National Assessment of Long-Term Groundwater Response to Pesticide Regulation.

Quantitative assessments of long-term, national-scale responses of groundwater quality to pesticide applications are essential to evaluate the effectiveness of pesticide regulations. Retardation time in the unsaturated zone (Ru) was estimated for selected herbicides (atrazine, simazine, and bentazon) and degradation products (desethylatrazine (DEA), desisopropylatrazine (DIA), desethyldesisopropylatrazine (DEIA), and BAM) using a multidecadal time series of groundwater solute chemistry (∼30 years) and herbicide sales (∼60 years). The sampling year was converted to recharge year using groundwater age. Then, Ru was estimated using a cross-correlation analysis of the sales and the frequencies of detection and exceedance of the drinking water standard (0.1 µg/L) of each selected compound. The results showed no retardation of the highly polar, thus mobile, parent compounds (i.e., bentazon), while Ru of the moderately polar compounds (i.e., simazine) was about a decade, and their degradation products showed even longer Ru. The temporal trends of the degradation products did not mirror those of the sale data, which were attributed to the various sale periods of the parent compounds, sorption of the parent compounds, and complex degradation pathways. The longer Ru in clayey/organic sediments than in sandy sediments further confirmed the role of soil-specific retardation as an important factor to consider in groundwater protection.

Leaching of 1,2,4-triazole from commercial barley seeds coated with tebuconazole and prothioconazole.

Seed dressing with fungicide or insecticide is a standard procedure for growing major crops, but very little is known about the leaching risk and the general fate of pesticides from coated seeds. Triazole fungicides are commonly used seed dressing fungicides and recently, there has been increasing concern that 1,2,4-triazole, a major degradation product of several triazole fungicides, may leach to groundwater in concentrations exceeding the 0.1 µg/L threshold limit of the European Union. We therefore carried out a laboratory column experiment with commercial barley seeds coated with the triazole fungicides tebuconazole and prothioconazole to study the fate of the fungicides and their degradation products, especially 1,2,4-triazole. Our experiment showed that the fungicides themselves were relatively immobile in the soil columns, but also that leaching of 1,2,4-triazole will occur no matter if tebuconazole or prothioconazole is used as seed dressing. Relatively high 1,2,4-triazole concentrations (up to 0.8 µg/L) were measured in the column leachates, but when the experiment was terminated after 63 days, a total of only 1 % of the fungicides was recovered as 1,2,4-triazole in the leachate. Our results suggest that seed dressing pesticides should be considered together with spray applications when estimating the total 1,2,4-triazole load from agriculture and that seed dressing pesticides and their degradation products should be included when evaluating leaching risks from pesticide applications in agriculture.

Super-complex mixtures of aliphatic- and aromatic acids may be common degradation products after marine oil spills: A lab-study of microbial oil degradation in a warm, pre-exposed marine environment.

When assessing oil spills in marine environments, focus has often been on describing degradation and removal of hydrocarbons. However, more and more attention is now given to the formation of mineral oil transformation products, and their potential toxicity and persistency in the environment. The aim of this study was to investigate the formation of dissolved acidic degradation products from crude oil in sea water from the Persian Gulf in a lab-experiment. A super-complex mixture of acidic degradation products was formed in the water phase and compound groups of aliphatic acids, monocyclic aromatic acids-, and polycyclic aromatic acids were identified. More specifically, alkylated PAHs were biodegraded to a high number of aromatic, carboxylic acids by hydroxylation of the alkyl side chains. These degradation products are more bioavailable than their parent compounds, and may therefore constitute a new group of contaminants that should be considered in oil spill assessments.

Full-scale bioremediation of diesel-polluted soil in an Arctic landfarm.

A full-scale, experimental landfarm was tested for the capacity to biodegrade oil-polluted soil under high-Arctic tundra conditions in northeast Greenland at the military outpost 9117 Station Mestersvig. Soil contaminated with Arctic diesel was transferred to the landfarm in August 2012 followed by yearly addition of fertilizer and plowing and irrigation to optimize microbial diesel biodegradation. Biodegradation was determined from changes in total petroleum hydrocarbons (TPH), enumeration of specific subpopulations of oil-degrading microorganisms (MPN), and changes in selected classes of alkylated isomers and isomer ratios. Sixty-four percent of the diesel was removed in the landfarm within the first year, but a recalcitrant fraction (18%) remained after five years. n-alkanes and naphthalenes were biodegraded as demonstrated by changing isomer ratios. Dibenzothiophenes and phenanthrenes showed almost constant isomer ratios indicating that their removal was mostly abiotic. Oil-degrading microorganisms were present for the major components of diesel (n-alkanes, alkylbenzenes and alkylnaphthalenes). The degraders showed very large population increases in the landfarm with a peak population of 1.2 × 109 cells g-1 of total diesel degraders. Some diesel compounds such as cycloalkanes, hydroxy-PAHs and sulfur-heterocycles had very few or no specific degraders, these compounds may consequently be degraded only by slow co-metabolic processes or not at all.

Test of vegetation-based surface exploration for detection of Arctic mineralizations: The deep buried Kangerluarsuk Zn-Pb-Ag anomaly.

The aim of our study was to test whether surficial geochemical techniques are applicable under arctic conditions where pedogenesis is slow or absent, and where the vegetation is arctic dwarf shrub tundra. To this end, we sampled vegetation and topsoil at a known Zn-Pb-Ag anomaly at Kangerluarsuk, northwest Greenland. This Zn-Pb-Ag mineralization surfaces in part of the test area and is deeply buried in other parts. The surface mineralization could readily be identified by element analysis of the omnipresent plant Salix glauca. The strongest signal came from the pathfinder element Tl. The target elements Pb and Ag gave only weak signals and Zn gave no signal, probably because the cellular concentration of these elements is actively regulated by the plant. The use of regulated plant micronutrients as reference elements gave a small reduction of analytical noise in Tl/Cu and Tl/B concentration ratios at low Tl concentrations which improved identification of the deep mineralization. Pathfinder elements in plants may thus prove useful when combined with a detailed geophysical model. Tl, Zn, Pb and Ag concentrations in topsoil identified the surface mineralization but failed to identify the deep mineralization. This difference between samples of S. glauca and topsoil is probably because target elements from the deep mineralization must be mobile to reach the surface. Mobile elements may be more accessible for ion-exchange and uptake into the plants compared to the recalcitrant and crystalline fraction in the topsoil.

Fluispotter, a novel automated and wearable device for accurate volume serial dried blood spot sampling.

Aim: A novel automated serial dried blood spot (DBS) sampler, 'Fluispotter', was tested for its sampling performance. Materials & methods: An LC-MS/MS method was developed for the analysis of cortisol in DBS samples serially spotted by Fluispotter. The cortisol concentrations in 148 paired DBS and plasma samples were compared across a hematocrit (HCT) range of 22-55%. Results: The interassay accuracy and precision were <10%. Overall assay bias was negligible across the HCTs tested when analyzing the whole-spot DBS samples. The accuracy and precision of the blood volume in 10 µl DBS samples spotted by Fluispotters and micropipettes were within 3%. Deming regression and Bland-Altman analysis showed a good agreement of DBS-predicted and measured plasma cortisol. Conclusion: The Fluispotter performed serial sampling with high accuracy and precision of the sample blood volume.

Soil Domain and Liquid Manure Affect Pesticide Sorption in Macroporous Clay Till.

Pesticides frequently leach through clayey tills, even when they are expected to be strongly adsorbed. In this study, we observed that sorption of two strongly sorbing pesticides, tebuconazole and glyphosate, varied by more than an order of magnitude across soil domains in 5-m-deep clay till profiles with biopores and fractures. Eight soil domains were identified in each of the profiles: five matrix soils and three in the macropores. Tebuconazole sorption was controlled by soil organic matter content, except in the reduced matrix, which was low in organic matter, where there was surprisingly high sorption. Glyphosate showed high variation in sorption between fractures and matrix soil from the same depths. The domain-specific sorption of both tebuconazole and glyphosate was, however, overruled by dilute liquid manure. Sorption of tebuconazole was, as expected, decreased by liquid manure in several domains, but tebuconazole sorption increased in a few domains due to sorption of the manure-derived organic matter itself. Liquid manure unexpectedly had a greater effect on glyphosate sorption, which was strongly decreased by dissolved organic matter and phosphate in the manure. The variation in sorption across domains, as well as the effects of liquid manure, should be taken into account when assessing leaching risks.

Arctic and Subarctic Natural Soils Emit Chloroform and Brominated Analogues by Alkaline Hydrolysis of Trihaloacetyl Compounds.

There has been increasing recognition of the occurrence of natural, halogenated organic compounds in marine and terrestrial environments. Chloroform is an example of a halogenated organic compound with natural formation as its primary source. Chloroform emission from soil has been reported from diverse Arctic, temperate, and (sub)tropical ecosystems. The terrestrial environment is a significant source to the atmosphere, but little is known about the formation pathway of chloroform in soil. Here, we present evidence that chloroform is formed through the hydrolysis of trichloroacetyl compounds in natural, organic-rich soils. In situ emissions of chloroform from soil in nine Arctic and subarctic ecosystems were linked to soil trichloroacetyl turnover. The residence time from formation of the trichloroacetyl compounds in soil to the release of chloroform to the atmosphere varied between 1 and 116 active months in unfrozen topsoil, depending on soil pH. Nonspecific halogenation that leads to trihaloacetyl formation does not discriminate between chloride and bromide, and brominated analogues were formed alongside chloroform. Soil may therefore be a previously unrecognized, natural source of brominated haloforms. The formation pathway of haloforms through trihaloacetyl compounds can most likely be extended to other ecosystems with organic topsoils.

Biodegradation of crude oil in Arctic subsurface water from the Disko Bay (Greenland) is limited.

Biological degradation is the main process for oil degradation in a subsurface oil plume. There is, however, little information on the biodegradation potential of Arctic, marine subsurface environments. We therefore investigated oil biodegradation in microcosms at 2 °C containing Arctic subsurface seawater from the Disko Bay (Greenland) and crude oil at three concentrations of 2.5-10 mg/L. Within 71 days, the total petroleum hydrocarbon concentration decreased only by 18 ± 18% for an initial concentration of 5 mg/L. The saturated alkanes nC13-nC30 and the isoprenoids iC18-iC21 were biodegraded at all concentrations indicating a substantial potential for biodegradation of these compound classes. Polycyclic aromatic compounds (PACs) disappeared from the oil phase, but dissolution was the main process of removal. Analysis of diagnostic ratios indicated almost no PAC biodegradation except for the C1-naphthalenes. To conclude, the marine subsurface microorganisms from the Disko Bay had the potential for biodegradation of n-alkanes and isoprenoids while the metabolically complex and toxic PACs and their alkylated homologs remained almost unchanged.

Role of Conserved Disulfide Bridges and Aromatic Residues in Extracellular Loop 2 of Chemokine Receptor CCR8 for Chemokine and Small Molecule Binding.

Chemokine receptors play important roles in the immune system and are linked to several human diseases. The initial contact of chemokines with their receptors depends on highly specified extracellular receptor features. Here we investigate the importance of conserved extracellular disulfide bridges and aromatic residues in extracellular loop 2 (ECL-2) for ligand binding and activation in the chemokine receptor CCR8. We used inositol 1,4,5-trisphosphate accumulation and radioligand binding experiments to determine the impact of receptor mutagenesis on both chemokine and small molecule agonist and antagonist binding and action in CCR8. We find that the seven-transmembrane (TM) receptor conserved disulfide bridge (7TM bridge) linking transmembrane helix III (TMIII) and ECL-2 is crucial for chemokine and small molecule action, whereas the chemokine receptor conserved disulfide bridge between the N terminus and TMVII is needed only for chemokines. Furthermore, we find that two distinct aromatic residues in ECL-2, Tyr(184) (Cys + 1) and Tyr(187) (Cys + 4), are crucial for binding of the CC chemokines CCL1 (agonist) and MC148 (antagonist), respectively, but not for small molecule binding. Finally, using in silico modeling, we predict an aromatic cluster of interaction partners for Tyr(187) in TMIV (Phe(171)) and TMV (Trp(194)). We show in vitro that these residues are crucial for the binding and action of MC148, thus supporting their participation in an aromatic cluster with Tyr(187) This aromatic cluster appears to be present in a large number of CC chemokine receptors and thereby could play a more general role to be exploited in future drug development targeting these receptors.

Marine biodegradation of crude oil in temperate and Arctic water samples.

Despite increased interest in marine oil exploration in the Arctic, little is known about the fate of Arctic offshore oil pollution. Therefore, in the present study, we examine the oil degradation potential for an Arctic site (Disko Bay, Greenland) and discuss this in relation to a temperate site (North Sea, Denmark). Biodegradation was assessed following exposure to Oseberg Blend crude oil (100 mg L(-1)) in microcosms. Changes in oil hydrocarbon fingerprints of polycyclic aromatic hydrocarbons (PAHs), alkyl-substituted PAHs, dibenzothiophenes, n-alkanes and alkyltoluenes were measured by gas chromatography-mass spectrometry (GC-MS). In the Disko Bay sample, the degradation order was n-alkanes>alkyltoluenes (para->meta->ortho-isomers)>PAHs and dibenzothiophenes, whereas, the degradation order in the North Sea samples was PAHs and dibenzothiophenes>alkyltoluenes>n-alkanes. These differences in degradation patterns significantly affect the environmental risk of oil spills and emphasise the need to consider the specific environmental conditions when conducting risk assessments of Arctic oil pollution.

Mining the granule proteome: a potential source of endocrine biomarkers.

Proteomics of secretory granules is an emerging strategy for identifying secreted proteins, including potentially novel candidate biomarkers and peptide hormones. In addition, proteomics can provide information about the abundance, localization and structure (post-translational modification) of granule proteins and peptides. Analytical strategies within this research line include so-called 'subtractive proteomics', 'peptidomics' and granule purification by the use of multiple gradient centrifugations. Here we review the literature, and describe the challenges and opportunities in proteomics of secretory granules.

Cardiomyocyte expression and cell-specific processing of procholecystokinin.

Heart muscle cells produce peptide hormones such as natriuretic peptides. Developing hearts also express the gene for the classic intestinal hormone cholecystokinin (CCK) in amounts similar to those in the intestine and brain. However, cardiac expression of peptides other than natriuretic peptides has only been suggested using transcriptional measures or methods, with the post-translational phase of gene expression unaddressed. In this study, we examined the cardiac expression of the CCK gene in adult mammals and its expression at the protein level. Using quantitative PCR, a library of sequence-specific pro-CCK assays, peptide purification, and mass spectrometry, we demonstrate that the mammalian heart expresses pro-CCK in amounts comparable to natriuretic prohormones and processes it to a unique, triple-sulfated, and N-terminally truncated product distinct from intestinal and cerebral CCK peptides. Isoprenaline rapidly stimulated cardiac CCK gene expression in vitro and in vivo, which suggests that the cardiac-specific truncated pro-CCK may have pathophysiological relevance as a new marker of heart failure. The suggestion is confirmed by measurement of plasma from heart failure patients.

The potential for bioaugmentation of sand filter materials from waterworks using bacterial cultures degrading 4-chloro-2-methylphenoxyacetic acid.

BACKGROUND: The herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) is found frequently in Danish groundwater in concentrations exceeding the EU threshold limit of 0.1 µg L(-1) . Groundwater is used for drinking water, and one potential remediation strategy is bioaugmentation using inoculation of sand filters at affected waterworks with degrader bacteria. Numerous bacteria degrading phenoxyacetic acid herbicide have previously been isolated, and they may be candidates for bioaugmentation processes. Designing the optimum inoculum, however, requires knowledge of the capacity for degrading realistically low herbicide concentrations and the robustness of the bacteria when inoculated into sand filter materials. RESULTS: Testing a range of different MCPA-mineralising bacterial combinations, using a high-throughput microplate radiorespirometric mineralisation assay, highlighted three efficient cocultures for mineralising low MCPA concentrations. Cocultures demonstrating a shorter time delay before initiation of (14) C-ring-labelled MCPA mineralisation to (14) CO2 , and a more extensive mineralisation of MCPA, compared with those of single strains, were found. When inoculated into different sand filter materials, the coculture effect was diminished, but several single strains enhanced MCPA mineralisation significantly at low MCPA concentrations. CONCLUSION: This study shows that an increase in the potential for mineralisation of low herbicide concentrations in sand filter materials can be achieved by inoculating with bacterial degrader cultures. © 2014 Society of Chemical Industry.

Polar metabolites of polycyclic aromatic compounds from fungi are potential soil and groundwater contaminants.

This study investigated the sorption to soil of water-soluble metabolites from polycyclic aromatic compounds (PACs). The soil fungus Cunninghamella elegans was used to produce PAC metabolites from two un-substituted PACs (phenanthrene, pyrene), three alkyl-substituted PACs (2-methylnaphthalene, 1-methylphenanthrene, 1-methylpyrene), and one sulfur-containing heterocyclic PAC (dibenzothiophene). Fifty-eight metabolites were tentatively identified; metabolites from the un-substituted PACs were hydroxylated and sulfate conjugated, whereas metabolites from alkyl-substituted PACs were sulfate conjugated and either hydroxylated or oxidized to carboxylic acids at the methyl group. The metabolism of the sulfur-containing heterocyclic PAC resulted in sulfate conjugates. The sorption of the PAC metabolites to three soils was determined using a batch equilibrium method, and partition coefficients (Kd's) were calculated for fourteen representative metabolites. Sulfate conjugated metabolites displayed Kd's below 70 whereas the metabolites with both a sulfate and a carboxylic acid group had Kd's below 2.8. The low Kd's of water-soluble PAC metabolites indicate high mobility in soil and a potential for leaching to surface- and groundwaters.

Does microbial centimeter-scale heterogeneity impact MCPA degradation in and leaching from a loamy agricultural soil?

The potential for pesticide degradation varies greatly at the centimeter-scale in agricultural soil. Three dimensional numerical simulations were conducted to evaluate how such small-scale spatial heterogeneity may affect the leaching of the biodegradable pesticide 2-methyl-4-chlorophenoxyacetic acid (MCPA) in the upper meter of a variably-saturated, loamy soil profile. To incorporate realistic spatial variation in degradation potential, we used data from a site where 420 mineralization curves over 5 depths have been measured. Monod kinetics was fitted to the individual curves to derive initial degrader biomass values, which were incorporated in a reactive transport model to simulate heterogeneous biodegradation. Six scenarios were set up using COMSOL Multiphysics to evaluate the difference between models having different degrader biomass distributions (homogeneous, heterogeneous, or no biomass) and either matrix flow or preferential flow through a soil matrix with a wormhole. MCPA leached, within 250 days, below 1m only when degrader biomass was absent and preferential flow occurred. Both biodegradation in the plow layer and the microbially active lining of the wormhole contributed to reducing MCPA-leaching below 1m. The spatial distribution of initial degrader biomass within each soil matrix layer, however, had little effect on the overall MCPA-leaching.

Quantification of small-scale variation in the size and composition of phenanthrene-degrader populations and PAH contaminants in traffic-impacted topsoil.

Small-scale colocalisation of microbial polycyclic aromatic hydrocarbon (PAH) degraders and PAHs in contaminated soil is a prerequisite for efficient biodegradation of the PAHs. We therefore tested the hypothesis that phenanthrene-degrading bacteria are colocalised with PAHs at the millimetre-to-centimetre-scale. Microbial populations and PAH concentrations were determined for 40-mg samples from a 112-mm transect of a traffic-impacted topsoil. The spatial distribution of cultivable phenanthrene degraders (0.3 × 10(5) -7.2 × 10(5) cells g(-1) ) mirrored neither the distribution of PAHs, nor the distribution of the total cultivable heterotrophic populations. Quantitative real-time PCR (qPCR) analysis of PAH dioxygenase genes (2 × 10(6) -4 × 10(6) cells g(-1) ) from a second transect showed distributions similar to the cultivable phenanthrene degraders, but at a 20-fold higher level. The omnipresence of high densities of PAH degraders at the millimetre scale indicate that PAH persistence may not be caused by local lack of degrader cells. To the best of our knowledge, this is the first time that either MPN of pollutant degraders, qPCR of functional genes, CFU of heterotrophic micro-organisms, or the content of PAHs have been determined with such high spatial resolution.

The gompertz function can coherently describe microbial mineralization of growth-sustaining pesticides.

Mineralization of (14)C-labeled tracers is a common way of studying the environmental fate of xenobiotics, but it can be difficult to extract relevant kinetic parameters from such experiments since complex kinetic functions or several kinetic functions may be needed to adequately describe large data sets. In this study, we suggest using a two-parameter, sigmoid Gompertz function for parametrizing mineralization curves. The function was applied to a data set of 252 normalized mineralization curves that represented the potential for degradation of the herbicide MCPA in three horizons of an agricultural soil. The Gompertz function fitted most of the normalized curves, and trends in the data set could be visualized by a scatter plot of the two Gompertz parameters (rate constant and time delay). For agricultural topsoil, we also tested the effect of the MCPA concentration on the mineralization kinetics. Reduced initial concentrations lead to shortened lag-phases, probably due to reduced need for bacterial growth. The effect of substrate concentration could be predicted by simply changing the time delay of the Gompertz curves. This delay could to some extent also simulate concentration effects for 2,4-D mineralization in agricultural soil and aquifer sediment and 2,6-dichlorobenzamide mineralization in single-species, mineral medium.