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A practical method for the determination of total selenium in environmental samples using isotope dilution-hydride generation-inductively coupled plasma-mass spectrometry

A safe, practical, and accurate method for the determination of selenium (Se) in range of environmental samples was developed. Small sample masses, 5–20 mg, were amended with 82 Se enriched isotope for the isotope dilution (ID), preceding a multi-step wet digestion with nitric acid (HNO 3 ) and hydrogen peroxide (H 2 O 2 ). Samples were incubated in an autoclave for 3 h at 20 psi and 126°C. Digestates were subsequently reduced with concentrated hydrochloric acid to Se(IV) the most favorable valence for hydride generation (HG). The solutions were then analyzed on an ICP-MS equipped with Flow Injection system (FIAS-400). Polyatomic, isobaric, and background interferences were removed through the use of HG and ID with an 82 Se enriched isotope spike. Recoveries for certified reference materials were determined and averaged 96% for biological tissues (NRCC DOLT3, DOLT4, DORM2, TORT2, and TORT3, and NIST 2976) and 108% for estuarine sediment (NRCC PACS2) with an average coefficient of variation for replicate measurements of ∼ 3.5%. Limit of detection was 0.13 ng Se g −1 dry weight or 0.19 ng Se L −1 . This method can be broadly applied to biological tissues, sediments, suspended particulates, and water samples with minimal modifications making this method highly useful for assessing the ecotoxicology of total Se in aquatic ecosystems.

Limnology and Oceanography: Methods

Comparison of in vitro estrogenic activity and estrogen concentrations insource and treated waters from 25 U.S. drinking water treatment plants

In vitro bioassays have been successfully used to screen for estrogenic activity in wastewater and surface water, however, few have been applied to treated drinking water. Here, extracts of source and treated water samples were assayed for estrogenic activity using T47D-KBluc cells and analyzed by liquid chromatography-Fourier transform mass spectrometry (LC-FTMS) for natural and synthetic estrogens (including estrone, 17β-estradiol, estriol, and ethinyl estradiol). None of the estrogenswere detected above the LC-FTMS quantification limits in treated samples and only 5 source waters had quantifiable concentrations of estrone, whereas 3 treated samples and 16 source samples displayed in vitro estrogenicity. Estrone accounted for themajority of estrogenic activity in respective samples, however the remaining samples that displayed estrogenic activity had no quantitative detections of known estrogenic compounds by chemical analyses. Source water estrogenicity (max, 0.47 ng 17β-estradiol equivalents (E2Eq) L−1) was below levels that have been linked to adverse effects in fish and other aquatic organisms. Treated water estrogenicity (max, 0.078 ng E2Eq L−1) was considerably below levels that are expected to be biologically relevant to human consumers. Overall, the advantage of using in vitro techniques in addition to analytical chemical determinations was displayed by the sensitivity of the T47D-KBluc bioassay, coupled with the ability tomeasure cumulative effects of mixtures, specifically when unknown chemicals may be present.

Science of the Total Environment

A Carboniferous apex for the late Paleozoic icehouse

Icehouse climate systems occur across an abbreviated portion of Earth history, constituting c. 25% of the Phanerozoic record. The Late Paleozoic Ice Age (LPIA) was the most extreme and longest lasting glaciation of the Phanerozoic and is characterized by periods of acute continental-scale glaciation, separated by periods of ice minima or ice-free conditions on the order of <10 6 years. The late Paleozoic glaciogenic record of the Paraná and Kalahari basins of southern Gondwana form one of the largest, best-preserved and well-calibrated records of this glaciation. In the Carboniferous, the eastern and southern margins of the Paraná Basin and the Kalahari Basin were characterized by subglacial conditions, with evidence for continental and upland glaciers. In the latest Carboniferous, these basins transitioned from subglacial reservoirs to ice-free or ‘ice distal‘ conditions evidenced by the widespread deposition of marine deposits juxtaposed on subglacial bedforms. High-precision U–Pb zircon chemical abrasion thermal ionization mass spectrometry geochronological constraints from volcanic ash deposits in the deglacial marine black shales of the Kalahari Basin and from fluvial and coal successions, which overlie marine deposits in the Paraná Basin, indicate subglacial evidence in these regions is constrained to the Carboniferous. The loss of ice in these regions is congruent with a late Carboniferous peak in p CO 2 and widespread marine anoxia in the late Carboniferous. The permeant retreat of glaciers in basinal settings, despite an early Permian p CO 2 nadir, highlights the influence of short-term perturbations on the longer-term CO 2 record and suggests an ice threshold had been crossed in the latest Carboniferous. A definitive driver for greenhouse gases in the LPIA, such as abundant and sustained volcanic activity or an increased biological pump driven by ocean fertilization, is unresolved for this period. Lastly, the proposed Carboniferous apex for the high-latitude LPIA record is incongruent with observations from the low-latitude tropics where an early Permian peak is proposed.

Geological Society, London, Special Publications

Fluid inclusions in quartz crystals from South-West Africa

Quartz crystals from calcite veins of unknown age in Precambrian metasedimentary rocks at Geiaus No. 6 and Aukam farms in South-West Africa contain both primary and secondary inclusions filled with one or a variable combination of: organic liquid, moderately saline aqueous liquid, dark-colored solid, and vapor. Analysis of these materials by microscopy and by gas chromatography and mass spectrometry shows the presence of constituents of both low and high molecular weights. The former include CH 4 , C 2 H 6 , C 3 H 8 and possibly C 4 H 10 as well as CO, CO 2 , H 2 O, N 2 and H 2 . High molecular weight components are dominantly n-alkanes and isoprenoid hydrocarbons. The n-alkanes range from at least n-C 10 to n-C 33 . Concentrations of n-alkanes larger than n-C 17 decrease regularly with increasing carbon number. An homologous series of isoprenoid hydrocarbons ranging from at least C 14 to C 20 is present in unusually high concentrations. Pristane (C 19 ) is most abundant, and C 17 isoprenoid is least abundant. The molecular composition and distribution of hydrocarbons suggest biological precursors for these components. Consideration of data provided by freezing, crushing and heating experiments suggests that the pressures at the time these in part supercritical fluids were trapped probably exceeded 30–40 atm, and the minimum trapping temperature was about 120–160°C. Both primary and secondary inclusions apparently containing only organic materials were trapped by the growth of the host quartz from aqueous solution. The data obtained neither prove nor preclude Precambrian, Paleozoic or younger sources for the organic materials.

Geochimica et Cosmochimica Acta

Ferromanganese crusts as archives of deep water Cd isotope compositions

The geochemistry of Cd in seawater has attracted significant attention owing to the nutrient-like properties of this element. Recent culturing studies have demonstrated that Cd is a biologically important trace metal that plays a role in the sequestration of inorganic carbon. This conclusion is supported by recent isotope data for Cd dissolved in seawater and incorporated in cultured phytoplankton. These results show that plankton features isotopically light Cd while Cd-depleted surface waters typically exhibit complimentary heavy Cd isotope compositions. Seawater samples from below 900 m depth display a uniform and intermediate isotope composition of ε 114/110 Cd = +3.3 ± 0.5. This study investigates whether ferromanganese (Fe-Mn) crusts are robust archives of deep water Cd isotope compositions. To this end, Cd isotope data were obtained for the recent growth surfaces of 15 Fe-Mn crusts from the Atlantic, Pacific, Indian, and Southern oceans and two USGS Fe-Mn reference nodules using double spike multiple collector inductively coupled plasma mass spectrometry. The Fe-Mn crusts yield a mean ε 114/110 Cd of +3.2 ± 0.4 (2 SE, n = 14). Data for all but one of the samples are identical, within the analytical uncertainty of ±1.1ε 114/110 Cd (2 SD), to the mean deep water Cd isotope value. This indicates that Fe-Mn crusts record seawater Cd isotope compositions without significant isotope fractionation. A single sample from the Southern Ocean exhibits a light Cd isotope composition of ε 114/110 Cd = 0.2 ± 1.1. The origin of this signature is unclear, but it may reflect variations in deep water Cd isotope compositions related to differences in surface water Cd utilization or long-term changes in seawater ε 114/110 Cd. The results suggest that time series analyses of Fe-Mn crusts may be utilized to study changes in marine Cd utilization.

Geochemistry, Geophysics, Geosystems

Occurrence of triclosan, triclocarban, and its lesser chlorinated congeners in Minnesota freshwater sediments collected near wastewater treatment plants

The antimicrobial agents triclosan (TCS), triclocarban (TCC) and their associated transformation products are of increasing concern as environmental pollutants due to their potential adverse effects on humans and wildlife, including bioaccumulation and endocrine-disrupting activity. Analysis by tandem mass spectrometry of 24 paired freshwater bed sediment samples (top 10 cm) collected by the U.S. Geological Survey near 12 wastewater treatment plants (WWTPs) in Minnesota revealed TCS and TCC concentrations of up to 85 and 822 ng/g dry weight (dw), respectively. Concentrations of TCS and TCC in bed sediments collected downstream of WWTPs were significantly greater than upstream concentrations in 58% and 42% of the sites, respectively. Dichloro- and non-chlorinated carbanilides (DCC and NCC) were detected in sediments collected at all sites at concentrations of up to 160 and 1.1 ng/g dw, respectively. Overall, antimicrobial concentrations were significantly higher in lakes than in rivers and creeks, with relative abundances decreasing from TCC > TCS > DCC > NCC. This is the first statewide report on the occurrence of TCS, TCC and TCC transformation products in freshwater sediments. Moreover, the results suggest biological or chemical TCC dechlorination products to be ubiquitous in freshwater environments of Minnesota, but whether this transformation occurs in the WWTP or bed sediment remains to be determined.

Minnesota

Male lake char release taurocholic acid as part of a mating pheromone

The evolutionary origins of sexual preferences for chemical signals remain poorly understood, due, in part, to scant information on the molecules involved. In the current study, we identified a male pheromone in lake char ( Salvelinus namaycush ) to evaluate the hypothesis that it exploits a non-sexual preference for juvenile odour. In anadromous char species, the odour of stream-resident juveniles guides migratory adults into spawning streams. Lake char are also attracted to juvenile odour but have lost the anadromous phenotype and spawn on nearshore reefs, where juvenile odour does not persist long enough to act as a cue for spawning site selection by adults. Previous behavioural data raised the possibility that males release a pheromone that includes components of juvenile odour. Using metabolomics, we found that the most abundant molecule released by males was also released by juveniles but not females. Tandem mass spectrometry and nuclear magnetic resonance were used to identify the molecule as taurocholic acid (TCA), which was previously implicated as a component of juvenile odour. Additional chemical analyses revealed that males release TCA at high rates via their urine during the spawning season. Finally, picomolar concentrations of TCA attracted pre-spawning and spawning females but not males. Taken together, our results indicate that male lake char release TCA as a mating pheromone and support the hypothesis that the pheromone is a partial match of juvenile odour.

Journal of Experimental Biology

American eels produce and release bile acids that vary across life stage

The American eel ( Anguilla rostrata ) is an imperilled fish hypothesized to use conspecific cues, in part, to coordinate long‐distance migration during their multistage life history. Here, holding water and tissue from multiple American eel life stages was collected and analysed for the presence, profile and concentration of bile acids. Distinct bile acid profiles were identified in glass, elver, yellow eel and silver eel holding waters using ultraperformance liquid chromatography high‐resolution mass spectrometry and principal component analysis. Taurochenodeoxycholic acid, taurodeoxycholic acid, cholic acid, deoxycholic acid, taurolithocholic acid and taurocholic acid were detected in whole tissue of American glass eels and elvers, and in liver, intestine and gallbladder samples of late‐stage yellow eels. Bile acids were not a major component of silver eel washings or tissue. This study is novel because little was previously known about bile acids produced and emitted into the environment by American eels. Future behavioural studies could evaluate whether any bile acids produced by American eels influence conspecific migratory behaviour.

Journal of Fish Biology

The influence of soil development on the depth distribution and structure of soil microbial communities.

Although it has been shown that the interaction of climate and time shape the dynamics of soil organic matter (SOM) storage and preservation in soil, the role of soil microbial communities in this dynamic remains unclear. Microbial communities are present throughout soil profiles and likely play critical roles in SOM and nutrient cycling, however the influence of other factors such as soil development (i.e., age) and the composition SOM on microbial community variation with depth has yet to be quantified. Improving our understanding of the relationship between soil development, soil depth, and microbial communities may provide insight to the critical role they play in cycling and preservation of SOM, as well as more mechanistic predictions of the response of soil communities to change, such as landscape-scale changes in available moisture or temperature regimes. Here we compare soils spanning a soil age by climate gradient (i.e., climo-chronosequence) to better understand the mechanisms which influence soil microbial community structure and the molecular composition of SOM. While we observed little depth-dependence in metrics of microbial community structure (i.e., composition, diversity, dissimilarity) across the range of soil development under a wetter climate, we found significant depth-dependent changes in community metrics under a drier climate, which became more pronounced as soils became older. This shift in bacterial and archaeal community structure and diversity is most apparent below a clay-rich argillic horizon formed in the older, drier soils. The molecular composition of SOM as measured by high resolution mass spectrometry (i.e., FTICR MS) also exhibited similar shifts in composition with soil depth and age. Our results highlight how soil moisture shapes the interaction of soil development, SOM, and microbial community composition. Differences in the moisture regime between our two study sites drives differences in biogeochemical depth gradients and subsequent variation in soil microbes and SOM. This suggests that knowledge of not just the pedogenic trajectory of soil development, but also the spatial position relative to distinct pedogenic features, are important for explaining variations in the depth-dependencies of microbial communities and associated SOM.

California

Instability of seawater pH in the South China Sea during the mid-late Holocene: Evidence from boron isotopic composition of corals

We used positive thermal ionization mass spectrometry (PTIMS) to generate high precision ??11B records in Porites corals of the mid-late Holocene from the South China Sea (SCS). The ??11B values of the Holocene corals vary significantly, ranging from 22.2??? to 25.5???. The paleo-pH records of the SCS, reconstructed from the ??11B data, were not stable as previously thought but show a gradual increase from the Holocene thermal optimal and a sharp decrease to modern values. The latter is likely caused by the large amount of anthropogenic CO2 emissions since the Industrial Revolution but variations of atmospheric pCO2 cannot explain the pH change of the SCS before the Industrial Revolution. We suggest that variations of monsoon intensity during the mid-late Holocene may have driven the sea surface pH increase from the mid to late Holocene. Results of this study indicate that the impact of anthropogenic atmospheric CO2 emissions may have reversed the natural pH trend in the SCS since the mid-Holocene. Such ocean pH records in the current interglacial period can help us better understand the physical and biological controls on ocean pH and possibly predict the long-term impact of climate change on future ocean acidification. ?? 2008 Elsevier Ltd. All rights reserved.

Geochimica et Cosmochimica Acta

Assessing dorsal scute microchemistry for reconstruction of shortnose sturgeon life histories

The imperiled status of sturgeons worldwide places priority on the identification and protection of critical habitats. We assessed the micro-structural and micro-chemical scope for a novel calcified structure, dorsal scutes, to be used for reconstruction of past habitat use and group separation in shortnose sturgeon ( Acipenser brevirostrum ). Dorsal scutes contained a dual-layered structure composed of a thin multi-layered translucent zone lying dorsally above a thicker multi-layered zone. Banding in the thick multi-layered zone correlated strongly with pectoral fin spine annuli supporting the presence of chronological structuring that could contain a chemical record of past environmental exposure. Trace element profiles (Sr:Ca), collected using both wavelength dispersive electron microprobe analysis and laser ablation inductively coupled mass spectrometry, suggest scutes record elemental information useful for tracing transitions between freshwater and marine environments. Moreover, mirror-image like Sr:Ca profiles were observed across the dual-zone structuring of the scute that may indicate duplication of the microchemical profile in a single structure. Additional element:calcium ratios measured in natal regions of dorsal scutes (Ba:Ca, Mg:Ca) suggest the potential for further refinement of techniques for identification of river systems of natal origin. In combination, our results provide proof of concept that dorsal scutes possess the necessary properties to be used as structures for reconstructions of past habitat use in sturgeons. Importantly, scutes may be collected non-lethally and with less injury than current structures, like otoliths and fin spines, affording an opportunity for broader application of microchemical techniques.

Environmental Biology of Fishes

Evidence that watershed nutrient management practices effectively reduce estrogens in environmental waters

We evaluate the impacts of different nutrient management strategies on the potential for co-managing estrogens and nutrients in environmental waters of the Potomac watershed of the Chesapeake Bay. These potential co-management approaches represent agricultural and urban runoff, wastewater treatment plant effluent, and combined sewer overflow replacements. Twelve estrogenic compounds and their metabolites were analysed by gas chromatography–mass spectrometry. Estrogenic activity (E 2 Eq) was measured by in vitro bioassay. We detected estrone E 1 (0.05–6.97 ng L −1 ) and estriol E 3 (below detection-8.13 ng L −1 ) and one conjugated estrogen (estrone-3-sulfate E 1 -3S; below detection-8.13 ng L −1 ). E 1 was widely distributed and positively correlated with E 2 Eq, water temperature, and dissolved organic carbon (DOC). Among nonpoint sources, E 2 Eq, and concentrations of E 1 , soluble reactive phosphorus (SRP) and total dissolved nitrogen (TDN) decreased by 51–61%, 77–82%, 62–64%, 4–16% in restored urban and agricultural streams with best management practices (BMPs) relative to unrestored streams without BMPs. In a wastewater treatment plant (Blue Plains WWTP), >94% of E 1 , E 1 -3S, E 3 , E 2 Eq and TDN were removed while SRP increased by 305% during nitrification/denitrification as a part of advanced wastewater treatment. Consequently, E 1 and TDN concentrations in WWTP effluents were comparable or even lower than those observed in the receiving stream or river waters, and the effects of wastewater discharges on downstream E 1 and TDN concentrations were minor. Highest E 2 Eq value and concentrations of E1, E3, and TDN were detected in combined sewer overflow (CSO). This study suggests that WWTP upgrades with biological nutrient removal, CSO management, and certain agricultural and urban BMPs for nutrient controls have the potential to remove estrogens from point and nonpoint sources along with other contaminants in streams and rivers.

Maryland

Uranium quantification in semen by inductively coupled plasma mass spectrometry

In this study we report uranium analysis for human semen samples. Uranium quantification was performed by inductively coupled plasma mass spectrometry. No additives, such as chymotrypsin or bovine serum albumin, were used for semen liquefaction, as they showed significant uranium content. For method validation we spiked 2 g aliquots of pooled control semen at three different levels of uranium: low at 5 pg/g, medium at 50 pg/g, and high at 1000 pg/g. The detection limit was determined to be 0.8 pg/g uranium in human semen. The data reproduced within 1.4–7% RSD and spike recoveries were 97–100%. The uranium level of the unspiked, pooled control semen was 2.9 pg/g of semen ( n = 10). In addition six semen samples from a cohort of Veterans exposed to depleted uranium (DU) in the 1991 Gulf War were analyzed with no knowledge of their exposure history. Uranium levels in the Veterans’ semen samples ranged from undetectable (<0.8 pg/g) to 3350 pg/g. This wide concentration range for uranium in semen is consistent with known differences in current DU body burdens in these individuals, some of whom have retained embedded DU fragments.

Journal of Trace Elements in Medicine and Biology

Lipidomics reveals specific lipid molecules associated with cold stress syndrome in the Florida manatee (Trichechus manatus latirostris)

Cold stress syndrome (CSS) in the Florida manatee ( Trichechus manatus latirostris) results in perturbations to many physiological pathways, often leading to further illness or death. In this study, we applied a non-targeted lipidomics approach with ultra-high performance liquid chromatography and high-resolution tandem mass spectrometry to characterize changes related to CSS in the lipidomic profiles of manatee plasma. Lipidomic analyses were conducted on healthy manatee (control) and cold-exposed manatee plasma samples with varying concentrations of Serum Amyloid A (SAA), an acute-phase protein that is associated with inflammatory disease. Control manatees ( n = 10) were compared to all manatees exposed to cold temperatures ( n = 17), and a subset of those manatees with SAA values > 120 μg/mL ( n = 9). Increased SAA values were associated with higher levels of various acylcarnitine lipids, while several triacylglycerols and oxidized triacylglycerols were significantly lower in manatees with cold exposure. These identified lipids are critical molecules involved in the maintenance of energy homeostasis and could potentially be examined in conjunction with current physical parameters to characterize cold stress. The ability to detect such differences highlights the addition of lipidomics as a valuable tool in understanding cold stress and potentially other illnesses in manatees. Further investigation into the function of the altered lipids could greatly increase our understanding of lipid metabolism in physiologically stressed manatees as well as other marine mammals and inform future management recovery strategies.

Marine Biology

Analysis of iron, zinc, selenium and cadmium in paraffin-embedded prostate tissue specimens using inductively coupled plasma mass-spectrometry

Formalin-fixed paraffin-embedded (FFPE) tissue specimens represent a valuable and abundant resource of pathologic material for various biomedical studies. In the present study, we report the application of high-resolution inductively coupled mass-spectrometry (ICP-MS) for quantification of Fe, Zn, Se and Cd in FFPE prostate tissue. These elements have a possible role in the development of prostate diseases: while Zn and Se are needed for a healthy prostate, Cd shows multiple toxic and carcinogenic effects. Excessive accumulation of Fe induces the production of highly reactive hydroxyl radical species, which may play a role in cancer etiopathogenesis. To assess whether the levels of these metals in the FFPE prostate tissue represent their original content, we compared their levels with those in the fresh tissue (on dry weight basis) in samples obtained from 15 patients. We found that in FFPE tissue, the recoveries of Se, Fe, Cd and Zn were progressively decreased, 97??11% (r=0.88), 82??22% (r=0.86), 59??23% (r=0.69) and 24??11% (r=0.38), respectively. Thus, the use of correction factors, determined as k=0.16 for Se, k=0.20 for Fe, k=0.27 for Cd and k=0.67 for Zn, is required to estimate the retrospective levels of these elements in the parental non-processed fresh (wet) prostate tissue. The technique used in this study enables the analysis of archival FFPE prostate tissue for the concentrations of Fe, Zn, Se and Cd to study association between the levels of these metals and prostate disease. ?? 2008.

Journal of Trace Elements in Medicine and Biology

Network connectivity contributes to native small-bodied fish assemblages in the upper Mississippi River system

Effective management and conservation of fishes requires understanding habitat use across multiple life stages while ensuring necessary habitats are both available and accessible. Tributary habitats may play an important role in recruitment and dispersal of fishes in anthropogenically modified rivers such as the Mississippi and Illinois Rivers of the Midwest U.S.A. Identifying source locations that contribute to recruitment of fish populations can determine the importance of connectivity within river networks and pinpoint critical habitats that sustain fish populations. In the Upper Mississippi River System (UMRS), the environments that fish use in early life stages (i.e., natal origin) can be identified using otolith trace element analysis due to stability and distinctness in water chemistry (strontium: calcium [Sr:Ca]) among water bodies that is reflected in otoliths. Here, we used trace element analysis to determine natal origin of six small-bodied fishes including bullhead minnow ( Pimephales vigilax ), emerald shiner ( Notropis atherinoides ), gizzard shad ( Dorosoma cepedianum ), bluegill ( Lepomis macrochirus ), orangespotted sunfish ( Lepomis humilis ), and yellow perch ( Perca flavescens ) across six reaches of the UMRS (Pools 4, 8, 13, 26, the Open River of the Middle Mississippi River, and the La Grange Pool of the Illinois River). Otolith core Sr:Ca for fishes was quantified using laser ablation inductively coupled plasma mass spectrometry. Using the resulting Sr:Ca chemical signatures of otolith cores, natal origin (tributary, immigrant, or potential resident) was determined for individual fish based on family-specific relationships between otolith and water chemistries. We found that all species originated from tributaries and other reaches (i.e., were immigrants) to varying extents, which acted as evidence for network connectivity. Specifically, tributaries contributed up to 48% of individuals at a given reach. In certain reaches, Pool 26 and the Open River reach, up to 80% of individuals in a species immigrated from another mainstem river reach. Network connectivity was also important in both upstream and downstream directions. Contributions from network connectivity varied among species: bullhead minnow used less whereas orangespotted sunfish used more network connectivity than when all species were combined. Further, the use of network connectivity varied spatially where individuals captured in Pool 8 and the La Grange Pool less often and those from Pool 26 and the Open River more often originated from network connectivity compared to the whole assemblage across reaches. These results indicate that species' life history traits probably interacted with the physical environment, which differs spatially, to yield observed recruitment source patterns. Our results show that network connectivity contributes to established assemblages of native small-bodied fishes throughout the UMRS and underscore the role of interjurisdictional management in maintaining network connectivity to sustain fish populations.

Illinois, Iowa, Minnesota, Missouri, Wisconsin