Prochloraz residue levels in ginger submitted to Sportak 450 CE" postharvest treatment

A preliminary study was conducted to determine the residue levels of prochloraz in ginger samples treated with Sportak 450 CE® (prochloraz as active ingredient) under laboratory conditions and cold-storage for 15 days at 10°C and 89% RH. Sampling was carried out at 10 and 15 days after Sportak 450 CE® dip treatment (450 and 900 µg mL-1). Pesticide residues were determined by GCECD. During the study, residue levels in ginger ranged between 3.6 and 10.6 mg Kg-1 for prochloraz.

Tratamento térmico e prochloraz no controle da antracnose em pós-colheita de frutos de banana 'Prata Anã'

O controle químico, térmico e a refrigeração são os processos mais utilizados no tratamento pós-colheita das bananas. O objetivo deste trabalho foi avaliar o efeito do tratamento térmico, químico e da combinação dos dois métodos e estes associados à baixa temperatura de conservação no controle da antracnose na pós-colheita da banana. Para tanto os experimentos foram realizados em três épocas quando, bananas (Musa sp) da variedade 'Prata Anã' (AAB) no estádio pré-climatérico eram coletadas e suas pencas individualizadas. As pencas foram submetidas a quatro tratamentos com cinco repetições cada: 1. Tratamento térmico (imersão em água a 56ºC por seis minutos, seguido de resfriamento em água à temperatura ambiente); 2. Tratamento químico por seis minutos (imersão em calda fungicida (prochloraz 2,5 mL.L-1)); 3. Tratamento térmico seguido do químico; 4. Testemunha, imersão em água por seis minutos. Após os tratamentos, as pencas eram divididas em duas partes iguais, sendo que uma parte ficou em câmara fria (14ºC com variação de 2ºC) e a outra permaneceu à temperatura ambiente. O tratamento térmico não foi eficiente no controle da doença. O fungicida prochloraz a 2,5 mL.L-1 foi eficiente no controle da podridão pós-colheita. A refrigeração retardou o surgimento da doença em até 12 dias. Os resultados indicam que a baixa temperatura, associada ou não ao controle químico, é capaz de controlar a podridão pós-colheita dos frutos por 12 dias.

Sensitivity distributions and cross-resistance patterns of Mycosphaerella graminicola to fluquinconazole, prochloraz and azoxystrobin over a period of nine years

The sensitivity of 73 isolates of Mycosphaerella graminicola collected over the period 1993–2002 from wheat fields in South England was tested in vitro against the triazole fluquinconazole, the strobilurin azoxystrobin and to the imidazole prochloraz. Over the sampling period, sensitivity of the population to fluquinconazole and prochloraz decreased by factors of approximately 10 and 2, respectively, but there was no evidence of changes in sensitivity to azoxystrobin. There was no correlation between sensitivity to fluquinconazole and prochloraz, but there was a weak negative cross-resistance between fluquinconazole and azoxystrobin.

Prochloraz residue levels in ginger submitted to Sportak 450 CE postharvest treatment

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

An alternative LC-UV procedure for the determination of prochloraz residues in fruits

An alternative method using liquid chromatography with UV detection for the determination of prochloraz as 2,4,6-trichlorophenol in mango, papaya and orange is described. Ethyl acetate, acetone and dichloromethane were tested for extraction of prochloraz from the fruits. After extraction the residue of prochloraz was derivatized with pyridine hydrochloride. The analysis was carried out using liquid chromatography with UV detection and gas chromatography with electron-capture detection. Average recoveries of prochloraz from spiked fruits (0.1 and 0.2 mg kg-1) ranged from 80% to 94% with relative standard deviations between 5.6% and 12.6% (n=8). Detection and quantification limits were 0.05 and 0.1 mg kg-1, respectively. The LC-UV method was applied to mango and papaya samples submitted to dip treatment with a prochloraz formulation under laboratory conditions. In addition, fruit samples obtained from local markets were analysed. ©2005 Sociedade Brasileira de Química.

Absence of effects on the rat sperm quality after subacute exposure to low doses of fungicide prochloraz

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prochloraz causes irreversible masculinization of zebrafish (Danio rerio).

The aim of the present study was to investigate the persistence of endocrine effects by prochloraz, a fungicide known to have multiple effects on the endocrine system of vertebrates. Since discontinuous exposure is particularly relevant in aquatic ecosystems, an exposure scenario with an exposure phase and a subsequent recovery period was chosen to assess the potential for reversibility of effects by prochloraz on the sexual development of zebrafish (Danio rerio). Zebrafish were exposed to different concentrations of prochloraz (10-300 μg/L) until 60 days post hatch (dph), which includes the period of sexual differentiation. For the subsequent 40 days, fish were either held in clean water for depuration or under further continuous exposure. Histological investigations of the gonads revealed persistent effects on sexual differentiation. The sex ratio was skewed towards males and significantly more intersex individuals were found after exposure to prochloraz at 60 dph. No intersex fish, but masculinized sex ratios were still present after the depuration period, documenting that prochloraz irreversibly affects the sexual development of zebrafish.

Mango stem-end rot (Botryosphaeria dothidea) disease control by partial-pressure infiltration of fungicides

The in vitro efficacy of several fungicides against Botryosphaeria dothidea (syn. Dothiorella dominicana) and their in vivo efficacy in controlling mango cv. Kensington Pride stem-end rot on partial-pressure infiltration v. dip treatment of green mature fruit was evaluated. In vitro sensitivity of B. dothidea to Benlate (benomyl), Sportak (prochloraz) and Scala (pyrimethanil) at 10 dilutions of the manufacturer's recommended rate was first determined at typical cold (13degreesC) and shelf (23degreesC) storage temperatures. The effectiveness of partial-pressure infiltration and conventional hot (52degreesC) or cold (26degreesC) dipping of fruit after harvest was then evaluated using the commercially recommended rate for each fungicide. In vitro, Benlate and Sportak prevented the growth of B. dothidea at both storage temperatures and at all concentrations, while Scala partially controlled growth of the pathogen. Benlate was the most effective fungicide for stem-end rot control. Sportak and Scala resulted in stem-end rot control when applied by partial-pressure infiltration, but not as dips. Partial-pressure infiltration holds promise for enhancing the efficacy of otherwise less effective but alternative fungicides for control of stem-end rot diseases.

Postharvest quality of cocona (Solanum sessiliflorum Dunal) stored under ambient condition

Cocona (Solanum sessiliflorum Dunal) is an important genetic resource that has been traditionally used for a variety of purposes, including food, medical and cosmetics applications. The objective of this study was evaluated the quality and the period of postharvest shelf life of cocona 'Mosquet", through the physical, chemical and physiological characterization of fruits stored under the ambient conditions. Physiologically mature fruits were harvested from an orchard, washed with tap water and soaked in a solution of the fungicide Prochloraz (49.5 g/100 L of water) for 5 minutes. After air drying, the fruits were packed in plastic containers and stored at 24 ± 2 ºC and relative humidity 60 ± 5% for 15 days. The fruits developed a respiratory climacteric respiratory patern and remained fit for consumption up to day 6 of storage, that is, without visual symptoms of loss of water and firmness. At this stage, the fruits showed firmness of 117,42 kPa, soluble solids of 6.62º Brix and citric acid of 1.22 %.

Use of biofilm in the postharvest conservation of 'Pedro Sato' guava

This present work aimed to assess the effect of different concentrations of cassava starch, associated or not with prochloraz fungicide, on the postharvest conservation of 'Pedro Sato' guavas. Physiologically mature fruits were immersed in a solution of prochloraz (49.5 g/100 liters of water) for 5 min. Fruit treated with distilled water and air dried were used as control. They were immersed in cassava starch suspension at concentrations of 0, 20, 30 and 40 g/L, plus 0.5 mL/L of mineral oil. The fruits were stored at 21.0 ± 1.0 ºC and relative humidity of 85 ± 5%, for 12 days, and were examined at every three days. The combination of prochloraz and cassava starch resulted in delayed loss of firmness and yellowness and inhibited the incidence of lesions caused by Colletotrichum gloeosporioides during the 12 days of storage. Fruits treated with 40 g/L of starch, whether containing prochloraz or not, had unpleasant taste and odor, which suggests the occurrence of fermentation. Control fruits, with and without prochloraz, and those treated with starch with no prochloraz, had nearly 100% lesion occurrence within the 12 days. Starch suspension of 30 g/L, containing prochloraz, was the most effective in maintaining fruit quality.

Mechanisms of antifungal resistance in the opportunistic fungus Aspergillus fumigatus

ABSTRACT Aspergillus fumigatus is one of the most prevalent airbone fungal pathogen and can cause severe fatal invasive aspergillosis in immunocompromised patients. Several antifungal agents are available to treat these infections but with limited success. These agents include polyenes (amphotericin B), echinocandins (caspofungin) and azoles, which constitute the most important class with itraconazole (ITC) and voriconazole as major active compounds. Azole-derived antifungal agents target the ergosterol biosynthesis pathway via the inhibition of the lanosterol 14α-demethylase (cyp51/ERG1 1), a cytochrome P450 responsible for the conversion of lanosterol to ergosterol, which is the main component of cell membrane in fungi. A. fumigatus is also found in the environment as a contaminant of rotting plant or present in composting of organic waste. Among antifungal agents used in the environment for crop protection, the class of azoles is also widely used with propiconazole or prochloraz as examples. However, other agents such as dicarboximide (iprodione), phenylamide (benalaxyl) or strobilurin (azoxystrobin) are also used. Emergence of clinical azole-resistant isolates has been described in several European countries. However the incidence of antifungal resistance has not been yet reported in details in Switzerland. In this study, the status of antifungal resistance was investigated on A. fumigatus isolates collected from Swiss hospitals and from different environmental sites and. tested for their susceptibility to several currently used antifungal agents. The data showed a low incidence of resistance for all tested agents among clinical and environmental isolates. Only two azole-resistant environmental isolates were detected and none among the clinical tested isolates. In general, A. fumigatus was susceptible to all antifungals tested in our study, except to azoxystrobin which was the less active agent against all isolates. Since mechanisms of antifungal resistance have been poorly investigated until now in A. fumigatus, this work was aimed 1) to identify A. fumigatus genes involved in antifungal resistance and 2) to test their involvement in the development of resistance in sampled isolates. Therefore, this work proposed to isolate A. fumigatus genes conferring resistance to a drug-hypersusceptible Saccharomyces cerevisiae strain due to a lack of multidrug transporter genes. Several genes were recovered including three distinct efflux transporters (atrF, atrH and mdrA) and a bZip transcription factor, yapA. The inactivation of each transporter in A. fumigatus indicated that the transporters were involved in the basal level of azole susceptibility. The inactivation of YapA led to a hypersusceptibility to H2O2, thus confirming the involvement of this gene in the oxidative stress response of A. fumigatus. The involvement of the abovementioned transporters genes and of other transporters genes identified by genome analysis in azole resistance was tested by probing their expression in some ITC-resistant isolates. Even if upregulation of some transporters genes was observed in some investigated isolates, the correlation between azole resistance and expression levels of all these transporters genes could not be clearly established for all tested isolates. Given these results, the present work addressed 1) alteration in the expression of cyp51A encoding for the azole target enzyme, and 2) mutation(s) in the cyp51A sequence as potential mechanisms of azote resistance in A. . However, overexpression of cyp51A in the investigated isolates was not linked with azote resistance. Since it was reported that mutation(s) in cyp51A were participating in azote resistance in A. fumigatus, a functional complementation of cyp51A cDNAs from ITC-resistant A. fumigatus strains in S. cerevisiae ergl 1 Δ mutant strain was attempted. Expression in S. cerevisiae allowed the testing of these cDNAs with regards to their functionality and involvement in resistance to specific azote compounds. We could demonstrate that Cyp51A protein with a G54E or M220K mutations conferred resistance to specific azoles in S. cerevisiae, therefore suggesting that these mutations were important for the development of azote resistance in A. fumigatus. In conclusion, this work showed a correlation between ITC resistance and mechanisms involving overexpression of transporters and cyp51A mutations in A. fumigatus isolates. However, azole resistance of some isolates has not been solved and thus it will be necessary to approach the study of resistance mechanisms in this fungal species using alternative methodologies. RESUME Aspergillus fumigatus est un champignon opportuniste répandu et est la cause d'aspergilloses invasives le plus souvent fatales chez des patients immunodéprimés. Plusieurs antifongiques sont disponibles afin de traiter ces infections, cependant avec un succès limité. Ces agents incluent les polyènes (amphotericin B), les échinocandines (caspofungin) et les azoles, qui représentent la plus importante classe d'antifongiques avec l'itraconazole (ITC) et le voriconazole comme principaux agents actifs. Les dérivés azolés ciblent la voie de biosynthèse de l'ergostérol via l'inhibition de la lanostérol 14α-demethylase (cyp51/ERG11), un cytochrome P450 impliqué dans la conversion du lanostérol en ergostérol, qui est un composant important de la membrane chez les champignons. A. fumigatus est également répandu dans l'environnement. Parmi les antifongiques employés en agriculture afin de protéger les cultures, les azoles sont aussi largement utilisés. Cependant, d'autres agents tels que les dicarboximides (iprodione), les phenylamides (benalaxyl) et les strobilurines (azoxystrobin) peuvent être également utilisés. L'émergence de souches cliniques résistantes aux azoles a été décrite dans différents pays européens. Cependant, l'incidence d'une telle résistance aux azoles n'a pas encore été reportée en détails en Suisse. Dans ce travail, l'émergence de la résistance aux antifongiques a été étudiée par analyse de souches d'A. fumigatus provenant de milieux hospitaliers en Suisse et de différents sites et leur susceptibilité testée envers plusieurs antifongiques couramment utilisés. Les données obtenues ont montré une faible incidence de la résistance parmi les souches cliniques et environnementales pour les agents testés. Seulement deux souches environnementales résistantes aux azoles ont été détectées et aucune parmi les souches cliniques. Les mécanismes de résistance aux antifongiques ayant été très peu étudiés jusqu'à présent chez A. fumigatus , ce travail a eu aussi pour but 1) d'identifier les gènes d' A. fumigatus impliqués dans la résistance aux antifongiques et 2) de tester leur implication dans la résistance de certaines souches. Ainsi, il a été proposé d'isoler les gènes d' A. fumigatus pouvant conférer une résistance aux antifongiques à une souche de Saccharomyces cerevisiae hypersensible aux antifongiques. Trois transporteurs à efflux (atrF, atrH et mdrA) et un facteur de transcription appartenant à la famille des bZip (YapA) ont ainsi été isolés. L'inactivation, dans une souche d'A. fumigatus, de chacun des ces transporteurs a permis de mettre en évidence leur implication dans la susceptibilité d'A. fumigatus aux antifongiques. L'inactivation de YapA a engendré une hypersusceptibilité à l' H2O2, confirmant ainsi le rôle de ce gène dans la réponse au stress oxydatif chez A . fumigatus. La participation dans la résistance aux antifongiques des gènes codant pour des transporteurs ainsi que d'autres gènes identifiés par analyse du génome a été déterminée en testant leur niveau d'expression dans des souches résistantes à l'ITC. Bien qu'une surexpression de transporteurs ait été observée dans certaines souches, une corrélation entre la résistance à l'ITC et les niveaux d'expression de ces transporteurs n'a pu être clairement établie. Ce présent travail s'est donc porté sur l'étude de 2 autres mécanismes potentiellement impliqués dans la résistance aux azoles : 1) la surexpression de cyp51A codant pour l'enzyme cible et 2) des mutations dans cyp51A. Cependant, la surexpression de cyp51A dans les souches étudiées n'a pas été constatée. L'effet des mutations de cyp51A dans la résistance aux azoles a été testée par complémentation fonctionnelle d'une souche S. cerevisiae déletée dans son gène ERG11. L'expression de ces gènes chez S. cerevisiae a permis de démontrer que les protéines Cyp51Ap contenant une mutation G54E ou M220K pouvaient conférer une résistance spécifique à certains azoles, ainsi suggérant que ces mutations pourraient être importantes dans le développement d'une résistance aux azoles chez A. fumigatus. En conclusion, ce travail a permis de mettre en évidence, dans des souches d'A. fumigatus , une corrélation entre leur résistance à l' ITC et les mécanismes impliquant une surexpression de transporteurs et des mutations dans cyp51A. Cependant, ces mécanismes n'ont pu expliquer la résistance aux azoles de certaines souches et c'est pourquoi de nouvelles approches doivent être envisagées afin d'étudier ces mécanismes.

Controle da antracnose na pós-colheita de manga 'Ubá' com o uso de produtos alternativos

Foi avaliada a eficácia de produtos alternativos aos agroquímicos no controle da antracnose na pós-colheita de mangas 'Ubá'. Frutos fisiologicamente maduros foram pulverizados até o completo molhamento, com suspensão de conídios de Colletotrichum gloeosporioides, na concentração de 2,5 x 10(5) conídios/mL. Após a secagem ao ar, foram pulverizados com água destilada (testemunha), tween 20 (8 mL/L de solução), Prochloraz (1,10 mL de Sportak 450 EC/L de solução), óleo de alho (10 mL/L + 8 mL/L de tween 20), óleo de amêndoa de Acrocomia aculeata + leite em pó instantâneo (LPI) (25 mL/L+ 10 g LPI/L), óleo de amêndoa de A. aculeata + tween (25 mL/L + 8 mL/L de tween 20), biofertilizante agro-mos® (100 µL/L), óleo de neen (10 mL/L + 8 mL/L de tween 20), quitosana (10 mL/L + 8 mL/L de tween 20) e biomassa cítrica (10 mL/L + 8 mL/L tween 20).O solvente utilizado foi água destilada. Avaliaram-se o período de incubação, o período latente, a perda de massa fresca, a produção de CO² e, diariamente, a severidade e incidência da doença. Os períodos mais curtos de incubação da doença foram observados nos frutos tratados com óleo de neen, água + tween e biomassa cítrica, com aproximadamente cinco dias. O óleo de amêndoa de A. aculeata + LPI e agro-mos® foram os produtos que mais retardaram o aparecimento dos sintomas, impondo à doença o período de incubação de nove dias após a inoculação do patógeno. Quanto à severidade, o óleo de amêndoa de macaúba + LPI e o Prochloraz foram os mais eficientes em conter o crescimento do patógeno até o oitavo dia após a inoculação, sendo que, logo depois, os frutos tratados com óleo de amêndoa de A. aculeata + LPI se igualaram àqueles tratados com a maioria dos demais produtos. Os frutos tratados com óleo de amêndoa de A. aculeata + LPI e óleo de amêndoa de A. aculeata + tween manifestaram as estruturas do patógeno apenas após 13 e 14 dias de avaliação, respectivamente. As maiores perdas de massa foram observadas nos frutos tratados com óleo de alho e biomassa cítrica, com 8,31% e 8,44%, respectivamente, no dia 14. Quanto à produção de CO², o óleo de amêndoa de A. aculeata + LPI e óleo de amêndoa de A. aculeata + tween mantiveram a taxa respiratória crescente, sendo que, no dia 12 ocorreu um leve aumento na respiração. Dessa forma, conclui-se que, além do Prochloraz, o óleo de amêndoa de A. aculeata + LPI e óleo de amêndoa de A. aculeata + tween têm bom potencial para controle da antracnose em manga 'Ubá'.

Efeito de fungicidas em Colletotrichum acutatum e controle da antracnose do morangueiro

Onze fungicidas foram analisados in vitro e seus efeitos testados em campo para o controle de Colletotrichum acutatum, agente da flor-preta do morangueiro (Fragaria X ananassa). Os tratamentos (campo) foram (dosagens de i.a/100 l): iprodione (75 ml), benomil (100 g), tebuconazole (50 ml), tiofanato metílico (70 g), prochloraz (100 ml), propiconazole (50 ml), mancozeb (200 g), folpet (270 g), sulfato de cobre (200 g) e chlorotalonil (200 g), em pulverizações semanais (protetores) e quinzenais (sistêmicos). Foram avaliados a produção de frutos e o número de flores com sintoma de queima. Nos testes de fungitoxicidade in vitro, estudou-se o crescimento micelial, a germinação de conídios e a formação de apressórios. Nos testes in vitro foram utilizadas as concentrações de 1 e 10 ppm de i.a. para todos os fungicidas e posteriormente testadas as concentrações de 0,01, 0,1 e 1 ppm para os mais eficientes, e 10, 50 e 100 ppm para os menos eficazes. Os que resultaram em maior inibição micelial em baixas concentrações foram prochloraz e tebuconazole e os que menos inibiram o crescimento micelial foram sulfato de cobre, clorotalonil e folpet. Para inibir a germinação conidial mostraram-se mais eficientes chlorotalonil, tebuconazole, prochloraz e benomil, e para inibir a formação de apressórios, chlorotalonil e benomil. Os resultados de campo diferiram parcialmente dos testes in vitro: prochloraz e sulfato de cobre apresentaram os menores percentuais de flores doentes (53-55%), enquanto benomil apresentou 100% de ataque. Embora prochloraz, seguido de iprodione, folpet e mancozeb tenham resultado em maiores produções, nenhum fungicida controlou a doença satisfatoriamente.

Fungicidas e mistura de fungicidas no controle do oídio da soja

Dois experimentos foram conduzidos em Londrina/PR para avaliar a eficácia de fungicidas e suas misturas no controle do oídio (Microsphaera diffusa) da soja (Glycine max). Uma aplicação dos produtos foi feita no estádio R4 da soja cv. EMBRAPA 48. Os melhores resultados foram obtidos com os seguintes fungicidas: tebuconazol-1 e tebuconazol-2, carbendazim-1 e carbendazim-2 (produtos comerciais diferentes). Teste 1 - tebuconazol-2 (150 g i. a./ha), prochloraz + propiconazol (225 + 62,5), propiconazole + carbendazim-1 (50 + 175) e tebuconazol-1 + propiconazol (75 + 62,5); Teste 2 - carbendazim-1 (250), carbendazim-2 (250) e tebuconazol-2 (100).