Expression Levels of hgcAB Genes and Mercury Availability Jointly Explain Methylmercury Formation in Stratified Brackish Waters.

Neurotoxic methylmercury (MeHg) is formed by microbial methylation of inorganic divalent Hg (HgII) and constitutes severe environmental and human health risks. The methylation is enabled by hgcA and hgcB genes, but it is not known if the associated molecular-level processes are rate-limiting or enable accurate prediction of MeHg formation in nature. In this study, we investigated the relationships between hgc genes and MeHg across redox-stratified water columns in the brackish Baltic Sea. We showed, for the first time, that hgc transcript abundance and the concentration of dissolved HgII-sulfide species were strong predictors of both the HgII methylation rate and MeHg concentration, implying their roles as principal joint drivers of MeHg formation in these systems. Additionally, we characterized the metabolic capacities of hgc+ microorganisms by reconstructing their genomes from metagenomes (i.e., hgc+ MAGs), which highlighted the versatility of putative HgII methylators in the water column of the Baltic Sea. In establishing relationships between hgc transcripts and the HgII methylation rate, we advance the fundamental understanding of mechanistic principles governing MeHg formation in nature and enable refined predictions of MeHg levels in coastal seas in response to the accelerating spread of oxygen-deficient zones.

Explosives compounds from sea-dumped relic munitions accumulate in marine biota.

Relic munitions are a hazardous legacy of the two world wars present in coastal waters worldwide. The southwest Baltic Sea has an especially high prevalence of unexploded ordnance and dumped munition material, which represent a large potential source of toxic explosive chemicals (munition compounds, MC). In the current study, diverse biota (plankton, macroalgae, tunicate, sponge, mollusc, echinoderm, polychaete, anemone, crustacea, fish) were collected from the Kiel Bight and a munitions dumpsite at Kolberger Heide, Germany, to evaluate the potential bioaccumulation of explosives and their derivatives (2,4,6-trinitrotoluene, TNT; 2-amino-4,6-dinitrotoluene and 4-amino-2,6-dinitrotoluene, ADNT; 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene, DANT; 1,3-dinitrobenzene, DNB; and 1,3,5-trinitro-1,3,5-triazinane, RDX). One or more MCs were detected in >98% of organisms collected throughout the study region (n = 178), at a median level of 6 pmol/g (approximately 1 ng/g) and up to 2 × 107 pmol/g (TNT in Asterias rubens collected from Kolberger Heide). In most cases, TNT and its transformation product compounds ADNT and DANT were significantly higher in biota from the munitions dumpsite compared with other locations. Generally, DNB and RDX were detected less frequently and at lower concentrations than TNT, ADNT, and DANT. In commercially important fish species (plaice, flounder) from Kolberger Heide, TNT and ADNT were detected in 17 and 33% of samples, respectively. In contrast DANT was detected in every fish sample, including those outside the dumpsite. Dinitrobenzene was the second most prevalent MC in fish tissue. Fish viscera (stomach, kidney, liver) showed higher levels of DANT than edible muscle flesh, with highest DANT in liver, suggesting reduced risk to seafood consumers. This study provides some of the first environmental evidence for widespread bioaccumulation of MC in a coastal marine food web. Although tissue MC content was generally low, corrosion of munition housings may lead to greater MC release in the future, and the ecological risk of this exposure is unknown.

Deltaproteobacteria and Spirochaetes-Like Bacteria Are Abundant Putative Mercury Methylators in Oxygen-Deficient Water and Marine Particles in the Baltic Sea.

Methylmercury (MeHg), a neurotoxic compound biomagnifying in aquatic food webs, can be a threat to human health via fish consumption. However, the composition and distribution of the microbial communities mediating the methylation of mercury (Hg) to MeHg in marine systems remain largely unknown. In order to fill this knowledge gap, we used the Baltic Sea Reference Metagenome (BARM) dataset to study the abundance and distribution of the genes involved in Hg methylation (the hgcAB gene cluster). We determined the relative abundance of the hgcAB genes and their taxonomic identity in 81 brackish metagenomes that cover spatial, seasonal and redox variability in the Baltic Sea water column. The hgcAB genes were predominantly detected in anoxic water, but some hgcAB genes were also detected in hypoxic and normoxic waters. Phylogenetic analysis identified putative Hg methylators within Deltaproteobacteria, in oxygen-deficient water layers, but also Spirochaetes-like and Kiritimatiellaeota-like bacteria. Higher relative quantities of hgcAB genes were found in metagenomes from marine particles compared to free-living communities in anoxic water, suggesting that such particles are hotspot habitats for Hg methylators in oxygen-depleted seawater. Altogether, our work unveils the diversity of the microorganisms with the potential to mediate MeHg production in the Baltic Sea and pinpoint the important ecological niches for these microorganisms within the marine water column.

Quantification of total concentration of thiol functional groups in environmental samples by titration with monobromo(trimethylammonio)bimane and determination with tandem mass spectrometry.

Thiol compounds (R-SH) have many important biological functions and are principal controls of the speciation of several toxic metals in the environment. However, determining the concentration of thiols associated with environmental matrices is challenging due to the compounds' low abundance and interferences from non-thiol compounds for many available methods. Here a novel method has been developed and validated to quantify the total concentration of thiol functional groups in aqueous samples using derivatization with monobromo(trimethylammonio)bimane (qBBr) and quantification with tandem mass spectrometry. The thiol concentration was determined by titration of the sample with qBBr, which reacts selectively with thiols, and quantification of the residual qBBr. We systematically evaluated potential interferences from various organic compounds, inorganic ions (including sea water matrices), sulfide and mercury (Hg) species, and demonstrate that the method is highly sensitive, selective and robust. The limit of detection (LOD) for total thiols is in the nanomolar concentration range (~6 nM). The method performance was also demonstrated by determination of the total thiol concentration in different natural samples including boreal stream water (1.16 µM), wetland porewater (0.96 µM) and the Suwanee River natural organic matter (NOM) reference material SR101 N (7.9 µmol g-1). The developed method represents a combination of low LOD and high selectivity and robustness that is unsurpassed for total thiol concentration measurements.

Assessment of Hg contamination by a Chlor-Alkali Plant in riverine and coastal sites combining Hg speciation and isotopic signature (Sagua la Grande River, Cuba).

Chlor-alkali plants (CAP) are recognized as major sources of mercury (Hg) in the environment. In this work, Hg concentration, speciation and isotopic signature were determined in sediments and biota (fish and oyster) from Sagua La Grande River (SG River) and the adjacent coastal zone in the vicinity of a CAP (Cuba). High Hg concentrations in surface sediments (up to 5072 ng g-1), mainly occurring as inorganic Hg, decrease with the distance from the CAP along the SG River and seaward. Meanwhile, Hg concentration and speciation in riverine catfish (Claria gariepinus) muscle (1093 ± 319 ng g-1, ˜70% as MeHg) and coastal oysters (Crassostrea rizophorae) (596 ± 233 ng g-1, ˜50% as MeHg) indicate a direct impact from CAP. Hg isotopic signature in sediments, following both mass dependent (MDF) and mass independent fractionation (MIF), exhibits a clear binary mixing between CAP pollution (+0.42‰, δ202Hg; -0.18‰, Δ201Hg) and regional background end-member (˜ -0.49‰, δ202Hg; +0.01‰, Δ201Hg). The combination of speciation and isotopic information in biota and sediments allows to trace Hg contamination pathways from contaminated sediments to the biota, establishing the importance of both methylation and demethylation extent in both river and coastal sites before Hg species bioaccumulation.

Unraveling prevalence and public health risks of arsenic, uranium and co-occurring trace metals in groundwater along riverine ecosystem in Sindh and Punjab, Pakistan.

The current study focuses on the understanding of contamination status, distribution, source apportionment and health perspectives of arsenic (As), uranium (U) and other co-occurring trace metals in the groundwater samples collected along the major rivers in Sindh and Punjab provinces, Pakistan. ICP-MS analysis revealed that the concentrations of As in the groundwater in Sindh and Punjab ranged from 0.2 to 81.1 µg/L (n = 38) and 1.1 to 501.1 µg/L (n = 110), respectively. Importantly, this study is the first evidence of U contamination in the groundwater samples in Pakistan, which revealed the concentrations of U at from 0.8 to 59.0 and 0.1 to 556.0 µg/L respectively, in Sindh and Punjab. Moreover, the concentrations of Sr and Mn exceeded the WHO limits in the current study area. Anthropogenic activities such as urbanization, direct dispose of industrial, agricultural waste into waterways and extensive use of pesticides and fertilizers might be the main sources of elevated levels of total dissolved solids and electrical conductivity, which increased the mobilization of As, U and Sr in the groundwater samples. Human health risk assessment parameters such as average daily dose, hazard quotient (HQ) and cancer risk indicated severe risks of As and U in the study area. The HQ values of As and U in Punjab were observed at 69.6 and 7.7, respectively, implying the severity of the health risks associated with consumption of contaminated groundwater for drinking purposes. In a nutshell, proactive control and rehabilitation measures are recommended to eradicate trace metals associated groundwater contamination in the targeted areas to avoid future worst scenarios.

Specific Effects of Dietary Methylmercury and Inorganic Mercury in Zebrafish (Danio rerio) Determined by Genetic, Histological, and Metallothionein Responses.

A multidisciplinary approach is proposed here to compare toxicity mechanisms of methylmercury (MeHg) and inorganic mercury (iHg) in muscle, liver, and brain from zebrafish (Danio rerio). Animals were dietary exposed to (1) 50 ng Hg g(-1), 80% as MeHg; (2) diet enriched in MeHg 10000 ng Hg g(-1), 95% as MeHg; (3) diet enriched in iHg 10000 ng Hg g(-1), 99% as iHg, for two months. Hg species specific bioaccumulation pathways were highlighted, with a preferential bioaccumulation of MeHg in brain and iHg in liver. In the same way, differences in genetic pattern were observed for both Hg species, (an early genetic response (7 days) for both species in the three organs and a late genetic response (62 days) for iHg) and revealed a dissimilar metabolization of both Hg species. Among the 18 studied genes involved in key metabolic pathways of the cell, major genetic responses were observed in muscle. Electron microscopy revealed damage mainly because of MeHg in muscle and also in liver tissue. In brain, high MeHg and iHg concentrations induced metallothionein production. Finally, the importance of the fish origin in ecotoxicological studies, here the seventh descent of a zebrafish line, is discussed.

Specific Pathways of Dietary Methylmercury and Inorganic Mercury Determined by Mercury Speciation and Isotopic Composition in Zebrafish (Danio rerio).

An original approach is proposed to investigate inorganic (iHg) and methylmercury (MeHg) trophic transfer and fate in a model fish, Danio rerio, by combining natural isotopic fractionation and speciation. Animals were exposed to three different dietary conditions: (1) 50 ng Hg g(-1), 80% as MeHg; (2) diet enriched in MeHg 10,000 ng Hg g(-1), 95% as MeHg, and (3) diet enriched in iHg 10,000 ng Hg g(-1), 99% as iHg. Harvesting was carried out after 0, 7, 25, and 62 days. Time-dependent Hg species distribution and isotopic fractionation in fish organs (muscle, brain, liver) and feces, exhibited different patterns, as a consequence of their dissimilar metabolization. The rapid isotopic re-equilibration to the new MeHg-food source reflects its high bioaccumulation rate. Relevant aspects related to Hg excretion are also described. This study confirms Hg isotopic fractionation as a powerful tool to investigate biological processes, although its deconvolution and fully understanding is still a challenge.