Dental composite biodeterioration in the presence of oral Streptococci and extracellular metabolic products.

OBJECTIVE: Secondary caries is a primary cause of early restoration failure. While primary dental caries has been extensively researched, our knowledge about the impact of secondary caries on dental restorations is relatively limited. In this study, we examined how different clinically relevant microbially-influenced environments impact the degradation of nano-filled (FIL) and micro-hybrid (AEL) dental composites. METHODS: Material strength of two commercial dental composites was measured following incubation in aqueous media containing: i) cariogenic (Streptococcus mutans) and non-cariogenic bacteria (Streptococcus sanguinis) grown on sucrose or glucose, ii) abiotic mixtures of artificial saliva and sucrose and glucose fermentation products (volatile fatty acids and ethanol) in proportions known to be produced by these microorganisms, and iii) abiotic mixtures of artificial saliva and esterase, a common oral extracellular enzyme. RESULTS: Nano-filled FIL composite strength decreased in all three types of incubations, while micro-hybrid AEL composite strength only decreased significantly in biotic incubations. The strength of both composites was statistically significantly decreased in all biotic incubations containing both cariogenic and non-cariogenic bacteria beyond that induced by either abiotic mixtures of fermentation products or esterase alone. Finally, there were no statistically significant differences in composite strength decrease among the tested biotic conditions. CONCLUSIONS: The results show that conditions created during the growth of both cariogenic and non-cariogenic oral Streptococci substantially reduce commercial composite strength, and this effect warrants further study to identify the mechanism(s). CLINICAL SIGNIFICANCE: Dental biofilms of oral Streptococci bacteria significantly affect the mechanical strength of dental restorations.

Relationships between composite roughness and Streptococcus mutans biofilm depth under shear in vitro.

OBJECTIVE: To investigate the effect of substrate, surface roughness, and hydraulic residence time (HRT) on Streptococcus mutans biofilms growing on dental composites under conditions relevant to the oral cavity. METHODS: Dental composites were prepared with varying amounts of polishing and incubated in a CDC bioreactor with an approximate shear of 0.4 Pa. S. mutans biofilms developed in the bioreactors fed sucrose or glucose and at 10-h or 40-h HRT for one week. Biofilms were characterized by confocal laser microscopy (CLM). Composite surface roughness was characterized by optical profilometry, and pre- and post-incubation composite surface fine structure and elemental composition were determined using scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS). RESULTS: Polishing had a significant impact on surface roughness, varying by a factor of 15 between the polished samples and the unpolished control. S. mutans biofilms grew statistically significantly thicker on the unpolished composites. Biofilm thickness was greater at shorter 10-h HRT compared to 40-h HRT. In most cases, biofilm thickness was not statistically significantly greater in sucrose-fed bioreactors than in glucose-fed bioreactors. SEM-EDS analysis did not identify any significant change in elemental composition after aging. CONCLUSIONS: Accurate characterization of oral cavity biofilms must consider shear forces and the use of techniques that minimize alteration of the biofilm structure. Under shear, surface smoothness is the most important factor determining S. mutans biofilm thickness followed by HRT, while sucrose presence did not result in significantly greater biofilm thickness. CLINICAL SIGNIFICANCE: The obvious patterns of S. mutans growth along sub-micron scale grooving created by the polishing process suggested that initial biofilm attachment occurred in the shear-protected grooves. These results suggest that fine polishing may help prevent the initial formation of S. mutans biofilms compared to unpolished/coarse polished composites.

Stoichiometric models of sucrose and glucose fermentation by oral streptococci: Implications for free acid formation and enamel demineralization.

OBJECTIVES: The objective of this study is to develop stoichiometric models of sugar fermentation and cell biosynthesis for model cariogenic Streptococcus mutans and non-cariogenic Streptococcus sanguinis to better understand and predict metabolic product formation. METHODS: Streptococcus mutans (strain UA159) and Streptococcus sanguinis (strain DSS-10) were grown separately in bioreactors fed brain heart infusion broth supplemented with either sucrose or glucose at 37 °C. Cell mass concentration and fermentation products were measured at different hydraulic residence times (HRT) to determine cell growth yield. RESULTS: Sucrose growth yields were 0.080 ± 0.0078 g cell/g and 0.18 ± 0.031 g cell/g for S. sanguinis and S. mutans, respectively. For glucose, this reversed, with S. sanguinis having a yield of 0.10 ± 0.0080 g cell/g and S. mutans 0.053 ± 0.0064 g cell/g. Stoichiometric equations to predict free acid concentrations were developed for each test case. Results demonstrate that S. sanguinis produces more free acid at a given pH than S. mutans due to lesser cell yield and production of more acetic acid. Greater amounts of free acid were produced at the shortest HRT of 2.5 hr compared to longer HRTs for both microorganisms and substrates. SIGNIFICANCE: The finding that the non-cariogenic S. sanguinis produces greater amounts of free acids than S. mutans strongly suggests that bacterial physiology and environmental factors affecting substrate/metabolite mass transfer play a much greater role in tooth or enamel/dentin demineralization than acidogenesis. These findings enhance the understanding of fermentation production by oral streptococci and provide useful data for comparing studies under different environmental conditions.

Logistic Regression Analysis of LC-MS/MS Data of Monomers Eluted from Aged Dental Composites: A Supervised Machine-Learning Approach.

Compound identification by database searching that matches experimental with library mass spectra is commonly used in mass spectrometric (MS) data analysis. Vendor software often outputs scores that represent the quality of each spectral match for the identified compounds. However, software-generated identification results can differ drastically depending on the initial search parameters. Machine learning is applied here to provide a statistical evaluation of software-generated compound identification results from experimental tandem MS data. This task was accomplished using the logistic regression algorithm to assign an identification probability value to each identified compound. Logistic regression is usually used for classification, but here it is used to generate identification probabilities without setting a threshold for classification. Liquid chromatography coupled with quadrupole-time-of-flight tandem MS was used to analyze the organic monomers leached from resin-based dental composites in a simulated oral environment. The collected tandem MS data were processed with vendor software, followed by statistical evaluation of these results using logistic regression. The assigned identification probability to each compound provides more confidence in identification beyond solely by database matching. A total of 21 distinct monomers were identified among all samples, including five intact monomers and chemical degradation products of bisphenol A glycidyl methacrylate (BisGMA), oligomers of bisphenol-A ethoxylate methacrylate (BisEMA), triethylene glycol dimethacrylate (TEGDMA), and urethane dimethacrylate (UDMA). The logistic regression model can be used to evaluate any database-matched liquid chromatography-tandem MS result by training a new model using analytical standards of compounds present in a chosen database and then generating identification probabilities for candidates from unknown data using the new model.

Aqueous Geochemical Controls on the Sestonic Microbial Community in Lakes Michigan and Superior.

Despite being the largest freshwater lake system in the world, relatively little is known about the sestonic microbial community structure in the Laurentian Great Lakes. The goal of this research was to better understand this ecosystem using high-throughput sequencing of microbial communities as a function of water depth at six locations in the westernmost Great Lakes of Superior and Michigan. The water column was characterized by gradients in temperature, dissolved oxygen (DO), pH, and other physicochemical parameters with depth. Mean nitrate concentrations were 32 µmol/L, with only slight variation within and between the lakes, and with depth. Mean available phosphorus was 0.07 µmol/L, resulting in relatively large N:P ratios (97:1) indicative of P limitation. Abundances of the phyla Actinobacteria, Bacteroidetes, Cyanobacteria, Thaumarchaeota, and Verrucomicrobia differed significantly among the Lakes. Candidatus Nitrosopumilus was present in greater abundance in Lake Superior compared to Lake Michigan, suggesting the importance of ammonia-oxidating archaea in water column N cycling in Lake Superior. The Shannon diversity index was negatively correlated with pH, temperature, and salinity, and positively correlated with DO, latitude, and N2 saturation. Results of this study suggest that DO, pH, temperature, and salinity were major drivers shaping the community composition in the Great Lakes.

The impact of long-term aging in artificial saliva media on resin-based dental composite strength.

Although salivary liquid can degrade constituents in resin-based dental composites in short-term incubations, there is a knowledge gap on how longer-term aging impacts their bulk strength. We address this through extended aging studies with resin-based dental composites in different environments. Two commercial composites (FIL and AEL) were aged aseptically at 37°C in air (A, control), artificial saliva (AS), and esterase enzyme amended AS (EAS). Diametral and pushout strength were measured after periods of 120-180 days. At 120 days, the diametral strength of composites aged in air was 69.9 ± 11.0 and 57.7 ± 3.31 MPa in FIL and AEL, respectively. These were significantly greater compared to composites aged in AS (32.1 ± 7.01 and 46.2 ± 9.38 MPa in FIL and AEL, respectively) or EAS (36.7 ± 8.49 and 43.5 ± 5.51 MPa in FIL and AEL, respectively). In contrast, pushout strength for both composites were smaller in A compared to those aged in AS and EAS, results attributed to AS absorption and polymer expansion. No significant change in either diametral or pushout strength occurred after 120 days. There was no significant difference between aging in AS and EAS, suggesting that esterase did not significantly decrease the bulk material strength to a greater extent than AS under the test conditions. Aqueous diffusivities for the composites ranged from 8.4 to 11 × 10-13  m2 /s, with associated porosities ranging from 0.06% to 0.10%. These results indicate that saturation of a typical dental composite occurs over a time frame of 4-5 months, longer than typical aging studies. Together, the results demonstrate the importance of aging time on composite strength.

Polyhalogenated carbazoles in freshwater and estuarine sediment from China and the United States: A multi-regional study.

The present study represents a multi-regional investigation of polyhalogenated carbazoles (PHCZs) contamination in estuarine and freshwater systems from the United States and China. Although recent studies have suggested that PHCZs are persistent and bioaccumulative, available data are not sufficient to understand their large-scale spatial and temporal distributions in the environment. The present study investigated spatial distributions of PHCZs in surface sediment from multiple freshwater and estuarine systems located in China and the United States (U.S.) during the period of 2012-2017, as well as temporal distributions from vertical trends in selected sediment cores. The results demonstrated large variations of PHCZ contamination across regions, with median concentrations of ΣPHCZs in surface sediment ranging from 3.1 to 134 ng/g. Profiles of PHCZ congener composition also exhibited regional variations and estuarine-freshwater differences. These differences likely reflect the relative contributions of different natural and industrial sources among the locations. Vertical profiles of concentrations and compositions in one Chinese estuarine sediment core and two freshwater sediment cores from the U.S. all demonstrated clear anthropogenic influences to varying degrees. Toxic equivalents (TEQ) of PHCZs were estimated based on their dioxin-like activities, which ranged from <0.001 to 4.94 pg TEQ/g in all sites. The results suggest that PHCZs could add additional ecological risks to the benthos and other aquatic organisms. Our findings constitute an essential contribution to the knowledge body of PHCZ contamination in global aquatic systems and congener-specific contamination characterizations.

Halogenated flame retardants in sediments from the Upper Laurentian Great Lakes: Implications to long-range transport and evidence of long-term transformation.

Most hydrophobic halogenated flame retardants (HFRs) are highly accumulative and persistent in aquatic sediments. The objective of this study was to reveal spatial distributions, temporal trends, and transformation of selected legacy and emerging HFRs in sediments of Lakes Superior, Michigan, and Huron. We collected Ponar grab samples at 112 locations and sediment cores at 28 sites in the three lakes, and measured concentrations of 19 brominated FRs and 12 chlorinated FRs. Based on grab samples, concentrations were higher at southeastern and sites near Sleeping Bear Dunes of Lake Michigan, and Saginaw Bay and the North Channel of Lake Huron. The annual loadings of polybrominated diphenyl either (PBDEs) and Dechlorane Plus (DPs) to sediment have leveled off or been declining since 2000, while loadings of DBDPE and Dec604 have increased since the 1960s in most cores. The concentration ratio of BB101 to BB153 increased with sediment depth, suggesting the occurrence of in situ debromination of BB153. The ratio of dechlorinated anti-Cl11DP over anti-DP increases with the increasing latitude of sampling locations, suggesting the occurrence of dechlorination of anti-DP to anti-Cl11DP during transport. This ratio also increases with increasing sediment age in most cores, implying in situ dechlorination over time.

The effect of esterase enzyme on aging dental composites.

We measured the push-out and diametral tensile strength of dental restorative composites following aging under environmental conditions relevant to the oral cavity; air (A), artificial saliva (AS), acidified (50 mM CH3 COOH, pH = 4.7) artificial saliva (AS + HAc), and AS with esterase enzyme (AS + ENZ). Cylindrical test specimens (6.3 mm diameter by 5.1 mm long) were prepared by placing 0.3 g of nanofilled composite in an epoxy ring and cured. Twenty samples were aged in each environment for 163-186 days at 37°C. The push-out strengths (mean ± standard error of the mean [SEM], in MPa) for specimens were: A-2.4 ± 0.2, AS-7.3 ± 0.5, AS + HAc-7.2 ± 0.9, and AS + ENZ-6.0 ± 0.6. Following the push-out test, the diametral tensile strength and elasticity were immediately determined. The diametral tensile strengths (mean ± SEM, in MPa) for specimens were: A-54.0 ± 1.6, AS-31.4 ± 1.3, AS + HAc-34.3 ± 1.2, and AS + ENZ-22.5 ± 0.7. The push-out strength was lowest for the A environment due to shrinkage of the composite. The push-out strength increased significantly as water diffused into the specimens (AS and AS + HAc) but decreased significantly in the enzyme environment (AS + ENZ). The diametral tensile strength was highest for specimens in the A environment, which was significantly higher than both the AS and AS + HAc specimens and > 2× higher than the AS + ENZ specimens. The results indicated that a water environment (with or without acid) caused a significant decrease in the mechanical properties of this composite, but the greatest decrease was seen in water with esterase. This is the first study to demonstrate that esterase enzymes affect the bulk strength of a commonly used commercial dental composite. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2178-2184, 2019.

Organic contaminated sediments remediation with active caps: Nonlinear adsorption unveiled by combined isotherm and column transportation studies.

Bench-scale column studies were performed with four cap materials (sand, apatite, organoclay and granular activated carbon) for five target compounds (toluene, naphthalene, phenanthrene, pyrene and dichlorobiphenyl) to represent a range of cap materials and pollutants (volatile & semi-volatile compounds) commonly found in sediments. Two moment-derived methods were used to model cap performance. Rough agreement was observed between the column experiments and modeling data with the fronting and tailing effects identified from certain breakthrough curves indicating a high potential of non-linear adsorption. Distribution coefficients (kd) were experimentally determined with isotherm studies together with measurements of surface area and microporosity of the cap materials via nitrogen adsorption porisimetry. These studies unveiled the occurrence of nonlinear adsorption by Freundlich simulation. The effects of nonlinear adsorption of the cap were further explored via modeling. Results suggested better prediction of cap performance assuming nonlinear adsorption instead of linear adsorption results based upon the risk of release for a 30-year period.

Spatial and temporal trends in poly- and per-fluorinated compounds in the Laurentian Great Lakes Erie, Ontario and St. Clair.

The temporal and spatial trends in sediment of 22 poly- and perfluorinated (PFAS) compounds were investigated in the southern Great Lakes Erie and Ontario as well as Lake St. Clair. Surface concentrations measured by Ponar grab samples indicated a trend for greater concentrations near to urban sites. Mean concentrations ∑22PFAS were 15.6, 18.2 and 19 ng g-1 dm for Lakes St. Clair, Erie and Ontario, respectively. Perfluoro-n-butanoic acid (PFBA) and Perfluoro-n-hexanoic acid (PFHxA) were frequently determined in surface sediment and upper core samples indicating a shift in use patterns. Where PFBA was identified it was at relatively great concentrations typically >10 ng g-1 dm. However as PFBA and PFHxA are less likely to bind to sediment they may be indicative of pore water concentrations Sedimentation rates between Lake Erie and Lake Ontario differ greatly with greater rates observed in Lake Erie. In Lake Ontario, in general concentrations of PFAS observed in core samples closely follow the increase in use along with an observable change due to regulation implementation in the 1970s for water protection. However some of the more water soluble PFAS were observed in deeper core layers than the time of production could account for, indicating potential diffusion within the sediment. Given the greater sedimentation rates in Lake Erie, it was hoped to observe in greater resolution changes since the mid-1990s. However, though some decrease was observed at some locations the results are not clear. Many cores in Lake Erie had clearly observable gas voids, indicative of gas ebullition activity due to biogenic production, there were also observable mussel beds that could indicate mixing by bioturbation of core layers.

Current and historical concentrations of poly and perfluorinated compounds in sediments of the northern Great Lakes - Superior, Huron, and Michigan.

Current and historical concentrations of 22 poly- and perfluorinated compounds (PFASs) in sediment collected from Lake Superior and northern Lake Michigan in 2011 and Lake Huron in 2012 are reported. The sampling was performed in two ways, Ponar grabs of surface sediments for current spatial distribution across the lake and dated cores for multi-decadal temporal trends. Mean concentrations of the sum of PFASs (∑PFASs) were 1.5, 4.6 and 3.1 ng g-1 dry mas (dm) in surface sediments for Lakes Superior, Michigan and Huron, respectively. Of the five Laurentian Lakes, the watersheds of Superior and Huron are the less densely populated by humans, and concentrations observed were typically less and from more diffuse sources, due to lesser urbanization and industrialization. However, some regions of greater concentrations were observed and might indicate more local, point sources. In core samples concentrations ranged from 7 that include perfluoro-n-octane sulfonate (PFOS) bind more strongly to sediment, which resulted in more accurate analyses of temporal trends. Shorter-chain PFASs, such as perfluoro-n-butanoic acid which is the primary replacement for C8 PFASs that have been phased out, are more soluble and were identified in some core layers at depths corresponding to pre-production periods. Thus, analyses of temporal trends of these more soluble compounds in cores of sediments were less accurate. Total elemental fluorine (TF) and extractable organic fluorine (EOF) indicated that identified PFASs were not a significant fraction of fluorine containing compounds in sediment (<0.01% in EOF).

Organophosphate Esters in Sediment of the Great Lakes.

This is the first study on organophosphate ester (OPEs) flame retardants and plasticizers in the sediment of the Great Lakes. Concentrations of 14 OPEs were measured in three sediment cores and 88 Ponar surface grabs collected from Lakes Ontario, Michigan, and Superior of North America. The sum of these OPEs (Σ14OPEs) in Ponar grabs averaged 2.2, 4.7, and 16.6 ng g-1 dw in Lakes Superior, Michigan, and Ontario, respectively. Multiple linear regression analyses demonstrated statistically significant associations between logarithm concentrations of Σ14OPEs as well as selected congeners in surface grab samples and sediment organic carbon content as well as a newly developed urban distance factor. Temporal trends observed in dated sediment cores from Lake Michigan demonstrated that the recent increase in depositional flux to sediment is dominated by chlorinated OPEs, particularly tris(2-chloroisopropyl) phosphate (TCPP), which has a doubling time of about 20 years. Downward diffusion within sediment may have caused vertical fractionation of OPEs over time. Two relatively hydrophilic OPEs including TCPP had much higher concentrations in sediment than estimated based on equilibria between water and sediment organic carbon. Approximately a quarter (17 tonnes) of the estimated total OPE burden (63 tonnes) in Lake Michigan resides in sediment, which may act as a secondary source releasing OPEs to the water column for years to come.

Spatial and Temporal Trends of Polyhalogenated Carbazoles in Sediments of Upper Great Lakes: Insights into Their Origin.

Polyhalogenated carbazoles (PHCZs) have been increasingly detected in the environment. Their similarities in chemical structure with legacy pollutants and their potential toxicity have caused increasing concern. In this work, 112 Ponar grab and 28 core sediment samples were collected from Lakes Michigan, Superior, and Huron, and a total of 26 PHCZs were analyzed along with unsubstituted carbazole using gas chromatography coupled with single- or triple-quadrupole mass spectrometry. Our results show that the total accumulation of PHCZs in the sediments of the upper Great Lakes is >3000 tonnes, orders of magnitude greater than those of polychlorinated biphenyls (PCBs) and decabromodiphenyl ether (BDE209). The 27 individual analytes differ in spatial distribution and temporal trend. Our results showed that PHCZs with substitution patterns of -Br2-5, -Cl1-2Br2-4, or having iodine, were more abundant in sediment of Lake Michigan deposited before 1900 than those deposited more recently, implying a natural origin. Some "emerging" PHCZs have been increasingly deposited into the sediment in recent decades, and deserve further environmental monitoring and research. Other PHCZs with low halogen substitution may form from in situ dehalogenation of PHCZs having more halogens. Anthropogenic sources of PHCZs may exist, particularly for the emerging and low molecular mass congeners.

Untargeted Screening and Distribution of Organo-Iodine Compounds in Sediments from Lake Michigan and the Arctic Ocean.

The majority of halogenated organic compounds present in the environment remain unidentified. To address this data gap, we recently developed an untargeted method (data-independent precursor isolation and characteristic fragment; DIPIC-Frag) for identification of unknown organo-bromine compounds. In this study, the method was adapted to enable untargeted screening of natural and synthetic organo-iodine compounds (NSOICs) in sediments. A total of 4,238 NSOIC peaks were detected in sediments from Lake Michigan. Precursor ions and formulas were determined for 2,991 (71%) of the NSOIC peaks. These compounds exhibited variations in abundances (<10(3) to ∼10(7)), m/z values (206.9304-996.9474), retention times (1.0-29.7 min), and number of iodine atoms (1-4). Hierarchical cluster analysis showed that sediments in closer proximity exhibited similar profiles of NSOICs. NSOICs were screened in 10 samples of sediment from the Arctic Ocean to compare the profiles of NSOICs between freshwater and marine sediments. A total of 3,168 NSOIC peaks were detected, and profiles of NSOICs in marine sediments were clearly distinct from Lake Michigan. The coexistence of brominated and iodinated analogues indicated that some NSOICs are of natural origin. Different ratios of abundances of iodinated compounds to brominated analogues were observed and proposed as a marker to distinguish sources of NSOICs.

Occurrence of Atrazine and Related Compounds in Sediments of Upper Great Lakes.

Surface grab and core sediment samples were collected from Lakes Michigan, Superior, and Huron from 2010 to 2012, and concentrations of herbicides atrazine, simazine, and alachlor, as well as desethylatrazine (DEA), were determined. Concentrations of atrazine in surface grabs ranged from 0.01 to 1.7 ng/g dry weight and are significantly higher in the southern basin of Lake Michigan (latitude <44°) than other parts of the three lakes. The highest concentration of alachlor was found in sediments of Saginaw Bay in Lake Huron. The inventory and net fluxes of these herbicides were found to decline exponentially from the south to the north. The concentration ratio of DEA to atrazine (DEA/ATZ) increased with latitude, suggesting degradation of atrazine to DEA during atmospheric transport. DEA/ATZ also increased with sediment depth in the sediment cores. Diffusion of deposited herbicides from the upper sediment into deeper sediments has occurred, on the basis of the observed patterns of concentrations in dated sediment cores. Concentrations of atrazine in pore water were estimated and were higher than those reported for the bulk waters, suggesting the occurrence of solid-phase deposition of atrazine through the water column and that contaminated sediments act as a source releasing atrazine to the overlying water.

Debromination of PBDEs in Arkansas Water Bodies Analyzed by Positive Matrix Factorization.

A previously generated data set for polybrominated diphenyl ethers (PBDEs) in dated sediment cores of West Lake of El Dorado (AED), Calion Lake (ACL), and the lagoon of Magnolia Wastewater Treatment Facility (AMW) from Southern Arkansas is examined by a weighted chemical mass balance (CMB) model and positive matrix factorization (PMF) in order to quantify PBDE sources and debromination. DNA extraction and pyrosequencing were done on several core sections in order to investigate microbial debromination. CMB and PMF analyses indicate that deca technical mixtures are the dominant PBDE input (>99% in mole fraction in AED and ACL, and 94.7% in AMW). Minor contributions of penta and octa technical mixtures were found in all three water bodies (<1% in AED and ACL; and 1.1% and 4.1% in AMW, respectively). Results suggest that debromination takes place in all three lakes, but is more intense in AMW. In-situ microbial debromination was supported by the microorganism analysis. The PMF results are validated by PBDE manufacturing records, and the operating history of AMW. Despite the high PBDE concentrations in these sediments near former manufacturing facilities, the extent of debromination is limited, possibly due to sorption to natural organic matter of the sediment.

Untargeted Screening and Distribution of Organo-Bromine Compounds in Sediments of Lake Michigan.

Previously unreported natural and synthetic organo-bromine compounds (NSOBCs) have been found to contribute more than 99% of total organic bromine (TOB) in environmental matrices. We recently developed a novel untargeted method (data-independent precursor isolation and characteristic fragment, DIPIC-Frag) and identified ∼2000 NSOBCs in two sediments from Lake Michigan. In this study, this method was used to investigate the distributions of these NSOBCs in 23 surficial samples and 24 segments of a sediment core from Lake Michigan. NSOBCs were detected in all 23 surficial samples and exhibited 10- to 100-fold variations in peak abundance among locations. The pattern of distributions of NSOBCs was correlated with depth of the water column (r(2) = 0.61, p < 0.001). Hierarchical cluster analysis showed that sediments in close proximity exhibited similar profiles of NSOBCs. Distributions of NSOBCs in 24 segments of a sediment core dated from 1766 to 2008 were investigated, and samples from similar depths exhibited similar profiles of NSOBCs. NSOBCs were grouped into four clusters (soft-cluster analysis) with different temporal trends of abundances. 515 and 768 of the NSOBCs were grouped into cluster 1 and cluster 3 with increasing temporal trends, especially since 1950, indicating that abundances of these compounds might have been affected by human activities.

Untargeted Identification of Organo-Bromine Compounds in Lake Sediments by Ultrahigh-Resolution Mass Spectrometry with the Data-Independent Precursor Isolation and Characteristic Fragment Method.

While previous studies have found that unknown natural and synthetic organo-bromine compounds (NSOBCs) contributed more than 99% of the total organic bromine (Br) in the environment, there was no efficient method for untargeted screening to identify NSOBCs in environmental matrixes. A novel untargeted method for identifying NSOBCs, based on ultrahigh-resolution mass spectrometry (UHRMS) with the Q Exactive instrument was developed. This method included a data-independent precursor isolation and characteristic fragment (DIPIC-Frag) procedure to identify NSOBCs. A total of 180 successive 5-m/z-wide windows were used to isolate precursor ions. This resulted in a sufficient dynamic range and specificity to identify peaks of Br fragment ions for analysis. A total of 2520 peaks of NSOBC compounds containing Br were observed in sediments from Lake Michigan, United States. A new chemometric strategy which combined chromatographic profiles, isotopic peaks, precursor isolation window information, and intensities was used to identify precursor ions and chemical formulas for detecting NSOBCs. Precursor ions for 2163 of the 2520 NSOBCs peaks (86%) were identified, and chemical formulas for 2071 NSOBCs peaks (82%) were determined. After exclusion of isotopic peaks, 1593 unique NSOBCs were identified and chemical formulas derived for each. Most of the compounds identified had not been reported previously and had intensities which were 100- to 1000-fold greater than the congeners of polybrominated diphenyl ethers (PBDEs). In extracts of sediments, these compounds exhibited variations in intensities (<10(3) to ∼10(8)), m/z values (170.9438-997.5217), retention times on a C18 column (1.0-29.3 min), and the number of Br atoms (1-8). Generally, compounds with greater m/z values had longer retention times and greater numbers of Br atoms. Three compounds were used in a proof-of-concept experiment to demonstrate that structures of some of the screened NSOBCs could be further predicted by combining searching of database libraries and high-resolution MS(2) spectra.

Geochemical influences and mercury methylation of a dental wastewater microbiome.

The microbiome of dental clinic wastewater and its impact on mercury methylation remains largely unknown. Waste generated during dental procedures enters the sewer system and contributes a significant fraction of the total mercury (tHg) and methyl mercury (MeHg) load to wastewater treatment facilities. Investigating the influence of geochemical factors and microbiome structure is a critical step linking the methylating microorganisms in dental wastewater (DWW) ecosystems. DWW samples from a dental clinic were collected over eight weeks and analyzed for geochemical parameters, tHg, MeHg and bacterio-toxic heavy metals. We employed bacterial fingerprinting and pyrosequencing for microbiome analysis. High concentrations of tHg, MeHg and heavy metals were detected in DWW. The microbiome was dominated by Proteobacteria, Actinobacteria, Bacteroidetes, Chloroflexi and many unclassified bacteria. Significant correlations were found between the bacterial community, Hg levels and geochemical factors including pH and the predicted total amount (not fraction) of neutral Hg-sulfide species. The most prevalent known methylators included Desulfobulbus propionicus, Desulfovibrio desulfuricans, Desulfovibrio magneticus and Geobacter sulfurreducens. This study is the first to investigate the impact of high loads of Hg, MeHg and other heavy metals on the dental clinic wastewater microbiome, and illuminates the role of many known and unknown sulfate-reducing bacteria in Hg methylation.