Imazalil Research Articles

Comparison of Imazalil Removal onto Montmorillonite and Nanomontmorillonite and Adsorption Surface Sites Involved: An Approach for Agricultural Wastewater Treatment

To remove fungicide Imazalil (IMZ) (commercial formulation) from agricultural wastewater, adsorption was performed and compared among montmorillonite (Mt) and four organo-Mts (OMts), with different octadecyltrimethylammonium (ODTMA) loadings. The agglomerates size increase of OMts with respect to Mt found encourages the use of the former in agricultural wastewater treatment. Characterization of the OMts adsorbents by powder X-ray diffraction, infrared spectroscopy, and thermogravimetric analyses, indicated that ODTMA was present in the interlayer with different molecular arrangements, a diminution of the thermal stability of the OMts and an increase of ODTMA loading being observed at the same time. Zeta potential measurements showed electric surface charge reversion from negative to positive when the surfactant loading exceeded 100% of the clay cation exchange capacity (CEC). The IMZ adsorption on the raw Mt showed high pH dependence and affinity toward the fungicide, while OMts showed best IMZ adsorption capacity at high IMZ concentrations inversely related with the ODTMA loading. The characterization of IMZ–adsorbed samples indicated that for IMZ–Mt the interaction between IMZH+ and the siloxane surface was electrostatic. For IMZ–OMt samples the presence of new XRD and FTIR peaks suggested different IMZ interlayer arrangements and direct interaction with the surface, respectively. Analysis of zeta potential measurements indicated synergetic effects between the ODTMA and IMZ molecules at the external surface.

Rapid Determination of Seven Fungicides in Citrus Fruits

A rapid and simple determination method of seven fungicides, thiabendazole (TBZ), pyrimethanil (PYR), o-phenylphenol (OPP), fludioxonil (FLD), azoxystrobin (AZX), imazalil (IMZ) and diphenyl (DP) in citrus fruits by LC-MS and HPLC-FL was developed. The seven fungicides were extracted with acetonitrile from citrus fruits and cleaned up with Z-Sep/C18 cartridges. The LC separation was performed on a phenyl-hexyl column with methanol-acetonitrile-10 mmol/L ammonium formate (10 : 35 : 55) as a mobile phase. The recoveries from citrus fruits fortified with the compounds at the MRLs and at 0.1 μg/g ranged from 85.4 to 106.3% and from 75.8 to 99.7%, respectively. The quantitation limits (S/N=10) were 0.03-0.07 μg/g.

Curative control of citrus green mould by imazalil as influenced by infection age, wound size, fruit exposure time, solution pH and fruit brushing after treatment

Green mould (Penicillium digitatum) is a major cause of postharvest losses in citrus. Wounds and infection can be inflicted during the harvest process and should be controlled during postharvest stages to prevent decay. Imazalil (IMZ) in the sulphate formulation is currently applied by the majority of South African packhouses through an aqueous dip treatment. In this study the effects of incubation time (infection age), exposure time, solution pH, wounds size and fruit brushing after dip treatments on residue loading and curative green mould control were investigated. Exposure time did not have a significant effect on residue loading on fruit dipped in pH 3 solutions of IMZ (<2.00μgg−1). Increasing the pH to 6 resulted in significantly increased residue loading, which increased with longer exposure time, but mostly to levels below the maximum residue level of 5μgg−1 after 180s. Post-dip treatment brushing reduced residue levels obtained in IMZ pH 3 solutions by up to 90% to levels <0.5μgg−1; however, curative control of the IMZ sensitive (S) isolate was mostly unaffected, but with poor sporulation inhibition. At pH 6, post-dip brushing reduced residues to ≈60%; again curative control of the sensitive isolate was unaffected, but with better sporulation inhibition than the pH 3 treatments. Wounded rind sections loaded higher residue levels compared with intact rind sections, and large wounds loaded higher levels than small wounds (≈10.19, ≈9.06 and ≈7.91μgg−1 for large, small and no wound, respectively). Curative control of infections originating from large wounds was significantly better than those from small wounds. The ability of IMZ to control sensitive green mould infections declined from 6 and 12h after inoculation on Clementine mandarin fruit of infections inflicted by small and large wounds, respectively; on navel orange fruit, curative control declined 18 and 36h after inoculation for the respective wound size treatments. This work shows the importance of timely fungicide treatment after harvest especially on more susceptible citrus types. Results also indicate that excess residues can be stripped from the fruit, retaining residues necessary for curative control in the wound sites. However, reduced residue loading compromised the sporulation inhibition activity of IMZ.

Antifungal activity of sodium propylparaben alone or in combination with low doses of imazalil against Penicillium decay on citrus fruit

The performance of postharvest treatments with sodium propylparaben (SPP), alone or combined with low doses of the fungicide imazalil (IMZ), against citrus green (GM) and blue (BM) molds was evaluated on several citrus species and cultivars artificially inoculated with Penicillium digitatum and P. italicum, respectively, and incubated at 20 °C or cold-stored at 5 °C. Effectiveness of 100 mM SPP dips at 20 °C for 60 s was higher on oranges than on mandarins, with GM and BM incidence reductions of up to 60–90 % after 7 days at 20 °C. Irrespective of citrus cultivar and storage condition, SPP generally improved the curative action of 25 μl l−1 IMZ to control Penicillium molds. In additional tests, 100 mM SPP dips at 20 °C for 60 s only prevented GM on ‘Valencia’ oranges inoculated 24 h after treatment when combined with IMZ. It can be concluded that postharvest SPP treatments show promise as an effective alternative to be considered in citrus postharvest disease control programs.

Phenomenological study and application of the combined influence of iron concentration and irradiance on the photo-Fenton process to remove micropollutants

The presence of low concentrations of persistent pollutants in waters (μg/L or ng/L), also called micropollutants, brings as a consequence the need to apply advanced oxidation treatments for their removal. The successful application of solar-driven photo-Fenton to treat highly polluted wastewaters (g/L and mg/L of pollutants) has prompted its application to lowly polluted effluents. However, a decrease in contaminant concentration may involve an alteration in the intrinsic process phenomenon, which until now has only been widely studied at the milligram-per-litre level or higher with this process. The aim of this research was to study the combined influence of the operating variable (iron concentration) and the environmental variable (irradiance) and application on the photo-Fenton process at pH2.8 when removing micropollutants. For this purpose, experimentation was carried out at laboratory and pilot plant scales with a biocide mixture of acetamiprid (ACTM), thiabendazole (TBZ) and imazalil (IMZ) (100μg/L each) as the model pollutant. Results indicated that above 15WUV/m2 and a light path length of 5cm (the most commonly used path for this type of application) iron concentration limited the process and there was irradiance excess under these conditions. On the other hand, and given the circumstances of irradiance excess, a higher light path length (10cm) was assessed. Results showed that path lengths wider than 5cm are recommended since more wastewater volume could be treated with a higher process rate per surface unit.

Effects of citrus wax coating and brush type on imazalil residue loading, green mould control and fruit quality retention of sweet oranges

Wax application plays an important role in prolonging fruit quality, and the addition of imazalil (IMZ) furthermore protects fruit against green mould caused by Penicillium digitatum. The objectives of this study were to evaluate green mould control and quality preservation effects of carnauba or polyethylene citrus coatings supplemented with IMZ, as well as the effect of synthetic or horsehair brush types used on sweet orange fruit. Single applications of IMZ at 3000μgmL−1 at rates of 0.6, 1.2 and 1.8Lt−1 resulted in residues that increased with increasing coating loads on navel oranges (1.31 to 3.32μgg−1) and Valencia oranges (3.22 to 6.00μgg−1). Coating with IMZ generally provided poorer curative control (≈14%) than protective control (≈58%), with less sporulation in treatments using horsehair (≈59%) than synthetic brushes (≈64%). More fruit weight and firmness losses were found in fruit treated with the polyethylene coating (≈1.18 and ≈0.93 ratios of treated vs. untreated, respectively) and lower in carnauba treated fruit (≈0.76 and ≈0.74 ratios, respectively). However, polyethylene coatings resulted in shinier fruit before (≈10.85 shine ratio) and after storage (11.60), whereas carnauba coatings resulted in lower shine ratios (≈7.45 and 10.15, respectively). Gas (CO2) exchange ratios remained similar for both waxes (≈0.67). Higher polyethylene coating loads (1.8Lt−1) resulted in off-tastes similar to uncoated control fruit (≈2.21 rating on a 5-point scale) and higher than the rating for carnauba coated fruit (≈1.82) at this rate. Scanning electron micrographs showed an amorphous crystallised natural wax layer with uncovered stomatal pores on the surface of uncoated fruit. The thickness of the applied wax layer increased with increasing coating load. A single application of IMZ in wax provided good protective green mould control and sporulation inhibition, with differing effects on some fruit quality parameters due to coating and brush types.

A new method for the postharvest application of imazalil fungicide to citrus fruit

Fungal infections contribute significantly to postharvest losses of citrus fruits. Imazalil (IMZ) is currently the most commonly used fungicide that is effective for controlling postharvest fungal pathogens in citrus. IMZ is currently used in Italy as a wax emulsion for application by spraying. This system simplifies treatment, but it requires high IMZ concentrations to be effective, which result in large amounts of fungicide per unit mass of citrus fruit being applied.

Use of Phosphite Salts in Laboratory and Semicommercial Tests to Control Citrus Postharvest Decay.

Potassium phosphite (KP) concentrations that inhibited the germination of 50% of Penicillium digitatum conidia were 229, 334, 360, 469, 498, or 580 mg/liter at pH 3, 4, 5, 6, 7, or 8, respectively. Increasing phosphate content in media reduced phosphite toxicity. To control green or blue mold, fruit were inoculated with P. digitatum or P. italicum, then immersed 24 h later in KP, calcium phosphite (CaP), sodium carbonate, sodium bicarbonate, or potassium sorbate for 1 min at 20 g/liter for each at 25 or 50°C. Mold incidence was lowest after potassium sorbate, CaP, or KP treatments at 50°C. CaP was often more effective than KP but left a white residue on fruit. KP was significantly more effective when fruit were stored at 10 or 15°C after treatment compared with 20°C. Acceptable levels of control were achieved only when KP was used in heated solutions or with fungicides. KP was compatible with imazalil (IMZ) and other fungicides and improved their effectiveness. KP increased thiabendazole or IMZ residues slightly. Phosphite residues did not change during storage for 3 weeks, except they declined when KP was applied with IMZ. KP caused no visible injuries or alteration in the rate of color change of citrus fruit in air or ethylene at 5 μl/liter.

Evaluation of curative and protective control of Penicillium digitatum following imazalil application in wax coating

Imazalil (IMZ) is widely used in citrus packhouses to manage green mould, caused by Penicillium digitatum. The aim of this study was to investigate green mould control efficacy of IMZ applied in a wax coating, and the combination of aqueous dip and coating IMZ applications. Single application of IMZ at 3000μgmL−1 in carnauba wax coating at rates of 0.6, 1.2 and 1.8Ltonne−1 of fruit gave better protective (mean 13% infection) than curative (mean 70% infection) control of the sensitive isolate. Imazalil residue levels increased (0.85 to 1.75μgg−1) with increasing coating load. However, the resistant isolate could not be controlled (>74% infection). Dip only treatment (IMZ sulphate at 500μgmL−1 for 45s and 90s) gave good curative control (≈77%) of the sensitive isolate at residue loading of 0.12–0.73μgg−1. Wax coating only treatment (IMZ at 3000μgmL−1 at 1.8L wax tonne−1) gave good protective control and improved sporulation inhibition (≈80%) at residue loading of 1.32–7.09μgg−1. The MRL of 5μgg−1 was exceeded at higher wax loads on navels and clementines. Double application with dip (45s in IMZ sulphate at 500μgmL−1) followed by 2000μgmL−1 IMZ in wax coating at 0.6, 1.2 and 1.8L wax tonne−1 resulted in residue loading of 1.42 to 2.83μgg−1, increased protective control (≈69%) as well as curative control (≈83%). In all treatments, poor curative and protective control of the resistant isolate was observed (<46% and <55%, respectively). Double application demonstrated superior green mould control by giving good curative and protective control and sporulation inhibition.

Imazalil residue loading and green mould control on citrus fruit as affected by formulation, solution pH and exposure time in aqueous dip treatments

Green mould, caused by Penicillium digitatum, is responsible for major postharvest fruit losses on the South African fresh citrus export market. Some of these losses as well as fungicide resistance development can be attributed to sub-optimal imazalil (IMZ) residue loading on citrus fruit (<2μgg−1), which is commonly the case in South African packhouses. This will result in loss of control and sporulation inhibition on decayed fruit. IMZ formulation [IMZ sulphate and emulsifiable concentrate (EC)], solution pH (IMZ sulphate at 500μgmL−1 buffered with NaHCO3 or NaOH to pH 6 and 8) and exposure time (15–540s) were investigated in order to improve IMZ residue loading and the green mould control on Clementine mandarin, ‘Eureka’ lemon, and navel and Valencia orange fruit. Exposure time had no significant effect on residue loading in the unbuffered IMZ sulphate solution (pH 3). No differences were observed between the pH buffers used, but residue loading improved with increase in pH. The maximum residue limit (MRL) of 5.0μgg−1 was exceeded following dip treatment in the IMZ EC (after 75s exposure time), and IMZ sulphate at pH 8 using NaHCO3 (77s) or NaOH (89s) as buffer. The MRL was exceeded after 161s in IMZ sulphate solutions buffered at pH 6 with either NaHCO3 or NaOH. An IMZ residue-loading curve was prepared from which residue levels can be predicted for the control of IMZ-sensitive and IMZ-resistant isolates of P. digitatum. From this model the benchmark residue level for 95% control of an IMZ-sensitive isolate and of an IMZ-resistant isolate were predicted to be 0.81 and 2.64μgg−1, respectively. Residue loading can be improved by adjusting the pH level of an IMZ sulphate solution to 6 or by using the IMZ EC formulation, but exposure time should be restricted to 45s so as not to exceed the MRL. Conversely, sufficient exposure time of ≈90s in an unbuffered IMZ sulphate solution (pH 3) will result to improved green mould control, but with residue loading below 2μgg−1. The resistant isolate could not be controlled adequately with residue levels below the MRL, therewith indicating the practical relevance of IMZ resistance.

Use of High-Intensity Ultrasound to Increase the Efficiency of Imazalil in Postharvest Storage of Citrus Fruits

Imazalil (IMZ) plays a key role in the storage and marketing of Citrus fruits. It represents a very important tool in the fight against postharvest pathogens and is an efficient system for reducing economic losses due to the development of rots from Penicillium digitatum and Penicillium italicum. However, the current technologies employed to apply the fungicide by drenching, spraying, or dipping are ever much less suitable and progressive restrictions by law limit the residue level on the fruit and in the waste-water. A critical analysis of the factors affecting imazalil efficacy in postharvest preservation of Citrus fruit suggests that ultrasound could be useful to overcome these drawbacks. Data obtained confirmed a faster and deeper uptake of the fungicide in mandarins and lemons when the fruits were dipped in sonicated imazalil mixtures compared to the un-sonicated ones, allowing reduction of the dose of application and duration of the treatment. Indeed dipping for 1 min in 500 mg L−1 IMZ sonicated mixture produced in mandarins the same residue level of dipping for 5 min in 1,000 mg L−1 IMZ mixture without ultrasound. Moreover, imazalil concentration in fruit after sonication did not exceed the maximum residue limit allowed by the law. Sonication did not affect the chemical parameters, weight loss, and rind color of the fruit. No damage on the fruit surface was observed after scanning electron microscopy analyses of the rind of sonicated fruit. The data obtained open new interesting perspectives in the development of new postharvest treatment technologies for Citrus fruit.

An antidote for imazalil-induced genotoxicityin vitro: The lichen,Dermatocarpon intestiniforme(Körber) hasse

Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce serious toxic effects in vertebrates. In recent years, a number of studies have suggested that lichens might be easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being performed to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. In this context, the antigenotoxic effect of aqueous Dermatocarpon intestiniforme (Körber) Hasse. extract (DIE) was studied against the genotoxic damage induced by IMA on cultured human lymphocytes (n = 6) using chromosomal aberration (CA) and micronucleus (MN) as cytogenetic endpoints. Human peripheral lymphocytes were treated in vitro with varying concentrations of DIE (0, 25, 50 and 100 μg/ml), tested in combination with IMA (336 μg/ml). DIE alone were not genotoxic and when combined with IMA treatment, it reduced the frequency of CAs and the rate of MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of DIE. The results of the present study suggest that this plant extract per se does not have a genotoxic potential, but can alleviate the genotoxicity of IMA on cultured human lymphocytes. In conclusion our findings may have an important application for the protection of cultured human lymphocyte from the genetic damage and side effects induced by medical and agricultural chemicals hazardous for people.

Role of aqueous Bryoria capillaris (Ach.) extract as a genoprotective agent on imazalil-induced genotoxicity in vitro

In recent years, a number of studies have suggested that lichens might be the easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being carried out to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. On the other hand, imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce very serious toxic effects in vertebrates. In this context, the antigenotoxic effect of aqueous Bryoria capillaris (Ach.) extract (BCE) was studied against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberrations (CA) and micronucleus (MN) as cytogenetic parameters. Human peripheral lymphocytes were treated in vitro with varying concentrations of BCE (5, 10, 25, 50 and 100µg/mL), tested in combination with IMA (336µg/mL). BCE alone was not genotoxic, and when combined with IMA treatment, it reduced the frequency of CAs and the rates of MN. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of BCE. The results of the present study suggest that this plant extract per se do not have genotoxic potential, but can modulate the genotoxicity of IMA on peripheral human lymphocytes in vitro. In conclusion, our findings may have an important application in the protection of cultured human lymphocyte from the genetic damage and side effects induced by agricultural and medical chemicals that are hazardous to people.

Anti-genotoxic role of eicosapentaenoic acid against imazalil-induced DNA damage in vitro

Eicosapentaenoic acid (EPA) is a polyunsaturated n-3 fatty acid and is essential to the health of mammals. Recent data show that EPA can act as anti-mutagenic agent. On the other hand, pesticides comprise a new and important class of environmental pollutants nowadays. Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains is suspected to produce very serious toxic effects in vertebrates. The present study investigated the anti-genotoxic effect of EPA against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberration (CA) and micronucleus (MN) tests as cytogenetic endpoints. Peripheral blood cells were treated in vitro with varying concentrations of EPA (2.5, 5, 10, 20 and 40 μg/ml), tested in combination with IMA (336 μg/ml). Our results revealed that the rates of CAs and MNs in lymphocytes were significantly (p < 0.05) increased by IMA as compared to the controls. The results also showed that EPA alone was not genotoxic. Moreover, when combined with IMA treatment, EPA reduced the frequencies of CAs and MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of EPA. In conclusion, our data may have an important application for the protection of cultured human lymphocyte from the genetic damage and repercussions induced by agricultural and industrial chemicals hazardous in people.

The protective role of ascorbic acid on imazalil-induced genetic damage assessed by the cytogenetic tests

Ascorbic acid (AA), known as vitamin C, has important antioxidant and metabolic functions, making its incorporation into the human diet essential. On the other hand, imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains is suspected to produce very serious toxic effects in vertebrates. In this study, the antigenotoxic effects of AA were studied against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberration (CA) and sister chromatid exchange (SCE) as genetic end points. Human peripheral lymphocytes were treated in vitro with varying concentrations of AA (25, 50, 100, 200, and 400 μg/ml), tested in combination with IMA (336 mg/L). AA alone was not genotoxic and when combined with IMA treatment, reduced the frequencies of CAs and SCEs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of AA. In conclusion, the preventive role of AA in alleviating IMA-induced DNA damage was indicated for the first time in the present study.

Role of Peltigera rufescens (Weis) Humb. (a lichen) on imazalil-induced genotoxicity: analysis of micronucleus and chromosome aberrations in vitro

Imazalil (IMA), a commonly used fungicide in both agricultural and clinical domains, is suspected to produce very serious toxic effects on vertebrates. On the other hand, in recent years, a number of studies have suggested that lichens might be easily accessible sources of natural drugs that could be used as a possible food supplement. Extensive research is being carried out to explore the importance of lichen species, which are known to contain a variety of pharmacological active compounds. In this context, the anti-genotoxic effects of aqueous Peltigera rufescens (Weis) Humb. extracts (PREs) were studied against the genotoxic damage induced by IMA on cultured human lymphocytes using chromosomal aberrations (CAs) and micronucleus (MN) as cytogenetic parameters. Human peripheral lymphocytes were treated in vitro with varying concentrations of PREs (0, 5, 10, 25, 50 and 100 mg/L), tested in combination with IMA (336 mg/L). PREs alone were not genotoxic and when combined with IMA treatment, reduced the frequency of CAs and the rates of MNs. A clear dose-dependent decrease in the genotoxic damage of IMA was observed, suggesting a genoprotective role of P. rufescens extract. The results of the present study indicate that this plant extract per se do not have genotoxic potential but can minimize the genotoxicity of IMA on human lymphocytes in vitro. In conclusion our findings may have an important application for the protection of human lymphocyte from the genetic damage and side effects induced by agricultural and medical chemicals hazardous in people.

Potential of an in vitro toolbox combined with exposure data as a first step for the risk assessment of dietary chemical contaminants

In vitro risk assessment of dietary contaminants has become a priority in human food safety. This paper proposes an in vitro approach associating different complementary tools in an original toolbox and aims to improve the assessment of the toxicological impact of dietary contaminants at realistic human exposure levels, with a special focus on the intestinal compartment. The system is based on the use of four complementary cellular tools, namely stress gene induction in transgenic strains of Escherichia coli, modulation of the activity of key biotransformation enzymes (cytochrome P-450 (CYP) 1A1 and 3A4) in a human intestinal cell line, and activation of aryl hydrocarbon receptor (AhR) and oestrogenic receptor (ER)-dependent genes in agonistic and antagonistic assays with luciferase reporter cells. It was applied to four chosen model molecules: ochratoxin A (OTA) and deoxynivalenol (DON), two common food-borne mycotoxins, and imazalil (IMA) and benomyl (BEN), two fungicides widely occurring in foodstuffs. All these assays were performed at or around a realistic intestinal concentration, determined through a deterministic approach based on the calculation of a theoretical maximum daily intake (TMDI). Using the four model molecules, it is clearly highlighted that induction of CYP1A1 activity and inhibition of CYP3A4 activity occurred in Caco-2 cells at a realistic intestinal concentration of IMA. Furthermore, some bacterial stress genes were induced in a range of realistic concentrations, following exposure to DON and IMA. In addition, BEN clearly provoked an ER agonistic activity in a human oestrogen sensitive reporter cell line. All these results are in accordance with the literature, suggesting that the in vitro toolbox constitutes an interesting approach in order to obtain a first ‘fingerprint’ of dietary contaminants at realistic human exposure for further risk assessment.

Imazalil residue loading and green mould control in citrus packhouses

Imazalil (IMZ) is commonly applied in South African citrus packhouses for the control of green mould, caused by Penicillium digitatum, yet the disease still causes significant postharvest losses. The maximum residue limit (MRL) for IMZ on citrus fruit is 5 μg g −1, whereas 2–3 μg g −1 is a biologically effective residue level that should at least inhibit green mould sporulation. Standard compliance auditing of residue levels of citrus fruit, however, indicate that fruit from the majority of packhouses have residues of ≈1 μg g −1. Poor disease control from insufficient residue loading might further be compounded by the presence of IMZ-resistant isolates of P. digitatum in packhouses. This study was conducted to assess the current status of IMZ application in South African packhouses, to determine the adequate residue levels needed to control green mould and inhibit its sporulation using both IMZ sensitive and resistant isolates, to investigate IMZ application methods and resultant residue levels in commercial citrus packhouses, and to study optimisation of modes of IMZ application in citrus packhouses. Factors studied were IMZ concentration, application type (spray vs. dip and drench), exposure time, solution temperature and pH, as well as curative and protective control of P. digitatum. The packhouse survey showed that the majority of packhouses applied IMZ in a sulphate salt formulation through a fungicide dip tank, and loaded an IMZ residue of ≈1 μg g −1. In dip applications, IMZ had excellent curative and protective activity against Penicillium isolates sensitive to IMZ. However, curative control of IMZ resistant isolates was substantially reduced and protective control was lost, even at twice the recommended concentration, nor was sporulation inhibited. The use of sodium bicarbonate (2%) buffered imazalil sulphate solutions at pH ±8, compared with pH ±3 of the unbuffered solutions, markedly increased IMZ residue loading on Navel and Valencia oranges and improved curative and protective control of IMZ resistant isolates. Exposure time did not affect IMZ residue loading in IMZ sulphate solutions at pH 3, although the MRL was exceeded after 45 s exposure in pH 8 solutions. Imazalil applied through spray or drench application improved residue loading, but green mould control was less effective than after dip application.

Imazalil Degradation upon Applying Ozone—Transformation Products, Kinetics, and Toxicity of Treated Aqueous Solutions

The elimination of the pesticide imazalil (IMZ) spiked into ultrapure water as well as into wastewater applying ozone (O3) and the identification of transformation products was investigated. O3 under hydroxyl radical suppression conditions reacted rapidly with the aliphatic double bond or the imidazole ring in IMZ, yielding several transformation products by partial oxidation. The structures of four oxidation products not yet described were characterized and identified after liquid chromatography coupled with high resolution, high mass accuracy, mass and tandem mass spectrometry (LC/MS and -MSn) in ultrapure water. For two identified transformation products, generated via direct ozone attack on IMZ, formation pathways were proposed. In wastewater, only two of those transformation products were observed. Kinetics studies for the reaction of IMZ with O3, evaluated by the competition kinetic method, resulted in a second-order rate constant kO3,IMZ ∼ (1.02 ± 0.03) × 105 M−1 s−1 at pH 6.6 ± 0.2, indicating that IMZ is completely transformed during the ozonation process. Tests of acute toxicity were performed applying a solution of IMZ in ultrapure water or treated wastewater to Daphnia magna. In both cases the decrease of toxicity was observed after ozone treatment.

Imazalil concentration for in vitro monitoring of imazalil resistant isolates of Penicillium digitatum in citrus packinghouses

The extensive use of imazalil (IMZ) in Uruguayan citrus packinghouses to control Penicillium spp. favored the selection and proliferation of resistant isolates. With the aim of detecting Penicillium digitatum biotypes that are not controlled by commercial doses of IMZ, the IMZ concentration within amended potato dextrose agar (PDA) plates was adjusted to 1.0 mg L −1 IMZ. This concentration allowed the detection of resistant isolates that were not controlled by commercial applications of 3.0 g L −1 IMZ. These isolates were able to grow in fruit with IMZ residues of 0.92 and 3.08 mg kg −1. Therefore, environmental monitoring of facilities where commercial dip applications containing 3.0 g L −1 IMZ are employed, should be done with 1.0 mg L −1 of IMZ in Petri dishes. We can conclude that use of IMZ in postharvest applications of 3.0 g L −1 remains effective to control wild Uruguayan P. digitatum isolates but not for resistant ones. A survey of 26 P. digitatum isolates collected in Uruguay indicated IMZ resistant isolates occurred in packinghouses and not in citrus groves.