Importance of sulfate reducing bacteria in mercury methylation and demethylation in periphyton from Bolivian Amazon region

Sulfate reducing bacteria (SRB) are important mercury methylators in sediments, but information on mercury methylators in other compartments is ambiguous. To investigate SRB involvement in methylation in Amazonian periphyton, the relationship between Hg methylation potential and SRB (Desulfobacteraceae, Desulfobulbaceae and Desulfovibrionaceae) abundance in Eichhornia crassipes and Polygonum densiflorum root associated periphyton was examined. Periphyton subsamples of each macrophyte were amended with electron donors (lactate, acetate and propionate) or inhibitors (molybdate) of sulfate reduction to create differences in SRB subgroup abundance, which was measured by quantitative real-time PCR with primers specific for the 16S rRNA gene. Mercury methylation and demethylation potentials were determined by a stable isotope tracer technique using ²⁰⁰HgCl and CH₃²⁰²HgCl, respectively. Relative abundance of Desulfobacteraceae (<0.01-12.5%) and Desulfovibrionaceae (0.01-6.8%) were both highly variable among samples and subsamples, but a significant linear relationship (p < 0.05) was found between Desulfobacteraceae abundance and net methylmercury formation among treatments of the same macrophyte periphyton and among all P. densiflorum samples, suggesting that Desulfobacteraceae bacteria are the most important mercury methylators among SRB families. Yet, molybdate only partially inhibited mercury methylation potentials, suggesting the involvement of other microorganisms as well. The response of net methylmercury production to the different electron donors and molybdate was highly variable (3-1104 pg g⁻¹ in 12 h) among samples, as was the net formation in control samples (17-164 pg g⁻¹ in 12 h). This demonstrates the importance of community variability and complexity of microbial interactions for the overall methylmercury production in periphyton and their response to external stimulus.     

Levels of methylmercury and controlling factors in surface sediments of the Carson River system, Nevada

Spatial and temporal distribution of methylmercury (MeHg) was determined in surficial sediments collected from a river-reservoir system impacted by Hg-contaminated mine wastes. Despite the fact that total mercury concentrations (HgT) in surface sediments of the Carson River system were in the microg.g(-1) range, levels of MeHg varied from about 2 to 28 ngHg.g(-1) dry weight, representing less than 3% of Hg(T). Concentrations of MeHg were well correlated with both the biotic (r=0.95) and abiotic activity (r=0.85) of the sediment, determined as the ability of each compartment to specifically reduce an alternative electron acceptor. However, the positive relationship between the two measured activities suggests that the abiotic activity may be due to reductant substances produced by micro-organisms. When sediments collected from the Carson River were used in laboratory assays for the determination of potential rates of MeHg production, the addition of inorganic Hg (added as HgCl2) resulted in increased rates of methylation when the spike concentration was lower or equal to 15.3 microg.g(-1) dry weight. This trend was reversed for spike concentration of inorganic Hg above 15.3 microg.g(-1). The reduction of methylation rate was associated with an inhibition of microbial activity. These observations suggest that seasonal inputs into the river of significant amounts of inorganic Hg eroded from mill tailings during winter and spring flooding events could have an inhibiting effect on Hg-methylating micro-organisms. This observation could explain the low [MeHg]/[HgT] ratios previously documented in waters of the Carson River system.     

Mercury methylation in aquatic systems affected by acid deposition

Recently, it has been noted that fish in acidified lakes may contain elevated levels of mercury. While there is correlation among lakes between depressed pH and high mercury concentrations in fish, the cause of this problem is unknown. A number of hypotheses have been advanced in explanation, including increased mercury deposition, changes in mercury mobility due to acidification, pH dependent changes in mercury uptake by biota, and alterations in population size and/or structure which result in increased bioaccumulation in fish. Because fish accumulate mercury mainly in an organic form, methylmercury, changes in the biogeochemical cycling of this compound might account for elevated bioaccumulation. Mercury methylation is predominantly a microbial process which occurs in situ in lakes. This review focuses on microbiological and biogeochemical changes that may lead to increased levels of methylmercury in fresh waters impacted by acid-deposition. In particular, we focus on the hypothesis that sulfate-reducing bacteria are important mediators of metal methylation in aquatic systems and, moreover, that sulfate-deposition may stimulate methylmercury production by enhancing the activity of sulfate-reducing bacteria in sediments.     

Chiral signature of venlafaxine as a marker of biological attenuation processes

The chiral signature of the antidepressant venlafaxine was used in this study to gain insight into biological attenuation processes and to differentiate abiotic and biotic transformation processes in water. Laboratory scale experiments revealed that sorption and phototransformation processes were not enantioselective while venlafaxine was enantioselectively biotransformed into O-desmethylvenlafaxine. The enantiomeric fraction (EF) variations of venlafaxine appeared to be proportional to its microbial fractional conversion. Enantioselective biotransformation of venlafaxine was also investigated in a eutrophic French river. Venlafaxine was found to be racemic at the output of the main wastewater treatment plant discharging into the river, independently of the sampling date during the year. An analysis of EF variations might provide evidence of biodegradation along a 30 km river stretch.     

干式厌氧消化过程挥发酸对硫酸盐还原菌的影响

在牛粪干式厌氧消化过程中,通过添加不同挥发酸(乙酸、丙酸、丁酸),考察消化稳定阶段,挥发性脂肪酸的分布特征,挥发性脂肪酸酸组成变化对硫酸盐还原菌(SRB)的影响,微生物种群组成和种群间关系。实验结果表明,挥发性脂肪酸对SRB还原速率的贡献依次为:丙酸丁酸乙酸。相比乙酸和丁酸,添加一定量的丙酸,更有利于激活SRB的活性,从而加强SRB与产甲烷菌(MB)的种间协同,保证厌氧系统的稳定运行。     

Mercury removal, methylmercury formation, and sulfate-reducing bacteria profiles in wetland mesocosms

A pilot-scale model was constructed to determine if a wetland treatment system (WTS) could effectively remove low-level mercury from an outfall located at the Department of Energy's Savannah River Site. Site-specific hydrosoil was planted with giant bulrush, Scirpus californicus, and surface amended with gypsum (CaSO4) prior to investigating the biogeochemical dynamics of sediment-based sulfur and mercury speciation. On average, the pilot WTS decreased total mercury concentrations in the outfall stream by 50%. Transformation of mercury to a more "bioavailable" species, methylmercury, was also observed in the wetland treatment system. Methylmercury formation in the wetland was ascertained with respect to sediment biogeochemistry and S. californicus influences. Differences in sulfate-reduction rates (SRRs) were observed between mesocosms that received additional decomposing Scirpus matter and mesocosms that were permitted growth of the submerged macrophyte, Potamogeton pusillus. Relative abundance measurements of sulfate-reducing bacteria (SRB) as characterized using oligonucleotide probes were also noticeably different between the two mesocosms. A positive correlation between increased sulfide, dissolved total mercury, and dissolved methylmercury concentrations was also observed in porewater. The data suggest that soluble mercury-sulfide complexes were formed and contributed, in part, to a slight increase in mercury solubility. Observed increases in methylmercury concentration also suggest that soluble mercury-sulfide complexes represent a significant source of mercury that is "available" for methylation. Finally, a volunteer macrophyte, Potamogeton pusillus, is implicated as having contributed additional suspended particulate matter in surface water that subsequently reduced the pool of dissolved mercury while also providing an environment suitable for demethylation.     

Alkane biodegradation and dynamics of phylogenetic subgroups of sulfate-reducing bacteria in an anoxic coastal marine sediment artificially contaminated with oil

For 503 days, unoiled control and artificially oiled sediments were incubated in situ at 20m water depth in a Mediterranean coastal area. Degradation of the aliphatic fraction of the oil added was followed by GC-MS. At the same time, terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA encoding genes was used to detect dynamics in the sulfate-reducing bacteria (SRB) community in response to the oil contamination. Specific polymerase chain reaction (PCR) primer sets for five generic or suprageneric groups of SRB were used for PCR amplification of DNA extracted from sediments. During the experiment, hydrocarbons from C(17) to C(30) were significantly degraded even in strictly anoxic sediment layers. Of the five SRB groups, only two groups were detected in the sediments (control and oiled), namely the Desulfococcus-Desulfonema-Desulfosarcina-like group and the Desulfovibrio-Desulfomicrobium-like group. Statistical analysis of community patterns revealed dynamic changes over time within these two groups following the contamination. Significant differences in community patterns were recorded in artificially oiled compared with control sediments. Cloning and sequencing of 16S rRNA encoding genes performed after 503 days showed that many of the most abundant sequences were closely related to hydrocarbonoclastic SRB which could have played an active role in the observed biodegradation of aliphatic hydrocarbons. Results from the present study provide useful information on the dynamics of dominant SRB in heavily oil-contaminated sediments and their potential for anaerobic biodegradation for the treatment of spilled oil in anoxic marine environments.     

Humic acids enhance the microbially mediated release of sedimentary ferrous iron

Iron (Fe) is an essential element for many organisms, but high concentrations of iron can be toxic. The complex relation between iron, arsenic (As), bacteria, and organic matter in sediments and groundwater is still an issue of environmental concern. The present study addresses the effects of humic acids and microorganisms on the mobilization of iron in sediments from an arsenic-affected area, and the microbial diversity was analyzed. The results showed that the addition of 50, 100, and 500 mg/L humic acids enhanced ferrous iron (Fe(II)) release in a time-dependent and dose-dependent fashion under anaerobic conditions. A significant increase in the soluble Fe(II) concentrations occurred in the aqueous phases of the samples during the first 2 weeks, and aqueous Fe(II) reached its maximum concentrations after 8 weeks at the following Fe(II) concentrations: 28.95?±?1.16 mg/L (original non-sterilized sediments), 32.50?±?0.71 mg/L (50 mg/L humic acid-amended, non-sterilized sediments), 37.50?±?1.85 mg/L (100 mg/L humic acid-amended, non-sterilized sediments), and 39.00?±?0.43 mg/L (500 mg/L humic acid-amended, non-sterilized sediments). These results suggest that humic acids can further enhance the microbially mediated release of sedimentary iron under anaerobic conditions. By contrast, very insignificant amounts of iron release were observed from sterilized sediments (the abiotic controls), even with the supplementation of humic acids under anaerobic incubation. In addition, the As(III) release was increased from 50?±?10 μg/L (original non-sterilized sediments) to 110?±?45 μg/L (100 mg/L humic acid-amended, non-sterilized sediments) after 8 weeks of anaerobic incubation. Furthermore, a microbial community analysis indicated that the predominant class was changed from Alphaproteobacteria to Deltaproteobacteria, and clearly increased populations of Geobacter sp., Paludibacter sp., and Methylophaga sp. were found after adding humic acids along with the increased release of iron and arsenic. Our findings provide evidence that humic acids can enhance the microbially mediated release of sedimentary ferrous iron in an arsenic-affected area. It is thus suggested that the control of anthropogenic humic acid use and entry into the environment is important for preventing the subsequent iron contamination in groundwater.     

Linkage between community diversity of sulfate-reducing microorganisms and methylmercury concentration in paddy soil

Sulfate-reducing microorganisms (SRM) have been thought to play a key role in mercury (Hg) methylation in anoxic environments. The current study examined the linkage between SRM abundance and diversity and contents of methylmercury (MeHg) in paddy soils collected from a historical Hg mining area in China. Soil profile samples were collected from four sites over a distance gradient downstream the Hg mining operation. Results showed that MeHg content in the soil of each site significantly decreased with the extending distance away from Hg mine. Soil MeHg content was correlated positively with abundance of SRM and the contents of organic matter (OM), NH₄ ⁺, SO₄ ^2?, and Hg. The abundances of SRM based on dissimilatory (bi) sulfite reductase (dsrAB) gene at 0–40 cm depths were higher than those at 40–80 cm depth at all sites. The SRM community composition varied in the soils of different sampling sites following terminal restriction fragment length polymorphism (T-RFLP) and phylogenetic analyses, which appeared to be correlated with contents of MeHg, OM, NH₄ ⁺, and SO₄ ^2? through canonical correspondence analysis. The dominant groups of SRM in the soils examined belonged to Deltaproteobacteria and some unknown SRM clusters that could have potential for Hg methylation. These results advance our understanding of the relationship between SRM and methylmercury concentration in paddy soil.     

Diversity of active microbial communities subjected to long-term exposure to chemical contaminants along a 40-year-old sediment core

In estuarine ecosystems, metallic and organic contaminants are mainly associated with fine grain sediments which settle on mudflats. Over time, the layers of sediment accumulate and are then transformed by diagenetic processes mainly controlled by microbial activity, recording the history of the estuary’s chemical contamination. In an environment of this specific type, we investigated the evolution of the chemical contamination and the structure of both total and active microbial communities, based on PhyloChip analysis of a 4.6-m core corresponding to a 40-year sedimentary record. While the archaeal abundance remained constant along the core, a decrease by one order of magnitude in the bacterial abundance was observed with depth. Both total and active microbial communities were dominated by Proteobacteria, Actinobacteria, and Firmicutes in all sediment samples. Among Proteobacteria, alpha-Proteobacteria dominated both total (from 37 to 60 %) and metabolically active (from 19.7 to 34.6 %) communities, including the Rhizobiales, Rhodobacter, Caulobacterales, and Sphingomonadales orders. Co-inertia analysis revealed a relationship between polycyclic aromatic hydrocarbons, zinc and some polychlorobiphenyls concentrations, and the structure of total and active microbial communities in the oldest and most contaminated sediments (from 1970 to 1975), suggesting that long-term exposure to chemicals shaped the structure of the microbial community.     

Mercury cycling in surface water, pore water and sediments of Mugu Lagoon, CA, USA

Mugu Lagoon is an estuary in southern California, listed as impaired for mercury. In 2005, we examined mercury cycling at ten sites within at most four habitats. In surface water (unfiltered and filtered) and pore water, the concentration of total mercury was correlated with methylmercury levels (R2=0.29, 0.26, 0.27, respectively, p<0.05), in contrast to sediments, where organic matter and reduced iron levels were most correlated with methylmercury content (R2=0.37, 0.26, respectively, p<0.05). Interestingly, levels for percent methylmercury of total mercury in sediments were higher than typical values for estuarine sediments (average 5.4%, range 0.024-38%, n=59), while pore water methylmercury Kd values were also high (average 3.1, range 2.0-4.2l kg(-1), n=39), and the estimated methylmercury flux from sediments was low (average 1.7, range 0.14-5.3ng m(-2) day(-1), n=19). Mercury levels in predatory fish tissue at Mugu are >0.3ppm, suggesting biogeochemical controls on methylmercury mobility do not completely mitigate methylmercury uptake through the food web.     

Characterization of bacterial communities at heavy-metal-contaminated sites

The microbial community in soil samples from two long-term contaminated sites was characterized by using culture-dependent and culture-independent methods. The two sites investigated contained high amounts of heavy metals and were located in the upper Silesia Industrial Region in southern Poland. The evaluation of the aerobic soil microbial population clearly demonstrated the presence of considerable numbers of viable, culturable bacteria at both sites. A high fraction of the bacterial population was able to grow in the presence of high amounts of metals, i.e. up to 10 mM Zn²⁺, 3 mM Pb²⁺ or 1 mM Cu²⁺. Site 1 contained significantly (P < 0.05) lower bacterial numbers growing in the presence of 10 mM Zn²⁺ than site 2, while the opposite was observed for bacteria tolerating 1 mM Cu²⁺. This coincided with the contents of these two metals at the two sites. Ecophysiological (EP) indices for copiotrophs (r-strategists) and oligotrophs (K-strategists) pointed to high bacterial diversity at both sites. Fluorescence in situ hybridization (FISH) analysis indicated that Actinobacteria and Proteobacteria represent the physiologically active fraction of bacteria at the two sites. Shannon diversity (H′) indices for FISH-detected bacterial phylogenetic groups were not significantly different at the two sites.     

Effects of sterile storage,cation saturation and substrate additions on the degradability and extractability of nonylphenol and phenanthrene in soil

The main objective of this study was to determine the effects of long-term abiotic processes during aging of organic pollutants in soil on their microbial degradability and formation of non-extractable residues. The specific aims of our study were to investigate how the fate of p353-nonylphenol (NP) and phenanthrene (Phe) in soils might be affected by: (i) saturation of soil by cations with different valency (Na⁺, Ca²⁺ or Al³⁺), (ii) addition of organic substrate (wood flour) during incubation period and (iii) different soil moisture levels.     

Antibiotic resistant bacteria/genes dissemination in lacustrine sediments highly increased following cultural eutrophication of Lake Geneva (Switzerland)

This study investigates faecal indicator bacteria (FIB), multiple antibiotic resistant (MAR), and antibiotic resistance genes (ARGs), of sediment profiles from different parts of Lake Geneva (Switzerland) over the last decades. MARs consist to expose culturable Escherichia coli (EC) and Enterococcus (ENT) to mixed five antibiotics including Ampicillin, Tetracycline, Amoxicillin, Chloramphenicol and Erythromycin. Culture-independent is performed to assess the distribution of ARGs responsible for, β-lactams (blaTEM; Amoxicillin/Ampicillin), Streptomycin/Spectinomycin (aadA), Tetracycline (tet) Chloramphenicol (cmlA) and Vancomycin (van). Bacterial cultures reveal that in the sediments deposited following eutrophication of Lake Geneva in the 1970s, the percentage of MARs to five antibiotics varied from 0.12% to 4.6% and 0.016% to 11.6% of total culturable EC and ENT, respectively. In these organic-rich bacteria-contaminated sediments, the blaTEM resistant of FIB varied from 22% to 48% and 16% to 37% for EC and ENT respectively, whereas the positive PCR assays responsible for tested ARGs were observed for EC, ENT, and total DNA from all samples. The aadA resistance gene was amplified for all the sediment samples, including those not influenced by WWTP effluent water. Our results demonstrate that bacteria MARs and ARGs highly increased in the sediments contaminated with WWTP effluent following the cultural eutrophication of Lake Geneva. Hence, the human-induced changing limnological conditions highly enhanced the sediment microbial activity, and therein the spreading of antibiotic resistant bacteria and genes in this aquatic environment used to supply drinking water in a highly populated area. Furthermore, the presence of the antibiotic resistance gene aadA in all the studied samples points out a regional dissemination of this emerging contaminant in freshwater sediments since at least the late nineteenth century.     

Influence of Chironomus riparius (Diptera, Chironomidae) and Tubifex tubifex (Annelida, Oligochaeta) on oxygen uptake by sediments. Consequences of uranium contamination

The diffusive oxygen uptake (DOU) of sediments inhabited by Chironomus riparius and Tubifex tubifex was investigated using a planar oxygen optode device, and complemented by measurements of bioturbation activity. Additional experiments were performed within contaminated sediments to assess the impact of uranium on these processes. After 72 h, the two invertebrate species significantly increased the DOU of sediments (13-14%), and no temporal variation occurred afterwards. Within contaminated sediments, it was already 24% higher before the introduction of the organisms, suggesting that uranium modified the sediment biogeochemistry. Although the two species firstly reacted by avoidance of contaminated sediment, they finally colonized it. Their bioturbation activity was reduced but, for T. tubifex, it remained sufficient to induce a release of uranium to the water column and an increase of the DOU (53%). These results highlight the necessity of further investigations to take into account the interactions between bioturbation, microbial metabolism and pollutants.     

Distinct diversity of the czcA gene in two sedimentary horizons from a contaminated estuarine core

In estuarine ecosystems, trace metals are mainly associated with fine grain sediments which settle on mudflats. Over time, the layers of sediments accumulate and are then transformed by diagenetic processes, recording the history of the estuary’s chemical contamination. In such a specific environment, we investigated to what extent a chronic exposure to contaminants could affect metal-resistant sedimentary bacteria in subsurface sediments. The occurrence and diversity of cadmium resistance genes (cadA, czcA) was investigated in 5- and 33-year-old sediments from a highly contaminated estuary (Seine France). Primers were designed to detect a 252-bp fragment of the czcA gene, specifically targeting a transmembrane helice domain (TMH IV) involved in the proton substrate antiport of this efflux pump. Although the cadA gene was not detected, the highest diversity of the sequence of the czcA gene was observed in the 5-year-old sediment. According to the percentage of identity at the amino acid level, the closest CzcA relatives were identified among Proteobacteria (α, β, γ, and δ), Verrucomicrobia, Nitrospirae, and Bacteroidetes. The most abundant sequences were affiliated with Stenotrophomonas. In contrast, in the 33-year-old sediment, CzcA sequences were mainly related to Rhodanobacter thiooxydans and Stenotrophomonas, suggesting a shaping of the metal-resistant microbial communities over time by both diagenetic processes and trace metal contamination.     

Degradation of sulfonamides antibiotics in lake water and sediment

Degradation of three sulfonamides (SAs), namely sulfamethoxazole (SMX), sulfamethazine (SMZ), and sulfadimethoxine (SDM) in surface water and sediments collected from Taihu Lake and Dianchi Lake, China was investigated in this study. The surface water (5–10 cm) was collected from the east region of Taihu Lake, China. Two sets of degradation experiments were conducted in 3-L glass bottles containing 2 L of fresh lake water and 100 μg/L of individual SAs aerated by bubbling air at a rate of approximately 1.2 L/min, one of which was sterilized by the addition of NaN₃ (0.1 %). Sediment samples were taken from Taihu Lake and Dianchi Lake, China. For the sediment experiment, 5 g of sediment were weighed into a 50-mL glass tube, with 10 mg/kg of individual SAs. Different experimental conditions including the sediment types, sterilization, light exposure, and redox condition were also considered in the experiments. The three SAs degraded in lake water with half-lives (t _1/2) of 10.5–12.9 days, and the half-lives increased significantly to 31.9–49.8 days in the sterilized water. SMZ and SDM were degraded by abiotic processes in Taihu and Dianchi sediments, and the different experimental conditions and sediments characteristics had no significant effect on their declines. SMX, however, was mainly transformed by facultative anaerobes in Taihu and Dianchi sediments under anaerobic conditions, and the degradation rate of SMX in non-sterile sediment (t _1/2 of 9.6–16.7 days) were higher than in sterilized sediment (t _1/2 of 18.7–135.9 days). Under abiotic conditions, degradation of SMX in Dianchi sediment was faster than in Taihu sediment, probably due to the higher organic matter content and inorganic photosensitizers concentrations in Dianchi sediment. High initial SAs concentration inhibited the SAs degradation, which was likely related to the inhibition of microorganism activities by high SAs levels in sediments. Results from this study could provide information on the persistence of commonly used sulfanomides antibiotics in lake environment.     

The flame retardant tetrabromobisphenol-A and its metabolite found in river and marine sediments in Japan

Tetrabromobisphenol-A (TBBP), used as a flame retardant, was detected in river sediments collected in Osaka, Japan, at a concentration range from 22 to 140 μg/kg (ppb), as determined on a dry weight basis. TBBP was also found in marine sediments collected at Osaka Bay, though the TBBP levels of these samples were much lower than those from river sediments. Dimethyl ether derivative of TBBP (TBBP-DM), thought to be a decomposition product from microbial methylation, was found in the river sediments but in none of the marine sediments. TBBP-DM residues were about one-hundredth of the TBBP levels.     

反硝化抑制石油集输系统中硫酸盐还原菌的现场实验

针对长庆油田采油四厂艾家湾作业区集输系统中硫化氢气体浓度较高的问题,采用生物竞争抑制技术对该石油集输系统中H2S进行了处理和现场实验研究。研究结果表明,采用单井生物前端抑制可以使集输系统中沉降罐、污水罐中H2S气体浓度由最初的268mg／m^3降低至《工作场所有害因素职业接触限值》（GBZ2—2002）要求的10mg／m^3以下。在实验周期内,系统中的反硝化细菌的数量随加药时间的延长逐渐增加,从最初的3000cfu／100mL增加至600000cfu／100mL,而此时系统内的ORP值由硫酸盐还原菌生存的最佳微环境（-300～-350mV）升高至反硝化细菌生存的最佳微环境（-50--100mV）。16SrDNA测序结果表明,反硝化微生物种群和数量增长很快,并得到适宜于艾家湾作业区集输系统中硫酸盐还原菌抑制的反硝化微生物主要是耐盐芽孢杆菌、产碱假单胞菌和奈瑟菌。完善了集输系统中硫酸盐还原菌次生H2S的生物抑制技术,证实了处理技术与实施工艺的可行性。