Decreased Immunoreactivity of Hepatitis E Virus Antigen Following Treatment with Sakhalin Spruce (Picea glehnii) Essential Oil.

The hepatitis E virus (HEV) causes a common infectious disease that infects pigs, wild boars, deer, and humans. In most cases, humans are infected by eating raw meat. Some essential oils have been reported to exhibit antiviral activities. In this study, in order to investigate the anti-HEV properties of essential oils, the immunoreactivities of HEV antigen proteins against the relevant antibodies were analyzed after the HEV antigens underwent treatment with various essential oils. The essential oils extracted from the tea tree, which was previously reported to exhibit antiviral activity, lavender, and lemon had strongly reduced activity. We found that treatment with the essential oil prepared from Sakhalin spruce was associated with the strongest reduction in immunoreactivity of HEV antigen protein(s) among the tested substances. The main volatile constituents of Sakhalin spruce essential oil were found to be bornyl acetate (32.30 %), α-pinene (16.66 %), camphene (11.14 %), camphor (5.52 %), ß-phellandrene (9.09 %), borneol (4.77 %), and limonene (4.57 %). The anti-HEV properties of the various components of the essential oils were examined: treatment with bornyl acetate, the main component of Sakhalin spruce oil, α-pinene, the main component of tea tree oil, and limonene, the main component of lemon oil, resulted in a strong reduction in HEV antigen immunoreactivity. These results indicate that each main component of the essential oils plays an important role in the reduction of the immunoreactivity of HEV antigen protein(s); they also suggest that Sakhalin spruce essential oil exhibits anti-HEV activity. In a formulation with the potential to eliminate the infectivity of HEV in foodborne infections, this essential oil can be applied as an inactivating agent for meat processing and cooking utensils, such as knives and chopping boards.

Selective capillary electrophoresis separation of mono and divalent cations within a high-surface area-to-volume ratio multi-lumen capillary.

In this study, the separation of inorganic mono and divalent cations using multi-lumen silica capillaries (MLCs) of 126 channels, each with either 4 or 8 µm inner diameter, was investigated using capillary electrophoresis and on-capillary capacitively coupled contactless conductivity detection (CE-C4D). MLCs provided sufficiently high surface area-to-volume ratios to generate significant wall ion-exchange interactions for the divalent cations, which significantly affected resultant selectivity, whereas monovalent cations were predominantly separated by simple electrophoresis. The resultant hybrid selectivity was seen for both 4 and 8 µm channel multi-lumen capillaries, without any preconditioning or capillary wall modification. Remarkably, the electrophoretic mobilities for the divalent cations Mg2+ and Ca2+ were reduced 7.5 times compared to those determined using a single channel open tubular capillary of 50 µm i.d., providing much improved selectivity. Apparent electrophoretic mobilities of divalent cations increased as the concentration of BGE increased, while those of monovalent cations decreased parallel to electroosmotic mobility. These results show the electrostatic interaction between the divalent cations and the silica wall. At least, this specific separation of mono- and divalent cations were clearly observed with a mixture standards solution of less than 200 µmol L-1. Using a MLC with 126 × 8 µm i.d. channels and 49.1 cm in length, together with a 20 mmol L-1 MES/His BGE, containing 2 mmol L-1 18-crown-6, monovalent cations (NH4+, K+ Na+ and Li+) and divalent cations (Ca2+ and Mg2+) could be completely separated within 4 min. For monovalent cations, on-capillary detection using C4D provided calibration curve (0-200 µmol L-1) correlation coefficients in the range R2 = 0.995-0.999, and limits of detection of 2.2-6.6 µmol L-1. Relative standard deviations for migration times were less than 0.6%, and recoveries ranged from the 93.8%-105.4%. The new method was applied to the separation and quantitative determination of monovalent and divalent cations in drinking waters and soil extracts.

Single injection ion-exclusion/cation-exchange chromatography for simultaneous determination of organic/inorganic anions, inorganic cations, and ethanol in beer samples.

Multicomponent simultaneous analysis is important for management programs, which are required in beer industries because the beer constituents measure by combining several methods in the present. In response to a requirement, our research group developed single sample injection ion chromatography systems, which comprise ion-exclusion/cation-exchange chromatography (IEC/CEC) and post-column derivatization and show promise for simultaneously determining concentrations of organic and inorganic species and alcohol commonly found in beer. Optimal chromatographic resolutions for determining 17 different species in beer samples by IEC/CEC were obtained on a H+-formed weakly acidic cation-exchange resin column with an eluent comprising 2 mM phthalic acid and 1 mM 18-crown-6. Consequently, the usefulness of developed method for monitoring beer samples was demonstrated in terms of beneficial information such as dependency of K+ concentration on the malt amount, influences of organic anion concentrations on different types of bottling methods, and validation of ethanol concentrations displayed in the ingredient table.

Rapid screening of inorganic and organic anions in liquid by-products from hydrothermal treatment of biomass by capillary electrophoresis.

A simple and rapid capillary electrophoresis method with capacitively coupled contactless conductivity detection (CE-C4 D) for the simultaneous determination of inorganic and organic anions in liquid product obtained from the hydrothermal treatment of biomass residues is presented. Under the optimal analytical conditions, limits of detection ranged from 1.8 to 9.4 µM for most target solutes and 53 µM for citrate. Relative standard deviations were below 0.5% for migration times and within 0.6-6.5% for peak areas for all solutes. The proposed method was successfully applied for the rapid determination and screening of inorganic and organic anions in liquid product produced following differing hydrothermal treatment temperatures for banana and pineapple biomass, and the contribution of organic acid formation to acidity in the liquid was evaluated. CE-C4 D could be a suitable method for the optimization or tailoring of HTT conditions for desired liquid product composition, and additionally for determination of the best variety(s) of biomass to use in such processes.

Simultaneous analysis of silicon and boron dissolved in water by combination of electrodialytic salt removal and ion-exclusion chromatography with corona charged aerosol detection.

Selective separation and sensitive detection of dissolved silicon and boron (DSi and DB) in aqueous solution was achieved by combining an electrodialytic ion isolation device (EID) as a salt remover, an ion-exclusion chromatography (IEC) column, and a corona charged aerosol detector (CCAD) in sequence. DSi and DB were separated by IEC on the H(+)-form of a cation exchange resin column using pure water eluent. DSi and DB were detected after IEC separation by the CCAD with much greater sensitivity than by conductimetric detection. The five-channel EID, which consisted of anion and cation acceptors, cathode and anode isolators, and a sample channel, removed salt from the sample prior to the IEC-CCAD. DSi and DB were scarcely attracted to the anion accepter in the EID and passed almost quantitatively through the sample channel. Thus, the coupled EID-IEC-CCAD device can isolate DSi and DB from artificial seawater and hot spring water by efficiently removing high concentrations of Cl(-) and SO4(2-) (e.g., 98% and 80% at 0.10molL(-1) each, respectively). The detection limits at a signal-to-noise ratio of 3 were 0.52µmolL(-1) for DSi and 7.1µmolL(-1) for DB. The relative standard deviations (RSD, n=5) of peak areas were 0.12% for DSi and 4.3% for DB.

Spatial distributions of and diurnal variations in low molecular weight carbonyl compounds in coastal seawater, and the controlling factors.

We studied the spatial distributions of and the diurnal variations in four low molecular weight (LMW) carbonyl compounds, formaldehyde, acetaldehyde, propionaldehyde, and glyoxal, in coastal seawater. The samples were taken from the coastal areas of Hiroshima Bay, the Iyo Nada, and the Bungo Channel, western Japan. The formaldehyde, acetaldehyde, and glyoxal concentrations were higher in the northern part of Hiroshima Bay than at offshore sampling points in the Iyo Nada and the Bungo Channel. These three compounds were found at much higher concentrations in the surface water than in deeper water layers in Hiroshima Bay. It is noteworthy that propionaldehyde was not detected in any of the seawater samples, the concentrations present being lower than the detection limit (1 nanomole per liter (nM)) of the high performance liquid chromatography (HPLC) system we used. Photochemical and biological experiments were performed in the laboratory to help understand the characteristic distributions and fates of the LMW carbonyl compounds. The primary process controlling their fate in the coastal environment appears to be their biological consumption. The direct photo degradation of propionaldehyde, initiated by ultraviolet (UV) absorption, was observed, although formaldehyde and acetaldehyde were not degraded by UV irradiation. Our results suggest that the degradation of the LMW carbonyl compounds by photochemically formed hydroxyl radicals is relatively insignificant in the study area. Atmospheric deposition is a possible source of soluble carbonyl compounds in coastal surface seawater, but it may not influence the carbonyl concentrations in offshore waters.

Indirect UV detection-ion-exclusion/cation-exchange chromatography of common inorganic ions with sulfosalicylic acid eluent.

Herein, we describe indirect UV detection-ion-exclusion/cation-exchange chromatography (IEC/CEC) on a weakly acidic cation-exchange resin in the H(+)-form (TSKgel Super IC-A/C) using sulfosalicylic acid as the eluent. The goal of the study was to characterize the peaks detected by UV detector. The peak directions of analyte ions in UV at 315 nm were negative because the molar absorbance coefficients of analyte anions and cations were lower than that of the sulfosalicylic acid eluent. Good chromatographic resolution and high signal-to-noise ratios of analyte ions were obtained for the separations performed using 1.1 mM sulfosalicylic acid and 1.5 mM 18-crown-6 as the eluent. The relative standard deviations (RSDs) of the peak areas ranged from 0.6 to 4.9%. Lower detection limits of the analytes were achieved using indirect UV detection at 315 nm (0.23 - 0.98 µM) than those obtained with conductometric detection (CD) (0.61 - 2.1 µM) under the optimized elution conditions. The calibration curves were linear in the range from 0.01 to 1.0 mM except for Cl(-), which was from 0.02 to 2.0 mM. The present method was successfully applied to determine common inorganic ions in a pond water sample.

Water quality monitoring system for determination of ionic nutrients by ion-exclusion chromatography with spectrophotometric detection on cation- and anion-exchange resin columns using water eluent.

A unified ion-exclusion chromatography (IEC) system for monitoring anionic and cationic nutrients like NH4+, NO2-, NO3-, phosphate ion, silicate ion and HCO3- was developed and applied to several environmental waters. The IEC system consisted of four IEC methodologies, including the IEC with ultraviolet (UV) form connected with detection at 210 nm for determining NH4+ on anion-exchange separation column in OH anion-exchange UV-conversion column in I- form in tandem, the IEC with UV-detection at 210 nm for determining simultaneously NO3- and NO3- on cation-exchange separation column in H+ form, the IEC with UV-detection at 210 nm for determining HCO3- on cation-exchange separation column in H+ form connected with anion-exchange UV-conversion column in I- form in tandem, and the IEC with visible-detection based on molybdenum-blue reaction for determining simultaneously silicate and phosphate ions on cation-exchange separation column in H+ form. These IEC systems were combined through three manually-driven 6-port column selection valves to select each separation column to determine selectively the ionic nutrients. Using this sequential water quality monitoring system, the analytical performances such as calibration linearity, reproducibility, detection limit and recovery were also tested under the optimized chromatographic conditions. This novel water quality monitoring system has been applied successfully for the determination of the ionic eutrophication components in sub-urban river waters.

Utilization of a diol-stationary phase column in ion chromatographic separation of inorganic anions.

We describe the ion chromatographic separation of inorganic anions using a diol-stationary phase column (-CH(OH)CH(2)OH; diol-column) without charged functional groups. Anions were separated using acidic eluent as in typical anion-exchange chromatography. The retention volumes of anions on the diol-column increased with increasing H(+) concentration in the eluent. The anion-exchange capacities of diol-columns in the acidic eluent (pH 2.8) were larger than that of zwitterionic stationary phase column but smaller than that of an anion-exchange column. The separation of anions using the diol-column was strongly affected by the interaction of H(+) ions with the diol-functional groups and by the types of the eluents. In particular, the selection of the eluent was very important for controlling the retention time and resolution. Good separation was obtained using a diol-column (HILIC-10) with 5 mM phthalic acid as eluent. The limits of detection at a signal-to-noise ratio of 3 ranged from 1.2 to 2.7 µM with relative standard deviations (RSD, n=5) of 0.04-0.07% for the retention time and 0.4-2.0% for the peak areas. This method was successfully applied to the determination of H(2)PO(4)(-), Cl(-), and NO(3)(-) in a liquid fertilizer sample.

Recent progress and applications of ion-exclusion/ion-exchange chromatography for simultaneous determination of inorganic anions and cations.

One of the ultimate goals of ion chromatography is to determine both anions and cations found in samples with a single chromatographic run. In the present article, recent progress in ion-exclusion/ion-exchange chromatography for the simultaneous determinations of inorganic anions and cations are reviewed. Firstly, the principle and the control for the simultaneous separation and detection of analyte ions using ion-exclusion/cation-exchange chromatography with a weakly acidic cation-exchange column are outlined. Then, advanced chromatographic techniques in terms of analytical time, selectively and sensitivity are summarized. As a related method, ion-exclusion/anion-exchange chromatography with an anion-exchange column could be used for the simultaneous determination of inorganic nitrogen species, such as ammonium, nitrite and nitrate ions. Their usefulness and applications to water-quality monitoring and related techniques are also described.

Simultaneous determination of NH4+, NO2(-) and NO3(-) by ion-exclusion/anion-exchange chromatography on a strongly basic anion-exchange resin with basic eluent.

Ion-exclusion/anion-exchange chromatography (IEC/AEC) on a combination of a strongly basic anion-exchange resin in the OH(-)-form with basic eluent has been developed. The separation mechanism is based on the ion-exclusion/penetration effect for cations and the anion-exchange effect for anions to anion-exchange resin phase. This system is useful for simultaneous separation and determination of ammonium ion (NH4+), nitrite ion (NO2(-)), and nitrate ion (NO3(-)) in water samples. The resolution of analyte ions can be manipulated by changing the concentration of base in eluent on a polystyrene-divinylbenzene based strongly basic anion-exchange resin column. In this study, several separation columns, which consisted of different particle sizes, different functional groups and different anion-exchange capacities, were compared. As the results, the separation column with the smaller anion-exchange capacity (TSKgel Super IC-Anion) showed well-resolved separation of cations and anions. In the optimization of the basic eluent, lithium hydroxide (LiOH) was used as the eluent and the optimal concentration was concluded to be 2 mmol/L, considering the resolution of analyte ions and the whole retention times. In the optimal conditions, the relative standard deviations of the peak areas and the retention times of NH4+, NO2(-), and NO3(-) ranged 1.28% - 3.57% and 0.54% - 1.55%, respectively. The limits of detection at signal-to-noise of 3 were 4.10 micromol/L for NH4+, 1.87 micromol/L for NO2(-) and 2.83 micromol/L for NO3(-).

Enhanced conductivity detection of common inorganic anions in electrostatic ion chromatography using water eluent.

To enhance the conductivity detection sensitivity of common anions (Na-anions) in electrostatic ion chromatography (EIC) by elution with water, a conductivity enhancement column packed with strong acid cation exchange resin in the H-form was inserted between an octadecyl silane (ODS)-silica separation column modified with zwitterionic surfactant (CHAPS: 3-{(3-cholamidopropyl)-dimethylammonio} propanesulfonate) and a conductivity detector. Specifically, the Na-anion pairing is converted to H-anion pairing after the EIC separation and then detected sensitively by the conductivity detector. The effects of conductivity enhancement and suppression in the EIC by the enhanced conductivity detection were characterized for the common strong acid anions such as SO4(2-), Cl(-), NO3(-), I(-) and ClO4(-) and weak acid anions such as F(-), NO2(-), HCOO(-), CH3COO(-) and HCO3(-). For the conductivity enhancement effect in the EIC, it is found that the conductivity of measured for all strong acid anions (Na-anions) was enhanced according to the theoretical conductivity predicted for H-anions and that of the measured for weak acid anions was suppressed depending on their pKa of H-anions. For the calibration linearity in the EIC, the strong acid anions were linear (r2 = 0.99 - 1.00) because the degree of dissociation is almost 1.0 over all the concentration range and that of the weak acid anions was non-linear because the degree of dissociation decreased by increasing the concentration of the weak acid anions. In conclusion, the EIC by enhanced conductivity detection was recognized to be useful only for the strong acid anions in terms of conductivity detection and calibration linearity.

[Application of simultaneous determination of inorganic ionic species by advanced ion chromatography for water quality monitoring of river water and wastewater].

In this study, our recent work on advanced ion chromatographic methods for the simultaneous determination of inorganic ionic species such as common anions (SO4(2-), Cl(-) and NO3(-)) and cations (Na+, NH4+, K+, Mg2+, and Ca2+), nutrients (phosphate and silicate) and hydrogen ion/alkalinity are summarized first. Then, the applications using these methods for monitoring environmental water quality are also presented. For the determination of common anions and cations with nutrients, the separation was successfully performed by a polymethacrylate-based weakly acidic cation-exchange column of TSKgel Super IC-A/C (Tosoh, 150 mm x 6.0 mm i. d.) and a mixture solution of 100 mmol/L ascorbic acid and 4 mmol/L 18-crown-6 as acidic eluent with dual detection of conductivity and spectrophotometry. For the determination of hydrogen ion/alkalinity, the separation was conducted by TSKgel ODS-100Z column (Tosoh, 150 mm x 4.5 mm i. d.) modified with lithium dodecylsulfate and an eluent of 40 mmol/L LiCl/0.1 mmol/L lithium dodecylsulfate/0.05 mmol/L H2SO4 with conductivity detector. The differences of ion concentration between untreated and treated wastewater showed the variation of ionic species during biological treatment process in a sewage treatment plant. Occurrence and distribution of water-quality conditions were related to the bioavailability and human activity in watershed. From these results, our advanced ion chromatographic methods have contributed significantly for water quality monitoring of environmental waters.

Automobile exhaust gas as a source of aqueous phase OH radical in the atmosphere and its effects on physiological status of pine trees.

Free radical generation potential of automobile exhaust gas was examined by measuring hydroxyl (OH) radical photo-formation rates in exhaust gas-scrubbing water. Effects of automobile exhausts on physiological status of Japanese red pine trees (Pinus densiflora Sieb. et Zucc.) were also investigated to elucidate the mechanism how the free radicals derived from exhaust gas damage higher plants. Gasoline and diesel exhaust gases were scrubbed into pure water. Potential photo-formation rates of OH radical in aqueous phase (normalized to sun light intensity of clear sky midday on May 1 at 34°N) for gasoline and diesel cars were ave. 51 and 107 µ Mh⁻¹ m⁻³ of exhaust gas, respectively. Nitrite was a dominant source (ca. 70-90%) of photochemical formation of OH radical in both gasoline and diesel car exhausts. The scrubbed solution of diesel car exhaust gas was sprayed for six times per week to needles of pine tree seedlings in open top chambers. Control, exhaust+mannitol (added as OH radical scavenger), and nitrite+nitrate standard solution (equivalent levels existed in the exhaust gas) were also sprayed. Two months sprays indicated that the sprayed solutions of diesel exhaust and nitrite+nitrate caused a decrease of maximum photosynthetic rate and stomata conductance in pine needles while the control and exhaust+mannitol solution showed no effects on photosynthetic activities of pine needles. These results indicated that OH radicals generated mainly from photolysis of nitrite occurring in the scrubbing solution of exhaust gas are responsible for the decrease of photosynthetic activities of pine needles.

Ion-exclusion/cation-exchange chromatography with dual detection of the conductivity and spectrophotometry for the simultaneous determination of common inorganic anionic species and cations in river and wastewater.

Simultaneous determinations of common inorganic anionic species (SO(4)(2-), Cl(-), NO(3)(-), phosphate and silicate) and cations (Na(+), NH(4)(+), K(+), Mg(2+) and Ca(2+)) were conducted using an ion-chromatography system with dual detection of conductivity and spectrophotometry in tandem. The separation of ionic species on a weakly acidic cation-exchange resin was accomplished using a mixture of 100 mM ascorbic acid and 4 mM 18-crown-6 as an acidic eluent (pH 2.6), after which the ions were detected using a conductivity detector. Subsequently, phosphate and silicate were analyzed based on derivatization with molybdate and spectrophotometry at 700 nm. The detection limits at S/N = 3 ranged from 0.11 to 2.9 µM for analyte ionic species. This method was applied to practical river water and wastewater with acceptable criteria for the anion-cation balance and comparisons of the measured and calculated electrical conductivity, demonstrating the usefulness of the present method for water quality monitoring.

Use of a polystyrene-divinylbenzene-based weakly acidic cation-exchange resin column and propionic acid as an eluent in ion-exclusion/adsorption chromatography of aliphatic carboxylic acids and ethanol in food samples.

We developed an ion-exclusion/adsorption chromatography (IEAC) method employing a polystyrene-divinylbenzene-based weakly acidic cation-exchange resin (PS-WCX) column with propionic acid as the eluent for the simultaneous determination of multivalent aliphatic carboxylic acids and ethanol in food samples. The PS-WCX column well resolved mono-, di-, and trivalent carboxylic acids in the acidic eluent. Propionic acid as the eluent gave a higher signal-to-noise ratio, and enabled sensitive conductimetric detection of analyte acids. We found the optimal separation condition to be the combination of a PS-WCX column and 20-mM propionic acid. Practical applicability of the developed method was confirmed by using a short precolumn with a strongly acidic cation-exchange resin in the H(+)-form connected before the separation column; this was to remove cations from food samples by converting them to hydrogen ions. Consequently, common carboxylic acids and ethanol in beer, wine, and soy sauce were successfully separated by the developed method.

Simultaneous spectrophotometric determination of orthophosphate and silicate ions in river water using ion-exclusion chromatography with an ascorbate solution as both eluent and reducing agent, followed by postcolumn derivatization with molybdate.

Ion-exclusion chromatography was examined for the simultaneous spectrophotometric determinations of orthophosphate and silicate ions in river water using an ascorbate solution as both an eluent and a reducing agent, followed by postcolumn derivatization using molybdate. The detector responses for both ions increased with increased ascorbic acid concentration in the eluent, but peak tailing was observed for the orthophosphate ion. This suggests that the amounts of undissociated orthophosphate ions increased with decreased eluent pH, resulting in the penetration of the phosphate to the Donnan's membrane formed on the resin surface. Using a neutral sodium ascorbate solution as an eluent, the peak shape was improved. With optimized separation and derivatization conditions (eluent, 20 mM sodium ascorbate; color-forming reagent, 10 mM sodium molybdate-60 mM sulfuric acid; flow rates of eluent and color-forming reagent, 0.4 and 0.2 mL min(-1); coil length, 6 m), the detection limits of orthophosphate and silicate ions were 0.9 and 1.0 microg L(-1), respectively. This method was successfully applied to the determination of orthophosphate and silicate ions in Kurose River water and the quantitative evaluations of the effects of water intake to a reservoir and discharge from a biological sewage treatment plant on the fluxes of these ions in the river.

Transesterification of soybean oil using combusted oyster shell waste as a catalyst.

Transesterification of soybean oil catalyzed by combusted oyster shell, which is waste material from shellfish farms, was examined. Powdered oyster shell combusted at a temperature above 700 degrees C, at which point the calcium carbonate of oyster shell transformed to calcium oxide, acted as a catalyst in the transesterification of soybean oil. On the basis of factorial design, the reaction conditions of catalyst concentration and reaction time were optimized in terms of the fatty acid methyl ester concentration expressed as biodiesel purity. Under the optimized reaction conditions of a catalyst concentration and reaction time of 25wt.%. and 5h, respectively, the biodiesel yield, expressed relative to the amount of soybean oil poured into the reaction vial, was more than 70% with high biodiesel purity. These results indicate oyster shell waste combusted at high temperature can be reused in biodiesel production as a catalyst.

Simultaneous spectrophotometric determination of phosphate and silicate ions in river water by using ion-exclusion chromatographic separation and post-column derivatization.

The simultaneous spectrophotometric determination of phosphate and silicate ions in river water was examined by using ion-exclusion chromatography and post-column derivatization. Phosphate and silicate ions were separated by the ion-exclusion column packed with a polymethacrylate-based weakly acidic cation-exchange resin in the H(+)-form (TSKgel Super IC-A/C) by using ultra pure water as an eluent. After the post-column derivatization with molybdate and ascorbic acid, so-called molybdenum-blue, both ions were determined simultaneously by spectrophotometry. The effects of sulfuric acid, sodium molybdate and ascorbic acid concentrations and reaction coil length, which have relation to form the reduced complexes of molybdate and ions, on the detector response for phosphate and silicate ions were investigated. Under the optimized conditions (color-forming reactant, 50 mM sulfuric acid-10 mM sodium molybdate; reducing agent, 50 mM ascorbic acid; reaction coil length, 6 m), the calibration curves of phosphate and silicate ions were linear in the range of 50-2000 microg L(-1) as P and 250-10,000 microg L(-1) as Si. This method was successfully applied to water quality monitoring of Kurose-river watershed and it suggested that the effluent from a biological sewage treatment plant was significant source of phosphate ion in Kurose-river water.

Phytotoxicities of fluoranthene and phenanthrene deposited on needle surfaces of the evergreen conifer, Japanese red pine (Pinus densiflora Sieb. et Zucc.).

Polycyclic aromatic hydrocarbons (PAHs) have been widely studied with respect to their carcinogenic and mutagenic effects on animals and human cells. Phenanthrene (PHE) and fluoranthene (FLU) effects on the needle photosynthetic traits of 2-year-old Japanese red pine (Pinus densiflora Sieb. et. Zucc.) seedlings were investigated. Three months after fumigation of foliage with solutions containing these PAHs (10 microM each), FLU had negative effects on net photosynthesis at near-saturating irradiance, stomatal conductance, initial chlorophyll fluorescence, and the contents of total chlorophyll, magnesium, and ribulose 1,5-bisphosphate carboxylase (rubisco) of current-year needles. PHE had similar negative effects to FLU but in lesser magnitude. The effects of the PAHs were mitigated by the addition of an OH-radical scavenger (mannitol) into the PAH solutions. PAHs deposited on the surface of pine needles may induce the generation of reactive oxygen species in the photosynthetic apparatus, a manner closely resembling the action of the herbicide paraquat.