Nitrogen concentration affects amphotericin B and fluconazole tolerance of pathogenic cryptococci.

Environmental stress often causes phenotypic changes among pathogenic cryptococci, such as altered antifungal susceptibility, changes in capsule and melanin formation, as well as altered levels of the membrane sterol and antifungal target, ergosterol. We therefore hypothesised that nitrogen limitation, a prevalent environmental stress in the natural habitat of these yeasts, might affect virulence and antifungal susceptibility. We tested the effect of different nitrogen concentrations on capsule, melanin and ergosterol biosynthesis, as well as amphotericin B (AmB) and fluconazole (FLU) susceptibility. This was achieved by culturing cryptococcal strains representing Cryptococcus neoformans and Cryptococcus gattii in media with high (0.53 g/l), control (0.42 g/l) and low (0.21 g/l) NH4Cl concentrations. India ink staining was used to determine capsule thickness microscopically, while melanin and ergosterol content were determined spectrophotometrically. We found that lower nitrogen concentrations enhanced both ergosterol and capsule biosynthesis, while a variable effect was observed on melanisation. Evaluation of drug tolerance using time-kill methodology, as well as tests for FLU heteroresistance, revealed that the low nitrogen cultures had the highest survival percentages in the presence of both AmB and FLU, and showed the highest frequency of FLU heteroresistance, suggesting that nitrogen concentration may indeed influence drug tolerance.

Effects of light, microorganisms, farming chemicals and water content on the degradation of microcystin-LR in agricultural soils.

An experiment was conducted to investigate the effect of farming activities on microcystin-LR (MC-LR) degradation in soils. Three farming activities were assessed: 1) fertilization via addition of different nitrogen sources and organic matter; 2) pesticide application by addition of different commercial pesticides; and 3) irrigation by addition of different amount of water. The contribution of the two major degradation processes of MC-LR in soils, photodegradation and biodegradation, were also evaluated. MC-LR was added into the soil samples to create a concentration of 500 µg kg-1 for each treatment. Results showed that natural degradation of MC-LR in soils was mainly by biodegradation rather than photodegradation. MC-degradation was stimulated by the addition of NaNO3 and humic acid, whereas degradation was inhibited by addition of NH4Cl, glucose, and glycine. Application of high concentrations of glyphosate and chlorothalonil significantly inhibited the degradation of MC-LR in soils and the half-life was almost twice as long as the control. No significant effect was found by addition of CO(NH2)2 and dimethoate. Both low (10%) and high water content (60%) could lead to inhibition of MC-LR degradation. Results from our study help to inform farm practices that could alleviate contamination by MC-LR in agroecosystems.

An infrared spectroscopy method to detect ammonia in gastric juice.

Ammonia in gastric juice is considered a potential biomarker for Helicobacter pylori infection and as a factor contributing to gastric mucosal injury. High ammonia concentrations are also found in patients with chronic renal failure, peptic ulcer disease, and chronic gastritis. Rapid and specific methods for ammonia detection are urgently required by the medical community. Here we present a method to detect ammonia directly in gastric juice based on Fourier transform infrared spectroscopy. The ammonia dissolved in biological liquid samples as ammonium ion was released in air as a gas by the shifting of the pH equilibrium of the ammonium/ammonia reaction and was detected in line by a Fourier transform infrared spectroscopy system equipped with a gas cell for the quantification. The method developed provided high sensitivity and selectivity in ammonia detection both in pure standard solutions and in a simulated gastric juice matrix over the range of diagnostic concentrations tested. Preliminary analyses were also performed on real gastric juice samples from patients with gastric mucosal injury and with symptoms of H. pylori infection, and the results were in agreement with the clinicopathology information. The whole analysis, performed in less than 10 min, can be directly applied on the sample without extraction procedures and it ensures high specificity of detection because of the ammonia fingerprint absorption bands in the infrared spectrum. This method could be easily used with endoscopy instrumentation to provide information in real time and would enable the endoscopist to improve and integrate gastroscopic examinations.

Receptor model source attributions for Utah's Salt Lake City airshed and the impacts of wintertime secondary ammonium nitrate and ammonium chloride aerosol.

UNLABELLED: Communities along Utah's Wasatch Front are currently developing strategies to reduce daily average PM2.5 levels to below National Ambient Air Quality Standards during wintertime persistent stable atmospheric conditions, or cold-air pools. Speciated PM2.5 data from the Wasatch Front airshed indicate that wintertime exceedances of the PM2.5 standard are mainly driven by high levels of ammonium nitrate. Stable wintertime conditions foster the formation of ammonium nitrate aerosol when sufficient sources of NO(x), ammonia, and oxidative capacity exist. However this work demonstrates that secondary ammonium chloride aerosol can also be a significant source of secondary wintertime PM2.5 if sufficient sources of atmospheric chlorine exist. Two factor analysis techniques, positive matrix factorization (PMF) and Unmix, were used to identify contributors to PM2.5 at three monitoring stations along Utah's Wasatch Front: Bountiful, Lindon, and Salt Lake City. The monitoring data included chemically speciated PM2.5 data for 227, 227, and 429 days at each location, respectively, during the period from May 2007 through May 2011. PMF identified 10-12 factors and Unmix identified 4-5 factors for each of the locations. The wintertime PMF and Unmix results showed large contributions from secondary PM2.5 when PM2.5 concentrations exceeded 20 microg/m3. PMF identified both ammonium nitrate and ammonium chloride aerosol as significant secondary contributors to PM2.5 (10-15% of total PM2.5 from ammonium chloride) during wintertime pollution episodes. Subsequent ion balance analysis of the monitoring data confirmed the presence of significant ammonium chloride aerosol on these highly polluted days at all three monitoring sites. The directly emitted primary PM2.5 portions of the source attribution results were further compared to county-level emissions inventories and showed generally good agreement for Salt Lake City and Lindon during wintertime except for wood smoke and fugitive dust, which have higher contributions in the receptor modeling results than in the emissions inventories. IMPLICATIONS: The study suggests that secondary ammonium chloride aerosol can be a significant source ofwintertime PM2.5 in an ammonia-rich environment, like the Wasatch Front airshed, if sufficient sources of atmospheric chlorine exist. During wintertime, cold-air-pool events, the source attribution results generally agree with the county emission inventories with the exception of wood smoke and cooking sources. At the Salt Lake City monitoring station, the estimated contributions from wood smoke and cooking are nearly double those of the corresponding inventory, suggesting that they are nearly as important as gasoline emissions.

Impact of prevalent chlorine quenchers on phenolic disinfection byproducts in drinking water and potential reaction mechanisms.

To prevent the reactions of disinfection byproducts (DBPs) or natural organic matters with residual chlorine in drinking water in the course of the water store, residual chlorine is quenched by chlorine quenchers, while some chlorine quenchers may result in dechlorination of DBPs. Phenolic compounds are a group of highly toxic DBPs compared to regulated aliphatic DBPs (trihalomethanes (THMs) and haloacetic acids (HAAs)), which might be a great threat to drinking water safety. Nevertheless, impact of popular chlorine quenchers on phenolic DBPs is less understanding. In this study, the influences of ammonium chloride, ascorbic acid, sodium thiosulfate, and sodium sulfite on phenolic DBPs are assessed. Total concentration of 19 phenolic DBPs in drinking water from 7 Chinese cities was 145-1821 ng/L, suggesting a widely occurrence of these pollutants. Four assessed chlorine quenchers have not impacts on mass spectra of studied phenolic DBPs. Additionally, when the storage time ≤24 h, recoveries of 19 phenolic DBPs using four assessed chlorine quenchers are within the accept levels (70-130 %). However, when the storage time increased to 168 h, ascorbic acid and sodium thiosulfate satisfied the recovery requirement of phenolic DBPs during the sample analysis, and ammonium chloride and sodium sulfite showed a unacceptable impact on bromo-chloro-phenols. In general, ascorbic acid and sodium thiosulfate are recommended to be the ideal chlorine quenchers of phenolic DBPs. Mechanism study indicated that sodium sulfite induced the dechlorination of 2-chloro-4-bromophenol via nucleophilic reaction. This study is the first attempt to provide the impact of chlorine quenchers on phenolic DBPs and corresponding reaction mechanism.

A single chamber packed bed microbial fuel cell biosensor for measuring organic content of wastewater.

This study reports an investigation of the effect of the anode surface area on the performance of a single chamber microbial fuel cell (SCMFC) based biosensor for measuring the organic content of wastewater. A packed bed of graphite granules was used as the anode. The surface area of the anode was changed by altering the granule bed thickness (0.3 cm and 1 cm). The anode surface area was found to play a role in the dynamic response of the system. For a granule bed thickness of 1 cm and with an external resistance of 500 Omega, the response time (defined as the time required to achieve 95% of the steady-state current) was reduced by approximately 65% in comparison to a SCMFC biosensor with a carbon cloth anode.

The dctA gene of Pseudomonas chlororaphis O6 is under RpoN control and is required for effective root colonization and induction of systemic resistance.

Transcription from the dctA gene, which encodes an organic acid transporter in the root-colonizing bacterium Pseudomonas chlororaphis O6, is under complex regulatory control. Promoter sequence analysis revealed an RpoN binding site. The regulation of transcript accumulation by the level of ammonium ions in the growth medium confirmed RpoN regulation, even in the presence of glucose. A dctA mutant colonized tobacco roots to a lesser extent than the wild-type mutant during early seedling development. Colonization by the dctA mutant, as compared to the wild type, also reduced the level of systemically induced resistance against the soft rot pathogen Erwinia carotovora SCC1. We ascribe this reduced colonization to the inability of the mutant to utilize certain organic acid components in the root exudates. The dctA mutant failed to grow on succinate and fumarate, and showed reduced growth on malate. All altered properties of the mutant were complemented by the full-length dctA gene. We propose that organic acids in root exudates may provide important nutrient sources for the beneficial root-colonizing pseudomonad.

Development of EPA method 535 for the determination of chloroacetanilide and other acetamide herbicide degradates in drinking water by solid-phase extraction and liquid chromatography/tandem mass spectrometry.

U.S. Environmental Protection Agency (EPA) Method 535 has been developed in order to provide a method for the analysis of "Alachlor ESA and other acetanilide degradation products," which are listed on EPA's 1998 Drinking Water Contaminant Candidate List. Method 535 uses solid-phase extraction with a nonporous graphitized carbon sorbent to extract the ethane sulfonic acid (ESA) and oxanilic acid degradates of propachlor, flufenacet, dimethenamid, alachlor, acetochlor, and metolachlor from finished drinking water matrixes. Separation and quantitation of the target analytes are achieved with liquid chromatography/tandem mass spectrometry. Dimethachlor ESA and butachlor ESA were chosen during the method development as the surrogate and internal standard. Drinking water samples were dechlorinated with ammonium chloride without adversely affecting the analyte recoveries. Typical mean recoveries of 92-116% in deionized water and 89-116% in ground water were observed with relative standard deviations of <5%.

On-line cation-exchange preconcentration and capillary electrophoresis coupled by tee joint interface.

An on-line preconcentration method based on ion exchange solid phase extraction was developed for the determination of cationic analytes in capillary electrophoresis (CE). The preconcentration-separation system consisted of a preconcentration capillary bonded with carboxyl cation-exchange stationary phase, a separation capillary for zone electrophoresis and a tee joint interface of the capillaries. Two capillaries were connected closely inside a 0.3 mm i.d. polytetrafluoroethylene tube with a side opening and fixed together by the interface. The preparations of the preconcentration capillaries and interface were described in detail in this paper. The on-line preconcentration and separation procedure of the analysis system included washing and conditioning the capillaries, loading analytes, filling with buffer solution, eluting analytes and separating by capillary zone electrophoresis (CZE). Several analysis parameters, including sample loading flow rate and time, eluting solution and volume, inner diameter and length of preconcentration capillary etc., were investigated. The proposed method enhanced the detection sensitivity of CE-UV about 5000 times for propranolol and metoprolol compared with normally electrokinetic injection. The detection limits of propranolol and metoprolol were 0.02 and 0.1 microg/L with the proposed method respectively, whereas those were 0.1 and 0.5 mg/L with conventional electrokinetic injection. The experiment results demonstrate that the proposed technique can increase the preconcentration factor evidently.

Effect of electrolyte and solvent composition on capillary electrophoretic separation of some pharmaceuticals in non-aqueous media.

Non-aqueous capillary electrophoresis was used to study the separation selectivity of positively charged drug substances and negatively charged diuretics. Study was made of the effects of organic solvent composition and the background electrolyte on the separation. The separation selectivity could be altered considerably by varying the methanol/acetonitrile composition. In addition, the migration order and the resolution of the pharmaceuticals could be altered merely by changing the electrolyte cation or the anion. The electrolytes tested were alkali metal acetates, ammonium acetate, ammonium chloride and ammonium bromide. As with aqueous background electrolyte solutions, the electroosmotic flow was decreased with increasing size of the alkali metal cation of the electrolyte in methanol/acetonitrile 50:50 (v/v).

Determination of N-chloramines in As-samra chlorinated wastewater and their effect on the disinfection process.

Chlorine is the most widely used disinfectant of wastewater due to its capacity to inactivate most pathogenic microorganisms quickly. However, chlorine reacts with natural organic compounds present in wastewater to produce some undesirable organic byproducts. One such class of compounds is the nitrogenous compounds. The reaction between chlorine and compounds containing a nitrogen atom with one or more hydrogen atoms attached to it will form chloramines which have lower disinfection efficiency. Eighty percent of the wastewater generated in Jordan is treated at the Khirbet As-Samra wastewater treatment plant for eventual use in agriculture. In this study efficiency of chlorination was studied by collecting samples from the effluent of the treatment plant. The yield concentration of N-chloramines upon chlorination was determined. Efficiency of disinfection process as a function of contact time, concentration of chlorine dosage, concentration of nitrogenous compound and pH were studied. In this study, it has been found that at a chlorine dosage of 15 mg/L and contact time of 15 min, the percentage total coliform kill in As-samra wastewater sample was 100%. After addition of histidine, glycine and phenylalanine (15 mg/L in each case) to the wastewater sample, the percentage of total coliform kill dropped to 58, 78 and 79% respectively. When chlorine dosage was increased to 24 mg/L the percentage total coliform kill reached 96, 99 and 100% in wastewater samples treated with 5 mg/L histidine, glycine and phenylalanine, respectively. The percentage total coliform kill dropped to zero in wastewater samples treated with histidine, glycine and phenylalanine at a concentration of 30 mg/L each. This work supports the theory that amino acids and ammonia preferentially react with chlorine to form N-chloramine which significantly reduces the disinfection efficiency of the chlorination process.

Investigation of the chemical equivalence of the trypanocidal products, Samorin and Veridium.

A procedure for the evaluation of chemical equivalence of proprietary formulations of isometamidium is described. The method combines the analysis of the principal component (isometamidium), HPLC profiling of related substances and determination of the inorganic impurity, ammonium chloride, using a modification of the Berthelot (Indophenol) reaction. Application of these procedures to analyses of commercially available sachets from four different batches of Samorin and four different batches of Veridium has demonstrated that there are marked qualitative and quantitative differences between batches from these two sources. Whilst Samorin samples showed inter-batch consistency of composition, there was considerable inter-batch variation between the samples of Veridium.

"Reagentless" flow injection determination of ammonia and urea using membrane separation and solid phase basification.

Flow injection analysis instrumentation and methodology for the determination of ammonia and ammonium ions in an aqueous solution are described. Using in-line solid phase basification beds containing crystalline media. the speciation of ammoniacal nitrogen is shifted toward the un-ionized form. which diffuses in the gas phase across a hydrophobic microporous hollow fiber membrane into a pure-water-containing analytical stream. The two streams flow in a countercurrent configuration on opposite sides of the membrane. The neutral pH of the analytical stream promotes the formation of ammonium cations, which are detected using specific conductance. The methodology provides a lower limit of detection of 10 microgram/L and a dynamic concentration range spanning three orders of magnitude using a 315-microliters sample injection volume. Using immobilized urease to enzymatically promote the hydrolysis of urea to produce ammonia and carbon dioxide, the technique has been extended to the determination of urea.

Determination of volatile bases in seafood using the ammonia ion selective electrode: collaborative study.

Nine collaborating laboratories tested a combination of 23 seafood samples for volatile bases using an ammonia ion selective electrode. Results were reported as mg NH3/100 g fish, but the method reflected levels of both ammonia and trimethylamine, which permeated the ammonia membrane. The 23 samples were broken down into 8 blind duplicate pairs, 2 Youden matched pairs, and 3 single samples covering fresh to spoiled product ranging from 8 to 82 mg NH3/100 g. Seven species were evaluated: Atlantic cod, squid, Atlantic halibut, gray sole, monkfish, dogfish, and Atlantic mackerel. The ammonia electrode assay was performed on an aqueous homogenate consisting of 95 mL distilled water and 5.0 g sample tissue. Alkaline ion strength adjusting solution (2 mL) was added to the homogenate to liberate ammonia that was sensed by the ion specific electrode and measured on a precalibrated portable meter. Repeatability standard deviations (RSDr) ranged from 4.2 to 17%; reproducibility standard deviations (RSDR) ranged from 8.8 to 21%. A standard ammonium chloride solution was provided to all laboratories to spike 3 different samples at 10 mg NH3/100 g. Recoveries of added ammonia as ammonium chloride for fresh, borderline, and spoiled samples were 88.6, 107, and 128%, respectively.

Ion chromatography studies of quaternary ammonium halide solutions and the determination of pharmaceuticals.

The feasibility of using ion chromatography (IC) in solutions containing quaternary ammoniums is studied. The results establish the applicability of IC for the determination of halide ions in a pharmaceutical sample without interferences by various excipient ions. For IC measurements with suitable methodical parameters, linearity is maintained over the range of 20 to 200 mg/L and 10.0 to 250 mg/L for the chloride and bromide of quaternary ammoniums, respectively. A comparison of the anion results obtained from differential pulse voltammetry shows good agreement.

Quick tests for the determination of ammonia in poultry litter.

The objective of this work was to test the ability of various available quick tests to determine ammonia concentration of poultry litters. A total of 136 samples was collected from brood chambers of poultry houses. Samples were equally divided between surface samples (top 25 mm) and core samples. Samples were frozen until analysis but received no further processing. Samples were analyzed for ammonia by autoanalyzer (standard) and several quick tests (conductivity, Quantofix N-Volumeter, and Reflectoquant). In addition, samples were analyzed by near-infrared spectroscopy by scanning samples using a large-sample transport device on a FOSS-NIRSystems model 6500 (64 co-added scans from 400 to 2,498 nm). Results showed that, although ammonia could be determined with reasonable accuracy by near-infrared spectroscopy using data in the 1,100 to 2,498 nm spectral range (final calibration R2 of approximately 0.90), none of the quick tests, including near-infrared, worked as well as previously found with dairy manures. The best results were found using the Quantofix or Reflectoquant (R2 of approximately 0.75), and conductivity worked only with the core samples (R2 of approximately 0.75). It is believed that interferences due to the presence of uric acid (spectroscopy, Quantofix, and Reflectoquant) and sodium bisulfate used to treat the litter (conductivity) were the cause of the decreased accuracies as compared to results achieved previously with dairy manures.

Commonly used metal and crystalline salts in South African traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional medicines in the form of plants, animals and/or minerals are used by millions of South Africans. There is currently no data regarding the commonly used mineral elements thus the potential benefits or hazards of such products remain unclear. MATERIALS AND METHODS: Metal and crystalline salts were purchased from a rural market (Nongoma, Zululand, South Africa). Information regarding the colloquial name, price and weight was recorded. Energy dispersive X-ray spectroscopy (EDX) was used to quantatively determine the unknown salts. RESULTS: Six widely available salts were analyzed. Ndonya, as it is colloquially known, refers to two products which look identical to the untrained eye-one is dyed table salt and the other is hexavalent chromium. A further product used medicinally, although not widely available, was identified as iron chromite ore. CONCLUSIONS: The array of substances documented, ranging from benign to carcinogenic, stresses the importance of documenting components used in traditional medicine and confirms the necessity to regulate South Africa traditional medicine. Healthcare workers should be aware of the complexities of using such metal salt.

Behaviour and dynamics of di-ammonium phosphate in bauxite processing residue sand in Western Australia--II. Phosphorus fractions and availability.

BACKGROUND, AIM AND SCOPE: The production of alumina involves its extraction from bauxite ore using sodium hydroxide under high temperature and pressure. This process yields a large amount of residue wastes, which are difficult to revegetate due to their inherent hostile properties--high alkalinity and sodicity, poor water retention and low nutrient availability. Although phosphorus (P) is a key element limiting successful ecosystem restoration, little information is available on the availability and dynamics of P in rehabilitated bauxite-processing residue sand (BRS). The major aim of this experiment was to quantify P availability and behaviour as affected by pH, source of BRS and di-ammonium phosphate (DAP) application rate. MATERIALS AND METHODS: This incubation experiment was undertaken using three sources of BRS, three DAP application rates (low, without addition of DAP; medium, 15.07 mg P and 13.63 mg N of DAP per jar, 100 g BRS; and high, 30.15 mg P and 27.26 mg N per jar, 100 g BRS), and four BRS pH treatments (4, 7, 9 and 11 (original)). The moisture content was adjusted to 55% water holding capacity and each BRS sample was incubated at 25 degrees C for a period of 119 days. After this period, Colwell P and 0.1 M H(2)SO(4) extractable P in BRS were determined. In addition, P sequential fractionation was carried out and the concentration of P in each pool was measured. RESULTS AND DISCUSSION: A significant proportion (37% recovered in Colwell P and 48% in 0.1 M H(2)SO(4) extraction) of P added as DAP in BRS are available for plant use. The pH did not significantly affect 0.1 M H(2)SO(4) extractable P, while concentrations of Colwell P in the higher initial pH treatments (pH 7, 9 and 11) were greater than in the pH 4 treatments. The labile fractions (sum of NH(4)Cl (AP), bicarbonate and first sodium hydroxide extractable P (N(I)P)) consisted of 58-64% and 70-72% of total P in the medium and high DAP rate treatments, respectively. This indicates that most P added as DAP remained labile or moderately labile in BRS, either in solution, or in adsorbed forms on the surface of more crystalline P compounds, sesquioxides and carbonate, or associated with amorphous and some crystalline Al and Fe hydrous oxides. In addition, differences in the hydrochloric acid extractable P and the residual-P fractions among the treatments with and without DAP addition were relative small comparing with other P pools (e.g., NaOH extractable P pools), further indicating the limited capacity of BRS for fixing P added in Ca-P and other most recalcitrant forms. CONCLUSIONS: P availability in the original BRS without addition of DAP was very low, mostly in recalcitrant form. It has been clearly demonstrated that significant proportions of P added as DAP could remain labile or moderately labile for plant use during the rehabilitation of bauxite-processing residue disposal areas. There was limited capacity of BRS for fixing P in more recalcitrant forms (e.g., Ca-P and residual-P). Concentrations of most P pools in BRS increased with the DAP application rate. The impact of the pH treatment on P availability varied with the type of P pools and the DAP rate. RECOMMENDATION AND PERSPECTIVES: It is recommended that the development of appropriate techniques for more accurate estimation of P availability in BRS and the quantification of the potential leaching loss of P in BRS are needed for the accurate understanding of P availability and dynamics in BRS. In addition, application of organic matters (e.g., biosolids and biochar, etc.) to BRS may be considered for improving P availability and buffering capacity.