Impacts of Mercury on Freshwater Fish-Eating Wildlife and Humans

This paper reviews the current state of knowledge of the toxic effects of mercury on fish-eating birds, mammals, and humans associated with freshwater ecosystems, including new information on the relative risk of elevated methyl Hg exposure for fish-eating birds inhabiting aquatic ecosystems impacted by mining/smelting activities and areas characterized by high geological sources of Hg. The influence of various environmental conditions such as lake pH, DOC, and chemical speciation of Hg, on fish-Hg concentrations and Hg exposure in fish-eating wildlife, are discussed. Although a continuing global effort to decrease the release of this nonessential metal into the environment is warranted, Hg methylation and biomagnification may be limited in some environments due to chemical speciation of mercury in soils and sediments (e.g., HgS) and water quality conditions (e.g., high alkalinity and pH) that do not facilitate high methylation rates. We have shown such limitations for a lake where historic Hg mining greatly increased sediment-Hg loadings, yet Hg increases in small fish of various species are currently lower than expected, and top predators (bald eagles), despite having elevated concentrations of Hg in their blood compared with individuals from nearby lakes, exhibit no Hg-related reproductive impairment or other signs of MeHg intoxication. Recent epidemiological studies have shown that fish-eating human populations may be exposed to Hg sufficient to cause significant developmental effects. However, for humans, we conclude that the current USEPA reference dose for MeHg may be too restrictive, particularly for the less sensitive adult. The health status of indigenous peoples relying on the subsistence harvest of wild foods may be negatively affected by such restrictions.     

Binding of manganese in human and rat plasma

Albumin, transferrin and 'transmanganin' have all been proposed as the major Mn-binding ligand in plasma. The present investigations were initiated in order to resolve these discrepancies. Compared to other metals tested (109 Cd2+, 65Zn2+, 59Fe3+), 54Mn2+ bound poorly to purified albumin. The addition of exogenous albumin to plasma did not result in an increased 54Mn radioactivity associated with this protein. Also, incubation of 65Zn-albumin in the presence of excess Mn2+ (1 mM) did not result in the displacement of Zn from albumin or Mn binding. In contrast to these results, 54Mn was bound to purified transferrin, not as readily as Fe3+, but better than Zn2+ or Cd2+. Saturation of transferrin with Fe3+ (1.6 micrograms Fe/mg) prevented the binding of 54Mn indicating that Mn probably binds to Fe-binding sites on the protein. Polyacrylamide gel electrophoresis further demonstrated the association of 54Mn with transferrin rather than with albumin in both human and rat plasma. The amount of 54Mn radioactivity recovered with transferrin increased as incubation time was increased, probably due to oxidation of Mn2+ to Mn3+. Mn binding to transferrin reached a maximum within 5 and 12 h of incubation. About 50% of 54Mn migrated with transferrin, whereas only 5% was associated with albumin. A significant portion (20-55%) of the 54Mn radioactivity migrated with electrophoretically slow plasma components whose identity was not determined. Possibilities include alpha 2-macroglobulin, heavy gamma-globulins and/or heavy lipoproteins.     

Factors affecting the determination of total mercury in biological samples by continuous-flow cold vapor atomic absorption spectrophotometry

The acidic reduction of Hg using a continuous-flow analytical system was evaluated. With 25% SnCl₂ as the reductant, characteristic concentrations (sensitivities) of 0.44 μg/L (open cell) and 0.29 μg/L (flow-through cell) were obtained using inorganic Hg²⁺ standards in 1.5% HCl. When CH₃Hg⁺ standards were used, absorption signals were an order of magnitude lower, indicating that Sn(II) is incapable of producing Hg° from organic Hg in this acidic reduction system. Addition of CdCl₂ to the SnCl₂ reductant, as suggested by Magos (1) for the reduction of organomercurials under alkaline conditions, was without beneficial effect. Similarly, combining Sn, with another reducing agent (hydroxylamine hydrochloride), or a strong alkaline solution (40% NaOH), in the reaction coil of the flow-through system did not significantly enhance the Hg absorption signal for either inorganic or organic Hg. Recovery of Hg from spiked liver homogenates digested at 70–80°C using a HNO₃/H₂SO₄/HCl procedure and stabilized with 0.5 mM K₂Cr₂O₇ was >85%, using either inorganic Hg²⁺ or CH₃Hg⁺, indicating that this digestion procedure successfully breaks the C-Hg bond to form readily reducible Hg species. Usingl-cysteine to stabilize standards of inorganic Hg²⁺ in HCl caused significant depressions of the Hg absorption signal atl-cysteine concentrations >0.001% (≈0.5 mM); 0.1%l-cysteine caused total suppression of the Hg signal. These results indicate that: (1) acidic reduction of Hg by Sn in this continuous-flow system requires breakdown of organomercurials prior to analysis; (2) tissue digestion using HNO₃/H₂SO₄/HCl followed by the addition of K₂Cr₂O₇ to stabilize Hg²⁺ achieves this breakdown and allows good recovery of total Hg; and (3) use ofl-cysteine to complex and prevent losses of Hg should be avoided in systems using acidic reduction of Hg. Concentrations of endogenous tissue sulfhydryls are generally lower than those associated with depressed absorbance signals during the acidic reduction of Hg.     

Effects of methylmercury on epigenetic markers in three model species: Mink,chicken and yellow perch

We previously reported that methylmercury (MeHg) exposure is associated with DNA hypomethylation in the brain stem of male polar bears. Here, we conveniently use archived tissues obtained from controlled laboratory exposure studies to look for evidence that MeHg can disrupt DNA methylation across taxa. Brain (cerebrum) tissues from MeHg-exposed mink (Neovison vison), chicken (Gallus gallus) and yellow perch (Perca flavescens) were analyzed for total Hg levels and global DNA methylation. Tissues from chicken and mink, but not perch, were also analyzed for DNA methyltransferase (DNMT) activity. In mink we observed significant reductions in global DNA methylation in an environmentally-relevant dietary exposure group (1 ppm MeHg), but not in a higher group (2 ppm MeHg). DNMT activity was significantly reduced in all treatment groups. In chicken or yellow perch, no statistically significant effects of MeHg were observed. Dose-dependent trends were observed in the chicken data but the direction of the change was not consistent between the two endpoints. Our results suggest that MeHg can be epigenetically active in that it has the capacity to affect DNA methylation in mammals. The variability in results across species may suggest inter-taxa differences in epigenetic responses to MeHg, or may be related to differences among the exposure scenarios used as animals were exposed to MeHg through different routes (dietary, egg injection), for different periods of time (19–89 days) and at different life stages (embryonic, juvenile, adult).     

In vitro and whole animal evidence that methylmercury disrupts GABAergic systems in discrete brain regions in captive mink

The effects of mercury (Hg) on key components of the GABAergic system were evaluated in discrete brain regions of captive juvenile male American mink (Neovison vison) using in vitro and in vivo (whole animal) experimental approaches. In vitro studies on cortical brain tissues revealed that inorganic Hg (HgCl₂; IC50 = 0.5 ± 0.2 µM) and methyl Hg (MeHgCl; IC50 = 1.6 ± 0.2 µM) inhibited glutamic acid decarboxylase (GAD; EC 4.1.1.15) activity. There were no Hg-related effects on [³H]-muscimol binding to GABA(A) receptors (IC50s > 100 µM). HgCl₂ (IC50 = 0.8 ± 0.3 µM) but not MeHgCl (IC50 > 100 µM) inhibited GABA-transaminase (GABA-T; EC 2.6.1.19) activity. In a whole animal study, neurochemical indicators of GABAergic function were measured in brain regions (occipital cortex, cerebellum, brain stem, and basal ganglia) of captive mink fed relevant levels of MeHgCl (0 to 2 µg/g feed, ppm) daily for 89 d. No effects on GAD activity were measured. Concentration-dependent decreases in [³H]-muscimol binding to GABA(A) receptors and GABA-T activity were found in several brain regions, with reductions as great as 94% (for GABA(A) receptor levels) and 71% (for GABA-T activity) measured in the brain stem and basal ganglia. These results show that chronic exposure to environmentally relevant levels of MeHg disrupts GABAergic signaling. Given that GABA is the main inhibitory neurotransmitter in the mammalian nervous system, prolonged disruptions of its function may underlie the sub-clinical impacts of MeHg at relevant levels to animal health.     

Potential risk to common loons (Gavia immer) from methylmercury exposure in acidified lakes

Piscivorous birds and mammals in areas remote from point sources of Hg contamination may be exposed to dietary methylmercury concentrations that are sufficiently high to cause reproductive impairment. Common loons (Gavia immer) were observed to show aberrant nesting behavior and low overall reproductive success when Hg concentrations in prey (small fish and crayfish) averaged > 0.3 µg g^–1 wet weight (Barr, 1986), levels known to occur in fish from many lakes in central Ontario. We used data on Hg in Ontario fish to estimate the proportion of lakes where fish small enough for loons to eat (< 250 g) had Hg concentrations that exceeded estimated thresholds for reproductive impairment. Up to 30 % of lakes exceeded thresholds for reproductive impairment, depending on the species of fish and the threshold Hg concentrations chosen. There was a significant negative correlation between fish-Hg concentration and lake pH in most fish species examined. For these species, reductions in sulfate deposition rates are predicted to result in a corresponding reduction of lakes in Ontario having fish with potentially toxic concentrations of Hg.