmercury (MeHg) exposure is known to adversely affect the reproductive health of laboratory fish, but its impacts on the sexual development of wild fishes are not well studied. Kejimkujik National Park and National Historic Site (KNPNHS) region of Nova Scotia, Canada, has been identified as a biological mercury (Hg) hotspot. To determine whether Hg was adversely affecting the reproductive health of wild yellow perch (Perca flavescens), sexually mature male and female perch were collected from 12 lakes within KNPNHS (mean muscle total Hg: 0.28-0.54 μg/g ww). Gonadosomatic index and germ cell development of male and female perch were measured, as well as the plasma 17 β-estradiol concentrations of females. These endpoints were compared between lakes, and were related to Hg concentrations measured in perch muscle and liver tissues. Our results indicate that the reproductive health of male and female perch was not adversely impacted by Hg, although a positive relationship existed between the proportions of primary spermatocytes in male testes and muscle total Hg concentrations. Perch were sampled at an early stage of recrudescence, and the tissue Hg concentrations in these perch were generally lower than those in laboratory studies reporting impacts on reproductive health, both of which may explain the absence of effects. Based on the measured endpoints, it appears that reproduction in perch was not affected at Hg concentrations known to affect fish eating wildlife.

Embryotoxicity of maternally‐transferred methylmercury to fathead minnows (Pimephales promelas)

Mercury (Hg) is a ubiquitous environmental contaminant and potent neurotoxin. In aquatic environments, Hg can be transformed into methylmercury (MeHg), which bioaccumulates in aquatic food webs, including fish. Methylmercury has been shown to transfer from female fish to developing eggs; however, relatively little is known regarding the effects of maternally transferred MeHg on fish embryos. The present study evaluated the effects of maternally transferred MeHg on fathead minnow (Pimephales promelas) embryos. Embryos were collected from adult fatheads exposed for 30 d to 1 of 3 diets spiked with MeHg: a control diet (0.02 ppm Hg dry wt), a low diet (0.87 ppm Hg dry wt), or a high diet (5.5 ppm Hg dry wt). No effects on spawning frequency, clutch size, or total egg output were observed. In embryos, Hg concentration was a function of female diet and the duration (number of days) of female exposure. Compared with controls, embryos from the low-diet treatment displayed altered embryonic movement patterns (hyperactivity) and decreased time to hatch. Embryos from the high-diet treatment had delayed hatching and increased mortality compared with the other treatments. Collectively, these results suggest that maternally transferred Hg may impact survival, behavior, and developmental milestones of the embryo-larval stages of fish. Environ Toxicol Chem 2016;35:1436-1441.