A novel framework for discharge uncertainty quantification applied to 500 UK gauging stations.

A generalized framework for discharge uncertainty estimation is presentedAllows estimation of place-specific discharge uncertainties for many catchmentsLocal conditions dominate in determining discharge uncertainty magnitudes.

New insights in the dihydroxybenzenes-driven Fenton reaction: electrochemical study of interaction between dihydroxybenzenes and Fe(III).

It has been reported that the dihydroxybenzene (DHB) driven Fenton reaction is more efficient to degrade recalcitrant substrates than the simple Fenton reaction. The enhanced reactivity of the DHB driven Fenton reaction is not clear, but it could be explained by the formation of oxidant species different from the ones formed by classical Fenton reaction or by the shift of the redox potential of the complex formed by DHB and Fe(III). The redox reaction between Fe(III) and the DHBs 1,2-dihydroxybenzene (catechol, CAT), 2,3-dihydroxybenzoic acid (2,3-DHBA), 3,4-dihydroxybenzoic acid (3,4-DHBA), and 1,2-dihydroxy-3,5-benzenedisulfonate (TIRON) was studied by cyclic voltammetry to better understand the enhanced reactivity of the DHB driven Fenton reaction. It was determined that the amount of Fe(II) produced by the redox reaction between Fe(III) and DHBs was insufficient to explain the enhanced reactivity. Cyclic voltammograms (CV) of the DHBs/Fe(III) systems show a quasi-reversible or irreversible behavior and also shifting and splitting the anodic peaks. This effect can be related to DHBs oxidation by Fe(III), but not to a real interaction.

Assessing catchment-scale erosion and yields of suspended solids from improved temperate grassland.

This paper quantifies the yields of suspended solids (SS) from a headwater catchment managed as improved temperate grassland, providing the first direct, catchment-scale evidence of the rates of erosion from this land-use in the UK and assessing the threat posed to aquatic ecosystems. High-resolution monitoring of catchment hydrology and the concentrations of SS and volatile organic matter (VOM) were carried out in the first-order channel of the Den Brook headwater catchment in Devon (UK) during the 2006-2007 hydrological season. The widely used 'rating curve' (discharge-concentration) approach was employed to estimate yields of SS, but as demonstrated by previous researchers, this study showed that discharge is a poor predictor of SS concentrations and therefore any yields estimated from this technique are likely to be highly uncertain. Nevertheless, for the purpose of providing estimates of yields that are comparable to previous studies on other land uses/sources, this technique was adopted albeit in an uncertainty-based framework. The findings suggest that contrary to the common perception, grasslands can be erosive landscapes with SS yields from this catchment estimated to be between 0.54 and 1.21 t ha(-1) y(-1). In terms of on-site erosion problems, this rate of erosion does not significantly exceed the commonly used 'tolerable' threshold in the UK ( approximately 1 t ha(-1) y(-1)). In terms of off-site erosion problems, it is argued here that the conventional expression of SS yield as a bulk annual figure has little relevance to the water quality and ecological status of surface waters and therefore an alternative technique (the concentration-frequency curve) is developed within this paper for the specific purpose of assessing the ecological threat posed by the delivery of SS into surface waters. This technique illustrates that concentrations of SS recorded at the catchment outlet frequently exceed the water quality guidelines, such as those of the EU Freshwater Fisheries Directive (78/659/EC), and pose a serious threat to aquatic organisms. It is suggested that failure to recognise improved temperate grasslands as a potential source of particulate material could result in the non-compliance of surface waters to water quality guidelines, deterioration of ecological status and failure of water quality remediation measures.

Aerial strategies advance volcanic gas measurements at inaccessible, strongly degassing volcanoes.

Volcanic emissions are a critical pathway in Earth's carbon cycle. Here, we show that aerial measurements of volcanic gases using unoccupied aerial systems (UAS) transform our ability to measure and monitor plumes remotely and to constrain global volatile fluxes from volcanoes. Combining multi-scale measurements from ground-based remote sensing, long-range aerial sampling, and satellites, we present comprehensive gas fluxes-3760 ± [600, 310] tons day-1 CO2 and 5150 ± [730, 340] tons day-1 SO2-for a strong yet previously uncharacterized volcanic emitter: Manam, Papua New Guinea. The CO2/ST ratio of 1.07 ± 0.06 suggests a modest slab sediment contribution to the sub-arc mantle. We find that aerial strategies reduce uncertainties associated with ground-based remote sensing of SO2 flux and enable near-real-time measurements of plume chemistry and carbon isotope composition. Our data emphasize the need to account for time averaging of temporal variability in volcanic gas emissions in global flux estimates.

Genetic Diversity of Prunus necrotic ringspot virus Isolates Within a Cherry Orchard in New York.

A survey for Prunus necrotic ringspot virus (PNRSV) in an orchard of Prunus cerasus cv. Montmorency and Prunus avium cv. Hedelfingen in New York by enzyme-linked immunosorbent assay indicated an eightfold higher infection rate in sour cherry (33%, 32 of 96) than in sweet cherry (4%, 6 of 136) trees. The presence of PNRSV was confirmed by reverse transcription-polymerase chain reaction and amplification of the coat protein (CP) gene in total RNA from infected leaf tissue. Latent infection was prevalent in the majority of trees infected (87%, 33 of 38), while a few of them exhibited shock symptoms or had severely reduced growth (13%, 5 of 38). Asymptomatic PNRSV-infected trees clustered in spatial proximity to symptomatic trees. Sequence analysis of the CP gene (675 bp) indicated a population structure consisting of one predominant molecular variant for 10 isolates and six minor molecular variants for seven isolates. A high sequence identity was found between the CP gene of PNRSV isolates from cherry trees and other isolates from diverse hosts and various geographic origins at the nucleotide and amino acid levels (88 to 100%). Phylogenetic analyses showed a clustering of PNRSV isolates from cherry trees in New York in the predominant group PV-96.

Determining the sources of nutrient flux to water in headwater catchments: Examining the speciation balance to inform the targeting of mitigation measures.

Diffuse water pollution from agriculture (DWPA) is a major environmental concern, with significant adverse impacts on both human and ecosystem health. However, without an appropriate understanding of the multiple factors impacting on water, mitigation measures cannot be targeted. Therefore, this paper addresses this gap in understanding, reporting the hydrochemical monitoring evidence collected from the UK Government's Demonstration Test Catchments (DTC) programme including contrasting chalk and clay/mudstone catchments. We use data collected at daily and sub-daily frequency over multiple sites to address: (1) How does the behaviour of the full range of nitrogen (N) species and phosphorus (P) fractions vary? (2) How do N species and P fractions vary inter- and intra-annually? (3) What do these data indicate about the primary pollution sources? And (4) which diffuse pollution mitigation measures are appropriate in our study landscapes? Key differences in the rates of flux of nutrients were identified, dependent on catchment characteristics. Full N speciation and P fractionation, together with dissolved organic carbon (DOC) enabled identification of the most likely contributing sources in each catchment. Nitrate (NO3-N) was the dominant N fraction in the chalk whereas organic and particulate N comprised the majority of the load in the clay/mudstone catchments. Despite current legislation, orthophosphate (PO4-P) was not found to be the dominant form of P in any of the catchments monitored. The chalk sub-catchments had the largest proportion of inorganic/dissolved organic P (DOP), accompanied by episodic delivery of particulate P (PP). Contrastingly, the clay/mudstone sub-catchments loads were dominated by PP and DOP. Thus, our results show that by monitoring both the inorganic and organic fractions a more complete picture of catchment nutrient fluxes can be determined, and sources of pollution pin-pointed. Ultimately, policy and management to bring nutrient impacts under control will only be successful if a multi-stressor approach is adopted.

Rethinking the contribution of drained and undrained grasslands to sediment-related water quality problems.

Grass vegetation has been recommended for use in the prevention and control of soil erosion because of its dense sward characteristics and stabilizing effect on the soil. A general assumption is that grassland environments suffer from minimal soil erosion and therefore present little threat to the water quality of surface waters in terms of sediment and sorbed contaminant pollution. Our data question this assumption, reporting results from one hydrological year of observations on a field-experiment monitoring overland flow, drain flow, fluxes of suspended solids, total phosphorus (TP), and molybdate-reactive phosphorus (<0.45 mum) in response to natural rainfall events. During individual rainfall events, 1-ha grassland lysimeters yield up to 15 kg of suspended solids, with concentrations in runoff waters of up to 400 mg L(-1). These concentrations exceed the water quality standards recommended by the European Freshwater Fisheries Directive (25 mg L(-1)) and the USEPA (80 mg L(-1)) and are beyond those reported to have caused chronic effects on freshwater aquatic organisms. Furthermore, TP concentrations in runoff waters from these field lysimeters exceeded 800 mug L(-1). These concentrations are in excess of those reported to cause eutrophication problems in rivers and lakes and contravene the ecoregional nutrient criteria in all of the USA ecoregions. This paper also examines how subsurface drainage, a common agricultural practice in intensively managed grasslands, influences the hydrology and export of sediment and nutrients from grasslands. This dataset suggests that we need to rethink the conceptual understanding of grasslands as non-erosive landscapes. Failure to acknowledge this will result in the noncompliance of surface waters to water quality standards.

Photocatalyzed degradation of flumequine by doped TiO2 and simulated solar light.

Titanium dioxide was obtained in its pure form (TiO2) and in the presence of urea (u-TiO2) and thiourea (t-TiO2) using the sol-gel technique. The obtained powders were characterized by BET surface area analysis, Infrared Spectroscopy, Diffuse Reflectance Spectroscopy and the Rietveld refinement of XRD measurements. All the prepared catalysts show high anatase content (>99%). The a and b-cell parameters of anatase increase in the order TiO2u-TiO2>TiO2. The photocatalytic activities of the samples were determined on flumequine under solar-simulated irradiation. The most active catalysts were u-TiO2 and t-TiO2, reaching values over 90% of flumequine degradation after 15 min irradiation, compared with values of 55% for the pure TiO2 catalyst. Changing simultaneously the catalyst amount (t-TiO2) and pH, multivariate analysis using the response surface methodology was used to determine the roughly optimal conditions for flumequine degradation. The optimized conditions found were pH below 7 and a catalyst amount of 1.6 g L(-1).

Coupling of photocatalytic and biological reactors to remove EDTA-Fe from aqueous solution.

Degradation of 2.5 mM EDTA-Fe solution was performed in a coupled photocatalytic-biological reactor. The system consists of a photochemical annular reactor filled with TiO2 immobilized on glass Raschig rings coupled with an activated sludge continuous reactor. Around 50% of EDTA degradation was reached after 150 min irradiation. Simultaneously a four-fold increase in biodegradability, measured as BOD5/COD ratio, was observed. The activated sludge is not capable to degrade the complex EDTA-Fe but it removed partially the COD and efficiently the BOD5 of the photochemically treated solution.

Major agricultural changes required to mitigate phosphorus losses under climate change.

Phosphorus losses from land to water will be impacted by climate change and land management for food production, with detrimental impacts on aquatic ecosystems. Here we use a unique combination of methods to evaluate the impact of projected climate change on future phosphorus transfers, and to assess what scale of agricultural change would be needed to mitigate these transfers. We combine novel high-frequency phosphorus flux data from three representative catchments across the UK, a new high-spatial resolution climate model, uncertainty estimates from an ensemble of future climate simulations, two phosphorus transfer models of contrasting complexity and a simplified representation of the potential intensification of agriculture based on expert elicitation from land managers. We show that the effect of climate change on average winter phosphorus loads (predicted increase up to 30% by 2050s) will be limited only by large-scale agricultural changes (e.g., 20-80% reduction in phosphorus inputs).The impact of climate change on phosphorus (P) loss from land to water is unclear. Here, the authors use P flux data, climate simulations and P transfer models to show that only large scale agricultural change will limit the effect of climate change on average winter P loads in three catchments across the UK.

Using hysteresis analysis of high-resolution water quality monitoring data, including uncertainty, to infer controls on nutrient and sediment transfer in catchments.

A large proportion of nutrients and sediment is mobilised in catchments during storm events. Therefore understanding a catchment's hydrological behaviour during storms and how this acts to mobilise and transport nutrients and sediment to nearby watercourses is extremely important for effective catchment management. The expansion of available in-situ sensors is allowing a wider range of water quality parameters to be monitored and at higher temporal resolution, meaning that the investigation of hydrochemical behaviours during storms is increasingly feasible. Studying the relationship between discharge and water quality parameters in storm events can provide a valuable research tool to infer the likely source areas and flow pathways contributing to nutrient and sediment transport. Therefore, this paper uses 2 years of high temporal resolution (15/30 min) discharge and water quality (nitrate-N, total phosphorus (TP) and turbidity) data to examine hysteretic behaviour during storm events in two contrasting catchments, in the Hampshire Avon catchment, UK. This paper provides one of the first examples of a study which comprehensively examines storm behaviours for up to 76 storm events and three water quality parameters. It also examines the observational uncertainties using a non-parametric approach. A range of metrics was used, such as loop direction, loop area and a hysteresis index (HI) to characterise and quantify the storm behaviour. With two years of high resolution information it was possible to see how transport mechanisms varied between parameters and through time. This study has also clearly shown the different transport regimes operating between a groundwater dominated chalk catchment versus a surface-water dominated clay catchment. This information, set within an uncertainty framework, means that confidence can be derived that the patterns and relationships thus identified are statistically robust. These insights can thus be used to provide information regarding transport processes and biogeochemical processing within river catchments.

Interaction of Clostridium perfringens theta-haemolysin, a contaminant of commercial phospholipase C, with erythrocyte ghost membranes and lipid dispersions. A morphological study.

Commercially available preparations of phospholipase C from Clostridium perfringens are commonly contaminated with theta haemolysin, one of a group of bacterial haemolysins called oxygen labile (O-labile) haemolysins. Treatment of erythrocyte ghosts and a mixed lipid dispersion containing cholesterol with commercially available phospholipase C in the absence of Ca-2+ and the presence of phosphate buffer and/or EDTA resulted in the formation and release of ring or arc-shaped structures. Highly purified phospholipase C, free of theta-haemolysin, produced no changes in the morphology of erythrocyte ghosts or lipid dispersions in the presence of phosphate or EDTA, but caused the formation of typical diglyceride droplets in the presence of Ca-2+ in the absence of these inhibitors. Ring structures, identical to those caused by commercial phospholipase C, were formed on addition of highly purified theta-haemolysin to erythrocyte ghost membranes, lipid dispersions containing cholesterol and cholesterol dispersions, but not on treatment of membranes from Micrococcus lysodeikticus. Heat-inactivated O-haemolysin (60 degrees C for 10 min) produced no such effects. The dimensions of rings and arcs displayed heterogeneity. The outside diameters in various preparations varied from approx. 27-58 nm with border thickness of 4.1-7.8 nm.

Analysis of enoxaparin dose titration at a large, tertiary teaching facility.

Therapeutic drug monitoring of enoxaparin with antifactor Xa levels (AXALs) is recommended in some populations; however, the approach to dose titration is poorly described. Our study at a large, tertiary teaching facility examined the dose response to titration of enoxaparin based on AXAL. Patients from 2008 to 2012 receiving enoxaparin were included, provided 2 or more steady state AXAL were obtained within 30 days and that the enoxaparin was prescribed for treatment rather than prophylaxis. The primary outcome was the percentage of dose change required to obtain goal range AXAL following dose titration. Eighty-seven patients were available for analysis with the following key characteristics: renal dysfunction during treatment 72%, obesity 8%, and solid organ transplant 26%. Initial goal AXAL was attained in 27 (31%) patients, and ultimately 54 (62%) patients achieved goal AXAL. Of the 31 patients who had initial AXAL above goal, 13 (42%) patients reached goal with a median dose decrease of 24%. In the 29 patients who had an initial AXAL below goal, 11 (38%) achieved therapeutic AXAL with a median dose increase of 16%. The AXAL monitoring can guide enoxaparin titration with subtherapeutic or supratherapeutic AXAL and an increase or decrease of roughly 20% is suggested as an initial change.

Evaluation of different methods for the detection of outer membrane proteins and lipopolysaccharides of Pasteurella haemolytica by immunoblotting.

The optimal conditions for the detection of outer membrane proteins (OMPs) and lipopolysaccharide (LPS) of Pasteurella haemolytica by immunoblotting were evaluated. The variables examined included the equilibration time of the gels before transfer, composition of the transfer buffer, type of blotting membrane, blocking agent, effect of the zwitterionic detergent Empigen BB on protein renaturation, and the development reagent. The composition of the transfer buffer and time of gel equilibration significantly affected the efficiency of transfer of both OMPs and LPS. However, the optimal conditions for the transfer of OMPs were not the same as those for LPS. Thus, optimal transfer of OMPs occurred in Tris-glycine buffer, with prior equilibration of the gels to allow for expansion, whereas optimal transfer of LPS was achieved in Tris-glycine-methanol buffer with no equilibration of the gels. In Tris-glycine-methanol buffer, gel equilibration resulted in a significantly reduced transfer of both OMPs and LPS, probably due to the removal of SDS from these components. The use of Zeta-Probe blotting membrane which, unlike nitrocellulose, does not require methanol for optimal protein binding, did not result in improved binding of OMPs or LPS in the absence of methanol and, even after prolonged blocking (> 2 h), gave higher background staining than did nitrocellulose. Effective blocking of nitrocellulose was achieved with 3% (w/v) gelatin, 2.5% (w/v) skimmed milk or 0.3% (v/v) Tween 20, whereas increased background staining occurred with 1% (w/v) bovine serum albumin or 1% (w/v) ovalbumin. The incorporation of Empigen BB in the primary antibody buffer did not improve antibody recognition of proteins as a result of their renaturation. For the horseradish-peroxidase enzyme development system, the substrate 3,3'-diaminobenzidine tetrahydrochloride was more sensitive, and developed more quickly, than 4-chloro-1-naphthol, but faded more rapidly after drying of the membrane. 4-chloro-1-naphthol was more suitable for identifying OMPs because less background staining occurred, whereas 3,3'-diaminobenzidine tetrahydrochloride was more suitable for the detection of LPS due to its greater sensitivity.

Kidney function during naproxen therapy in patients at risk for renal insufficiency.

The effects of naproxen on renal function in 34 patients with minimally elevated serum creatinine (Scr) or subnormal creatinine clearance (Ccr) were evaluated in a parallel-design study. All patients received open-label naproxen 375 mg twice daily for 2 weeks (phase I); patients were then randomly assigned to receive naproxen 750 mg twice daily (n = 26) or to continue naproxen 375 mg twice daily (n = 8) double-blind for an additional 2 weeks (phase II). Renal function was assessed by Scr, Ccr, and BUN measurements at baseline and at the end of each treatment phase. Neither treatment group had a clinically meaningful change in median laboratory values between baseline and the end of phase I, or between baseline and the end of phase II. During the first 2 weeks of treatment with naproxen 375 mg twice daily, there was no change in Scr. At the time of the first Scr measurement following the increase in naproxen dose to 750 mg twice daily, 13 of 26 patients had Scr levels of 1.1 mg/dL or higher, but four days later, only three patients had Scr levels of 1.1 mg/dL or higher, suggesting that a transient increase in Scr may accompany dosage increase. Chronic administration of naproxen 375 mg twice daily in patients at risk for renal insufficiency based on laboratory evidence of renal impairment was not associated with further deterioration in renal function. An increase in dosage to 750 mg twice daily in such patients appeared to be associated with only small, transient changes in laboratory measures of renal function.

Optimal conditions for the analysis of Pasteurella haemolytica lipopolysaccharide by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.

The optimal conditions for the analysis of the lipopolysaccharide (LPS) of two serotype A1 isolates and a serotype A2 isolate of Pasteurella haemolytica by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining were determined. The LPS of the A1 isolates possessed O side chains, consisting of high molecular mass bands with the appearance of a ladder-like pattern, as well as a low molecular mass core-oligosaccharide region; the LPS of the A2 isolate consisted only of the core-oligosaccharide region. Furthermore, the LPS of the two A1 isolates differed in the core-oligosaccharide region. Optimal resolution of low molecular mass LPS components was obtained in a 15% acrylamide resolving gel containing 4 M urea whereas optimal resolution of high molecular mass components was obtained when urea was omitted. Conventional silver staining resulted in excellent visualisation of LPS bands, whereas a modified staining method did not detect additional bands, as has been demonstrated with the LPS of Pseudomonas aeruginosa. Proteinase K digestion of outer membranes gave more clearly defined LPS profiles than did similar digestions of whole cells, and more closely resembled the profiles of purified LPS. With the exception of slight variation in the average molecular mass of a group of O side chains between logarithmic and stationary phases there were no differences in LPS profiles at various stages of the growth cycle; freezing and thawing of LPS samples had no effect on the profiles.

Accumulation of a precursor of subunit S1 of pertussis toxin in cell envelopes of Bordetella pertussis in response to the membrane perturbant phenethyl alcohol.

Exponential cultures of Bordetella pertussis strain 18334 were treated with the membrane-perturbing agent phenethyl alcohol which, at a concentration of 0.075% v/v, blocked the synthesis of mature subunit S1 of pertussis toxin as revealed by Western blotting. It also caused the accumulation of a precursor, pS1, with an estimated mol. wt of 32 X 10(3), that was located in the cytoplasmic membrane. These findings suggested that subunit S1 of pertussis toxin was exported in a signal peptide-dependent manner.

Binding of 125I-alpha toxin of Staphylococcus aureus to erythrocytes.

Alpha toxin purified from Staphylococcus aureus strain Wood 46 and radioiodinated by the lactoperoxidase method retained full haemolytic activity and was used to study factors affecting binding to rabbit and horse erythrocytes. A relatively fixed percentage of added toxin bound to both cell types; the percentage bound was independent of temperature, pH, cell concentration and toxin concentration. Neither a 50-fold excess of native toxin nor Concanavalin A inhibited the binding of iodinated toxin to erythrocytes. The results suggest that differences in the sensitivity of erythrocytes to haemolysis do not reflect the abundance of high affinity toxin receptors on sensitive cells, but are more probably the result of differences in the intrinsic stability of the membrane and its sensitivity to perturbation by amphiphilic agents.

Inhibition of secretion of staphylococcal alpha toxin by cerulenin.

Secretion of alpha toxin by Staphylococcus aureus strain Wood 46 was preferentially inhibited by cerulenin, an antibiotic that stops fatty-acid synthesis by inhibiting beta-keto acyl acyl carrier-protein synthetase. At the concentrations used, cerulenin had a negligible effect on cell growth and total protein synthesis, but reduced lipid synthesis by 50%. Extracellular and membrane-associated alpha toxin was absent in cultures treated with cerulenin, but toxin formation was resumed after either removal of the antibiotic or addition of exogenous fatty acids. The apparent absence of toxin precursor in membranes of inhibited cells favours inhibition at an earlier stage in toxin synthesis.

Assay of Bordetella pertussis heat-labile toxin with human embryonic lung cells.

An assay has been developed for Bordetella pertussis heat-labile toxin (HLT) based on morphological alterations in certain human embryonic lung (HEL) cell lines. Eighteen cell lines from human and other sources were tested but only two, MRC-5 and HELu2, were responsive to HLT. Confluent monolayers of the cells contracted within 24 h of exposure to the toxin, but without loss of viability during incubation for a further 3 days. The effect of HLT was quantitated by scoring the extent of morphological change, and by the decrease in Giemsa staining of the cell monolayers, as measured on an ELISA plate reader. This cell culture assay for HLT was more sensitive than lethality titration in mice but the dose-response curve had a lower slope. The specificity of the response was established by comparing unheated HLT with HLT heated at 56 degrees C, and with extracts from transposon-insertion mutants of B. pertussis which were deficient in HLT. Purified preparations of pertussis toxin and B. pertussis lipopolysaccharide gave no morphological response even at high doses.