Using species-specific enriched stable isotopes to study the effect of fresh mercury inputs in soil-earthworm systems.

The fate of mercury (Hg) in the soil-earthworm system is still far from being fully understood, especially regarding recurrent and challenging questions about the importance of the reactivity of exogenous Hg species. Thus, to predict the potential effect of Hg inputs in terrestrial ecosystems, it is necessary to evaluate separately the reactivity of the endogenous and exogenous Hg species and, for this purpose, the use of enriched stable isotope tracers is a promising tool. In the present work, earthworms (Lumbricus terrestris) were exposed to historically Hg contaminated soils from the Almadén mining district, Spain. The soils were either non-spiked, which contain only endogenous or native Hg naturally occurring in the soil, or spiked with isotopically enriched inorganic Hg (199IHg), representing exogenous or spiked Hg apart from the native one. The differential reactivity of endogenous and exogenous Hg in the soil conditioned the processes of methylation, mobilization, and assimilation of inorganic Hg by earthworms. Both endogenous and exogenous Hg species also behave distinctly regarding their bioaccumulation in earthworms, as suggested by the bioaccumulation factors, being the endogenous methylmercury (MeHg) the species more readily bioaccumulated by earthworms and in a higher extent. To the best of our knowledge, this work demonstrates for the first time the potential of enriched stable isotopes to study the effects of fresh Hg inputs in soil-earthworm systems. The findings of this work can be taken as a case study on the dynamics of Hg species in complex terrestrial systems and open a new door for future experiments.

Identical Hg isotope mass dependent fractionation signature during methylation by sulfate-reducing bacteria in sulfate and sulfate-free environment.

Inorganic mercury (iHg) methylation in aquatic environments is the first step leading to monomethylmercury (MMHg) bioaccumulation in food webs and might play a role in the Hg isotopic composition measured in sediments and organisms. Methylation by sulfate reducing bacteria (SRB) under sulfate-reducing conditions is probably one of the most important sources of MMHg in natural aquatic environments, but its influence on natural Hg isotopic composition remains to be ascertained. In this context, the methylating SRB Desulfovibrio dechloracetivorans (strain BerOc1) was incubated under sulfate reducing and fumarate respiration conditions (SR and FR, respectively) to determine Hg species specific (MMHg and IHg) isotopic composition associated with methylation and demethylation kinetics. Our results clearly establish Hg isotope mass-dependent fractionation (MDF) during biotic methylation (-1.20 to +0.58‰ for δ(202)Hg), but insignificant mass-independent fractionation (MIF) (-0.12 to +0.15‰ for Δ(201)Hg). During the 24h of the time-course experiments Hg isotopic composition in the produced MMHg becomes significantly lighter than the residual IHg after 1.5h and shows similar δ(202)Hg values under both FR and SR conditions at the end of the experiments. This suggests a unique pathway responsible for the MDF of Hg isotopes during methylation by this strain regardless the metabolism of the cells. After 9 h of experiment, significant simultaneous demethylation is occurring in the culture and demethylates preferentially the lighter Hg isotopes of MMHg. Therefore, depending on their methylation/demethylation capacities, SRB communities in natural sulfate reducing conditions likely have a significant and specific influence on the Hg isotope composition of MMHg (MDF) in sediments and aquatic organisms.

Tracking the fate of mercury in the fish and bottom sediments of Minamata Bay, Japan, using stable mercury isotopes.

Between 1932 and 1968, industrial wastewater containing methylmercury (MeHg) and other mercury (Hg) compounds was discharged directly into Minamata Bay, Japan, seriously contaminating the fishery. Thousands of people who consumed tainted fish and shellfish developed a neurological disorder now known as Minamata disease. Concentrations of total mercury (THg) in recent fish and sediment samples from Minamata Bay remain higher than those in other Japanese coastal waters, and elevated concentrations of THg in sediments in the greater Yatsushiro Sea suggest that Hg has moved beyond the bay. We measured stable Hg isotope ratios in sediment cores from Minamata Bay and the southern Yatsushiro Sea and in archived fish from Minamata Bay dating from 1978 to 2013. Values of δ(202)Hg and Δ(199)Hg in Yatsushiro Sea surface sediments were indistinguishable from those in highly contaminated Minamata Bay sediments but distinct from and nonoverlapping with values in background (noncontaminated) sediments. We conclude that stable Hg isotope data can be used to track Minamata Bay Hg as it moves into the greater Yatsushiro Sea. In addition, our data suggest that MeHg is produced in bottom sediments and enters the food web without substantial prior photodegradation, possibly in sediment porewaters or near the sediment-water interface.

Assessment of mercury exposure among small-scale gold miners using mercury stable isotopes.

Total mercury (Hg) concentrations in hair and urine are often used as biomarkers of exposure to fish-derived methylmercury (MeHg) and gaseous elemental Hg, respectively. We used Hg stable isotopes to assess the validity of these biomarkers among small-scale gold mining populations in Ghana and Indonesia. Urine from Ghanaian miners displayed similar Δ(199)Hg values to Hg derived from ore deposits (mean urine Δ(199)Hg=0.01‰, n=6). This suggests that urine total Hg concentrations accurately reflect exposure to inorganic Hg among this population. Hair samples from Ghanaian miners displayed low positive Δ(199)Hg values (0.23-0.55‰, n=6) and low percentages of total Hg as MeHg (7.6-29%, n=7). These data suggest that the majority of the Hg in these miners' hair samples is exogenously adsorbed inorganic Hg and not fish-derived MeHg. Hair samples from Indonesian gold miners who eat fish daily displayed a wider range of positive Δ(199)Hg values (0.21-1.32‰, n=5) and percentages of total Hg as MeHg (32-72%, n=4). This suggests that total Hg in the hair samples from Indonesian gold miners is likely a mixture of ingested fish MeHg and exogenously adsorbed inorganic Hg. Based on data from both populations, we suggest that total Hg concentrations in hair samples from small-scale gold miners likely overestimate exposure to MeHg from fish consumption.

Organ-specific mercury stable isotopes, speciation and particle measurements reveal methylmercury detoxification processes in Atlantic Bluefin Tuna.

Identifying metabolism and detoxification mechanisms of Hg in biota has important implications for biomonitoring, ecotoxicology, and food safety. Compared to marine mammals and waterbirds, detoxification of MeHg in fish is understudied. Here, we investigated Hg detoxification in Atlantic bluefin tuna Thunnus thynnus using organ-specific Hg and Se speciation data, stable Hg isotope signatures, and Hg and Se particle measurements in multiple tissues. Our results provide evidence for in vivo demethylation and biomineralization of HgSe particles, particularly in spleen and kidney. We observed a maximum range of 1.83‰ for δ202Hg between spleen and lean muscle, whereas Δ199Hg values were similar across all tissues. Mean percent methylmercury ranged from 8% in spleen to 90% in lean muscle. The particulate masses of Hg and Se were higher in spleen and kidney (Hg: 61% and 59%, Se: 12% and 6%, respectively) compared to muscle (Hg: 2%, Se: 0.05%). Our data supports the hypothesis of an organ-specific, two-step detoxification of methylmercury in wild marine fish, consisting of demethylation and biomineralization, like reported for waterbirds. While mass dependent fractionation signatures were highly organ specific, stable mass independent fractionation signatures across all tissues make them potential candidates for source apportionment studies of Hg using ABFT.

In vivo mercury methylation and demethylation in freshwater tilapia quantified by mercury stable isotopes.

In vivo methylation and demethylation processes were simultaneously investigated in freshwater tilapia after dietary exposure to mercury ((198)Hg(II) and methyl(200)Hg). During one month dietary exposure followed by two month depuration, both MeHg and THg increased continuously in muscle tissues but decreased in liver during depuration, indicating the inter-organ transportation of MeHg from liver toward muscle. Direct evidence of in vivo net methylation process in freshwater tilapia was observed. Specifically, 0.67-1.60% of the ingested Hg(198)(II) was converted into Me(198)Hg and deposited in fish muscle at the end of depuration. The methylation potential in terms of methylated fraction was elevated at higher temperature and decreased at higher dosage. However, no direct evidence of MeHg demethylation was observed. In contrast to some previous reports of dose-dependent demethylation, the percentage of MeHg in the liver decreased significantly with increasing THg concentrations, likely due to the faster inter-organ MeHg transportation from liver toward muscle. Our study demonstrates the important role of organ- and species-specific biodynamics in understanding mercury transformation and speciation in fish. The observed in vivo methylation process in tilapia was slow, suggesting that the high %MeHg in fish should be mainly derived from MeHg ingestion instead of in vivo transformation.

Investigating uptake and translocation of mercury species by sawgrass ( Cladium jamaicense ) using a stable isotope tracer technique.

The role of macrophytes in mercury (Hg) cycling in the Florida Everglades ecosystem has not been fully understood. In this study, a stable isotope ((199)Hg(2+)) addition technique was used to trace the methylation, uptake, and translocation of Hg by sawgrass ( Cladium jamaicense ) and quantitatively evaluate the contribution of atmospheric and soil Hg to Hg in sawgrass leaves and below-ground biomass. The results showed that spiked (199)Hg(2+) could be rapidly methylated to monomethylmercury (Me(199)Hg) in the soil of the sawgrass pots. Only small portions of total Hg (THg) and monomethylmercury (MeHg) in the soil could be taken up by sawgrass, indicated by the ratios of T(199)Hg and Me(199)Hg (tracer) concentrations in the sawgrass below-ground biomass (BGBM) over that in the soil (6.50 ± 1.9% and 12.8 ± 3.6% for THg and MeHg, respectively). Concentrations of T(199)Hg (tracer) and Me(199)Hg (tracer) in sawgrass leaves only accounted for 5.50 ± 2.8% and 15.6 ± 4.0%, respectively, of that in the BGBM, implying that the fractions of mercury species transported upward by sawgrass were also small. Statistical analysis (t test) showed that sawgrass preferred MeHg over THg in both uptake and upward translocation. The majority (>90%) of THg in sawgrass leaves were estimated to be obtained from atmospheric Hg, rather than from soil, suggesting that assimilation of atmospheric Hg could increase the overall Hg stock in the Florida Everglades ecosystem. The finding about foliar uptake of Hg is especially important for a better understanding of mercury cycling in the Everglades, given the large amount of sawgrass biomass in this ecosystem.

Mercury isotope compositions in seawater and marine fish revealed the sources and processes of mercury in the food web within differing marine compartments.

Anthropogenic activities and climate change have significantly increased mercury (Hg) levels in seawater. However, the processes and sources of Hg in differing marine compartments (e.g. estuary, marine continental shelf (MCS) or pelagic area) have not been well studied, which makes it difficult to understand Hg cycling in marine ecosystems. To address this issue, the total Hg (THg) concentration, methylmercury (MeHg) concentration and stable Hg isotopes were determined in seawater and fish samples collected from differing marine compartments of the South China Sea (SCS). The results showed that the estuarine seawater exhibited substantially higher THg and MeHg concentrations than those in the MCS and pelagic seawater. Significantly negative δ202Hg (-1.63‰ ± 0.42‰) in estuarine seawater compared with that in pelagic seawater (-0.58‰ ± 0.08‰) may suggest watershed input and domestic sewage discharge of Hg in the estuarine compartment. The Δ199Hg value in estuarine fish (0.39‰ ± 0.35‰) was obviously lower than that in MCS (1.10‰ ± 0.54‰) and pelagic fish (1.15‰ ± 0.46‰), which showed that relatively little MeHg photodegradation occurred in the estuarine compartment. The Hg isotope binary mixing model based on Δ200Hg revealed that approximately 74% MeHg in pelagic fish is derived from atmospheric Hg(II) deposition, and over 60% MeHg in MCS fish is derived from sediments. MeHg sources for estuarine fish may be highly complex (e.g. sediment or riverine/atmospheric input) and further investigations are warranted to clarify the contribution of each source. Our study showed that Hg stable isotopes in seawater and marine fish can be used to identify the processes and sources of Hg in different marine compartments. This finding is of great relevance to the development of marine Hg food web models and the management of Hg in fish.

Mercury isotopes link mercury in San Francisco Bay forage fish to surface sediments.

Identification of sources of biologically accessible Hg is necessary to fully evaluate Hg exposure in aquatic ecosystems. This study assesses the relationship between Hg in forage fish and Hg in surface sediments throughout San Francisco Bay (SF Bay) and evaluates processes influencing the incorporation of Hg into the aquatic food web. We measured the Hg stable isotope compositions of two nearshore fish species and compared them with previously reported analyses of colocated intertidal surface sediments. Fish δ(202)Hg values (mass-dependent fractionation) demonstrated a distinct spatial gradient within SF Bay that ranged from 0.60‰ in the south to -0.25‰ in the north. Fish δ(202)Hg values were consistently higher than sediment δ(202)Hg values by 0.73‰ (±0.16‰, 1SD). Fish and sediment δ(202)Hg values in SF Bay proper were well correlated (r(2) = 0.83), suggesting that sediment is a primary source of Hg to the nearshore aquatic food web. Fish Δ(199)Hg values (mass-independent fractionation) ranged from 0.46‰ to 1.55‰, did not correlate with sediment values, and yielded a Δ(199)Hg/Δ(201)Hg ratio of 1.26 (±0.01, 1SD; r(2) = 0.99). This mass-independent fractionation is consistent with photodegradation of MeHg to varying degrees at each site prior to incorporation into the food web.

Tracing sources and bioaccumulation of mercury in fish of Lake Baikal--Angara River using Hg isotopic composition.

This study presents the determination and comparison of isotopic compositions of Hg in sediments, plankton, roach, and perch of two freshwater systems in the Lake Baikal-Angara River aquatic ecosystem: the man-made Bratsk Water Reservoir contaminated by Hg from a chlor-alkali factory and the noncontaminated Lake Baikal. Isotopic ratios of biota exhibit both significant mass-independent fractionation (MIF) (Δ(199)Hg from 0.20 to 1.87‰) and mass-dependent fractionation (MDF) (δ(202)Hg from -0.97 to -0.16‰), whereas sediments exhibit high MDF (δ(202)Hg from -1.99 to -0.83‰) but no MIF. δ(15)N and δ(13)C are correlated with methylmercury in organisms from both sites, indicating bioaccumulation and biomagnification through food webs of both regions. Combining this with isotopic composition of samples shows that δ(202)Hg increases with the trophic level of organisms and also with methylmercury in fish from Lake Baikal. This study demonstrates that MIF in fish samples from Bratsk Water Reservoir allow to trace anthropogenic Hg, since fish with the highest levels of Hg in muscle have the same isotopic composition as the sediment in which anthropogenic Hg was deposited. Less contaminated fish do not exhibit this anthropogenic signature accumulating relatively lower Hg amount from the contaminated sediments. This work reveals that Hg isotopic composition can be used to track the contribution of anthropogenic sources in fish from a contaminated lake.

Understanding sources of methylmercury in songbirds with stable mercury isotopes: Challenges and future directions.

Mercury (Hg) stable isotope analysis is an emerging technique that has contributed to a better understanding of many aspects of the biogeochemical cycling of Hg in the environment. However, no study has yet evaluated its usefulness in elucidating the sources of methylmercury (MeHg) in songbird species, a common organism for biomonitoring of Hg in forested ecosystems. In the present pilot study, we examined stable mercury isotope ratios in blood of 4 species of songbirds and the invertebrates they are likely foraging on in multiple habitats in a small watershed of mixed forest and wetlands in Acadia National Park in Maine (USA). We found distinct isotopic signatures of MeHg in invertebrates (both mass-dependent fractionation [as δ202 Hg] and mass-independent fractionation [as Δ199 Hg]) among 3 interconnected aquatic habitats. It appears that the Hg isotopic compositions in bird blood cannot be fully accounted for by the isotopic compositions of MeHg in lower trophic levels in each of the habitats examined. Furthermore, the bird blood isotope results cannot be simply explained by an isotopic offset as a result of metabolic fractionation of δ202 Hg (e.g., internal demethylation). Our results suggest that many of the birds sampled obtain MeHg from sources outside the habitat they were captured in. Our findings also indicate that mass-independent fractionation is a more reliable and conservative tracer than mass-dependent fractionation for identifying sources of MeHg in bird blood. The results demonstrate the feasibility of Hg isotope studies of songbirds but suggest that larger numbers of samples and an expanded geographic area of study may be required for conclusive interpretation. Environ Toxicol Chem 2018;37:166-174. © 2017 SETAC.

Using mercury isotopes to understand the bioaccumulation of Hg in the subtropical Pearl River Estuary, South China.

Coastal and estuarine regions are important areas of mercury pollution. Therefore, it is important to properly characterize the sources and bioaccumulation processes of mercury in these regions. Here, we present mercury stable isotopic compositions in 18 species of wild marine fish collected from the Pearl River Estuary (PRE), south China. Our results showed variations in mass-independent fractionation (Δ(199)Hg: +0.05 ± 0.10‰ to +0.59 ± 0.30‰) with a Δ(199)Hg/Δ(201)Hg of ∼1.26, suggesting that aqueous MeHg underwent photo-degradation prior to incorporation into the food chain. For the results, we discovered small but significant differences of Δ(199)Hg values among herbivorous, demersal, and carnivorous fish, indicating that different feeding guilds of fish may have incorporated MeHg with various degrees of photo-demethylation. The consistent mercury isotope compositions between fish feeding habitat and mercury sources in the estuary provide potentially important findings on the transformation and bioaccumulation of this toxic metal in subtropical coastal environments.

Natural Hg isotopic composition of different Hg compounds in mammal tissues as a proxy for in vivo breakdown of toxic methylmercury.

In the last decade, specific attention has been paid to total mercury (HgT) stable isotopic composition, especially in natural samples such as aquatic organisms, due to its potential to track the cycle of this toxic element in the environment. Here, we investigated Hg Compound Specific stable Isotopic Composition (CSIC) of natural inorganic Hg (iHg) and methylmercury (MMHg) in various tissues of aquatic mammals (Beluga whale from the Arctic marine environment and seals from the freshwater lake Baikal, Russia). In seals' organs the variation in mass dependent fractionation (MDF, δ(202)Hg) for total Hg was significantly correlated to the respective fraction of iHg and MMHg compounds, with MMHg being enriched by ∼ 3‰ in heavier isotopes relative to iHg. On the other hand, we observe insignificant variation in Hg mass independent isotope fractionation (MIF, Δ(199)Hg) among iHg and MMHg in all organs for the same mammal species and MMHg in prey items. MIF signatures suggest that both MMHg and iHg in aquatic mammals have the same origin (i.e., MMHg from food), and are representative of Hg photochemistry in the water column of the mammal ecosystem. MDF signatures of Hg compounds indicate that MMHg is demethylated in vivo before being stored in the muscle, and the iHg formed is stored in the liver, and to a lesser extent in the kidney, before excretion. Thus, Hg CSIC analysis in mammals can be a powerful tool for tracing the metabolic response to Hg exposure.

Inter-individual variations of human mercury exposure biomarkers: a cross-sectional assessment.

BACKGROUND: Biomarkers for mercury (Hg) exposure have frequently been used to assess exposure and risk in various groups of the general population. We have evaluated the most frequently used biomarkers and the physiology on which they are based, to explore the inter-individual variations and their suitability for exposure assessment. METHODS: Concentrations of total Hg (THg), inorganic Hg (IHg) and organic Hg (OHg, assumed to be methylmercury; MeHg) were determined in whole blood, red blood cells, plasma, hair and urine from Swedish men and women. An automated multiple injection cold vapour atomic fluorescence spectrophotometry analytical system for Hg analysis was developed, which provided high sensitivity, accuracy, and precision. The distribution of the various mercury forms in the different biological media was explored. RESULTS: About 90% of the mercury found in the red blood cells was in the form of MeHg with small inter-individual variations, and part of the IHg found in the red blood cells could be attributed to demethylated MeHg. THg in plasma was associated with both IHg and MeHg, with large inter-individual variations in the distribution between red blood cells and plasma. THg in hair reflects MeHg exposure at all exposure levels, and not IHg exposure. The small fraction of IHg in hair is most probably emanating from demethylated MeHg. The inter-individual variation in the blood to hair ratio was very large. The variability seemed to decrease with increasing OHg in blood, most probably due to more frequent fish consumption and thereby blood concentrations approaching steady state. THg in urine reflected IHg exposure, also at very low IHg exposure levels. CONCLUSION: The use of THg concentration in whole blood as a proxy for MeHg exposure will give rise to an overestimation of the MeHg exposure depending on the degree of IHg exposure, why speciation of mercury forms is needed. THg in RBC and hair are suitable proxies for MeHg exposure. Using THg concentration in plasma as a measure of IHg exposure can lead to significant exposure misclassification. THg in urine is a suitable proxy for IHg exposure.

Tracing mercury pathways in Augusta Bay (southern Italy) by total concentration and isotope determination.

The mercury (Hg) pollution of sediments is the main carrier of Hg for the biota and, subsequently, for the local fish consumers in Augusta Bay area (SE Sicily, Italy), a coastal marine system affected by relevant sewage from an important chlor-alkali factory. This relationship was revealed by the determination of Mass Dependent (MDF) and Mass Independent Fractionation (MIF) of Hg isotopes in sediment, fish and human hair samples. Sediments showed MDF but no MIF, while fish showed MIF, possibly due to photochemical reduction in the water column and depending on the feeding habitat of the species. Benthic and demersal fish exhibited MDF similar to that of sediments in which anthropogenic Hg was deposited, while pelagic organisms evidenced higher MDF and MIF due to photoreduction. Human hair showed high values of δ(202)Hg (offset of +2.2‰ with respect to the consumed fish) and Δ(199)Hg, both associated to fish consumption.

Mercury stable isotopes in sediments and largemouth bass from Florida lakes, USA.

Humans and wildlife can be exposed to mercury (Hg) through the consumption of fish with elevated concentrations of methylmercury (MeHg). Studies have shown that increased atmospheric deposition of Hg often leads to increased MeHg concentrations in aquatic organisms. However, depending on the ecosystem characteristics, reductions in Hg emissions may not always lead to immediate decreases in fish MeHg concentrations. Measurements of natural abundance Hg stable isotope ratios may enable a better understanding of these complex relationships. To gain insight into the sources of Hg to sport fish in central Florida, we measured the Hg isotopic compositions of surface sediments and largemouth bass from freshwater lakes. We found that fish collected from lakes located near the large Crystal River coal-fired power plant did not display evidence of anomalous negative δ(202)Hg values that were observed in nearby precipitation. This suggests that Hg recently deposited from the atmosphere is not preferentially methylated and bioaccumulated in these lakes relative to previously deposited Hg accumulated in the lake sediments. We also observed significant positive Δ(199)Hg values in the fish that were correlated with light penetration depth in the lakes from which they were collected. This indicates that a significant amount of photochemical degradation of MeHg (up to ~40%) occurred prior to uptake of the remaining MeHg into the food webs. These results suggest that depending on physical lake characteristics and biogeochemical factors, decreased atmospheric Hg deposition may not lead to immediate short-term reductions in fish MeHg concentrations. Instead, recovery of some freshwater fish populations to baseline MeHg concentrations may take decades to centuries.

Stable isotope (N, C, Hg) study of methylmercury sources and trophic transfer in the northern gulf of Mexico.

We combined N, C, and Hg stable isotope measurements to identify the most important factors that influence MeHg accumulation in fish from the northern Gulf of Mexico (nGOM), and to determine if coastal species residing in the Mississippi River (MR) plume and migratory oceanic species derive their MeHg from the same, or different, sources. In six coastal species and two oceanic species (blackfin and yellowfin tuna), trophic position as measured by delta(15)N explained most of the variance in log[MeHg] (r(2) approximately 0.8), but coastal species and tuna fell along distinct, nearly parallel lines with significantly different intercepts. The tuna also had significantly higher delta(202)Hg (0.2-0.5 per thousand) and Delta(201)Hg ( approximately 1.5 per thousand) than the coastal fish (delta(202)Hg = 0 to -1.0 per thousand; Delta(201)Hg approximately 0.4 per thousand). The observations can be best explained by largely disconnected food webs rooted in different baseline delta(15)N signatures (MR-plume vs oceanic) and isotopically distinct MeHg sources, with oceanic MeHg having undergone substantial photodegradation ( approximately 50%) before entering the base of the food web. Given the MR's large, productive footprint in the nGOM and the potential for exporting prey and MeHg to the adjacent oligotrophic GOM, the disconnected food webs and different MeHg sources are consistent with recent evidence in other systems of important oceanic MeHg sources.