Persistence, dissipation, and risk assessment of a combination formulation of trifloxystrobin and tebuconazole fungicides in/on tomato.

Multi-locational supervised field trials were conducted in different agro-climatic regions in India to study dissipation of trifloxystrobin and tebuconazole in tomato after spraying a combination formulation (trifloxystrobin 25% + tebuconazole 50%, 75WG) at recommended doses: (i) single (trifloxystrobin 87.5 g a.i. ha-1 + tebuconazole 175 g a.i. ha-1) and (ii) double (trifloxystrobin 175 g a.i. ha-1 + tebuconazole 350 g a.i. ha-1). Fruit samples were extracted with ethyl acetate using a modified QuEChERS method. The residues (parent fungicides + metabolite) were analyzed and confirmed by GC-ECD and GC-MS, respectively. The half-life (t1/2) of trifloxystrobin and tebuconazole in tomato varied from 1.08 to 1.72 and 1.13 -to 1.64 days at single; and 1.27 to 2.13 and 1.24 to 1.96 days at double dose, respectively. Since maximum residue limit (MRL) at pre-harvest interval (PHI) of 5 days is impractical, as tomato is usually harvested and consumed almost everyday after the last spray, the risk assessment was performed at minimum PHI of 1 day. Accordingly, on the basis of supervised field trial data and using OECD MRL calculator, MRL of 0.5 and 1.5 mg kg-1 at single dose were proposed for trifloxystrobin and tebuconazole in/on tomato, respectively.

Persistence, dissipation and consumer risk assessment of a combination formulation of flubendiamide and deltamethrin on cucumber.

Multi-location supervised field trials were conducted in India at four locations of the All India Network Project (AINP) on Pesticide Residues to study the persistence, dissipation and risk assessment of flubendiamide and deltamethrin on cucumber (Cucumis sativus). Residues of flubendiamide and deltamethrin on cucumber resulting from three spray applications of a combination formulation (flubendiamide 90% + deltamethrin 60%, 150 SC) at recommended (22.5 + 15 g a.i./ha) and double the recommended (45 + 30 g a.i./ha) dose were analysed. On the basis of persistence and dissipation studies, the half- life (T1/2) of flubendiamide on cucumber varied from 1.40 to 2.98 (recommended dose) and 1.55 to 2.76 days (double the recommended dose), while that of deltamethrin ranged from 2.5 to 4.9 (recommended dose) and 2.7 to 3.9 days (double the recommended dose) at the four locations. On the basis of supervised field trial data and using OECD calculator, MRLs in the combination product of 3 mg kg-1 for flubendiamide and 1.5 mg kg-1 for deltamethrin has been proposed for consideration by the Food Safety and Standards Authority of India (FSSAI). Codex, EU and EPA have fixed MRL of 0.2 mg kg-1 for flubendiamide and deltamethrin.

Development of chitosan nanocapsules for the controlled release of hexaconazole.

Accelerated use of pesticides in cutting edge agriculture prompted us to explore smart nanoformulations to subside the consumption of these perilous chemicals. Polymer nanocapsules carrying a fungicide, hexaconazole were developed through ionotropic gelation method utilizing chitosan and tripolyphosphate (TPP). The nanocapsules were characterized by photon correlation spectroscope (PCS), transmission electron microscope (TEM), and Fourier transform infra-red (FTIR) spectroscope. Nanocapsules were optimized for size and high encapsulation efficiency using central composite design (CCD) software. The encapsulation efficiency of nanocapsules for hexaconazole was 73% as assessed by gas chromatography (GC). Nanocapsules were analysed and compared with commercial formulation for controlled release in vitro at three different pH values. Release of hexaconazole from nanocapsules was fastest at pH 4 in comparison to pH 7 and pH 10. Release study in soil was also conducted and revealed a controlled pattern for nanoformulation. The fungicidal activity of the prepared nanoformulation was evaluated against R. solani and was compared with commercial formulation of hexaconazole. The cytotoxicity assay performed on vero cell lines by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay confirmed that nanoformulation is less toxic than commercial formulation of pesticide.

Effect of Nanohexaconazole on Nitrogen Fixing Blue Green Algae and Bacteria.

Nanohexaconazole is a highly efficient fungicide against Rhizoctonia solani. Nanoparticles are alleged to adversely affect the non-target organisms. In order to evaluate such concern, the present study was carried out to investigate the effect of nanohexaconazole and its commercial formulation on sensitive nitrogen fixing blue green algae (BGA) and bacteria. Various activities of algae and bacteria namely growth, N-fixation, N-assimilation, Indole acetic acid (IAA) production and phosphate solubilization were differently affected in the presence of hexaconazole. Although, there was stimulatory to slightly inhibitory effect on the growth measurable parameters of the organisms studied at the recommended dose of nanohexaconazole, but its higher dose was inhibitory to all these microorganisms. On the other hand, the recommended as well as higher dose of commercial hexaconazole showed much severe inhibition of growth and metabolic activity of these organisms as compared to the nano preparation. The uses of nanohexazconazole instead of hexaconazole as a fungicide will not only help to control various fungal pathogens but also sustain the growth and activity of these beneficial microorganisms for sustaining soil fertility and productivity.

Development and evaluation of alginate-chitosan nanocapsules for controlled release of acetamiprid.

Smart formulations based on nanomaterials have the capability to reduce the consumption of hazardous pesticides and their impact on human health and environment. Nanoformulations of agrochemicals have the potential to improve food productivity without compromising with the ecosystem. In the present work, controlled release nanocapsules containing acetamiprid were prepared by polyelectrolyte complexation of two natural macromolecules, i.e. alginate and chitosan. The size, morphology and chemical interaction studies of the prepared nanocapsules were investigated by Dynamic Light Scattering (DLS), Transmission Electron Microscopy (TEM), and Fourier Transform Infrared Spectroscopy (FTIR). The zetapotential studies revealed stability of the nanocapsules. TEM results show spherical morphology of the nanocapsules. The encapsulation efficiency was found to be 62% as quantified by Ultra High Pressure Liquid Chromatography (UHPLC). Nanocapsules were analysed for controlled release in vitro at three different pH. Maximum release was observed at pH 10 followed by pH 7 and 4, respectively. A non-Fickian release mechanism was found to be followed by the nanoformulation. A controlled release pattern was also found from nanoformulation as compared to commercial formulation in soil. Thus this formulation can reduce the frequency of application of pesticides by controlling the release and will subsequently reduce their side effects.

Development and Quality Control of Nanohexaconazole as an Effective Fungicide and Its Biosafety Studies on Soil Nitifiers.

The study was aimed to develop a nano form of an existing fungicide for improving plant protection and reducing crop losses caused by fungal pathogens. The protocol for the preparation and estimation of nanohexaconazole was developed. Technically pure hexaconazole was converted into its nanoform using polyethyleneglycol-400 (PEG) as the surface stabilizing agent. Nanohexaconazole was characterized using Scanning Electron Microscopy (SEM) and Dynamic Light Scattering (DLS) studies. The average particle size of nanohexaconazole was about 100 nm. An analytical method was also developed for quality control of the nanofungicide by GLC fitted with flame ionization detector. Its limit of detection was 2.5 ppm. Fungicidal potential of nanohexaconazole was better in comparison to that of conventional hexaconazole. Hydrolytic and thermal stability studies confirmed its stability at par with the conventional formulation of fungicide. Impact of nanohexaconazole on soil nitrifiers was tested in vitro and there were no significant adverse effect in their numbers observed as compared to conventional registered formulation, proving the safety of the nanofungicide.

Bioefficacy evaluation and dissipation pattern of nanoformulation versus commercial formulation of pyridalyl in tomato (Solanum lycopersicum).

A study was undertaken to evaluate the decline of the residues of pyridalyl in tomatoes grown in two different cultivation systems: open field (conventional cultivation) and net house (pot experiment). Field experiment was conducted with commercial (10 EC) and nanoformulation of pyridalyl on tomato crop. Tomato plants were subjected to pesticide spray, when fruits were close to ripeness. Fruit samples were taken periodically and cleaned up using QuEChERS methodology, and the residue of pyridalyl was analyzed by ultrahigh-performance liquid chromatography (UHPLC). It dissipated in tomato fruit following the first-order kinetics. In field, average initial deposit of pyridalyl in tomato was observed to be 0.222 and 0.371 µg g(-1) at recommended and double the recommended application rate, respectively, using nanoformulation while it was 0.223 and 0.393 µg g(-1) on using commercial formulation, respectively. The half-life (t1/2) value of nanopyridalyl in tomato fruit was 2.8 and 3.2 days while for commercial formulation, it was 2.5 and 2.6 days for recommended and double the recommended dose, respectively. In India, maximum residue limit (MRL) on tomato has not been fixed for pyridalyl but its residues were always below European MRLs (5 µg g(-1)) on tomato at both application rates. The results of terminal residue showed that pyridalyl residues were below the available MRL. Low residues in tomatoes suggested that this pesticide is safe to use under the recommended dosage. No statistical differences were observed between the cultivation systems in relation to the residue levels of pyridalyl.

Residue, dissipation, and safety evaluation of pyridalyl nanoformulation in Okra (Abelmoschus esculentus [L] Moench).

A comparative study on residues and rate of dissipation of a new nanoformulation of pyridalyl and commercial pyridalyl was carried out on okra under net house and field conditions. Okra crop was sprayed with commercial and nanoformulation of pyridalyl at recommended (75 g a. i./ha) and double the recommended dose (150 g a. i./ha) at the time of fruiting. Quantitation of residues of pyridalyl in okra was done by ultra high performance liquid chromatography over a period of 15 days, and recovery of the method ranged from 79 to 87 %. The half life calculated from the dissipation pattern of pyridalyl on okra for commercial and developed nanoformulation proved that residues of nanopyridalyl did not persist much longer than that of conventional formulation in net house as well as in field trials. The risk quotient value of pyridalyl in okra was significantly lower than 1, implying its negligible risk to the humans.

Degradation of sulphonated azo dye Red HE7B by Bacillus sp. and elucidation of degradative pathways.

Bacteria capable of degrading the sulfonated azo dye Red HE7B were isolated from textile mill effluent contaminated soil. The most efficient isolate was identified as Bacillus sp. Azo1 and the isolate could successfully decolorize up to 89% of the dye. The decolorized cultural extract analyzed by HPLC confirmed degradation. Enzymatic analysis showed twofold and fourfold increase in the activity of azoreductase and laccase enzymes, respectively, indicating involvement of both reductive and oxidative enzymes in biodegradation of Red HE7B. Degraded products which were identified by GC/MS analysis included various metabolites like 8-nitroso 1-naphthol, 2-diazonium naphthalene. Mono azo dye intermediate was initially generated from the parent molecule. This mono azo dye was further degraded by the organism, into additional products, depending on the site of cleavage of R-N=N-R molecule. Based on the degradation products identified, three different pathways have been proposed. The mechanism of degradation in two of these pathways is different from that of the previously reported pathway for azo dye degradation. This is the first report of a microbial isolate following multiple pathways for azo dye degradation. Azo dye Red HE7B was observed to be phytotoxic, leading to decrease in root development, shoot length and seedling fresh weight. However, after biotreatment the resulting degradation products were non-phytotoxic.

Sensitive and rapid determination of elemental nanosulfur/sulfur by liquid chromatography.

In order to identify the most suitable method for the estimation of nanosulfur for studying its residue dynamics, the present work was taken up. HPLC and GC methods were explored for its analysis. A comparative study of the existing analytical methods for the quality control of nanosulfur was undertaken. UV spectrophotometry and HPLC methods were superior with lower LOD when compared to GC-MS, which was not satisfactory due to breakage of catenated S20 into S6 and S8 . The method has been validated by analyzing various nanosulfur formulations of known concentrations. The recovery of the UV and HPLC methods ranged from 80.71 to 109.51% and 82.31 to 109.84%, respectively. The LOD of UV, GC-MS, and HPLC is 4, 20, and 1 ppm, respectively. The retention time of sulfur was 13.77 (HPLC), 2.89 (ultra high performance liquid chromatography), and 12.715 + 21.524 min (GC-MS). The method was successfully utilized for estimating sulfur in natural samples such as water from a sulfur hot spring and wastewater. The method has been validated by following the method recommended by the American Society for Testing and Materials. The HPLC method emerged as the best analytical method for the estimation of elemental sulfur.

Comparative persistence of thiacloprid in Bt-transgenic cabbage (Brassica oleracea cv. capitata) vis-à-vis non-transgenic crop and its decontamination.

Thiacloprid is a systemic neonicotinoid. The study hypothesized that difference may be seen in the rate of dissipation of thiacloprid when applied on non-transgenic and transgenic cabbage. Thiacloprid was estimated by HPLC. Half life of thiacloprid in transgenic as well as in normal cabbage ranged between 12.3-13.1 days in two doses of application. Under field condition, after 15 days, 59.2% and 54.3% dissipation was recorded at lower and higher rates of application in transgenic cabbage, where as the insecticide dissipated 57.5% and 59.1% for single dose and double dose application, respectively in non-transgenic cabbage. The study establishes that there is no significant difference in dissipation of a systemic pesticide in transgenic versus non-transgenic cabbage. Decontamination of thiacloprid contaminated cabbage was carried out by different chemical treatments. The application of 0.5% NaHCO(3) (an edible alkali) may be recommended for decontamination. Thiacloprid residues in the day-3 field samples of cabbage could be reduced below Japanese MRL (1.0 mg kg(-1)) by treating with 0.5% NaHCO(3) solution for 1 h.

Synthesis of 2-furyl-4-arylidene-5(4H)-oxazolones as new potent antibacterial agents against phyto-pathogenic and nitrifying bacteria.

Crop losses due to bacterial pathogens are a major global concern. Most of the available pesticides for these pathogens suffer from various drawbacks such as complicated synthesis, high cost, high toxicity, pesticide resistance and environmental hazards. To overcome these drawbacks, the present study was undertaken to find a potent bactericide. Therefore, a series of compounds comprising bioactive furyl and oxazolone rings was synthesized under microwave irradiation and screened for in vitro antibacterial activity. The reactions were completed in fewer than 2 minutes with minimal use of solvents and resulted in high yields. These compounds were screened for antibacterial activity against plant pathogens, Xanthomonas oryzae, Ralstonia solanacearum and nitrifying bacteria, Nitrosomonas species under laboratory conditions. Five compounds were active as antibacterial agents against Xanthomonas oryzae and Ralstonia solanacearum. However, all compounds were effective against the Nitrosomonas species and the best one was 2-furyl-4-(3-methoxy-4-hydroxybenzylidene)-5(4H)-oxazolone. The study revealed the fast and environmentally friendly synthesis of bioactive title compounds, which also hold promise to be used as prototypes for the discovery of potent analogues.

Isolation and characterization of alkalotolerant Pseudomonas sp. strain ISTDF1 for degradation of dibenzofuran.

An alkalotolerant Pseudomonas strain was enriched and isolated from effluent of the pulp and paper industry. This strain was able to degrade dibenzofuran and utilize it as a sole source of energy and carbon. The GC-MS based detection of various intermediary metabolites of biodegradation suggested the involvement of angular as well as lateral pathway of dibenzofuran biodegradation. The GC-MS based detection of various intermediary metabolites of biodegradation suggested the involvement of angular as well as lateral pathway of dibenzofuran biodegradation. This diverse dioxygenation property of the strain allowed it to utilize various recalcitrant chlorinated xenobiotics and PAHs compounds. This strain showed optimum utilization (~85%) of dibenzofuran (200 mg l⁻¹) within 36 h at pH 10 at 40 °C. The growth of the strain was supported by a wide range of environmental conditions such as temperature, pH, and concentration of dibenzofuran, suggesting that it can be used for in situ bioremediation of dioxin-like compound.

Evaluation of performance and community dynamics of microorganisms during treatment of distillery spent wash in a three stage bioreactor.

The ability of Emericella nidulans var. lata, Neurospora intermedia and Bacillus sp. to treat distillery spent wash in a three stage bioreactor was investigated. Process parameters were optimized in shake flask cultures with the individual strains before treatment of the effluent in a 15-l bioreactor. Treatment was first carried out by the fungi followed by bacteria. The treated effluent showed significant reduction in color (82%) and COD (93%) after 30 h. Metabolites formed after degradation of complex polymers in distillery effluent were assayed by gas chromatography-mass spectroscopy and included furan, simple acid types and organic compounds. Denaturing gradient gel electrophoresis of 16S rDNA and 18S rDNA sequences amplified from DNA isolated from the reactor communities indicated the presence of other organisms besides those introduced initially. The microbial communities were able to carry out bioremediation of distillery effluent and produce discharge that conforms to safety standards.

Determination of pesticide residues in integrated pest management and nonintegrated pest management samples of apple (Malus pumila Mill.).

Studies were undertaken to analyze the residues of commonly used pesticides viz. chlorpyrifos, endosulfan, dicofol, cypermethrin, fenvalerate, propargite, malathion, phorate, carbendazim, carbosulfan, thiamethoxam, and mancozeb in apple of integrated pest management (IPM) and non-IPM samples collected from the IPM and non-IPM fields of Shimla. We also present a method for the determination of these pesticides in apple samples. Residues of chlorpyrifos, endosulfan, dicofol, cypermethrin, fenvalerate, and propargite were analyzed by gas chromatography, while residues of carbendazim, carbosulfan, and thiamethoxam were analyzed by high-performance liquid chromatography. Residues of mancozeb were determined by a colorimetric method. Recoveries of all of the pesticides ranged from 61.30 to 95.46% at 0.1, 0.2, and 1.0 microg g(-1) levels of fortification with relative standard deviations ranging between 0.8 and 8.7. Apples from IPM and non-IPM orchards were analyzed for these pesticides using a developed method. Except for carbendazim and chlorpyrifos, the residues of all of the pesticides analyzed were below detectable limits. Although residues of carbendazim and chlorpyrifos were below the prescribed limits of maximum residue levels in both IPM and non-IPM orchards, residues were lower in apples from IPM orchards.

An analysis of the persistence and potency of film-coated seed protectant as influenced by various storage parameters.

BACKGROUND: An efficient delivery system for seed-protectant chemicals is needed in light of several disadvantages of conventional seed treatment methods. This study evaluates the efficacy of film-coat application in maintaining the persistence and potency of imidacloprid on Lycopersicon esculentum (L.) Mill. seeds after simultaneous storage under ambient and regulated environment in paper and aluminium packages. RESULTS: High-performance liquid chromatography (HPLC) revealed 0.135 mg kg(-1) of herbage material to be the threshold value beyond which absolute control was obtained, and with film coating the latter was achieved even with half-dosage seed treatment, irrespective of the storage condition. The technique provided early protection to the crop and also nullified the deleterious effects of ambient storage on the persistence and potency of the pesticide. CONCLUSION: Film coating enabled superior pesticide dosage as well as higher biological efficacy to be achieved. Hence, in addition to being an ecofriendly alternative, the technique would be a more economically viable option for storage of treated seeds.

Application of indoxacarb for managing shoot and fruit borer of eggplant (Solanum melongena L.) and its decontamination.

Indoxacarb was applied at 75 and 150 g a.i. ha(-1) for two years to an eggplant (Solanum melongena L.) crop grown in the field plots in order to evaluate its efficacy for management of the lepidopteron pest, shoot and fruit borer. The residues of the insecticide were quantified by high-pressure liquid chromatography (HPLC). The mean initial deposits of indoxacarb on eggplant fruits were found to be 2.60-2.634 mg kg(-1) and 3.64-3.68 mg kg(-1) from the two rates of applications, respectively. They declined with time and reached to non-detectable (< 0.02 mg kg(-1)) after 15-20 d. Residues dissipated with a half-life of 3.0-3.8 d from both first and second-year application. A 3 d waiting period for harvest of fruits after insecticide application and processing resulted in the residue levels that were below the Codex maximum residue limit (MRL) of 0.5 mg kg(-1) thereby achieving a maximum safety and minimum risk to consumers. The best combination of chemicals for decontamination of indoxacarb was found to be by washing with a mixture of alkali and potassium permanganate (KMnO(4)) thereby resulting in the removal of 67.5% and 59.2 % residues for 5 and 10 microg g(-1) spiking doses, respectively. Major products formed on reaction of indoxacarb with alkali were identified by electron spray ionization chromatography/mass spectrometry (ESI/MS). The per cent reduction on the weight and number basis of treated eggplant plots were compared to those observed in control plots to demonstrate the effectiveness of indoxacarb treatment on shoot and fruit borer population.

Risk assessment of thiacloprid and its chemical decontamination on eggplant, Solanum melongena L.

BACKGROUND: Thiacloprid [(Z)-3-(6-chloro-3-pyridylmethyl)-1,3-thiazolidin-2-ylidenecyanamide; Calypso] is a systemic insecticide having persistence in the plant system. It was chosen for the management of the eggplant shoot and fruit borer, Leucinodes orbonalis Guen. Management of this insect pest is difficult because it harbours inside the shoot and fruit portions of eggplant. The persistence of thiacloprid on eggplant has not been studied in India. The Food and Agriculture Organisation (FAO) has proposed its maximum residue limit (MRL) on eggplant as 0.7 mg kg(-1), and there is a need to validate this value. Since residues were found to be above this level, five different decontamination agents were tested for the decontamination of thiacloprid from eggplant. RESULTS: The half-life of thiacloprid was 11.1 and 11.6 days from trials in 2 years. Safety factors such as theoretical maximum daily intake (TMDI) and maximum permissible intake (MPI) were used to arrive at a risk assessment to human health from the analytical data obtained from the field trials. Thiacloprid at the doses tested (30 and 60 g AI ha(-1)) was not effective in managing eggplant fruit borer. A waiting period of 3 days before harvest of the fruits after insecticide application and a processing factor (PF) could not ensure a sufficient margin of safety (MOS). Subjecting the data to a processing factor of 60% could not bring the residues below the proposed MRL. CONCLUSION: Thiacloprid is not found to be an appropriate and effective agent for application to eggplant. Either the proposed MRL needs to be revised or good agricultural practice involving thiacloprid for plant protection in eggplant cultivation is required.

Analytical method for estimation of a new insecticide flubendiamide and its safety evaluation for usage in rice crop.

The paper presents a method for residue analysis of flubendiamide in rice (Oryza sativa), which includes improved extraction, cleanup and determination of flubendiamide in rice seeds, husk and straw by using LC with UV detection. Safety evaluation of this insecticide in rice has been carried out after applying its soluble concentrate (SC) formulation at recommended dose (30 g a.i. ha(-1)) and double of the recommended dose (60 g a.i. ha(-1)) on rice crop. MRL of flubendiamide on rice grain can be proposed as 0.2 mg kg(-1).

Determination of pesticide residues in IPM and non-IPM samples of mango (Mangifera indica).

Studies were conducted to analyze the residue of commonly used pesticides viz. methyl parathion, chloropyrifos, endosulfan, cypermethrin, fenvalerate, carbendazim, imidacloprid and carbaryl in mango, Dashehari variety, integrated pest management (IPM) and non-IPM samples were collected from the IPM and non-IPM orchards, Lucknow, India. We also present a method for the simultaneous determination of these pesticides in mango samples. Residues of methyl parathion, chloropyriphos, endosulfan, cypermethrin, fenvalerate were extracted from the samples with acetone: cyclohexane: ethyl acetate in the ratio 2:1:1 followed by cleanup using neutral alumina. Analysis was performed by gas chromatography-electron capture detector (GC-ECD) with a megabore column (OV-1). Residues of carbendazim, imidacloprid and carbaryl were extracted with acetone and after cleanup, analysis was performed by high performance liquid chromatography (HPLC) using photo diode array (PDA) detector. Recoveries of all the pesticides ranged between 72.7-110.6%, at 0.1 and 1.0 microg g(-1) level of fortification. The residues detected in non-IPM samples of mango were found to be below the prescribed limits of maximum residue limit (MRL) while IPM samples were free from pesticide residues.