The preparation and properties of a stable mercury derivative of transfer RNAs from Escherichia coli

Further investigations into the properties of the mercury derivative formed by the reaction of 4-thiouridine-containing tRNAs and pentafluorophenylmercury chloride have been carried out. tRNA^fMet (which contains only one 4-thiouridine residue) has been isolated by a one-step column Chromatographic procedure from unfractionated Escherichia coli tRNA and has been shown to react with the mercury compound to give a derivative which has similar properties to those previously reported for the corresponding mercury derivative of tRNA^Tyr which contains two adjacent 4-thiouridine residues. The mercury derivative of tRNA^Tyr appears to be a competitive inhibitor of tRNA^Tyr in the aminoacylation reaction (tRNA^TyrK_m = 0.42 μM, mercury derivative of tRNA^TyrK_i = 0.11 μM). The mercury derivative of Tyr-tRNA^Tyr can be made, but only by the reaction of the mercury compound with the aminoacylated tRNA.     

Mercury and osmoregulation in the euryhaline crab,Eriocheir sinensis

This review will focus on cases where it might be possible that the toxicity of the heavy metal mercury results from an interaction with osmoregulatory mechanisms. It is shown that mercury-induced impairment of osmoregulatory capability in the sense of severe modifications of the blood osmotic concentration is more pronounced in brachyuran decapod species adapted to dilute waters. The rationale for considering these effects is based on a comparison between mercury effects on three species of decapod crustaceans exhibiting various degrees of osmoregulation capability: the strong regulator crab Eriocheir sinensis, the weak regulator Carcinus maenas, both of them being euryhaline, and the stenohaline osmoconformer Cancer pagurus. It is established that a synergistic effect exists between salinity and HgCl₂ toxicity in euryhaline species which are hyperregulators in dilute media, that is, E. sinensis and C. maenas only. Depth study of E. sinensis as a model demonstrates that Na⁺ and Cl⁻ permeabilities of the gill epithelium is affected by mercury, as well as the Na⁺ and Cl⁻ active transport processes located at the same level. Evidences are brought showing that mercury drastically disturbs the Na⁺/K⁺ pump and the Cl⁻ channels located in the serosal baso-lateral membranes of the posterior gills.     

Bioaccumulation of Trace Mercury in Trophic Levels of Benthic, Benthopelagic, Pelagic Fish Species, and Sea Birds from Arvand River, Iran

In this study, concentration of mercury was determined in the trophic levels of benthic, benthopelagic, pelagic fish species, and river birds from Arvand River, located in the Khuzestan province in the lowlands of southwestern Iran at the head of the Persian Gulf. The order of mercury concentrations in tissues of the fish species was as follows: liver>gill>muscle and in tissues of the kingfisher species was as follows: feather>liver>kidney>muscle. Therefore, liver in fish and feather in kingfisher exhibited higher mercury concentration than the other tissues. There was a positive correlation between mercury concentrations in fish and kingfisher species with size of its food items. We expected to see higher mercury levels in tissues of female species because they are larger and can eat larger food items. The results of this study show that the highest mean mercury level were found in the kingfisher (Anas crecca), followed by benthic (Epinephelus diacanthus), benthopelagic (Chanos chanos), and pelagic fish (Strongylura strongylura). Mean value of mercury in fish species, S. strongylura were (0.61 μg g^?1 dry weight), C. chanos (0.45 μg g^?1 dry weight), E. diacanthus (0.87 μg g^?1 dry weight), and in kingfisher species A. crecca was (2.64 μg g^?1 dry weight). Significant correlation between mercury concentration in fish and kingfisher may be related to high variability of mercury in the fish.     

Effects of methyl mercury at different dose regimes on eggshell formation and some biochemical characteristics of the eggshell gland mucosa of the domestic fowl

Eggshell formation and egg production in domestic fowl were studied following the administration of methyl mercury (two dose regimes: 5 mg daily for 6 consecutive days and 1 mg daily for 50 consecutive days). A daily oral dose of 5 mg of methyl mercury for 6 consecutive days induced significant eggshell thinning and deformation and inhibited egg production. Uptake of ⁴⁵Ca and synthesis of prostaglandins by a homogenate of eggshell gland mucosa from methyl-mercury-treated birds were significantly reduced, as was the calcium content of blood plasma. A daily oral dose of 1 mg of methyl mercury administered for 50 consecutive days also induced eggshell deformation and thinning and reduced egg production. This dose did not, however, have significant effects on the following: ⁴⁵Ca uptake and prostaglandin synthesis by a homogenate of the eggshell gland mucosa; ⁴⁵Ca uptake by a homogenate of duodenal mucosa; the Ca content of the blood plasma, shell gland mucosa or shell gland lumen; the HCO₃⁻ content of the shell gland lumen or the specific gravity of tibia. Methyl mercury added in vitro to a homogenate of eggshell gland mucosa significantly stimulated the synthesis of prostaglandins PGF_2α and PGE₂. Addition of mercury chloride to the same type of preparation stimulated the synthesis of PGF_2α at the expense of thromboxane (T × B₂) synthesis. Administration of 5 mg methyl mercury for 6 consecutive days seemed to reduce the availability of calcium for eggshell formation. This effect could have been due to a direct inhibitory effect of methyl mercury on calcium uptake from the gastrointestinal tract and/or to mobilization of medullary bone. The administration of 1 mg methyl mercury for 50 consecutive days probably induced the reproductive effects by another mechanism. The effects of methyl mercury on avian eggshell formation are quite different from the effects p,p′-DDE exerts on that process.     

Determination of the spatial characteristics of an RF electrodeless discharge by the method of emission tomography

The spatial distribution of the density of mercury atoms in the 7³ S state in a spherical RF electrode-less gas-discharge lamp is reconstructed by the method of emission tomography. The local values of the corresponding emission coefficients, which are proportional to the density of mercury atoms in the 7³ S state, are determined from integral (over the plasma volume) measurements of the lamp radiation at a wavelength of 546.1 nm with the help of an algorithm based on the maximum entropy method. The results obtained show that, for all of the operating modes under study, the profile of the density of mercury atoms in the 7³ S state has a minimum in the center of the lamp and a maximum near its wall. At a generator current of 100 mA and cold-spot temperature of 41°C, the density of mercury atoms in the 7³ S state is observed to drop substantially both in the center of the lamp and near its wall, the density in the center being reduced to almost zero. An explanation of this phenomenon is proposed.     

A Rhizosphere-Associated Symbiont,Photobacterium spp. Strain MELD1, and Its Targeted Synergistic Activity for Phytoprotection against Mercury

Though heavy metal such as mercury is toxic to plants and microorganisms, the synergistic activity between them may offer benefit for surviving. In this study, a mercury-reducing bacterium, Photobacterium spp. strain MELD1, with an MIC of 33 mg . kg^-1 mercury was isolated from a severely mercury and dioxin contaminated rhizosphere soil of reed (Phragmites australis). While the whole genome sequencing of MELD1 confirmed the presence of a mer operon, the mercury reductase MerA gene showed 99% sequence identity to Vibrio shilloni AK1 and implicates its route resulted from the event of horizontal gene transfer. The efficiency of MELD1 to vaporize mercury (25 mg . kg^-1, 24 h) and its tolerance to toxic metals and xenobiotics such as lead, cadmium, pentachlorophenol, pentachloroethylene, 3-chlorobenzoic acid, 2,3,7,8-tetrachlorodibenzo-p-dioxin and 1,2,3,7,8,9-hexachlorodibenzo-p-dioxin is promising. Combination of a long yard bean (Vigna unguiculata ssp. Sesquipedalis) and strain MELD1 proved beneficial in the phytoprotection of mercury in vivo. The effect of mercury (Hg) on growth, distribution and tolerance was examined in root, shoot, leaves and pod of yard long bean with and without the inoculation of strain MELD1. The model plant inoculated with MELD1 had significant increases in biomass, root length, seed number, and increased mercury uptake limited to roots. Biolog plate assay were used to assess the sole-carbon source utilization pattern of the isolate and Indole-3-acetic acid (IAA) productivity was analyzed to examine if the strain could contribute to plant growth. The results of this study suggest that, as a rhizosphere-associated symbiont, the synergistic activity between the plant and MELD1 can improve the efficiency for phytoprotection, phytostabilization and phytoremediation of mercury.     

Ferrous Iron-Dependent Volatilization of Mercury by the Plasma Membrane of Thiobacillus ferrooxidans

Of 100 strains of iron-oxidizing bacteria isolated, Thiobacillus ferrooxidans SUG 2-2 was the most resistant to mercury toxicity and could grow in an Fe²⁺ medium (pH 2.5) supplemented with 6 μM Hg²⁺. In contrast, T. ferrooxidans AP19-3, a mercury-sensitive T. ferrooxidans strain, could not grow with 0.7 μM Hg²⁺. When incubated for 3 h in a salt solution (pH 2.5) with 0.7 μM Hg²⁺, resting cells of resistant and sensitive strains volatilized approximately 20 and 1.7%, respectively, of the total mercury added. The amount of mercury volatilized by resistant cells, but not by sensitive cells, increased to 62% when Fe²⁺ was added. The optimum pH and temperature for mercury volatilization activity were 2.3 and 30°C, respectively. Sodium cyanide, sodium molybdate, sodium tungstate, and silver nitrate strongly inhibited the Fe²⁺-dependent mercury volatilization activity of T. ferrooxidans. When incubated in a salt solution (pH 3.8) with 0.7 μM Hg²⁺ and 1 mM Fe²⁺, plasma membranes prepared from resistant cells volatilized 48% of the total mercury added after 5 days of incubation. However, the membrane did not have mercury reductase activity with NADPH as an electron donor. Fe²⁺-dependent mercury volatilization activity was not observed with plasma membranes pretreated with 2 mM sodium cyanide. Rusticyanin from resistant cells activated iron oxidation activity of the plasma membrane and activated the Fe²⁺-dependent mercury volatilization activity of the plasma membrane.     

Sulfhydryl Group Involvement in Plasmalemma Transport of HCO(3) and OH in Chara corallina

The effect of the sulfhydryl reagents (—SH) p-chloromercuribenzene-sulfonic acid (PCMBS), N-ethylmaleimide (NEM), and inorganic mercury on H¹⁴CO₃⁻ assimilation in Chara corallina is reported. Commercial grade PCMBS caused severe inhibition of H¹⁴CO₃⁻ assimilation. Results obtained using purified PCMBS (stock solution passed through a chelating resin) indicated that inhibition observed using unpurified PCMBS was due predominantly to the presence of inorganic mercury (as a contaminant). The inhibitory role of inorganic mercury was verified using HgCl₂. This chemical caused a dramatic inhibition of H¹⁴CO₃⁻ assimilation, while it had little effect on cellular ¹⁴CO₂ fixation. Reversal of the Hg²⁺ inhibition of H¹⁴CO₃⁻ assimilation (in presence of 1.0 millimolar dithioerythritol) was extremely slow, requiring 2 to 3 hours for the reestablishment of control rates. This slow recovery may reflect de novo synthesis of transport proteins.     

Hair Mercury Concentrations of Lactating Mothers and Breastfed Infants in Iran (Fish Consumption and Mercury Exposure)

Coastal populations with high seafood consumption in the South Caspian Sea (Iran) have a significant exposure to dietary mercury. This study assesses the biomonitoring of mercury in mothers and breastfed infants in the South Caspian Sea. The mean of mercury concentration in the hair of 93 pairs of mothers and infants was obtained and was 3.55 and 1.89???g?g^?1, respectively. A statistically significant correlation (R?=?0.850, P?=?0.000) was seen between mercury concentration in the hair of mothers and infants. The results of this study indicate that hair mercury concentrations exceeded the USEPA reference dose of 1???g?g^?1 in 82.7?% of mothers and 61.2?% of infants. Also, 31?% of the mothers and 10.7?% of the infants had mercury concentrations more than the WHO ??threshold?? level (5???g?g^?1). The age and fish consumption of mothers were the factors that significantly affected the hair mercury concentration of mothers and infants. Number of dental amalgam fillings of mothers was the factor that only affected mercury in the hair of mothers. According to the results, we can conclude that the main determinant of mercury exposure was the intake of mercury through fish consumption of mothers.     

Mercury in Scalp Hair Near the Mid-Atlantic Ridge (MAR) in Relation to High Fish Consumption

The aim of this study was to evaluate the potential risk of mercury contamination near the Mid-Atlantic Ridge relating total mercury (THg) concentrations in the human scalp hair (n?=?110) and high fish consumption levels. THg was quantified in human scalp hair, and volunteers were questioned about age, gender, and smoking habits being subsequently grouped in categories based on the individual average intake of fish meals per week. THg concentrations ([THg]) in hair samples ranged from 0.05 to 2.24 μg g^?1, and significant differences were found according to age (p?<?0.05) and also among volunteers presenting different fish consumption rates (p?<?0.001) being the highest [THg] observed on the adult population and also on volunteers that indicated consuming five or more meals of fish per week. Results indicate a pattern of increased mercury accumulation with increasing fish consumption. Despite mercury availability and a potential mercury intake of up to seven times, the WHO provisional tolerable weekly intake of mercury value, in consequence of high fish consumption, mercury concentrations in scalp hair are comparatively low regarding recommended levels by WHO.     

Long-lived Ar-Hg plasma in the afterglow of a high-current pulsed discharge

High-density (n > 10¹² cm^?3) argon-mercury plasma produced by a short (t ～ 20 μs) high-power pulsed discharge in argon with an admixture of mercury vapor at a discharge current of ～50 A, an argon pressure of ～4 mm Hg, and a mercury vapor pressure of ～10^?3 mm Hg was studied using optical spectroscopy and radio physics methods. It is found that the lifetime of this plasma after the end of the discharge pulse is up to 10^?2 s. It is shown that such an abnormally long lifetime of such an afterglow plasma, as compared to the plasma of an argon discharge without an admixture of mercury vapor, is related to the long residence time of atoms and ions of both argon and mercury in highly excited states due to chemi-ionization processes involving long-lived metastable argon ions. It is suggested that dissociative recombination of highly excited molecular ions of argon play an important role in the transfer of excitation to argon atoms and ions that are close to autoionization states.     

Comparative Study of Uptake and Cellular Distribution of Hg203-Labeled Phenyl-Mercuric Acetate and Mercuric Acetate by Pea Roots

Uptake and cellular distribution of mercury²⁰³ from dilute mercuric acetate or phenylmercuric acetate solutions by excised pea roots (Pisum sativum) have been investigated. The time course of uptake showed that the amount of mercury uptake was increased with the time of incubation, and was similar for inorganic mercury or phenylmercuric acetate. The trend of mercury²⁰³ incorporation into cellular components from mercuric acetate and phenylmercuric acetate differed greatly as the time of incubation increased. The concentrations of mercuric acetate and phenylmercuric acetate solutions or the temperature of incubation also affected the mercury²⁰³ uptake as well as its cellular distribution. Longer time of exposure or higher concentration resulted in a greater mercury incorporation into mitochondrial fraction from phenylmercuric acetate than from inorganic mercury. This difference in intracellular distribution may be responsible for the degree of toxicity between inorganic mercury and phenylmercuric acetate in biological systems.     

Mercury inhibits the L170C mutant of aquaporin Z by making waters clog the water channel

We conduct in silico experiments of the L170C mutant of the Escherichia coli aquaporin Z (AQPZ) with and without mercury bonded to residue Cys 170. We find that bonding mercury to Cys 170 does not induce consequential structural changes to the protein. We further find that mercury does not stick in the middle of the water channel to simply occlude water permeation, but resides on the wall of the water pore. However, we observe that the water permeation coefficient of L170C-Hg⁺ (with one mercury ion bonded to Cys 170) is approximately half of that of the mercury-free L170C. We examine the interactions between the mercury ion and the waters in its vicinity and find that five to six waters are strongly attracted by the mercury ion, occluding the space of the water channel. Therefore we conclude that mercury, at low concentration, inhibits AQPZ-L170C mutant by making water molecules clog the water channel.     

Electrocatalytic four-electron reduction of oxygen at the cytochrome c3-adsorbed electrode

The electrocatalytic activity of cytochrome c₃ for the reduction of molecular oxygen was characterized from the studies of the adsorption of cytochrome c₃ and the co-adsorption of cytochrome c₃ with cytochrome c on the mercury electrode by the a.c. polarographic technique. The adsorption of cytochrome c₃ on the mercury electrode is irreversible and is diffusion-controlled. The maximum amount of cytochrome c₃ adsorbed was 0.92 · 10^?11 mol · cm^?2 at ?0.90 V. The amount of cytochrome c₃ in the mixed adsorbed layer with cytochrome c was determined from the differential capacitance measurement. It was shown that the fractional coverage of cytochrome c₃ can be estimated from its bulk concentration and the diffusion coefficient (1.05 · 10^?6 cm² · s^?1). Cytochrome c₃ catalyzes the electrochemical reduction of molecular oxygen from the two-electron pathways via hydrogen peroxide to the four-electron pathway at the mercury electrode in neutral phosphate buffer solution. The catalytic activity varies with the bulk concentration of cytochrome c₃. The highest catalytic activity for the oxygen reduction (no hydrogen peroxide formation) is attained when one-half of the mercury electrode surface is covered by cytochrome c₃. The addition of cytochrome c or bovine serum albumin to the cytochrome c₃ solution inhibits the catalytic activity of cytochrome c₃. The reversible polarographic behavior of cytochrome c₃ through the mixed adsorbed layer of cytochrome c₃ and cytochrome c was also investigated.     

Marine foraging ecology influences mercury bioaccumulation in deep-diving northern elephant seals

Mercury contamination of oceans is prevalent worldwide and methylmercury concentrations in the mesopelagic zone (200–1000 m) are increasing more rapidly than in surface waters. Yet mercury bioaccumulation in mesopelagic predators has been understudied. Northern elephant seals (Mirounga angustirostris) biannually travel thousands of kilometres to forage within coastal and open-ocean regions of the northeast Pacific Ocean. We coupled satellite telemetry, diving behaviour and stable isotopes (carbon and nitrogen) from 77 adult females, and showed that variability among individuals in foraging location, diving depth and δ¹³C values were correlated with mercury concentrations in blood and muscle. We identified three clusters of foraging strategies, and these resulted in substantially different mercury concentrations: (i) deeper-diving and offshore-foraging seals had the greatest mercury concentrations, (ii) shallower-diving and offshore-foraging seals had intermediate levels, and (iii) coastal and more northerly foraging seals had the lowest mercury concentrations. Additionally, mercury concentrations were lower at the end of the seven-month-long foraging trip (n = 31) than after the two-month- long post-breeding trip (n = 46). Our results indicate that foraging behaviour influences mercury exposure and mesopelagic predators foraging in the northeast Pacific Ocean may be at high risk for mercury bioaccumulation.     

Evidence of elevated mercury levels in carnivorous and omnivorous fishes downstream from an Amazon reservoir

Hydroelectric reservoirs can stratify, producing favorable conditions for mercury methylation in the hypolimnion. The methylmercury (MeHg) can be exported downstream, increasing its bioavailability below the dam. Our objective was to assess the mercury levels in plankton, suspended particulate matter (SPM) and fish collected upstream (UP) and downstream (DW) from the Reservatório de Samuel dam, an Amazonian reservoir that stratifies during half of the year. Mercury concentrations in both SPM and plankton were similar between the two sites, which could indicate there are no conditions favoring methylation at the moment of sampling (absence of stratification). Almost all mercury found in the muscle of fishes was in organic form, and differences of mercury levels between sites were dependent on the fishes trophic level. Herbivores showed similar mean organic mercury levels (UP = 117 μg g^?1; DW = 120 μg g^?1; n = 12), whereas omnivores (UP = 142 μg g^?1; DW = 534 μg g^?1; n = 27) and carnivores (UP = 545 μg g^?1; DW = 1,366 μg g^?1; n = 69) showed significantly higher values below the dam. The absence of a reservoir effect in herbivores is expected, since they feed on grassy vegetation, near the riverbanks, which is not much influenced by mercury in aquatic systems. On the other hand, the higher mercury levels below the dam observed for omnivores and carnivores suggest a possible influence of the reservoir since they feed on items that could be contaminated by MeHg exported from upstream. The results highlight the necessity of assessing areas downstream of reservoirs.     

The Binding of Mercury by the Yeast Cell in Relation to Changes in Permeability

Yeast cells exposed to mercuric chloride suffer irreversible damage to the membrane, resulting in a loss of potassium and cellular anions to the medium. The maximal loss of K⁺, but not the time course of K⁺ loss is related to the mercury concentration, the relationship following a normal curve on a graph of log-concentration versus effect. It is concluded that the response is all or none for individual cells, and that with increasing concentrations of metal, the threshold is exceeded in an increasing proportion of the cells. Parallel studies of the binding of mercury by the cells indicate two distinct phases, only one of which is associated with the physiological response. The binding process is relatively slow but reaches an equilibrium state. Desorption is markedly dependent on temperature. No simple stoichiometric relationship exists between the binding of mercury and the physiological response (K⁺ loss).     

Evaluation of the association of mercury(II) with some dicysteinyl tripeptides

The present study was undertaken to gain insight into the associations of mercury(II) with dicysteinyl tripeptides in buffered media at pH 7.4. We investigated the effects of increasing the distance between cysteinyl residues on mercury(II) associations and complex formations. The peptide–mercury(II) formation constants and their associated thermodynamic parameters in 3-(N-morpholino)propanesulfonic acid (MOPS) buffered solutions were evaluated by isothermal titration calorimetry. Complexes formed in different relative ratios of mercury(II) to cysteinyl peptides in ammonium formate buffered solutions were characterized by LTQ Orbitrap mass spectrometry. The results from these studies show that n-alkyl dicysteinyl peptides (CP 1–4), and an aryl dicysteinyl peptide (CP 5) can serve as effective “double anchors” to accommodate the coordination sites of mercury(II) to form predominantly one-to-one Hg(peptide) complexes. The aryl dicysteinyl peptide (CP 5) also forms the two-to-two Hg₂(peptide)₂ complex. In the presence of excess peptide, Hg(peptide)₂ complexes are also detected. Notably, increasing the distance between the ligating groups or “anchor points” in CP 1–5 does not significantly affect their affinity for mercury(II). However, the enthalpy change (ΔH) values (ΔH₁ ∼ −91 kJ mol⁻¹ and ΔH₂ ∼ −66 kJ mol⁻¹) for complex formation between CP 4 and 5 with mercury(II) are about one and a half times larger than the related values for CP 1, 2 and 3 (ΔH₁ ∼ −66 kJ mol⁻¹ and ΔH₂ ∼ 46 kJ mol⁻¹). The corresponding entropy change (ΔS) values (ΔS₁ ∼ −129 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −116 J K⁻¹ mol⁻¹) of the structurally larger dicysteinyl peptides CP 4 and 5 are less entropically favorable than for CP 1, 2 and 3 (ΔS₁ ∼ −48 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −44 J K⁻¹ mol⁻¹). Generally, these associations result in a decrease in entropy, indicating that these peptide–mercury complexes potentially form highly ordered structures. The results from this study show that dicysteinyl tripeptides are effective in binding mercury(II) and they are promising motifs for the design of multi-cysteinyl peptides for binding more than one mercury(II) ion per peptide.     

Short-term mercury exposure on Na+/K+-ATPase activity and ionoregulation in gill and brain of an Indian major carp,Cirrhinus mrigala

Recently mercury pollution has been increased considerably in aquatic resources throughout the world and it is a growing global concern. In this study, the 96 h LC50 value of waterborne mercuric chloride for Cirrhinus mrigala was found to be 0.34 mg/L (with 95% confidence limits). Fingerlings of C. mrigala were exposed to 0.068 and 0.034 mg/L of mercuric chloride for 96 h to assess the Na⁺/K⁺-ATPase activity and ionoregulation (Na⁺, K⁺ and Cl^?) in gill and brain. Results showed that Na⁺/K⁺-ATPase activity and ionic levels (Na⁺, K⁺ and Cl^?) in gill and brain of fish exposed to different concentrations of mercuric chloride were found to be significantly (p < 0.05) decreased throughout the study period. Mercury inactivates many enzymes by attaching to sulfur atoms in which the enzyme Na⁺/K⁺-ATPase is highly sensitive to mercury. The inhibition of gill and brain Na⁺/K⁺-ATPase activity might have resulted from the physicochemical alteration of the membrane due to mercury toxicity. Moreover, inhibition of Na⁺/K⁺-ATPase may affect the ion transport and osmoregulatory function by blocking the transport of substances across the membrane by active transport. The present study indicates that the alterations in these parameters can be used in environmental biomonitoring of mercury contamination in aquatic ecosystem.     

Characterization of the metabolically modified heavy metal-resistant Cupriavidus metallidurans strain MSR33 generated for mercury bioremediation

Background

Mercury-polluted environments are often contaminated with other heavy metals. Therefore, bacteria with resistance to several heavy metals may be useful for bioremediation. Cupriavidus metallidurans CH34 is a model heavy metal-resistant bacterium, but possesses a low resistance to mercury compounds.

Methodology/Principal Findings

To improve inorganic and organic mercury resistance of strain CH34, the IncP-1β plasmid pTP6 that provides novel merB, merG genes and additional other mer genes was introduced into the bacterium by biparental mating. The transconjugant Cupriavidus metallidurans strain MSR33 was genetically and biochemically characterized. Strain MSR33 maintained stably the plasmid pTP6 over 70 generations under non-selective conditions. The organomercurial lyase protein MerB and the mercuric reductase MerA of strain MSR33 were synthesized in presence of Hg²⁺. The minimum inhibitory concentrations (mM) for strain MSR33 were: Hg²⁺, 0.12 and CH₃Hg⁺, 0.08. The addition of Hg²⁺ (0.04 mM) at exponential phase had not an effect on the growth rate of strain MSR33. In contrast, after Hg²⁺ addition at exponential phase the parental strain CH34 showed an immediate cessation of cell growth. During exposure to Hg²⁺ no effects in the morphology of MSR33 cells were observed, whereas CH34 cells exposed to Hg²⁺ showed a fuzzy outer membrane. Bioremediation with strain MSR33 of two mercury-contaminated aqueous solutions was evaluated. Hg²⁺ (0.10 and 0.15 mM) was completely volatilized by strain MSR33 from the polluted waters in presence of thioglycolate (5 mM) after 2 h.

Conclusions/Significance

A broad-spectrum mercury-resistant strain MSR33 was generated by incorporation of plasmid pTP6 that was directly isolated from the environment into C. metallidurans CH34. Strain MSR33 is capable to remove mercury from polluted waters. This is the first study to use an IncP-1β plasmid directly isolated from the environment, to generate a novel and stable bacterial strain useful for mercury bioremediation.