Toxicity, Biodegradation, and Metabolic Fate of Organophosphorus Pesticide Trichlorfon on the Freshwater Algae Chlamydomonas reinhardtii.

This study investigated the toxicity of trichlorfon (TCF) to the freshwater algae Chlamydomonas reinhardtii, as well as its biodegradation and metabolic fate. The growth of C. reinhardtii decreased with increasing TCF concentration, and the maximum inhibition ratio was 51.3% at 200 mg L-1 TCF compared to the control. Analyses of pigment content, chlorophyll fluorescence, and antioxidant enzymes indicated that C. reinhardtii can produce resistance and acclimatize to the presence of TCF. The variations in pH during cultivation suggested that photosynthetic microalgae have innate advantages over bacteria and fungi in remediating TCF. A 100% biodegradation rate was achieved at a maximum concentration of 100 mg L-1 TCF. Ten metabolites were identified by GC-MS, and the degradation pathways of TCF by the algae were proposed. This research demonstrated that C. reinhardtii is highly tolerant to and can efficiently degrade TCF. Thus, C. reinhardtii can be used to remove traces of TCF from natural water environments and to treat TCF-contaminated wastewater.

Enhanced removal of trichlorfon and Cd(II) from aqueous solution by magnetically separable chitosan beads immobilized Aspergillus sydowii.

Simultaneous removal of heavy metals and organics from wastewater has always been an environmental problem with great concern. In this study, a novel ecofriendly bioborbent, magnetic chitosan beads immobilized Aspergillus sydowii (MCBAs) were synthesized and used to simultaneously remove trichlorfon (TCF) and Cd(II) from aqueous solution. MCBAs showed an increased special surface area (55.38 m2·g-1) through immobilizing A. sydowii and its saturation magnetization reached 14.62 emu·g-1. The equilibrium removal capacities of TCF and Cd(II) were 135.43 mg·g-1 and 56.40 mg·g-1 in the co-system with 200 mg·L-1 TCF and 50 mg·L-1 Cd(II), respectively. The removal capacities of TCF and Cd(II) were strongly depended on the immobilized A. sydowii spore concentration, initial concentrations of TCF and Cd(II), and MCBAs dose. TCF biodegradation intermediates were identified by gas chromatography-mass spectrometry system. Fourier transform infrared spectra displayed that -OH and -NH groups on MCBAs mainly participated in the Cd(II) sequestration and the CO stretching vibration was possibly related to the degradation intermediates of TCF. MCBAs exhibited excellent recyclability upto four cycles. Therefore, MCBAs are suitable and effective for the simultaneous removal of TCF and Cd(II) from wastewater.

Construction of Persistent Luminescence-Plastic Antibody Hybrid Nanoprobe for In Vivo Recognition and Clearance of Pesticide Using Background-Free Nanobioimaging.

We prepared a specific adsorptive nanocarrier for pesticide due to its challenge to cleanup and low detoxification in the treatment after intake, whether intentional or by mistake. We modified the plastic antibody (molecularly imprinted polymer (MIP)) on the surface of persistent luminescence nanoparticle (La3Ga5GeO14: Cr3+, Zn2+, LGGO) as the specific adsorptive nanocarrier for toxic molecules and realized the nanocarrier was widely distributed for absorbing pesticide and real-time in vivo bioimaging. We used LGGO as the core and trichlorphon as the template to prepare the plastic antibody nanocarrier. After in vivo bioimaging and biodistribution of mice, LGGO@MIP could be distributed evenly in the gastrointestinal tract, circulated in the blood for a long time, and finally excreted to achieve the adsorption and removal of pesticide in the body. The LGGO@MIP nanocarrier prepared in this study opens a new way for the treatment of poisoning.

A study on biomimetic immunoassay-capillary electrophoresis method based on molecularly imprinted polymer for determination of trace trichlorfon residue in vegetables.

Pesticide residue in vegetables is a serious problem that has adverse effects on human health. In our study, we designed and synthesized a molecularly imprinted polymer that can selectively recognize trichlorfon. Using the polymer material as biomimetic antibody, we developed a biomimetic immunoassay-capillary electrophoresis method with improved sensitivity for the detection of trichlorfon. We evaluated the competitive reactions between HRP labeled trichlorfon hapten and free trichlorfon with the biomimetic antibody. Factors that affected the sensitivity of our method were tested in detail. Under optimal conditions, the limit of detection (LOD, IC15) and the sensitivity (IC50) of this method were 0.16mgL-1 and 0.13µgL-1 for trichlorfon. We used this method to determine the trichlorfon spiked in the kidney bean and cucumber samples with recoveries ranging from 78.8% to 103%. We also detected residual trichlorfons in the leek samples, and these results were verified by gas chromatography method.

Sensitive inkjet printing paper-based colormetric strips for acetylcholinesterase inhibitors with indoxyl acetate substrate.

A new paper-based biosensing approach has been developed for sensitive and rapid detection of acetylcholinesterase (AChE) inhibitors. The biosensing zone of the paper strip is constructed with an inkjet printing method, and the biomolecule AChE is immobilized into two layers of biocompatible sol-gel-derived silica ink with a "sandwich" form. Indoxyl acetate (IDA) is used as a chromogenic substrate, which is colorless and can be catalytically hydrolyzed into blue-colored indigo dipolymer. When the enzymatic activity of AChE is inhibited after incubation with organophosphate pesticides (OPs), there is a decreased hydrolysis of IDA accompanying with a drop in color intensity. Paraoxon and trichlorfon are used as the representative OPs in the assay. Due to the low solubility and high molar absorption coefficient of the IDA dipolymer product, the paper-based strip can form a neat blue sensing zone and shows obviously improved sensitivity with a limit of detection (LOD) of 0.01ngmL-1 paraoxon and 0.04ngmL-1 trichlorfon (S/N=3) and the LODs for visual detection are 0.03ngmL-1 for paraoxon and 0.1ngmL-1 for trichlorfon comparing with the previously reported colorimetric methods. The concentrations of paraoxon in apple juice samples are also detected, and the results are in accord well with these results from high-performance liquid chromatography, showing great potential for on-site detection of OPs in practical application. The developed assay can be used to qualitatively and semiquantitatively estimate with naked eyes and quantitatively assess OPs through image analysis.

Biodegradation of the Organophosphate Trichlorfon and Its Major Degradation Products by a Novel Aspergillus sydowii PA F-2.

Trichlorfon (TCF) is an important organophosphate pesticide in agriculture. However, limited information is known about the biodegradation behaviors and kinetics of this pesticide. In this study, a newly isolated fungus (PA F-2) from pesticide-polluted soils was identified as Aspergillus sydowii on the basis of the sequencing of internal transcribed spacer rDNA. This fungus degraded TCF as sole carbon, sole phosphorus, and sole carbon-phosphorus sources in a mineral salt medium (MSM). Optimal TCF degradation conditions were determined through response surface methodology, and results also revealed that 75.31% of 100 mg/L TCF was metabolized within 7 days. The degradation of TCF was accelerated, and the mycelial dry weight of PA F-2 was remarkably increased in MSM supplemented with exogenous sucrose and yeast extract. Five TCF metabolic products were identified through gas chromatography-mass spectrometry. TCF could be initially hydrolyzed to dichlorvos and then be degraded through the cleavage of the P-C bond to produce dimethyl hydrogen phosphate and chloral hydrate. These two compounds were subsequently deoxidized to produce dimethyl phosphite and trichloroethanal. These results demonstrate the biodegradation pathways of TCF and promote the potential use of PA F-2 to bioremediate TCF-contaminated environments.

Dissipation kinetics, safety evaluation, and preharvest interval assessment of trichlorfon application on rice.

Nowadays, there is an urgent need for the investigation of the field dissipation and assessment of the preharvest interval for trichlorfon residues on rice. To protect consumers from potential health risks, this study can provide references for the safe application of trichlorfon in the rice fields. Results of the field dissipation study showed that the dissipation dynamic equations of trichlorfon were based on the first-order reaction dynamic equations and that the dissipation rates vary among rice plant, brown rice, rice bran, soil, and water. The 2-year field trials conducted in Yangzhou and Xiaogan suggested the interval of each application for trichlorfon on rice to be at least 7 days when 80 % trichlorfon SP was sprayed with a dose ranges between 80 and 160 a.i g/667 m(2). Additionally, the preharvest interval of the last application should be at least 15 days to ensure the amounts of residues below the maximum residue limits of trichlorfon on brown rice (0.1 mg/kg).

Investigation on the Gas-phase decomposition of trichlorfon by GC-MS and theoretical calculation.

The gas phase pyrolysis of trichlorfon was investigated by the on-line gas chromatography - mass spectrometry (GC-MS) pyrolysis and theoretical calculations. Two reaction channels were proposed in the pyrolytic reaction, by analyzing the detected pyrolytic products in the total ion chromatography, including 2,2,2-trichloroacetaldehyde, dimethyl phosphite, and dichlorvos. Theoretical calculations showed that there is an intramolecular hydrogen bond between the hydroxyl group and the phosphate O atom in trichlorfon, through which the hydroxyl H atom can be easily transferred to phosphate O atom to trigger two pyrolytic channels. In path-a, migration of H atom results in direct decomposition of trichlorfon to give 2,2,2-trichloroacetaldehyde and dimethyl phosphite in one step. In path-b, migration of H atom in trichlorfon is combined with formation of the O-P bond to give an intermediate, followed by HCl elimination to afford dichlorvos. Path-a is kinetically more favorable than path-b, which is consistent with the GC-MS results.

Destruction of halogen-containing pesticides by means of detonation combustion.

Pesticides that contain a halogen functional group have been destructed by means of detonative combustion. The following compounds were examined: (1) atrazine-2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine-herbicide; (2) bromophos-O,4-bromo-2,5-dichlorophenyl O,O-dimethyl phosphorothioate-insecticide; (3) chloridazon-5-amino-4-chloro-2-phenylopyridazin-3(2H)-one-herbicide; (4) linuron-3-(3,4-dichlorophenyl)-1-metoxy-1-methylurea-herbicide; (5) metoxychlor-1,1,1-trichloro-2,2-bis(4-metoxyphenyl)ethane-insecticide and acaricide; and (6) trichlorfon-dimethyl 2,2,2-trichloro-1-hydroxyethylphosphonate-insecticide. Explosive material has been produced on the basis of ammonium nitrate, which served as an oxidizer while the pesticides were used as fuels. Composition of the explosive was adjusted in such a way as to respect thermodynamic parameters. Detonative decomposition of the mixtures has been carried out in shot-holes pre-drilled in soil. Efficiency of the pesticide decomposition has been examined with gas chromatography in order to determine pesticides residues in the environment. It was found that for some, the amount of pesticides in some compounds in the analyzed samples after decomposition was below the determination threshold of the applied method.

Mechanism and kinetics study on the OH-initiated oxidation of organophosphorus pesticide trichlorfon in atmosphere.

Trichlorfon [O,O-dimethyl-(2,2,2-trichloro-1-hydroxy-ethyl) phosphonate] (TCF) is a kind of widely used organophosphorus pesticides. In this paper, the mechanism and possible oxidation products for the OH-initiated reactions of TCF are studied at the MPWB1K/6-311+G(3df,2p)//MPWB1K/6-31+G(d,p) level. The study shows that H abstraction reaction from the CH(3) group and the CH group as well as OH addition reaction to the P atom are energetically favorable for the reactions of TCF and the main products are (CH(3)O)(2)POOH (P1), CCl(3)CHOHPOOH(OCH(3)) (P2), CH(3)OPO(2) (P3), CCl(3)COPO(OCH(3))(2) (P6) and HCHO. On the basis of the quantum chemical information, the kinetic calculation is performed and the rate constants are calculated over a temperature range of 200-800K using the transition state theory and canonical variational transition state theory with small-curvature tunneling effect. The Arrhenius formulas of rate constants with the temperature are fitted and the lifetimes of the reaction species in the troposphere are estimated according to the rate constants, which can provide helpful information for the model simulation study.

Photoelectrocatalytic degradation of high COD dipterex pesticide by using TiO2/Ni photo electrode.

A TiO2 thin film electrode deposited on porous nickel net (TiO2/Ni) was prepared by the sol-gel method, and the surface morphology, crystal structure features and the grain size were characterized by Field emission scan electron microscopy (FESEM) and X-ray diffraction (XRD). The photoelectrocatalytic system was set up using a UV high-pressure mercury lamp as the light source, TiO2 coated on foamed nickel as photo anode, Pt sheet as counter electrode and the pesticide dipterex in synthetic wastewater. Various factors that influence the photoelectrocatalytic decomposition of dipterex pesticide have been studied, such as degradation time, the type of electrolyte, current density, original pH value and different degradation methods. The prepared catalysts were employed to photoelectrocatalytically degrade the pesticide dipterex under UV irradiation, comparing the results with photocatalytic degradation and electrochemical oxidation. The results indicated that under the optimal conditions of 0.02 mol/L NaCl as the supporting electrolyte, current density = 2.5 mA/cm2, pH 6.0 and dipterex pesticide 40 mg/L, and reaction time 2 hr, dipterex chemical oxygen demand (COD) removal rate and organophosphorous conversion of up to 82.6% and 83.5% were achieved, respectively. The method of photoelectrocatalytic degradation is more efficient than photocatalysis and electrochemical oxidation. The possible roles of the electrolytes on the reactions and probable mechanisms were also discussed.

Metrifonate alters antioxidant levels and caspase activity in cerebral cortex of Wistar rats.

Metrifonate (trichlorfon) is an inhibitor of acetylcholinesterase (AChE). It was used as an Alzheimer's disease (AD) drug; however, the application was withdrawn due to adverse effects. Implication of metrifonate for the antioxidant status and regulation of apoptotic processes was evaluated in the present study. Wistar rats (six per group) were exposed subcutaneously to either 60 or 120 mg/kg of body weight of metrifonate and compared with the controls treated with saline only. Cerebral cortex and liver tissues were collected from animals 40 min after exposure. Activities of AChE, glutathione reductase, glutathione-S-transferase, caspase 3, total protein level, thiobarbituric acid reactive substances, reduced glutathione level and ferric reducing antioxidant power (FRAP) were assayed in the tissue samples. Metrifonate had only lower impact on oxidative stress in the liver. Cerebral cortex tissues had decreased AChE and increased caspase 3 activities as well as the FRAP level. Owing to the novel findings, suitability of metrifonate for AD therapy is discussed.

Simultaneous determination of residues of trichlorfon and dichlorvos in animal tissues by LC-MS/MS.

Trichlorfon is a thermally unstable insecticide that can be easily decomposed to dichlorvos at high temperatures. In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of the residues of trichlorfon and dichlorvos in animal tissues. Trichlorfon and dichlorvos in animal tissues were extracted with dichloromethane, homogenized/shaken, concentrated, and determined by LC-MS/MS. The two insecticides can be effectively separated and individually determined. The limits of detection (LODs) of trichlorfon and dichlorvos were 0.04 and 0.07 microg kg(-1), respectively; the limits of quantification (LOQs) of the two insecticides were both 5 microg kg(-1). The average recoveries for three spiked blank samples at 10, 20 and 40 microg kg(-1) were in the range 85-106%, with relative standard deviations (RSDs) below 10.6%, which falls in a reasonable range for the analysis of the two insecticides in animal-derived foods. The method is fit-for-purpose for the simultaneous determination of residues of trichlorfon and dichlorvos in animal tissues.

High performance gold nanorods and silver nanocubes in surface-enhanced Raman spectroscopy of pesticides.

The behavior of Au nanorods and Ag nanocubes as analytical sensors was evaluated for three different classes of herbicides. The use of such anisotropic nanoparticles in surface-enhanced Raman scattering (SERS) experiments allows the one to obtain the spectrum of crystal violet dye in the single molecule regime, as well as the pesticides dichlorophenoxyacetic acid (2,4-D), trichlorfon and ametryn. Such metallic substrates show high SERS performance at low analyte concentrations making them adequate for use as analytical sensors. Density functional theory (DFT) calculations of the geometries and vibrational wavenumbers of the adsorbates in the presence of silver or gold atoms were used to elucidate the nature of adsorbate-nanostructure bonding in each case and support the enhancement patterns observed in each SERS spectrum.

In vitro reactivation of trichlorfon-inhibited butyrylcholinesterase using HI-6, obidoxime, pralidoxime and K048.

Trichlorfon is a specific inhibitor of cholinesterases. It was typically used as an insecticide; however, trichlorfon was described as useful for symptomatic treatment of Alzheimer's disease some years ago. The presented study is aimed at reactivation of trichlorfon-inhibited butyrylcholinesterase since this enzyme play an important role in Alzheimer's disease as deputy for acetylcholinesterase and furthermore it could be applied as a scavenger in case of overdosing. We used in vitro reactivation test for considering only reactivation efficacy of butyrylcholinesterase that is inhibited by trichlorfon and not reactivation of butyrylcholinesterase inhibited by trichlorfon metabolic products. Four reactivators were used: HI-6, pralidoxime, obidoxime, and K048. Although all of the reactivators seem to be effective at 1 mM concentration, a lower concentration was not able ensure sufficient reactivation. There was also an observed lowering of reactivation efficacy when butyrylcholinesterase was exposed to trichlorfon for a longer time interval.

Avaliação de risco de pesticidas aplicados no município de Arari, Maranhão, Brasil: base para programa de controle ambiental do Rio Mearim / Risk assessment of pesticides applied in Arari city, Maranhão, Brazil: basis for the environmental control program in Mearim river

Os objetivos deste trabalho foram avaliar a toxicidade aguda do sulfato de cobre e do triclorfon para três espécies de Daphnia (D. similiis, D. magna e D. laevis) na presença ou ausência de sedimento. Os valores de CE estimados para D. magna foram de 0,3496 mg de sulfato de cobre/L com sedimento e 0,0447 sem sedimento, para D. similiis 0,2859 com sedimento e 0,0426 sem sedimento e para D. laevis 0,1437 com sedimento e 0,1094 sem sedimento. A CE (50-48H) estimada para a D. magna foi de 299,70 ng de triclorfon/L com sedimento e 0,70 sem sedimento, para D. similiis 381,62 com sedimento e 0,52 sem sedimento e para D. laevis 282,72 com sedimento e 0,92 sem sedimento. O triclorfon mostrou-se mais tóxico que o sulfato de cobre para as tres espécies estudadas, sendo que a presença de sedimento diminuiu a toxicidade de ambos para as tres espécies na ausência de sedimento, a Daphnia laevis (espécie nativa do Brasil) pode ser usada como organismo-teste na avaliação da toxicidade aguda e crônica de inseticidas organofosforados e a base de cobre

Partitioning and persistence of trichlorfon and chlorpyrifos in a creeping bentgrass putting green.

Golf course putting greens typically receive high pesticide applications to meet high quality demands. Research on pesticide fate in turf ecosystems is important to better understand the potential impact of pesticide use on the environment and human health. This research was conducted to evaluate the environmental fate of two commonly used insecticides--trichlorfon (dimethyl 2,2,2-trichloro-1-hydroxyethylphosphonate) and chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridylphosphorothioate)--in a creeping bentgrass (Agrostis palustris Huds.) putting green under customary field management practices at the University of California-Riverside Turf Research Facility during 1996 and 1997. The two insecticides were chosen because of their difference in water solubility, persistence, adsorption, and vapor pressure. Volatilization, clipping removal, and soil residues of the insecticides were quantified and leaching was monitored using lysimeters installed in putting green plots. Results showed trichlorfon volatilization, clipping removal, and leaching loss was insignificant (in the range of 0.0001-0.06% of applied mass) both in 1996 and 1997. No significant difference in clipping removal of trichlorfon and chlorpyrifos was observed in both years (0.06 and 0.05% of applied mass for trichlorfon and 0.15 and 0.19% of applied mass for chlorpyrifos, respectively, in 1996 and 1997), but significantly lower cumulative leaching and lower soil concentration was observed in 1997 than in 1996. Volatilization loss of chlorpyrifos was not significantly different between 1996 (2.05%) and 1997 (2.71%). Volatilization loss of trichlorfon in 1996 (0.01%) was significantly higher than in 1997 (0.008%). This study demonstrated the fraction of applied insecticides leaving the turf putting greens was minimal.

Selective activity of butyrylcholinesterase in serum by a chemiluminescent assay.

In a previous study, we showed that purified commercial esterase activity can be detected in a chemiluminescent assay based on the hydrolysis of 2-methyl-1-propenylbenzoate (MPB) to 2-methyl-1-propenol, which is subsequently oxidized by the horseradish peroxidase (HRP)-H(2)O(2) system. The purpose of this study was to verify the applicability of this assay to human serum. The existence of an esterase activity capable of hydrolysing MPB is indicated by the fact that the MPB-serum-HRP-H(2)O(2) system consumes oxygen and emits light. Both signals were abolished by prior serum heat inactivation and were preserved when serum was stored at < or =4 degrees C. Addition of aliesterase inhibitors, such as fluoride ion and trichlorfon or the cholinesterase inhibitor eserine, totally prevents light emission. The butyrylcholinesterase-specific substrate benzoylcholine causes a delay in both O(2) uptake and light emission, while the specific acetylcholinesterase substrate, acetyl-beta-methylcholine, had practically no effect. Purified butyrylcholinesterase, but not acetylcholinesterase, triggered light emission. The finding that butyrylcholinesterase is responsible for the hydrolysis of MPB in serum should serve as the basis for the development of a specific chemiluminescent assay for this enzyme.

Residues of fenthion and trichloron in olives and olive oil after olive tree treatments.

The residue levels of trichlorfon, fenthion and its metabolites were determined in olives, olive oil and vegetation water after treatment of olive trees at different times before harvest. The highest residues of fenthion were detected in oil, while the highest level of trichlorfon was found in vegetation water. The time gap between treatment and harvest strongly influences the residue levels of fenthion in olives and olive products. The levels of fenthion, which were lower than the maximum residue level (MRL) established by the Italian regulations, were obtained only when the treatment was carried out 60 days before harvest, so it may be necessary to use the insecticide trichlorfon 30 or 10 days before harvest since it leaves low residue levels in oils.

Determination of metrifonate and dichlorvos in whole blood using gas chromatography and gas chromatography-mass spectrometry.

Analytical methods for determining metrifonate and dichlorvos in whole blood and a sampling procedure suitable for pharmacokinetic studies in man are described. Metrifonate concentrations were determined after chloroform extraction using gas chromatography-nitrogen-phosphorus detection. The within-assay coefficients of variation were 4 and 9% at 19.4 and 0.8 mumol/l (limits of determination), respectively. Dichlorvos was determined using gas chromatography-mass spectrometry of toluene extracts. The within-assay coefficients of variation were 2 and 5% at 225 and 50 nmol/l (limits of determination), respectively. Since both substances are chemically unstable, the blood was collected by dripping it directly from the vein into 0.74 M phosphoric acid.